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Interactions between Skeletal Muscle Myoblasts and their Extracellular Matrix Revealed by a Serum Free Culture System.
Chaturvedi, Vishal; Dye, Danielle E; Kinnear, Beverley F; van Kuppevelt, Toin H; Grounds, Miranda D; Coombe, Deirdre R
Decellularisation of skeletal muscle provides a system to study the interactions of myoblasts with muscle extracellular matrix (ECM). This study describes the efficient decellularisation of quadriceps muscle with the retention of matrix components and the use of this matrix for myoblast proliferation and differentiation under serum free culture conditions. Three decellularisation approaches were examined; the most effective was phospholipase A2 treatment, which removed cellular material while maximizing the retention of ECM components. Decellularised muscle matrices were then solubilized and used as substrates for C2C12 mouse myoblast serum free cultures. The muscle matrix supported myoblast proliferation and differentiation equally as well as collagen and fibronectin. Immunofluorescence analyses revealed that myoblasts seeded on muscle matrix and fibronectin differentiated to form long, well-aligned myotubes, while myoblasts seeded on collagen were less organized. qPCR analyses showed a time dependent increase in genes involved in skeletal muscle differentiation and suggested that muscle-derived matrix may stimulate an increased rate of differentiation compared to collagen and fibronectin. Decellularized whole muscle three-dimensional scaffolds also supported cell adhesion and spreading, with myoblasts aligning along specific tracts of matrix proteins within the scaffolds. Thus, under serum free conditions, intact acellular muscle matrices provided cues to direct myoblast adhesion and migration. In addition, myoblasts were shown to rapidly secrete and organise their own matrix glycoproteins to create a localized ECM microenvironment. This serum free culture system has revealed that the correct muscle ECM facilitates more rapid cell organisation and differentiation than single matrix glycoprotein substrates.
Serum-free media formulations are cell line-specific and require optimization for microcarrier culture.
Tan, Kah Yong; Teo, Kim Leng; Lim, Jessica F Y; Chen, Allen K L; Choolani, Mahesh; Reuveny, Shaul; Chan, Jerry; Oh, Steve Kw
Mesenchymal stromal cells (MSCs) are being investigated as potential cell therapies for many different indications. Current methods of production rely on traditional monolayer culture on tissue-culture plastic, usually with the use of serum-supplemented growth media. However, the monolayer culturing system has scale-up limitations and may not meet the projected hundreds of billions to trillions batches of cells needed for therapy. Furthermore, serum-free medium offers several advantages over serum-supplemented medium, which may have supply and contaminant issues, leading to many serum-free medium formulations being developed. We cultured seven MSC lines in six different serum-free media and compared their growth between monolayer and microcarrier culture. We show that (i) expansion levels of MSCs in serum-free monolayer cultures may not correlate with expansion in serum-containing media; (ii) optimal culture conditions (serum-free media for monolayer or microcarrier culture) differ for each cell line; (iii) growth in static microcarrier culture does not correlate with growth in stirred spinner culture; (iv) and that early cell attachment and spreading onto microcarriers does not necessarily predict efficiency of cell expansion in agitated microcarrier culture. Current serum-free media developed for monolayer cultures of MSCs may not support MSC proliferation in microcarrier cultures. Further optimization in medium composition will be required for microcarrier suspension culture for each cell line. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
A procedure for culturing rat neocortex explants in a serum-free nutrient medium
Romijn, H. J.; de Jong, B. M.; Ruijter, J. M.
A procedure is described for long-term culturing of rat neocortex explants in a serum-free growth medium. Slices spanning the entire cortical depth from pial to ventricular side are prepared from 6-day-old rat pups. After preincubation in Hanks' balanced salt solution with extra glucose, the
Fetal bovine serum and human constitutive androstane receptor: Evidence for activation of the SV23 splice variant by artemisinin, artemether, and arteether in a serum-free cell culture system
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Lau, Aik Jiang; Chang, Thomas K.H., E-mail: email@example.com
The naturally occurring SV23 splice variant of human constitutive androstane receptor (hCAR-SV23) is activated by di-(2-ethylhexyl)phthalate (DEHP), which is detected as a contaminant in fetal bovine serum (FBS). In our initial experiment, we compared the effect of dialyzed FBS, charcoal-stripped, dextran-treated FBS (CS-FBS), and regular FBS on the basal activity and ligand-activation of hCAR-SV23 in a cell-based reporter gene assay. In transfected HepG2 cells cultured in medium supplemented with 10% FBS, basal hCAR-SV23 activity varied with the type of FBS (regular > dialyzed > CS). DEHP increased hCAR-SV23 activity when 10% CS-FBS, but not regular FBS or dialyzed FBS, was used. With increasing concentrations (1–10%) of regular FBS or CS-FBS, hCAR-SV23 basal activity increased, whereas in DEHP-treated cells, hCAR-SV23 activity remained similar (regular FBS) or slightly increased (CS-FBS). Subsequent experiments identified a serum-free culture condition to detect DEHP activation of hCAR-SV23. Under this condition, artemisinin, artemether, and arteether increased hCAR-SV23 activity, whereas they decreased it in cells cultured in medium supplemented with 10% regular FBS. By comparison, FBS increased the basal activity of the wild-type isoform of hCAR (hCAR-WT), whereas it did not affect the basal activity of the SV24 splice variant (hCAR-SV24) or ligand activation of hCAR-SV24 and hCAR-WT by 6-(4-chlorophenyl)imidazo[2,1-b][1,3]thiazole-5-carbaldehyde O-(3,4-dichlorobenzyl)oxime (CITCO). The use of serum-free culture condition was suitable for detecting CITCO activation of hCAR-WT and hCAR-SV24. In conclusion, FBS leads to erroneous classification of pharmacological ligands of hCAR-SV23 in cell-based assays, but investigations on functional ligands of hCAR isoforms can be conducted in serum-free culture condition. - Highlights: • FBS leads to erroneous pharmacological classification of hCAR-SV23 ligands. • Artemisinin, artemether, and arteether activate h
Norepinephrine stimulates progesterone production in highly estrogenic bovine granulosa cells cultured under serum-free, chemically defined conditions
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Piccinato Carla A
Full Text Available Abstract Background Since noradrenergic innervation was described in the ovarian follicle, the actions of the intraovarian catecholaminergic system have been the focus of a variety of studies. We aimed to determine the gonadotropin-independent effects of the catecholamine norepinephrine (NE in the steroid hormone profile of a serum-free granulosa cell (GC culture system in the context of follicular development and dominance. Methods Primary bovine GCs were cultivated in a serum-free, chemically defined culture system supplemented with 0.1% polyvinyl alcohol. The culture features were assessed by hormone measurements and ultrastructural characteristics of GCs. Results GCs produced increasing amounts of estradiol and pregnenolone for 144h and maintained ultrastructural features of healthy steroidogenic cells. Progesterone production was also detected, although it significantly increased only after 96h of culture. There was a highly significant positive correlation between estradiol and pregnenolone production in high E2-producing cultures. The effects of NE were further evaluated in a dose–response study. The highest tested concentration of NE (10 (−7 M resulted in a significant increase in progesterone production, but not in estradiol or pregnenolone production. The specificity of NE effects on progesterone productio n was further investigated by incubating GCs with propranolol (10 (−8 M, a non-selective beta-adrenergic antagonist. Conclusions The present culture system represents a robust model to study the impact of intrafollicular factors, such as catecholamines, in ovarian steroidogenesis and follicular development. The results of noradrenergic effects in the steroidogenesis of GC have implications on physiological follicular fate and on certain pathological ovarian conditions such as cyst formation and anovulation.
Norepinephrine stimulates progesterone production in highly estrogenic bovine granulosa cells cultured under serum-free, chemically defined conditions.
Piccinato, Carla A; Montrezor, Luis H; Collares, Cristhianna A V; Vireque, Alessandra A; Rosa e Silva, Alzira A M
Since noradrenergic innervation was described in the ovarian follicle, the actions of the intraovarian catecholaminergic system have been the focus of a variety of studies. We aimed to determine the gonadotropin-independent effects of the catecholamine norepinephrine (NE) in the steroid hormone profile of a serum-free granulosa cell (GC) culture system in the context of follicular development and dominance. Primary bovine GCs were cultivated in a serum-free, chemically defined culture system supplemented with 0.1% polyvinyl alcohol. The culture features were assessed by hormone measurements and ultrastructural characteristics of GCs. GCs produced increasing amounts of estradiol and pregnenolone for 144h and maintained ultrastructural features of healthy steroidogenic cells. Progesterone production was also detected, although it significantly increased only after 96h of culture. There was a highly significant positive correlation between estradiol and pregnenolone production in high E2-producing cultures. The effects of NE were further evaluated in a dose-response study. The highest tested concentration of NE (10 (-7) M) resulted in a significant increase in progesterone production, but not in estradiol or pregnenolone production. The specificity of NE effects on progesterone production was further investigated by incubating GCs with propranolol (10 (-8) M), a non-selective beta-adrenergic antagonist. The present culture system represents a robust model to study the impact of intrafollicular factors, such as catecholamines, in ovarian steroidogenesis and follicular development. The results of noradrenergic effects in the steroidogenesis of GC have implications on physiological follicular fate and on certain pathological ovarian conditions such as cyst formation and anovulation.
In vitro maintenance of spermatogenesis in Xenopus laevis testis explants cultured in serum-free media
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Risley, M.S.; Miller, A.; Bumcrot, D.A.
Spermatogenesis has been maintained for extended periods in Xenopus laevis testis explants cultured in serum-free media supplemented with bovine serum albumin, insulin, transferrin, follicle-stimulating hormone, dihydrotestosterone, testosterone, retinol, ascorbate, and tocopherol. The organization of the testis fragments was maintained for 28 days, and all stages of development were present throughout the culture period. 3 H-Thymidine-labeled secondary (Type B) spermatogonia developed in 28 days into spermatids at the acrosomal vesicle stage whereas labeled zygotene spermatocytes became mature spermatids in 28 days. Spermatogonial proliferation also continued in vitro for 28 days. Germ cell differentiation was not dependent upon exogenous testosterone, ascorbate, or tocopherol since 3 H-labeled spermatogonia became mature spermatids in testes cultured 35 days in media lacking these supplements. Autoradiography demonstrated that 55% of the luminal sperm present in explants cultured 10 days had differentiated in vitro. Sperm from testes cultured 10-35 days were similar to sperm from freshly dissected testes with regard to motility and fecundity, and eggs fertilized with sperm from explant cultures developed normally into swimming tadpoles. The results demonstrate the feasibility of maintaining vertebrate spermatogenesis in culture and suggest that in vitro analysis of Xenopus spermatogenesis using defined media may provide important insights into the evolution of regulatory mechanisms in spermatogenesis
A novel serum-free monolayer culture for orderly hematopoietic differentiation of human pluripotent cells via mesodermal progenitors.
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Akira Niwa
Full Text Available Elucidating the in vitro differentiation of human embryonic stem (ES and induced pluripotent stem (iPS cells is important for understanding both normal and pathological hematopoietic development in vivo. For this purpose, a robust and simple hematopoietic differentiation system that can faithfully trace in vivo hematopoiesis is necessary. In this study, we established a novel serum-free monolayer culture that can trace the in vivo hematopoietic pathway from ES/iPS cells to functional definitive blood cells via mesodermal progenitors. Stepwise tuning of exogenous cytokine cocktails induced the hematopoietic mesodermal progenitors via primitive streak cells. These progenitors were then differentiated into various cell lineages depending on the hematopoietic cytokines present. Moreover, single cell deposition assay revealed that common bipotential hemoangiogenic progenitors were induced in our culture. Our system provides a new, robust, and simple method for investigating the mechanisms of mesodermal and hematopoietic differentiation.
Human dental pulp stem cells cultured in serum-free supplemented medium
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Virginie eBonnamain
Full Text Available Growing evidence show that human dental pulp stem cells (DPSCs could provide a source of adult stem cells for the treatment of neurodegenerative pathologies. In this study, DPSCs were expanded and cultured with a protocol generally used for the culture of neural stem/progenitor cells.Methodology: DPSC cultures were established from third molars. The pulp tissue was enzymatically digested and cultured in serum-supplemented basal medium for 12 hours. Adherent (ADH and non-adherent (non-ADH cell populations were separated according to their differential adhesion to plastic and then cultured in serum-free defined N2 medium with epidermal growth factor (EGF and basic fibroblast growth factor (bFGF. Both ADH and non-ADH populations were analyzed by FACS and/or PCR.Results: FACS analysis of ADH-DPSCs revealed the expression of the mesenchymal cell marker CD90, the neuronal marker CD56, the transferrin receptor CD71, and the chemokine receptor CXCR3, whereas hematopoietic stem cells markers CD45, CD133 and CD34 were not expressed. ADH-DPSCs expressed transcripts coding for the Nestin gene, whereas expression levels of genes coding for the neuronal markers β-III tubulin and NF-M, and the oligodendrocyte marker PLP-1 were donor dependent. ADH-DPSCs did not express the transcripts for GFAP, an astrocyte marker. Cells of the non-ADH population that grew as spheroids expressed Nestin, β-III tubulin, NF-M and PLP-1 transcripts. DPSCs migrated out of the spheroids exhibited an odontoblast-like morphology and expressed a higher level of DSPP and osteocalcin transcripts than ADH-DPSCs. Conclusion: Collectively, these data indicate that human DPSCs can be expended and cultured in serum-free supplemented medium with EGF and bFGF. ADH-DPSCs and non-ADH populations contained neuronal and/or oligodendrocyte precursors at different stages of commitment and interestingly, cells from spheroid structures seem to be more engaged into the odontoblastic lineage than the
Serum-free culture alters the quantity and protein composition of neuroblastoma-derived extracellular vesicles
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Jinghuan Li
Full Text Available Extracellular vesicles (EVs play a significant role in cell–cell communication in numerous physiological processes and pathological conditions, and offer promise as novel biomarkers and therapeutic agents for genetic diseases. Many recent studies have described different molecular mechanisms that contribute to EV biogenesis and release from cells. However, little is known about how external stimuli such as cell culture conditions can affect the quantity and content of EVs. While N2a neuroblastoma cells cultured in serum-free (OptiMEM conditions did not result in EVs with significant biophysical or size differences compared with cells cultured in serum-containing (pre-spun conditions, the quantity of isolated EVs was greatly increased. Moreover, the expression levels of certain vesicular proteins (e.g. small GTPases, G-protein complexes, mRNA processing proteins and splicing factors, some of which were previously reported to be involved in EV biogenesis, were found to be differentially expressed in EVs under different culture conditions. These data, therefore, contribute to the understanding of how extracellular factors and intracellular molecular pathways affect the composition and release of EVs.
The influence of serum-free culture conditions on skeletal muscle differentiation in a tissue-engineered model
Gawlitta, D.; Boonen, K.J.M.; Oomens, C.W.J.; Baaijens, F.P.T.; Bouten, C.V.C.
The influence of differentiation medium (DM) components on C2C12 murine myoblast differentiation has only been studied in monolayer cultures. Serum-free formulations have been applied that omit the use of sera with unknown composition. The goal of the present study was to compare the influence of
Immobilization of sericin molecules via amorphous carbon plasma modified-polystyrene dish for serum-free culture
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Tunma, Somruthai; Song, Doo-Hoon; Kim, Si-Eun; Kim, Kyoung-Nam; Han, Jeon-Geon; Boonyawan, Dheerawan
In this study, we focused on sericin hydrolysates, originating from silkworm used in serum-free human bone marrow-derived mesenchymal stem cells (hBM-MSCs) culture. We reported the effect of a covalent linkage between a bioactive protein molecule and polystyrene dish surface via a carbon intermediate layer which can slow down the release rate of protein compounds into the phosphate buffer saline (PBS) solution. Films of amorphous carbon (a-C) and functionalized-carbon were deposited on PS culture dish surfaces by using a DC magnetron sputtering system and RF PECVD system. We found that a-C based-films can increase the hydrophilicity and biocompatibility of polystyrene (PS) dishes, especially a-C films and a-C:N 2 films showed good attachment of hBM-MSCs at 24 h. However, in the case of silica surface (a-C:SiO x films), the cells showed a ragged and unattached boundary resulting from the presence of surface silanol groups. For the UV–vis absorbance, all carbon modified-PS dishes showed a lower release rate of sericin molecules into PBS solution than PS control. This revealed that the functionalized carbon could be enhanced by specific binding properties with given molecules. The carbon-coated PS dishes grafting with sericin protein were used in a serum-free condition. We also found that hBM-MSCs have higher percentage of proliferated cells at day 7 for the modified dishes with carbon films and coated with sericin than the PS control coated with sericin. The physical film properties were measured by atomic force microscopy (AFM), scanning electron microscope (SEM) and contact angle measurement. The presence of -NH 2 groups of sericin compounds on the PS dish was revealed by Fourier transform infrared spectroscopy (FTIR). The stability of covalent bonds of sericin molecules after washing out ungrafted sericin was confirmed by X-ray photoelectron spectroscopy (XPS).
Immobilization of sericin molecules via amorphous carbon plasma modified-polystyrene dish for serum-free culture
Tunma, Somruthai; Song, Doo-Hoon; Kim, Si-Eun; Kim, Kyoung-Nam; Han, Jeon-Geon; Boonyawan, Dheerawan
In this study, we focused on sericin hydrolysates, originating from silkworm used in serum-free human bone marrow-derived mesenchymal stem cells (hBM-MSCs) culture. We reported the effect of a covalent linkage between a bioactive protein molecule and polystyrene dish surface via a carbon intermediate layer which can slow down the release rate of protein compounds into the phosphate buffer saline (PBS) solution. Films of amorphous carbon (a-C) and functionalized-carbon were deposited on PS culture dish surfaces by using a DC magnetron sputtering system and RF PECVD system. We found that a-C based-films can increase the hydrophilicity and biocompatibility of polystyrene (PS) dishes, especially a-C films and a-C:N2 films showed good attachment of hBM-MSCs at 24 h. However, in the case of silica surface (a-C:SiOx films), the cells showed a ragged and unattached boundary resulting from the presence of surface silanol groups. For the UV-vis absorbance, all carbon modified-PS dishes showed a lower release rate of sericin molecules into PBS solution than PS control. This revealed that the functionalized carbon could be enhanced by specific binding properties with given molecules. The carbon-coated PS dishes grafting with sericin protein were used in a serum-free condition. We also found that hBM-MSCs have higher percentage of proliferated cells at day 7 for the modified dishes with carbon films and coated with sericin than the PS control coated with sericin. The physical film properties were measured by atomic force microscopy (AFM), scanning electron microscope (SEM) and contact angle measurement. The presence of sbnd NH2 groups of sericin compounds on the PS dish was revealed by Fourier transform infrared spectroscopy (FTIR). The stability of covalent bonds of sericin molecules after washing out ungrafted sericin was confirmed by X-ray photoelectron spectroscopy (XPS).
Immobilization of sericin molecules via amorphous carbon plasma modified-polystyrene dish for serum-free culture
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Tunma, Somruthai [The Graduate School, Chiang Mai University, 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand); Thailand Center of Excellence in Physics (ThEP), 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand); Song, Doo-Hoon [Research Center for Orofacial Hard Tissue Regeneration, College of Dentistry, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Kim, Si-Eun; Kim, Kyoung-Nam [Research Center for Orofacial Hard Tissue Regeneration, College of Dentistry, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Department and Research Institute of Dental Biomaterials and Bioengineering, College of Dentistry, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Han, Jeon-Geon [Center for Advanced Plasma Surface Technology, Sungkyunkwan University, 300 Chunchun-dong, Jangan-gu, Suwon 440-746 (Korea, Republic of); Boonyawan, Dheerawan [Thailand Center of Excellence in Physics (ThEP), 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand); Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand)
In this study, we focused on sericin hydrolysates, originating from silkworm used in serum-free human bone marrow-derived mesenchymal stem cells (hBM-MSCs) culture. We reported the effect of a covalent linkage between a bioactive protein molecule and polystyrene dish surface via a carbon intermediate layer which can slow down the release rate of protein compounds into the phosphate buffer saline (PBS) solution. Films of amorphous carbon (a-C) and functionalized-carbon were deposited on PS culture dish surfaces by using a DC magnetron sputtering system and RF PECVD system. We found that a-C based-films can increase the hydrophilicity and biocompatibility of polystyrene (PS) dishes, especially a-C films and a-C:N{sub 2} films showed good attachment of hBM-MSCs at 24 h. However, in the case of silica surface (a-C:SiO{sub x} films), the cells showed a ragged and unattached boundary resulting from the presence of surface silanol groups. For the UV–vis absorbance, all carbon modified-PS dishes showed a lower release rate of sericin molecules into PBS solution than PS control. This revealed that the functionalized carbon could be enhanced by specific binding properties with given molecules. The carbon-coated PS dishes grafting with sericin protein were used in a serum-free condition. We also found that hBM-MSCs have higher percentage of proliferated cells at day 7 for the modified dishes with carbon films and coated with sericin than the PS control coated with sericin. The physical film properties were measured by atomic force microscopy (AFM), scanning electron microscope (SEM) and contact angle measurement. The presence of -NH{sub 2} groups of sericin compounds on the PS dish was revealed by Fourier transform infrared spectroscopy (FTIR). The stability of covalent bonds of sericin molecules after washing out ungrafted sericin was confirmed by X-ray photoelectron spectroscopy (XPS).
Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1-6
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Mathiesen, Christian K; Jensen, Tanja B; Prentoe, Jannick
Recently, cell culture systems producing hepatitis C virus particles (HCVcc) were developed. Establishment of serum-free culture conditions is expected to facilitate development of a whole-virus inactivated HCV vaccine. We describe generation of genotype 1-6 serum-free HCVcc (sf-HCVcc) from Huh7.......5 hepatoma cells cultured in adenovirus expression medium. Compared to HCVcc, sf-HCVcc showed 0.6-2.1 log10 higher infectivity titers (4.7-6.2 log10 Focus Forming Units/mL), possibly due to increased release and specific infectivity of sf-HCVcc. In contrast to HCVcc, sf-HCVcc had a homogeneous single......-peak density profile. Entry of sf-HCVcc depended on HCV co-receptors CD81, LDLr, and SR-BI, and clathrin-mediated endocytosis. HCVcc and sf-HCVcc were neutralized similarly by chronic-phase patient sera and by human monoclonal antibodies targeting conformational epitopes. Thus, we developed serum-free culture...
Serum-free keloid fibroblast cell culture: an in vitro model for the study of aberrant wound healing.
Koch, R J; Goode, R L; Simpson, G T
The purpose of this study was to develop an in vitro serum-free keloid fibroblast model. Keloid formation remains a problem for every surgeon. Prior evaluations of fibroblast characteristics in vitro, especially those of growth factor measurement, have been confounded by the presence of serum-containing tissue culture media. The serum itself contains growth factors, yet has been a "necessary evil" to sustain cell growth. The design of this study is laboratory-based and uses keloid fibroblasts obtained from five patients undergoing facial (ear lobule) keloid removal in a university-affiliated clinic. Keloid fibroblasts were established in primary cell culture and then propagated in a serum-free environment. The main outcome measures included sustained keloid fibroblast growth and viability, which was comparable to serum-based models. The keloid fibroblast cell cultures exhibited logarithmic growth, sustained a high cellular viability, maintained a monolayer, and displayed contact inhibition. Demonstrating model consistency, there was no statistically significant difference between the mean cell counts of the five keloid fibroblast cell lines at each experimental time point. The in vitro growth of keloid fibroblasts in a serum-free model has not been done previous to this study. The results of this study indicate that the proliferative characteristics described are comparable to those of serum-based models. The described model will facilitate the evaluation of potential wound healing modulators, and cellular effects and collagen modifications of laser resurfacing techniques, and may serve as a harvest source for contaminant-free fibroblast autoimplants. Perhaps its greatest utility will be in the evaluation of endogenous and exogenous growth factors.
Evolution of triiodothyronine nuclear binding sites in hypothalamic serum-free cultures: evidence for their presence in neurons and astrocytes
International Nuclear Information System (INIS)
Puymirat, J.; Faivre-Bauman, A.
( 125 I)Triiodothyronine (T 3 ) nuclear binding was studied in hypothalamic cultures from fetal mouse grown in serum-free medium. In enriched neuronal cultures, the apparent dissociation constant of the binding does not change with time in vitro (7 x 10 -11 M), but the maximum binding capacity (MBC) doubles between day 7 and day 14 in vitro. We show here for the first time that homologous astrocyte cell cultures, devoid of neurons as checked by tetanus toxin binding, also display T 3 nuclear binding, with the same affinity as neuronal cultures. However, their MBC is 3 times lower than that of neurons after a week in vitro, and increases more quickly thereafter (Author)
Transient transfection of serum-free suspension HEK 293 cell culture for efficient production of human rFVIII
Background Hemophilia A is a bleeding disorder caused by deficiency in coagulation factor VIII. Recombinant factor VIII (rFVIII) is an alternative to plasma-derived FVIII for the treatment of hemophilia A. However, commercial manufacturing of rFVIII products is inefficient and costly and is associated to high prices and product shortage, even in economically privileged countries. This situation may be solved by adopting more efficient production methods. Here, we evaluated the potential of transient transfection in producing rFVIII in serum-free suspension HEK 293 cell cultures and investigated the effects of different DNA concentration (0.4, 0.6 and 0.8 μg/106 cells) and repeated transfections done at 34° and 37°C. Results We observed a decrease in cell growth when high DNA concentrations were used, but no significant differences in transfection efficiency and in the biological activity of the rFVIII were noticed. The best condition for rFVIII production was obtained with repeated transfections at 34°C using 0.4 μg DNA/106 cells through which almost 50 IU of active rFVIII was produced six days post-transfection. Conclusion Serum-free suspension transient transfection is thus a viable option for high-yield-rFVIII production. Work is in progress to further optimize the process and validate its scalability. PMID:22115125
The influence of serum substituents on serum-free Vero cell conditioned culture media manufactured from Dulbecco's modified Eagle medium in mouse embryo culture.
Lee, Jong-Seon; Kim, Ju-Hwan; Seo, Young-Seok; Yang, Jung-Bo; Kim, Yong-Il; Kim, Hye-Jin; Lee, Ki-Hwan
This study was conducted to examine the influences of supplementation of the serum substituents and available period of serum-free Vero cell conditioned media (SF-VCM) manufactured from Dulbecco's modified Eagle medium cultured with Vero cells for in vitro development of mouse preimplantation embryos. A total of 1,099 two-cell embryos collected from imprinting control region mice were cultured in SF-VCM with 10% and 20% human follicular fluid (hFF), serum substitute supplement (SSS), and serum protein substitute (SPS). Development of embryos was observed every 24 hours. Results between different groups were analyzed by chi-square test, and considered statistically significant when P-value was less than 0.05. The rates of embryonic development cultured in SF-VCM supplemented with serum substituents were significantly higher compare with serum-free group (P media up to 4 weeks did not affect on embryonic development.
Development of a serum-free co-culture of human intestinal epithelium cell-lines (Caco-2/HT29-5M21
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Schneider Yves-Jacques
Full Text Available Abstract Background The absorptive and goblet cells are the main cellular types encountered in the intestine epithelium. The cell lineage Caco-2 is a model commonly used to reproduce the features of the bowel epithelium. However, there is a strong debate regarding the value of Caco-2 cell culture to mimick in vivo situation. Indeed, some authors report in Caco-2 a low paracellular permeability and an ease of access of highly diffusible small molecules to the microvilli, due to an almost complete lack of mucus. The HT29-5M21 intestinal cell lineage is a mucin-secreting cellular population. A co-culture system carried out in a serum-free medium and comprising both Caco-2 and HT29-5M21 cells was developed. The systematic use of a co-culture system requires the characterization of the monolayer under a given experimental procedure. Results In this study, we investigated the activity and localization of the alkaline phosphatase and the expression of IAP and MUC5AC genes to determine a correlation between these markers and the cellular composition of a differentiated monolayer obtained from a mixture of Caco-2 and HT29-5M21 cells. We observed that the culture conditions used (serum-free medium did not change the phenotype of each cell type, and produced a reproducible model. The alkaline phosphatase expression characterizing Caco-2 cells was influenced by the presence of HT29-5M21 cells. Conclusion The culture formed by 75% Caco-2 and 25% HT29-5M21 produce a monolayer containing the two main cell types of human intestinal epithelium and characterized by a reduced permeability to macromolecules.
Development of a serum-free co-culture of human intestinal epithelium cell-lines (Caco-2/HT29-5M21)
Nollevaux, Géraldine; Devillé, Christelle; El Moualij, Benaïssa; Zorzi, Willy; Deloyer, Patricia; Schneider, Yves-Jacques; Peulen, Olivier; Dandrifosse, Guy
Background The absorptive and goblet cells are the main cellular types encountered in the intestine epithelium. The cell lineage Caco-2 is a model commonly used to reproduce the features of the bowel epithelium. However, there is a strong debate regarding the value of Caco-2 cell culture to mimick in vivo situation. Indeed, some authors report in Caco-2 a low paracellular permeability and an ease of access of highly diffusible small molecules to the microvilli, due to an almost complete lack of mucus. The HT29-5M21 intestinal cell lineage is a mucin-secreting cellular population. A co-culture system carried out in a serum-free medium and comprising both Caco-2 and HT29-5M21 cells was developed. The systematic use of a co-culture system requires the characterization of the monolayer under a given experimental procedure. Results In this study, we investigated the activity and localization of the alkaline phosphatase and the expression of IAP and MUC5AC genes to determine a correlation between these markers and the cellular composition of a differentiated monolayer obtained from a mixture of Caco-2 and HT29-5M21 cells. We observed that the culture conditions used (serum-free medium) did not change the phenotype of each cell type, and produced a reproducible model. The alkaline phosphatase expression characterizing Caco-2 cells was influenced by the presence of HT29-5M21 cells. Conclusion The culture formed by 75% Caco-2 and 25% HT29-5M21 produce a monolayer containing the two main cell types of human intestinal epithelium and characterized by a reduced permeability to macromolecules. PMID:16670004
Keratinocytes propagated in serum-free, feeder-free culture conditions fail to form stratified epidermis in a reconstituted skin model.
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Rebecca Lamb
Full Text Available Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification.
Effect of IL-6 on proliferation and IG production of human EBV-transformed cell lines in serum free culture media
Jochems, G. J.; Jordens, R.; van Lier, R. A.; Zeijlemaker, W. P.
To optimalize growth and Ig production of EBV transformed B cells for large scale tissue culture, we analyzed five stable monoclonal EBV-B cell lines for their responsiveness to interleukin (IL)-6 in standard medium with 5% FCS and in several serum-free media. As we previously demonstrated these
Enhancement of Human Prolactin Synthesis by Sodium Butyrate Addition to Serum-Free CHO Cell Culture
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Herbert Rodrigues Goulart
Full Text Available Sodium butyrate (NaBu has been used as a productivity enhancer for the synthesis of recombinant proteins in Chinese hamster ovary (CHO cells. Thus, the influence of NaBu on the production of recombinant human prolactin (hPRL from CHO cells was investigated for the first time. CHO cell cultures were submitted to a treatment with different concentrations of NaBu (0.25 to 4 mM. Quantitative and qualitative analyses by reverse-phase high-performance liquid chromatography (RP-HPLC and Western blot or SDS-PAGE, carried out directly on CHO-conditioned medium, showed that the highest hPRL expression was obtained with 1 mM NaBu. In vitro biological assays based on noble rat lymphoma (Nb2 and mouse pro-B lymphoma (Ba/F3-LLP cells were carried out on purified hPRL. Its bioactivity in the presence of NaBu was not apparently different from that of the First International Reference Reagent of recombinant hPRL (WHO 97/714. Our results show that NaBu increased the synthesis of recombinant hPRL in CHO cells, apparently without compromising either its structure or function.
Defined culture medium for stem cell differentiation: applicability of serum-free conditions in the mouse embryonic stem cell test.
Riebeling, Christian; Schlechter, Katharina; Buesen, Roland; Spielmann, Horst; Luch, Andreas; Seiler, Andrea
The embryonic stem cell test (EST) is a validated method to assess the developmental toxicity potency of chemicals. It was developed to reduce animal use and allow faster testing for hazard assessment. The cells used in this method are maintained and differentiated in media containing foetal calf serum. This animal product is of considerable variation in quality, and individual batches require extensive testing for their applicability in the EST. Moreover, its production involves a large number of foetuses and possible animal suffering. We demonstrate the serum-free medium and feeder cell-free maintenance of the mouse embryonic stem cell line D3 and investigate the use of specific growth factors for induction of cardiac differentiation. Using a combination of bone morphogenetic protein-2, bone morphogenetic protein-4, activin A and ascorbic acid, embryoid bodies efficiently differentiated into contracting myocardium. Additionally, examining levels of intracellular marker proteins by flow cytometry not only confirmed differentiation into cardiomyocytes, but demonstrated significant differentiation into neuronal cells in the same time frame. Thus, this approach might allow for simultaneous detection of developmental effects on both early mesodermal and neuroectodermal differentiation. The serum-free conditions for maintenance and differentiation of D3 cells described here enhance the transferability and standardisation and hence the performance of the EST. Copyright © 2011 Elsevier Ltd. All rights reserved.
Generation of hematopoietic stem cells from human embryonic stem cells using a defined, stepwise, serum-free, and serum replacement-free monolayer culture method.
Kim, So-Jung; Jung, Ji-Won; Ha, Hye-Yeong; Koo, Soo Kyung; Kim, Eung-Gook; Kim, Jung-Hyun
Embryonic stem cells (ESCs) can be expanded infinitely in vitro and have the potential to differentiate into hematopoietic stem cells (HSCs); thus, they are considered a useful source of cells for HSC production. Although several technical in vitro methods for engineering HSCs from pluripotent stem cells have been developed, clinical application of HSCs engineered from pluripotent stem cells is restricted because of the possibility of xenogeneic contamination resulting from the use of murine materials. Human ESCs (CHA-hES15) were cultured on growth factor-reduced Matrigel-coated dishes in the mTeSR1 serum-free medium. When the cells were 70% confluent, we initiated HSC differentiation by three methods involving (1) knockout serum replacement (KSR), cytokines, TGFb1, EPO, and FLT3L; (2) KSR, cytokines, and bFGF; or (3) cytokines and bFGF. Among the three differentiation methods, the minimal number of cytokines without KSR resulted in the greatest production of HSCs. The optimized method resulted in a higher proportion of CD34 + CD43 + hematopoietic progenitor cells (HPCs) and CD34 + CD45 + HPCs compared to the other methods. In addition, the HSCs showed the potential to differentiate into multiple lineages of hematopoietic cells in vitro . In this study, we optimized a two-step, serum-free, animal protein-free, KSR-free, feeder-free, chemically defined monolayer culture method for generation of HSCs and hematopoietic stem and progenitor cells (HSPCs) from human ESCs.
Bovine babesiosis: Cattle protected in the field with a frozen vaccine containing Babesia bovis and Babesia bigemina cultured in vitro with a serum-free medium.
Rojas-Martínez, Carmen; Rodríguez-Vivas, Roger Iván; Millán, Julio Vicente Figueroa; Bautista-Garfias, Carlos Ramón; Castañeda-Arriola, Roberto Omar; Lira-Amaya, José Juan; Urióstegui, Patricia Vargas; Carrasco, Juan José Ojeda; Martínez, Jesús Antonio Álvarez
An attenuated live vaccine containing Babesia bovis and B. bigemina cultured in vitro with a serum-free medium was assessed for its clinical protection conferred of naïve cattle, under natural tick-challenge in a high endemicity zone to Babesia spp. Three groups of six animals were treated as follows: group I (GI) received a vaccine derived from parasites cultured with a free-serum medium; group II (GII) were immunized with the standard vaccine, with parasites cultured in a medium supplemented with 40% (v/v) bovine serum; and a control group (GIII) inoculated with non-infected bovine erythrocytes. Inocula were administered by IM route. Experimental animals were kept during 23days after vaccination in a cattle farm free of ticks and Babesia spp. Thereafter, cattle were moved to a high endemicity farm for natural exposure to Babesia spp. transmitted by Rhipicephalus microplus ticks. Protection against clinical babesiosis was observed in bovines belonging to GI (100%) and GII (83.33%), while the control animals (GIII) were not protected, and showed severe clinical signs, closely related to babesiosis, were observed for at least three consecutive days during the challenge. These were fever, anemia, which were measured simultaneously, and circulating parasites were detected by optic light microscopy. All cattle showed B. bovis and B. bigemina in stained blood films during the challenge; B. bovis antibody titers were higher than those to B. bigemina in GI and GII, and lower titers were determined in GIII. The protective capacity of the vaccine derived from B. bovis and B. bigemina cultured in vitro in a serum-free medium was demonstrated. Copyright © 2017 Elsevier B.V. All rights reserved.
Prognostic value of the serum free light chain ratio in newly diagnosed myeloma: proposed incorporation into the international staging system.
Snozek, C L H; Katzmann, J A; Kyle, R A; Dispenzieri, A; Larson, D R; Therneau, T M; Melton, L J; Kumar, S; Greipp, P R; Clark, R J; Rajkumar, S V
To determine if the serum free light chain (FLC) ratio has prognostic value in patients with symptomatic multiple myeloma (MM), baseline serum samples from a well-characterized cohort of 790 newly diagnosed MM patients were tested with the FLC assay. FLC ratio was calculated as kappa/lambda (reference range 0.26-1.65). On the basis of the distribution of values, a cutpoint kappa/lambda FLC ratio of 32 was chosen for further analysis. Overall survival was significantly inferior in patients with an abnormal FLC ratio of 32 (n=479) compared with those with an FLC ratio between 0.03 and 32 (n=311), with median survival of 30 versus 39 months, respectively. We incorporated abnormal FLC ratio with the International Staging System (ISS) risk factors (that is, albumin or=3.5 g/l), to create a risk stratification model with improved prognostic capabilities. Patients with 0, 1, 2 or 3 adverse risk factors had significantly different overall survival, with median survival times of 51, 39, 30 and 22 months, respectively (P<0.001). These findings suggest that the serum FLC ratio at initial diagnosis is an important predictor of prognosis in myeloma, and can be incorporated into the ISS for improved risk stratification.
Specific estrogen-induced cell proliferation of cultured Syrian hamster renal proximal tubular cells in serum-free chemically defined media
International Nuclear Information System (INIS)
Oberley, T.D.; Lauchner, L.J.; Pugh, T.D.; Gonzalez, A.; Goldfarb, S.; Li, S.A.; Li, J.J.
It has long been recognized that the renal proximal tubular epithelium of the hamster is a bona fide estrogen target tissue. The effect of estrogens on the growth of proximal tubule cell explants and dissociated single cells derived from these explant outgrowths has been studied in culture. Renal tubular cells were grown on a PF-HR-9 basement membrane under serum-free chemically defined culture conditions. At 7-14 days in culture, cell number was enhanced 3-fold in the presence of either 17β-estradiol or diethylstilbestrol. A similar 3-fold increase in cell number was also seen at 1 nM 17β-estradiol in subcultured dissociated single tubular cells derived from hamster renal tubular explant outgrowths at 21 days in culture. Concomitant exposure of tamoxifen at 3-fold molar excess in culture completely abolished the increase in cell number seen with 17β-estradiol. The proliferation effect of estrogens on proximal tubular cell growth appears to be species specific since 17β-estradiol did not alter the growth of either rat or guinea pig proximal tubules in culture. In addition, at 7-10 days in culture in the presence of 17β-estradiol, [ 3 H]thymidine labeling of hamster tubular cells was enhanced 3-fold. These results clearly indicate that estrogens can directly induce primary epithelial cell proliferation at physiologic concentrations and provide strong additional evidence for an important hormonal role in the neoplastic transformation of the hamster kidney
Optimization of a serum-free culture medium for mouse embryonic stem cells using design of experiments (DoE) methodology.
Knöspel, Fanny; Schindler, Rudolf K; Lübberstedt, Marc; Petzolt, Stephanie; Gerlach, Jörg C; Zeilinger, Katrin
The in vitro culture behaviour of embryonic stem cells (ESC) is strongly influenced by the culture conditions. Current culture media for expansion of ESC contain some undefined substances. Considering potential clinical translation work with such cells, the use of defined media is desirable. We have used Design of Experiments (DoE) methods to investigate the composition of a serum-free chemically defined culture medium for expansion of mouse embryonic stem cells (mESC). Factor screening analysis according to Plackett-Burman revealed that insulin and leukaemia inhibitory factor (LIF) had a significant positive influence on the proliferation activity of the cells, while zinc and L: -cysteine reduced the cell growth. Further analysis using minimum run resolution IV (MinRes IV) design indicates that following factor adjustment LIF becomes the main factor for the survival and proliferation of mESC. In conclusion, DoE screening assays are applicable to develop and to refine culture media for stem cells and could also be employed to optimize culture media for human embryonic stem cells (hESC).
Effects of Macromolecular Crowding on Human Adipose Stem Cell Culture in Fetal Bovine Serum, Human Serum, and Defined Xeno-Free/Serum-Free Conditions.
Patrikoski, Mimmi; Lee, Michelle Hui Ching; Mäkinen, Laura; Ang, Xiu Min; Mannerström, Bettina; Raghunath, Michael; Miettinen, Susanna
Microenvironment plays an important role for stem cell proliferation and differentiation. Macromolecular crowding (MMC) was recently shown to assist stem cells in forming their own matrix microenvironment in vitro. The ability of MMC to support adipose stem cell (ASC) proliferation, metabolism, and multilineage differentiation was studied under different conditions: fetal bovine serum- (FBS-) and human serum- (HS-) based media and xeno- and serum-free (XF/SF) media. Furthermore, the immunophenotype of ASCs under MMC was evaluated. The proliferative capacity of ASCs under MMC was attenuated in each condition. However, osteogenic differentiation was enhanced under MMC, shown by increased deposition of mineralized matrix in FBS and HS cultures. Likewise, significantly greater lipid droplet accumulation and increased collagen IV deposition indicated enhanced adipogenesis under MMC in FBS and HS cultures. In contrast, chondrogenic differentiation was attenuated in ASCs expanded under MMC. The ASC immunophenotype was maintained under MMC with significantly higher expression of CD54. However, MMC impaired metabolic activity and differentiation capacity of ASCs in XF/SF conditions. Both the supportive and inhibitory effects of MMC on ASC are culture condition dependent. In the presence of serum, MMC maintains ASC immunophenotype and enhances adipogenic and osteogenic differentiation at the cost of reduced proliferation.
Short-term, serum-free, static culture of cord blood-derived CD34+ cells: effects of FLT3-L and MIP-1alpha on in vitro expansion of hematopoietic progenitor cells.
Capmany, G; Querol, S; Cancelas, J A; García, J
The use of ex vivo expanded cells has been suggested as a possible means to accelerate the speed of engraftment in cord blood (CB) transplantation. The aim of this study was to fix the optimal condition for the generation of committed progenitors without affecting the stem cell compartment. Analysis of the effects of FLT3-L and MIP-1alpha when combined with SCF, IL-3 and IL-6, in short-term (6 days), serum-free expansion cultures of CB-selected CD34+ cells. An important expansion was obtained that ranged between 8-15 times for CFU-GM, 21-51 times for the BFU-E/CFU-Mix population and 11 to 30 times for CD34+ cells assessed by flow cytometry. From the combinations tested, those in which FLT3-L was present had a significant increase in the expansion of committed progenitors, while the presence of MIP-1alpha had a detrimental effect on the generation of more differentiated cells. However, stem cell candidates assessed by week 5 CAFC assay could be maintained in culture when both MIP-1a and FLT3-L were present (up to 91% recovery). This culture system was also able to expand megakaryocytic precursors as determined by the co-expression of CD34 and CD61 antigens (45-70 times), in spite of the use of cytokines non-specific for the megakaryocytic lineage. The results obtained point to the combination of SCF, IL-3, IL-6, FLT3-L and MIP-1alpha as the best suited for a pre-clinical short-term serum-free static ex vivo expansion protocol of CB CD34+ cells, since it can generate large numbers of committed progenitor cells as well as maintaining week 5 CAFC.
Immunohistochemical expression of stem cell, endothelial cell, and chemosensitivity markers in primary glioma spheroids cultured in serum-containing and serum-free medium
DEFF Research Database (Denmark)
Christensen, Karina; Aaberg-Jessen, Charlotte; Andersen, Claus
To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas.......To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas....
Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.
Kaneko, Ai; Sankai, Yoshiyuki
The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months) primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4) cells/mL (8.9×10(3) cells/cm2) without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm), greater cell viability (≥30%) for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks).
Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.
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Ai Kaneko
Full Text Available The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4 cells/mL (8.9×10(3 cells/cm2 without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm, greater cell viability (≥30% for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks.
Growth in serum-free medium improves isolation of Chlamydia pneumoniae.
Maass, M; Essig, A; Marre, R; Henkel, W
Infectivity titers were determined for eight Chlamydia pneumoniae strains simultaneously grown in serum-free and serum-supplemented cell culture media. Use of serum-free medium resulted in a 10- to 50-fold increase in the susceptibility of HL cells to chlamydial infection. Comparative primary isolation of a wild-type strain also produced higher inclusion counts in a serum-free environment. Serum-free cultivation is recommended to increase the efficiency of C. pneumoniae isolation from clinica...
Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells.
Naskou, Maria C; Sumner, Scarlett M; Chocallo, Anna; Kemelmakher, Hannah; Thoresen, Merrilee; Copland, Ian; Galipeau, Jacques; Peroni, John F
Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) pooled from donor horses in place of FBS to culture equine MSCs. We hypothesized that ePL, produced following apheresis, will function as the sole media supplement to accelerate the expansion of equine bone marrow-derived MSCs without altering their phenotype and their immunomodulatory capacity. Platelet concentrate was obtained via plateletpheresis and ePL were produced via freeze-thaw and centrifugation cycles. Population doublings (PD) and doubling time (DT) of bone marrow-derived MSCs (n = 3) cultured with FBS or ePL media were calculated. Cell viability, immunophenotypic analysis, and trilineage differentiation capacity of MSCs were assessed accordingly. To assess the ability of MSCs to modulate inflammatory responses, E. coli lipopolysaccharide (LPS)-stimulated monocytes were cocultured with MSCs cultured in the two different media formulations, and cell culture supernatants were assayed for the production of tumor necrosis factor (TNF)-α. Our results showed that MSCs cultured in ePL media exhibited similar proliferation rates (PD and DT) compared with those cultured in FBS at individual time points. MSCs cultured in ePL showed a statistically significant increased viability following a single washing step, expressed similar levels of MSC markers compared to FBS, and were able to differentiate towards the three lineages. Finally, MSCs cultured in ePL efficiently suppressed
Inactivated Enterovirus 71 Vaccine Produced by 200-L Scale Serum-Free Microcarrier Bioreactor System Provides Cross-Protective Efficacy in Human SCARB2 Transgenic Mouse.
Wu, Chia-Ying; Lin, Yi-Wen; Kuo, Chia-Ho; Liu, Wan-Hsin; Tai, Hsiu-Fen; Pan, Chien-Hung; Chen, Yung-Tsung; Hsiao, Pei-Wen; Chan, Chi-Hsien; Chang, Ching-Chuan; Liu, Chung-Cheng; Chow, Yen-Hung; Chen, Juine-Ruey
Epidemics and outbreaks caused by infections of several subgenotypes of EV71 and other serotypes of coxsackie A viruses have raised serious public health concerns in the Asia-Pacific region. These concerns highlight the urgent need to develop a scalable manufacturing platform for producing an effective and sufficient quantity of vaccines against deadly enteroviruses. In this report, we present a platform for the large-scale production of a vaccine based on the inactivated EV71(E59-B4) virus. The viruses were produced in Vero cells in a 200 L bioreactor with serum-free medium, and the viral titer reached 10(7) TCID50/mL 10 days after infection when using an MOI of 10(-4). The EV71 virus particles were harvested and purified by sucrose density gradient centrifugation. Fractions containing viral particles were pooled based on ELISA and SDS-PAGE. TEM was used to characterize the morphologies of the viral particles. To evaluate the cross-protective efficacy of the EV71 vaccine, the pooled antigens were combined with squalene-based adjuvant (AddaVAX) or aluminum phosphate (AlPO4) and tested in human SCARB2 transgenic (Tg) mice. The Tg mice immunized with either the AddaVAX- or AlPO4-adjuvanted EV71 vaccine were fully protected from challenges by the subgenotype C2 and C4 viruses, and surviving animals did not show any degree of neurological paralysis symptoms or muscle damage. Vaccine treatments significantly reduced virus antigen presented in the central nervous system of Tg mice and alleviated the virus-associated inflammatory response. These results strongly suggest that this preparation results in an efficacious vaccine and that the microcarrier/bioreactor platform offers a superior alternative to the previously described roller-bottle system.
Purification and characterization of enterovirus 71 viral particles produced from vero cells grown in a serum-free microcarrier bioreactor system.
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Chia-Chyi Liu
Full Text Available BACKGROUND: Enterovirus 71 (EV71 infections manifest most commonly as a childhood exanthema known as hand-foot-and-mouth disease (HFMD and can cause neurological disease during acute infection. PRINCIPAL FINDING: In this study, we describe the production, purification and characterization of EV71 virus produced from Vero cells grown in a five-liter serum-free bioreactor system containing 5 g/L Cytodex 1 microcarrier. The viral titer was >10(6 TCID(50/mL by 6 days post infection when a MOI of 10(-5 was used at the initial infection. Two EV71 virus fractions were separated and detected when the harvested EV71 virus concentrate was purified by sucrose gradient zonal ultracentrifugation. The EV71 viral particles detected in the 24-28% sucrose fractions had an icosahedral structure 30-31 nm in diameter and had low viral infectivity and RNA content. Three major viral proteins (VP0, VP1 and VP3 were observed by SDS-PAGE. The EV71 viral particles detected in the fractions containing 35-38% sucrose were 33-35 nm in size, had high viral infectivity and RNA content, and were composed of four viral proteins (VP1, VP2, VP3 and VP4, as shown by SDS-PAGE analyses. The two virus fractions were formalin-inactivated and induced high virus neutralizing antibody responses in mouse immunogenicity studies. Both mouse antisera recognized the immunodominant linear neutralization epitope of VP1 (residues 211-225. CONCLUSION: These results provide important information for cell-based EV71 vaccine development, particularly for the preparation of working standards for viral antigen quantification.
Wnt3a protein reduces growth factor-driven expansion of human hematopoietic stem and progenitor cells in serum-free cultures
L.E. Duinhouwer (Lucia); N. Tüysüz (Nesrin); E.J. Rombouts (Elwin); M.N.D. Ter Borg (Mariëtte N. D.); E. Mastrobattista; J. Spanholtz (Jan); J.J. Cornelissen (Jan); D. ten Berge (Derk); E. Braakman (Eric)
textabstractAbstract Ex vivo expansion of hematopoietic stem and progenitor cells (HSPC) is a promising approach to improve insufficient engraftment after umbilical cord blood stem cell transplantation (UCB-SCT). Although culturing HSPC with hematopoietic cytokines results in
Development of serum-free media for lepidopteran insect cell lines.
Agathos, Spiros N
Lepidopteran insect cell culture technology has progressed to the point of becoming an essential part of one of the most successful eukaryotic expression systems and is increasingly used industrially on a large scale. Therefore, there is a constant need for convenient and low-cost culture media capable of supporting good insect cell growth and ensuring high yield of baculovirus as well as the strong expression of recombinant proteins. Vertebrate sera or invertebrate hemolymph were essential supplements in first-generation insect cell media. These supplements, however, are cumbersome and expensive for routine large-scale culture; thus, their use is now circumvented by substituting the essential growth factors present in these supplements with serum-free substances. Such non-serum supplements are typically of non-animal origin and include protein hydrolysates, lipid emulsions, and specialized substances (e.g., surfactants and shear damage protecting chemicals). These supplements need to complement the defined, synthetic basal medium to ensure that the fundamental nutritional needs of the cells are satisfied. Although there is a significant number of proprietary serum-free and low-protein or protein-free media on the market, the lack of information concerning their detailed composition is a drawback in their adoption for different applications, including their adaptation to the metabolic and kinetic analysis and monitoring of a given insect cell based bioprocess. Hence, there is wide appeal for formulating serum-free media based on a rational assessment of the metabolic requirements of the lepidopteran cells during both the growth and the production phases. Techniques such as statistical experimental design and genetic algorithms adapted to the cellular behavior and the bioreactor operation mode (batch, fed-batch, or perfusion) permit the formulation of versatile serum- and protein-free media. These techniques are illustrated with recent developments of serum-free
Wnt3a protein reduces growth factor-driven expansion of human hematopoietic stem and progenitor cells in serum-free cultures
Duinhouwer, Lucia E.; Tüysüz, Nesrin; Rombouts, Elwin W J C; Ter Borg, Mariette N D; Mastrobattista, Enrico; Spanholtz, Jan; Cornelissen, Jan J.; Berge, Derk Ten; Braakman, Eric
Ex vivo expansion of hematopoietic stem and progenitor cells (HSPC) is a promising approach to improve insufficient engraftment after umbilical cord blood stem cell transplantation (UCB-SCT). Although culturing HSPC with hematopoietic cytokines results in robust proliferation, it is accompanied with
Isolation and characterization of Wharton's jelly-derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum-free three-dimensional system
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Cardoso Tereza C
Full Text Available Abstract Background The possibility for isolating bovine mesenchymal multipotent cells (MSCs from fetal adnexa is an interesting prospect because of the potential for these cells to be used for biotechnological applications. Bone marrow and adipose tissue are the most common sources of MSCs derived from adult animals. However, little knowledge exists about the characteristics of these progenitors cells in the bovine species. Traditionally most cell cultures are developed in two dimensional (2D environments. In mammalian tissue, cells connect not only to each other, but also support structures called the extracellular matrix (ECM. The three-dimensional (3D cultures may play a potential role in cell biotechnology, especially in tissue therapy. In this study, bovine-derived umbilical cord Wharton's jelly (UC-WJ cells were isolated, characterized and maintained under 3D-free serum condition as an alternative of stem cell source for future cell banking. Results Bovine-derived UC-WJ cells, collected individually from 5 different umbilical cords sources, were successfully cultured under serum-free conditions and were capable to support 60 consecutive passages using commercial Stemline® mesenchymal stem cells expansion medium. Moreover, the UC-WJ cells were differentiated into osteocytes, chondrocytes, adipocytes and neural-like cells and cultured separately. Additionally, the genes that are considered important embryonic, POU5F1 and ITSN1, and mesenchymal cell markers, CD105+, CD29+, CD73+ and CD90+ in MSCs were also expressed in five bovine-derived UC-WJ cultures. Morphology of proliferating cells typically appeared fibroblast-like spindle shape presenting the same viability and number. These characteristics were not affected during passages. There were 60 chromosomes at the metaphase, with acrocentric morphology and intense telomerase activity. Moreover, the proliferative capacity of T cells in response to a mitogen stimulus was suppressed when
Shift of microRNA profile upon glioma cell migration using patient-derived spheroids and serum-free conditions
DEFF Research Database (Denmark)
Munthe, Sune; Halle, Bo; Boldt, Henning B
Glioblastoma multiforme (GBM) is the most frequent malignant primary brain tumor. A major reason for the overall median survival being only 14.6 months is migrating tumor cells left behind after surgery. Another major reason is tumor cells having a so-called cancer stem cell phenotype being...... therefore resistant towards traditional chemo- and radiotherapy. A group of novel molecular targets are microRNAs (miRNAs). MiRNAs are small non-coding RNAs exerting post-transcriptional regulation of gene expression. The aim of this study was to identify differentially expressed miRNAs in migrating GBM...... cells using serum-free stem cell conditions. We used patient-derived GBM spheroid cultures for a novel serum-free migration assay. MiRNA expression of migrating tumor cells isolated at maximum migration speed was compared with corresponding spheroids using an OpenArray Real-Time PCR System. The mi...
Growth regulation on human acute myeloid leukemia effects of five recombinant hematopoietic factors in a serum-free culture system
Delwel, E.; Salem, M.; Pellens, C.; Dorssers, L.; Wagemaker, G.; Clark, S.; Loewenberg, B
The response of human acute myeloid leukemia (AML) cells to the distinct hematopoietic growth factors (HGFs), ie, recombinant interleukin-3 (IL-3), granulocyte-macrophage-CSF (GM-CSF), granulocyte-CSF (G-CSF), macrophage-CSF (M-CSF), and erythropoietin (Epo) was investigated under well-defined
Serum-free Erythroid Differentiation for Efficient Genetic Modification and High-Level Adult Hemoglobin Production.
Uchida, Naoya; Demirci, Selami; Haro-Mora, Juan J; Fujita, Atsushi; Raines, Lydia N; Hsieh, Matthew M; Tisdale, John F
In vitro erythroid differentiation from primary human cells is valuable to develop genetic strategies for hemoglobin disorders. However, current erythroid differentiation methods are encumbered by modest transduction rates and high baseline fetal hemoglobin production. In this study, we sought to improve both genetic modification and hemoglobin production among human erythroid cells in vitro . To model therapeutic strategies, we transduced human CD34 + cells and peripheral blood mononuclear cells (PBMCs) with lentiviral vectors and compared erythropoietin-based erythroid differentiation using fetal-bovine-serum-containing media and serum-free media. We observed more efficient transduction (85%-93%) in serum-free media than serum-containing media (20%-69%), whereas the addition of knockout serum replacement (KSR) was required for serum-free media to promote efficient erythroid differentiation (96%). High-level adult hemoglobin production detectable by electrophoresis was achieved using serum-free media similar to serum-containing media. Importantly, low fetal hemoglobin production was observed in the optimized serum-free media. Using KSR-containing, serum-free erythroid differentiation media, therapeutic adult hemoglobin production was detected at protein levels with β-globin lentiviral transduction in both CD34 + cells and PBMCs from sickle cell disease subjects. Our in vitro erythroid differentiation system provides a practical evaluation platform for adult hemoglobin production among human erythroid cells following genetic manipulation.
A defined, feeder-free, serum-free system to generate in vitro hematopoietic progenitors and differentiated blood cells from hESCs and hiPSCs.
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Giorgia Salvagiotto
Full Text Available Human ESC and iPSC are an attractive source of cells of high quantity and purity to be used to elucidate early human development processes, for drug discovery, and in clinical cell therapy applications. To efficiently differentiate pluripotent cells into a pure population of hematopoietic progenitors we have developed a new 2-dimensional, defined and highly efficient protocol that avoids the use of feeder cells, serum or embryoid body formation. Here we showed that a single matrix protein in combination with growth factors and a hypoxic environment is sufficient to generate from pluripotent cells hematopoietic progenitors capable of differentiating further in mature cell types of different lineages of the blood system. We tested the differentiation method using hESCs and 9 iPSC lines generated from different tissues. These data indicate the robustness of the protocol providing a valuable tool for the generation of clinical-grade hematopoietic cells from pluripotent cells.
Rabbit chondrocytes maintained in serum-free medium. I. Synthesis and secretion of hydrodynamically-small proteoglycans
International Nuclear Information System (INIS)
Malemud, C.J.; Papay, R.S.
The biosynthesis of sulfated proteoglycan in vitro by rabbit articular chondrocytes in first passage monolayer culture maintained in fetal bovine serum (FBS) or in serum-free conditions was compared. Neosynthesized proteoglycan in the culture medium in the most dense fraction of an associative CsCl density gradient (fraction dAl) declined with increasing time under serum-free conditions, but not when cells were maintained in the presence of serum. After one day, the major peak of incorporated 35 SO 4 in medium fraction dAl eluted as a retarded peak on Sepharose CL-2B, whether cells were maintained under serum-free or serum-containing conditions. The hydrodynamic size of proteoglycan monomer fraction dAlDl obtained after one day of exposure to serum-free culture media was smaller than dAlDl from serum-containing cultures. The hydrodynamic size of dAlDl obtained from serum-free culture media became even progressively smaller after 2 and 3 days' exposure to these conditions. Hydrodynamically small sulfated proteoglycans were identified in the cell-associated dAlDl fraction as early as one day after switching chondrocytes from serum-containing to serum-free medium. Proteoglycan monomer from serum-containing medium reaggregated more efficiently under both conditions. No change in the size of glycosaminoglycan chains was seen in the smaller proteoglycan subpopulations, nor was there any indication of marked changes in the glycosaminoglycan types
Serum-Free Cryopreservation of Five Mammalian Cell Lines in Either a Pelleted or Suspended State
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Corsini Joe
Full Text Available Herein we have explored two practical aspects of cryopreserving cultured mammalian cells during routine laboratory maintenance. First, we have examined the possibility of using a serum-free, hence more affordable, cryopreservative. Using five mammalian lines (Crandell Feline Kidney, MCF7, A72, WI 38 and NB324K, we found that the serum-free alternative preserves nearly as efficiently as the serum-containing preservatives. Second, we compared cryostorage of those cells in suspended versus a pellet form using both aforementioned cryopreservatives. Under our conditions, cells were in general recovered equally well in a suspended versus a pellet form.
Serum-Free Media and the Immunoregulatory Properties of Mesenchymal Stem Cells In Vivo and In Vitro
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Mei Wu
Full Text Available Background: Mesenchymal stem cells are capable of self-renewal and multi-lineage differentiation. They are used extensively to treat several diseases. Traditionally, mesenchymal stem cells are cultured in serum-containing media, typically supplemented with fetal bovine serum (FBS. However, the variability of FBS is likely to skew experimental results. Although serum-free media used to expand mesenchymal stem cells has facilitated remarkable achievements, immunomodulation of these cells in under serum-free conditions is poorly understood. We hypothesized that mesenchymal stem cells expanded in serum-free media will retain powerful immunoregulatory functions in vitro and in vivo. Design and Methods: Immunosuppressive activity and the immunomodulatory cytokines produced by mesenchymal stem cells in serum-free media were characterized in vitro. Immunomodulation by serum-free mesenchymal stem cell expansion in monocrotaline-induced pulmonary hypertension was explored in vivo. Results: Similar to cells in serum-containing media, mesenchymal stem cells expanded in serum-free media inhibited proliferation and apoptosis of CD4+T cells. They also exhibited strong immunosuppressive activities and secreted high levels of immunomodulatory cytokines such as PGE2, IDO1, COX2, IL-6, and IL-1β, but not HGF. On the other hand, growth of mesenchymal stem cells in serum-free media attenuated pulmonary vascular remodeling and inhibited mRNA expression of proinflammatory cytokines TNF-α, IFN-γ, IL-6, IL-1β, and IL-18. Conclusions: Mesenchymal stem cells in serum-free media maintained powerful immunomodulatory function in vitro and in vivo; serum-free media may replace serum-containing media for basic research and clinical applications.
Production of high titer attenuated poliovirus strains on the serum-free PER.C6(®) cell culture platform for the generation of safe and affordable next generation IPV.
Sanders, Barbara P; Oakes, Isabel de los Rios; van Hoek, Vladimir; Liu, Ying; Marissen, Wilfred; Minor, Philip D; Wimmer, Eckard; Schuitemaker, Hanneke; Custers, Jerome H H V; Macadam, Andrew; Cello, Jeronimo; Edo-Matas, Diana
As poliovirus eradication draws closer, alternative Inactivated Poliovirus Vaccines (IPV) are needed to overcome the risks associated with continued use of the Oral Poliovirus Vaccine and of neurovirulent strains used during manufacture of conventional (c) IPV. We have previously demonstrated the susceptibility of the PER.C6(®) cell line to cIPV strains; here we investigated the suspension cell culture platform for growth of attenuated poliovirus strains. We examined attenuated Sabin strain productivity on the PER.C6(®) cell platform compared to the conventional Vero cell platform. The suitability of the suspension cell platform for propagation of rationally-attenuated poliovirus strains (stabilized Sabin type 3 S19 derivatives and genetically attenuated and stabilized MonoCre(X) strains), was also assessed. Yields were quantified by infectious titer determination and D-antigen ELISA using either serotype-specific polyclonal rabbit sera for Sabin strains or monoclonal cIPV-strain-specific antibodies for cIPV, S19 and MonoCre(X) strains. PER.C6(®) cells supported the replication of Sabin strains to yields of infectious titers that were in the range of cIPV strains at 32.5°C. Sabin strains achieved 30-fold higher yields (pSabin strain productivity on the PER.C6(®) cell platform was maintained at 10l scale. Yields of infectious titers of S19 and MonoCre(X) strains were 0.5-1 log10 lower than seen for cIPV strains, whereas D-antigen yield and productivities in doses/ml using rationally-attenuated strains were in line with yields reported for cIPV strains. Sabin and rationally-attenuated polioviruses can be grown to high infectious titers and D-antigen yields. Sabin strain infection shows increased productivity on the PER.C6(®) cell platform as compared to the conventional Vero cell platform. Novel cell platforms with the potential for higher yields could contribute to increased affordability of a next generation of IPV vaccines needed for achieving and
Attachment and growth of human keratinocytes in a serum-free environment.
Gilchrest, B A; Calhoun, J K; Maciag, T
Using a serum-free system, we have investigated the influence of human fibronectin (HFN) and selected growth factors (GF) on the attachment and growth of normal human keratinocytes in vitro. Single-cell suspensions of keratinocytes from near-confluent primary plates, plated on 5-10 microgram/cm2 HFN, showed approximately 30-40% attachment after 2-24 hours of incubation at 37 degrees C, compared with 4-6% attachment on uncoated platic plates. Percentage of attached cells was independent of seed density, tissue donor age, in vitro culture age, or medium composition, while subsequent cellular proliferation was strongly dependent on these factors. Keratinocytes grown on an adequate HFN matrix in a previously described hormone-supplemented medium (Maciag et al., 1981a) achieved four to eight population doubling over 7-12 days at densities greater than or equal to 104 cell/cm2. Removal of most GF individually from the medium had little or no effect on growth, while removal of epidermal growth factor (EGF) alone reduced growth by 30-35% and removal of bovine brain extract (BE) alone reduced growth by approximately 90%. Conversely, EGF alone in basal medium supported approximately 10% control growth, BE alone supported 30-40% control growth, and the combination of EGF and BE approximately 70%. In addition to its major effect on proliferation in this system, BE was necessary to preserve normal keratinocyte morphology and protein production. These findings expand earlier observations that HFN facilitates keratinocyte attachment in vitro and that a brain-derived extract can exert a major positive influence on cultured keratinocytes.
Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells
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Yan Mylene L
Full Text Available Abstract Background Influenza virus is a major health concern that has huge impacts on the human society, and vaccination remains as one of the most effective ways to mitigate this disease. Comparing the two types of commercially available Influenza vaccine, the live attenuated virus vaccine is more cross-reactive and easier to administer than the traditional inactivated vaccines. One promising live attenuated Influenza vaccine that has completed Phase I clinical trial is deltaFLU, a deletion mutant lacking the viral Nonstructural Protein 1 (NS1 gene. As a consequence of this gene deletion, this mutant virus can only propagate effectively in cells with a deficient interferon-mediated antiviral response. To demonstrate the manufacturability of this vaccine candidate, a batch bioreactor production process using adherent Vero cells on microcarriers in commercially available animal-component free, serum-free media is described. Results Five commercially available animal-component free, serum-free media (SFM were evaluated for growth of Vero cells in agitated Cytodex 1 spinner flask microcarrier cultures. EX-CELL Vero SFM achieved the highest cell concentration of 2.6 × 10^6 cells/ml, whereas other SFM achieved about 1.2 × 10^6 cells/ml. Time points for infection between the late exponential and stationary phases of cell growth had no significant effect in the final virus titres. A virus yield of 7.6 Log10 TCID50/ml was achieved using trypsin concentration of 10 μg/ml and MOI of 0.001. The Influenza vaccine production process was scaled up to a 3 liter controlled stirred tank bioreactor to achieve a cell density of 2.7 × 10^6 cells/ml and virus titre of 8.3 Log10 TCID50/ml. Finally, the bioreactor system was tested for the production of the corresponding wild type H1N1 Influenza virus, which is conventionally used in the production of inactivated vaccine. High virus titres of up to 10 Log10 TCID50/ml were achieved. Conclusions We describe for the
Serum-Free Media and the Immunoregulatory Properties of Mesenchymal Stem Cells In Vivo and In Vitro
Mei Wu; Zhi-Bo Han; Jun Feng Liu; You Wei Wang; Jian Zhong Zhang; Chun Tuan Li; Peng Liang Xin; Zhong Chao Han; Xiong Peng Zhu
Background: Mesenchymal stem cells are capable of self-renewal and multi-lineage differentiation. They are used extensively to treat several diseases. Traditionally, mesenchymal stem cells are cultured in serum-containing media, typically supplemented with fetal bovine serum (FBS). However, the variability of FBS is likely to skew experimental results. Although serum-free media used to expand mesenchymal stem cells has facilitated remarkable achievements, immunomodulation of these cells in un...
The B-domain of factor VIII reduces cell membrane attachement to host cells in serum free conditions
DEFF Research Database (Denmark)
Kolind, Mille Petersen; Nørby, Peder Lisby; Flintegaard, Thomas Veje
engineered extensively throughout the years to increase the low production yields that initially were obtained from mammalian cell cultures. The scope of this work was to investigate the interaction of rFVIII with the cell membrane surface of the producing cells in serum free medium. We wondered whether...... binding of rFVIII to the cell membrane could be a factor diminishing the production yield. We studied the contribution of the rFVIII B-domain to membrane attachment by transfecting several constructs containing increasing lengths of the B-domain into cells under serum free conditions. We found that 90......% of rFVIII is attached to the cell membrane of the producing cell when the rFVIII variant contains a short B-domain (21 aa). By increasing the length of the B-domain the membrane attached fraction can be reduced to 50% of the total expressed rFVIII. Further, our studies show that the N...
Newly-derived neuroblastoma cell lines propagated in serum-free media recapitulate the genotype and phenotype of primary neuroblastoma tumours
Bate-Eya, Laurel T.; Ebus, Marli E.; Koster, Jan; den Hartog, Ilona J. M.; Zwijnenburg, Danny A.; Schild, Linda; van der Ploeg, Ida; Dolman, M. Emmy M.; Caron, Huib N.; Versteeg, Rogier; Molenaar, Jan J.
Recently protocols have been devised for the culturing of cell lines from fresh tumours under serum-free conditions in defined neural stem cell medium. These cells, frequently called tumour initiating cells (TICs) closely retained characteristics of the tumours of origin. We report the isolation of
Prognostic Value of Serum Free Light Chain in Multiple Myeloma.
El Naggar, Amel A; El-Naggar, Mostafa; Mokhamer, El-Hassan; Avad, Mona W
The measurement of serum free light chain (sFLC) has been shown to be valuable in screening for the presence of plasma cell dyscrasia as well as for baseline prognosis in newly diagnosed patients. The aim of the present work was to study the prognostic value of sFLC in multiple myeloma in relation to other serum biomarkers, response to therapy and survival. Forty five newly diagnosed patients with MM were included in the study. Patients were divided into responders and non-responders groups according to response to therapy. sFLC and serum Amyloid A (SAA) were measured by immunonephelometry. The non-responders group showed a statistically significant higher kappa/lambda or lambda/kappa ratio and higher β2 microglobulin level, but lower albumin level at presentation, as compared to the responders group (P < 0.001). However, no statistically significant difference was detected between the two groups regarding SA A or calcium levels. Comparison between sFLC ratio obtained before and after therapy revealed significant decrease after treatment in the responders group (P = 0.05). Survival was significantly inferior in patients with an FLC ratio of ≥ 2.6 or ≤ 0.56 compared with those with an FLC ratio that was between 0.56 and 2.6 (P = 0.002).
Serum free light chain reference values: a critical approach.
Altinier, Sara; Seguso, Mara; Zaninotto, Martina; Varagnolo, Mariacristina; Adami, Fausto; Angeli, Paolo; Plebani, Mario
The clinical usefulness of serum free light chain (FLC) measurement in the management of patients with plasma cell proliferative disorders has been reported in several papers, and most clinical studies use the reference ranges declared by the manufacturer. The aim of the present study was to evaluate the reproducibility of FLCs immunoassay and to validate the reference range, before introducing it in routine setting. Internal quality control materials and a pool of fresh serum samples were used to evaluate imprecision; 162 fresh sera from healthy blood donors were analyzed to evaluate the reference range for FLCs. In order to verify the κ/λ FLC ratio, 43 sera from patients with polyclonal hypergammaglobulinemia were tested. The FLC immunoassay was performed using a nephelometer with the Freelite reagents. The imprecision studies performed using a serum pool tested with two different lots of reagents showed a mean CV of 16.09% for κFLC and of 16.72% for λFLC. Lower CV%s and different mean values were found by calculating the results from each specific lot separately, while different results were obtained using the control materials provided by the manufacturer. In reference subjects, the 2.5-97.5th percentiles were found to be 4.52-22.33 and 4.84-21.88mg/L for κFLC and λFLC, respectively. The range for κ/λ ratio (0.65-2.36) was validated with the values obtained from subjects with polyclonal hypergammaglobulinemia. In retesting 15 samples from blood donor subjects with a different lot of reagents, mean bias percentages of 17.60 for κFLC and 15.26 for λFLC were obtained. These findings confirm the lot-to-lot variability of the FLC assays also in the measurement of polyclonal light chains, as well as the need to carefully validate the reference values. Published by Elsevier Inc.
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Ranka Jeknić
Full Text Available This paper analyses one by one the neo-liberalism, social-democracy, radicalism and political-islamism, as four typical socio-political and economic attitudes toward individualism and collectivism as cultural dimensions in the contemporary socio-political and economic contex of globalization. The paper points out principal differences between these four standpoints, and after that, makes conclusions and points out some problematic questions in the conection with the cultural and systemic differences. Their comparative analysis is in the connection with new sociological theories of culture: functionalistic orientation, marxistic and postmodernistic.
Organotypic hippocampal slice culture from the adult mouse brain: a versatile tool for translational neuropsychopharmacology.
Kim, Hyunjeong; Kim, Eosu; Park, Minsun; Lee, Eun; Namkoong, Kee
One of the most significant barriers towards translational neuropsychiatry would be an unavailability of living brain tissues. Although organotypic brain tissue culture could be a useful alternative enabling observation of temporal changes induced by various drugs in living brain tissues, a proper method to establish a stable organotypic brain slice culture system using adult (rather than neonatal) hippocampus has been still elusive. In this study, we evaluated our simple method using the serum-free culture medium for successful adult organotypic hippocampal slice culture. Several tens of hippocampal slices from a single adult mouse (3-5 months old) were cultured in serum-free versus serum-containing conventional culture medium for 30 days and underwent various experiments to validate the effects of the existence of serum in the culture medium. Neither the excessive regression of neuronal viability nor metabolic deficiency was observed in the serum-free medium culture in contrast to the serum-containing medium culture. Despite such viability, newly generated immature neurons were scarcely detected in the serum-free culture, suggesting that the original neurons in the brain slice persist rather than being replaced by neurogenesis. Key structural features of in vivo neural tissue constituting astrocytes, neural processes, and pre- and post-synapses were also well preserved in the serum-free culture. In conclusion, using the serum-free culture medium, the adult hippocampal slice culture system will serve as a promising ex vivo tool for various fields of neuroscience, especially for studies on aging-related neuropsychiatric disorders or for high throughput screening of potential agents working against such disorders. Copyright © 2012 Elsevier Inc. All rights reserved.
Hematopoietic stem cell cytokines and fibroblast growth factor-2 stimulate human endothelial cell-pericyte tube co-assembly in 3D fibrin matrices under serum-free defined conditions.
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Annie O Smith
Full Text Available We describe a novel 3D fibrin matrix model using recombinant hematopoietic stem cell cytokines under serum-free defined conditions which promotes the assembly of human endothelial cell (EC tubes with co-associated pericytes. Individual ECs and pericytes are randomly mixed together and EC tubes form that is accompanied by pericyte recruitment to the EC tube abluminal surface over a 3-5 day period. These morphogenic processes are stimulated by a combination of the hematopoietic stem cell cytokines, stem cell factor, interleukin-3, stromal derived factor-1α, and Flt-3 ligand which are added in conjunction with fibroblast growth factor (FGF-2 into the fibrin matrix. In contrast, this tube morphogenic response does not occur under serum-free defined conditions when VEGF and FGF-2 are added together in the fibrin matrices. We recently demonstrated that VEGF and FGF-2 are able to prime EC tube morphogenic responses (i.e. added overnight prior to the morphogenic assay to hematopoietic stem cell cytokines in collagen matrices and, interestingly, they also prime EC tube morphogenesis in 3D fibrin matrices. EC-pericyte interactions in 3D fibrin matrices leads to marked vascular basement membrane assembly as demonstrated using immunofluorescence and transmission electron microscopy. Furthermore, we show that hematopoietic stem cell cytokines and pericytes stimulate EC sprouting in fibrin matrices in a manner dependent on the α5β1 integrin. This novel co-culture system, under serum-free defined conditions, allows for a molecular analysis of EC tube assembly, pericyte recruitment and maturation events in a critical ECM environment (i.e. fibrin matrices that regulates angiogenic events in postnatal life.
Generation of megakaryocytic progenitors from human embryonic stem cells in a feeder- and serum-free medium.
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Marjorie Pick
Full Text Available BACKGROUND: The production of human platelets from embryonic stem cells in a defined culture system is a prerequisite for the generation of platelets for therapeutic use. As an important step towards this goal, we report the differentiation of human embryonic stem cells (hESCs towards the megakaryocyte (Mk lineage using a 'spin embryoid body' method in serum-free differentiation medium. METHODOLOGY AND PRINCIPAL FINDINGS: Immunophenotypic analyses of differentiating hESC identified a subpopulation of cells expressing high levels of CD41a that expressed other markers associated with the Mk lineage, including CD110, CD42b and CD61. Differentiated cells were sorted on the basis of their expression of CD41a, CD34 and CD45 and assessed for Mk colony formation, expression of myeloid and Mk genes and ability to endoreplicate DNA. In a collagen-based colony assay, the CD41a⁺ cells sorted from these differentiation cultures produced 100-800 Mk progenitors at day 13 and 25-160 Mk progenitors at day 20 of differentiation per 100,000 cells assayed. Differentiated Mk cells produced platelet-like particles which expressed CD42b and were activated by ADP, similar to platelets generated from precursors in cord blood. These studies were complemented by real time PCR analyses showing that subsets of cells enriched for CD41a⁺ Mk precursors expressed high levels of Mk associated genes such as PF4 and MPL. Conversely, high levels of myeloid and erythroid related transcripts, such as GATA1, TAL1/SCL and PU.1, were detected in sorted fractions containing CD34⁺ and CD45⁺ cells. CONCLUSIONS: We describe a serum- and feeder-free culture system that enabled the generation of Mk progenitors from human embryonic stem cells. These cells formed colonies that included differentiated Mks that fragmented to form platelet-like particles. This protocol represents an important step towards the generation of human platelets for therapeutic use.
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Paul Ballonoff
Full Text Available A theory of cultural structures predicts the objects observed by anthropologists. We here define those which use kinship relationships to define systems. A finite structure we call a partially defined quasigroup (or pdq, as stated by Definition 1 below on a dictionary (called a natural language allows prediction of certain anthropological descriptions, using homomorphisms of pdqs onto finite groups. A viable history (defined using pdqs states how an individual in a population following such history may perform culturally allowed associations, which allows a viable history to continue to survive. The vector states on sets of viable histories identify demographic observables on descent sequences. Paths of vector states on sets of viable histories may determine which histories can exist empirically.
Production of inactivated influenza H5N1 vaccines from MDCK cells in serum-free medium.
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Alan Yung-Chih Hu
Full Text Available BACKGROUND: Highly pathogenic influenza viruses pose a constant threat which could lead to a global pandemic. Vaccination remains the principal measure to reduce morbidity and mortality from such pandemics. The availability and surging demand for pandemic vaccines needs to be addressed in the preparedness plans. This study presents an improved high-yield manufacturing process for the inactivated influenza H5N1 vaccines using Madin-Darby canine kidney (MDCK cells grown in a serum-free (SF medium microcarrier cell culture system. PRINCIPAL FINDING: The current study has evaluated the performance of cell adaptation switched from serum-containing (SC medium to several commercial SF media. The selected SF medium was further evaluated in various bioreactor culture systems for process scale-up evaluation. No significant difference was found in the cell growth in different sizes of bioreactors studied. In the 7.5 L bioreactor runs, the cell concentration reached to 2.3 × 10(6 cells/mL after 5 days. The maximum virus titers of 1024 Hemagglutinin (HA units/50 µL and 7.1 ± 0.3 × 10(8 pfu/mL were obtained after 3 days infection. The concentration of HA antigen as determined by SRID was found to be 14.1 µg/mL which was higher than those obtained from the SC medium. A mouse immunogenicity study showed that the formalin-inactivated purified SF vaccine candidate formulated with alum adjuvant could induce protective level of virus neutralization titers similar to those obtained from the SC medium. In addition, the H5N1 viruses produced from either SC or SF media showed the same antigenic reactivity with the NIBRG14 standard antisera. CONCLUSIONS: The advantages of this SF cell-based manufacturing process could reduce the animal serum contamination, the cost and lot-to-lot variation of SC medium production. This study provides useful information to manufacturers that are planning to use SF medium for cell-based influenza vaccine production.
A single-cell and feeder-free culture system for monkey embryonic stem cells.
Ono, Takashi; Suzuki, Yutaka; Kato, Yosuke; Fujita, Risako; Araki, Toshihiro; Yamashita, Tomoko; Kato, Hidemasa; Torii, Ryuzo; Sato, Naoya
Primate pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), hold great potential for research and application in regenerative medicine and drug discovery. To maximize primate PSC potential, a practical system is required for generating desired functional cells and reproducible differentiation techniques. Much progress regarding their culture systems has been reported to date; however, better methods would still be required for their practical use, particularly in industrial and clinical fields. Here we report a new single-cell and feeder-free culture system for primate PSCs, the key feature of which is an originally formulated serum-free medium containing FGF and activin. In this culture system, cynomolgus monkey ESCs can be passaged many times by single-cell dissociation with traditional trypsin treatment and can be propagated with a high proliferation rate as a monolayer without any feeder cells; further, typical PSC properties and genomic stability can be retained. In addition, it has been demonstrated that monkey ESCs maintained in the culture system can be used for various experiments such as in vitro differentiation and gene manipulation. Thus, compared with the conventional culture system, monkey ESCs grown in the aforementioned culture system can serve as a cell source with the following practical advantages: simple, stable, and easy cell maintenance; gene manipulation; cryopreservation; and desired differentiation. We propose that this culture system can serve as a reliable platform to prepare primate PSCs useful for future research and application.
Metaphase II oocytes from human unilaminar follicles grown in a multi-step culture system.
McLaughlin, M; Albertini, D F; Wallace, W H B; Anderson, R A; Telfer, E E
Can complete oocyte development be achieved from human ovarian tissue containing primordial/unilaminar follicles and grown in vitro in a multi-step culture to meiotic maturation demonstrated by the formation of polar bodies and a Metaphase II spindle? Development of human oocytes from primordial/unilaminar stages to resumption of meiosis (Metaphase II) and emission of a polar body was achieved within a serum free multi-step culture system. Complete development of oocytes in vitro has been achieved in mouse, where in vitro grown (IVG) oocytes from primordial follicles have resulted in the production of live offspring. Human oocytes have been grown in vitro from the secondary/multi-laminar stage to obtain fully grown oocytes capable of meiotic maturation. However, there are no reports of a culture system supporting complete growth from the earliest stages of human follicle development through to Metaphase II. Ovarian cortical biopsies were obtained with informed consent from women undergoing elective caesarean section (mean age: 30.7 ± 1.7; range: 25-39 years, n = 10). Laboratory setting. Ovarian biopsies were dissected into thin strips, and after removal of growing follicles were cultured in serum free medium for 8 days (Step 1). At the end of this period secondary/multi-laminar follicles were dissected from the strips and intact follicles 100-150 μm in diameter were selected for further culture. Isolated follicles were cultured individually in serum free medium in the presence of 100 ng/ml of human recombinant Activin A (Step 2). Individual follicles were monitored and after 8 days, cumulus oocyte complexes (COCs) were retrieved by gentle pressure on the cultured follicles. Complexes with complete cumulus and adherent mural granulosa cells were selected and cultured in the presence of Activin A and FSH on membranes for a further 4 days (Step 3). At the end of Step 3, complexes containing oocytes >100 μm diameter were selected for IVM in SAGE medium (Step 4) then
Active Stat3 is required for survival of human squamous cell carcinoma cells in serum-free conditions
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DiGiovanni John
Full Text Available Abstract Background Squamous cell carcinoma (SCC of the skin is the most aggressive form of non-melanoma skin cancer (NMSC, and is the single most commonly diagnosed cancer in the U.S., with over one million new cases reported each year. Recent studies have revealed an oncogenic role of activated signal transducer and activator of transcription 3 (Stat3 in many human tumors, especially in those of epithelial origin, including skin SCC. Stat3 is a mediator of numerous growth factor and cytokine signaling pathways, all of which activate it through phosphorylation of tyrosine 705. Results To further address the role of Stat3 in skin SCC tumorigenesis, we have analyzed a panel of human skin-derived cell lines ranging from normal human epidermal keratinocytes (NHEK, to non-tumorigenic transformed skin cells (HaCaT, to highly tumorigenic cells (SRB1-m7 and SRB12-p9 and observed a positive correlation between Stat3 phosphorylation and SCC malignancy. We next determined the role of Stat3 activity in cell proliferation and viability under serum-free culture conditions. This was accomplished by suppressing Stat3 activity in the SRB12-p9 cells through stable expression of a dominant negative acting form of Stat3β, which contains a tyrosine 705 to phenylalanine mutation (S3DN. The S3DN cells behaved similar to parental SRB12-p9 cells when cultured in optimal growth conditions, in the presence of 10% fetal calf serum. However, unlike the SRB12-p9 cells, S3DN cells underwent apoptotic cell death when cultured in serum-free medium (SFM. This was evidenced by multiple criteria, including accumulation of sub-G1 particles, induced PARP cleavage, and acquisition of the characteristic morphological changes associated with apoptosis. Conclusion This study provides direct evidence for a role for Stat3 in maintaining cell survival in the conditions of exogenous growth factor deprivation produced by culture in SFM. We also propose that delivery of the S3DN gene or
Changes in Serum Free Amino Acids and Muscle Fatigue Experienced during a Half-Ironman Triathlon.
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Francisco Areces
Full Text Available The aim of this study was to investigate the relationship between changes in serum free amino acids, muscle fatigue and exercise-induced muscle damage during a half-ironman triathlon. Twenty-six experienced triathletes (age = 37.0 ± 6.8 yr; experience = 7.4 ± 3.0 yr competed in a real half-ironman triathlon in which sector times and total race time were measured by means of chip timing. Before and after the race, a countermovement jump and a maximal isometric force test were performed, and blood samples were withdrawn to measure serum free amino acids concentrations, and serum creatine kinase levels as a blood marker of muscle damage. Total race time was 320 ± 37 min and jump height (-16.3 ± 15.2%, P 20%. However, neither the changes in serum free amino acids nor the tryptophan/BCAA ratio were related muscle fatigue or muscle damage during the race.
Development of a new mouse palate organ culture system and effect of X-irradiation on palatogenesis
International Nuclear Information System (INIS)
Hiranuma, Hiroko; Jikko, Akitoshi; Maeda, Takashi; Furukawa, Souhei
On the basis of an already established suspension system of organ culture for mouse palate explants, we developed a new culture system, which has several advantages over the previous methods. We used a 48-well culture plate in which the explants can be cultured individually, and only 300 μl of medium is needed for each well. In order to optimize the culture results, we systematically studied the influence of main ''culture conditions'' such as tilt degree of the culturing palate, rotation speed, and addition of ascorbic acid to the medium. This system allows culturing of palates from day 13.5 of gestation to day 16.5 under serum-free conditions using a chemically defined medium, which resulted in 78% of the palates growing fused. Utilizing this culture system, the direct effect of X-irradiation on palataogesis was analyzed. A 4 Gy dose of X-irradiation was administrated at the beginning of culture period. The incidence of palatal fusion was not significantly different from that of the non-irradiated group. (author)
Establishing elements of a synthetic biology platform for Vaccinia virus production: BioBrick™ design, serum-free virus production and microcarrier-based cultivation of CV-1 cells.
Liu, Shuchang; Ruban, Ludmila; Wang, Yaohe; Zhou, Yuhong; Nesbeth, Darren N
Vaccinia virus (VACV) is an established vector for vaccination and is beginning to prove effective as an oncolytic agent. Industrial production of VACV stands to benefit in future from advances made by synthetic biology in genome engineering and standardisation. The CV-1 cell line can be used for VACV propagation and has been used extensively with the CRISPR/Cas9 system for making precise edits of the VACV genome. Here we take first steps toward establishing a scalable synthetic biology platform for VACV production with CV-1 cells featuring standardised biological tools and serum free cell cultivation. We propose a new BioBrick™ plasmid backbone format for inserting transgenes into VACV. We then test the performance of CV-1 cells in propagation of a conventional recombinant Lister strain VACV, VACVL-15 RFP, in a serum-free process. CV-1 cells grown in 5% foetal bovine serum (FBS) Dulbecco's Modified Eagle Medium (DMEM) were adapted to growth in OptiPRO and VP-SFM brands of serum-free media. Specific growth rates of 0.047 h -1 and 0.044 h -1 were observed for cells adapted to OptiPRO and VP-SFM respectively, compared to 0.035 h -1 in 5% FBS DMEM. Cells adapted to OptiPRO and to 5% FBS DMEM achieved recovery ratios of over 96%, an indication of their robustness to cryopreservation. Cells adapted to VP-SFM showed a recovery ratio of 82%. Virus productivity in static culture, measured as plaque forming units (PFU) per propagator cell, was 75 PFU/cell for cells in 5% FBS DMEM. VP-SFM and OptiPRO adaptation increased VACV production to 150 PFU/cell and 350 PFU/cell respectively. Boosted PFU/cell from OptiPRO-adapted cells persisted when 5% FBS DMEM or OptiPRO medium was observed during the infection step and when titre was measured using cells adapted to 5% FBS DMEM or OptiPRO medium. Finally, OptiPRO-adapted CV-1 cells were successfully cultivated using Cytodex-1 microcarriers to inform future scale up studies.
Establishing elements of a synthetic biology platform for Vaccinia virus production: BioBrick™ design, serum-free virus production and microcarrier-based cultivation of CV-1 cells
Directory of Open Access Journals (Sweden)
Shuchang Liu
Full Text Available Vaccinia virus (VACV is an established vector for vaccination and is beginning to prove effective as an oncolytic agent. Industrial production of VACV stands to benefit in future from advances made by synthetic biology in genome engineering and standardisation. The CV-1 cell line can be used for VACV propagation and has been used extensively with the CRISPR/Cas9 system for making precise edits of the VACV genome. Here we take first steps toward establishing a scalable synthetic biology platform for VACV production with CV-1 cells featuring standardised biological tools and serum free cell cultivation. We propose a new BioBrick™ plasmid backbone format for inserting transgenes into VACV. We then test the performance of CV-1 cells in propagation of a conventional recombinant Lister strain VACV, VACVL-15 RFP, in a serum-free process. CV-1 cells grown in 5% foetal bovine serum (FBS Dulbecco's Modified Eagle Medium (DMEM were adapted to growth in OptiPRO and VP-SFM brands of serum-free media. Specific growth rates of 0.047 h−1 and 0.044 h−1 were observed for cells adapted to OptiPRO and VP-SFM respectively, compared to 0.035 h−1 in 5% FBS DMEM. Cells adapted to OptiPRO and to 5% FBS DMEM achieved recovery ratios of over 96%, an indication of their robustness to cryopreservation. Cells adapted to VP-SFM showed a recovery ratio of 82%. Virus productivity in static culture, measured as plaque forming units (PFU per propagator cell, was 75 PFU/cell for cells in 5% FBS DMEM. VP-SFM and OptiPRO adaptation increased VACV production to 150 PFU/cell and 350 PFU/cell respectively. Boosted PFU/cell from OptiPRO-adapted cells persisted when 5% FBS DMEM or OptiPRO medium was observed during the infection step and when titre was measured using cells adapted to 5% FBS DMEM or OptiPRO medium. Finally, OptiPRO-adapted CV-1 cells were successfully cultivated using Cytodex-1 microcarriers to inform future scale up studies.
[Diagnostic significance of serum free DNA human telomerase reverse transcriptase quantitative determination on spinal cord injury].
Yang, M K; Tang, J; Xiang, Z; Zhang, X; Wang, J; Li, Z; Li, Y; Sheng, W B
Objective: To investigate the relationship between the content of human telomerase reverse transcriptase (hTERT) and its clinical features in serum free DNA in patients with different degree of spinal cord injury. Methods: From December 2013 to December 2016, inpatients of the Central Hospital of Bazhong, Sichuan Province were enrolledand divided into the experimental group, the disease control group and the negative control group. For the experimental group: 46 patients with spinal cord injury were graded according to the criteria of the American Association of Spinal Cord Injury (ASIA), including 12 cases of grade A, 10 cases of grade B, 10 cases of grade C, 7 cases of grade D and 7 cases of grade E; for the disease control group: 15 patients with spinal fractures (without spinal cord injury) at the same period were included; and for the negative control group: 20 healthy adult volunteers aged 18-50 years were selected.Real-time fluorescence quantitative PCR and immunoblotting were performed to detect the content of hTERT in serum free DNA both in patients and healthy controls and to compare the difference between them. The results of the somatosensory evoked potential (SEP) of all patients were compared and analyzed.The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of hTERT content in serum free DNA in patients with spinal cord injury. Results: Comparison of serum free DNA hTERT content: in the experimental group, the serum free DNA hTERT content of grade A, B, C, D, E was (99.63±8.23), (76.24±4.37), (46.07±5.43), (16.30±0.95) and (15.74±1.12)μg/L, respectively.While it was (15.01±1.39)μg/L in the disease control group and (14.54±1.03)μg/L in the negative control group. The total difference was statistically significant between patients of each group and the control group ( F =857.917, P spinal cord injury has a certain guiding significance for the diagnosis of spinal cord injury and the degree of injury.
Development of a serum-free medium for in vitro expansion of human cytotoxic T lymphocytes using a statistical design
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Lee Gyun
Full Text Available Abstract Background Serum-containing medium (SCM, which has a number of poorly defined components with varying concentrations, hampers standardization of lymphocyte cultures. In order to develop a serum-free medium (SFM for the expansion of human lymphocytes from peripheral blood mononuclear cells (PBMCs, a statistical optimization approach based on a fractional factorial method and a response surface method was adopted. A basal medium was prepared by supplementing RPMI1640 medium with insulin, albumin, ferric citrate, ethanolamine, fatty acids, glutamine, sodium pyruvate, 2-mercaptoethanol, 1-thioglycerol, nonessential amino acids, and vitamins. We identified additional positive determinants and their optimal concentrations for cell growth through a statistical analysis. Results From a statistical analysis using the fractional factorial method, cholesterol and polyamine supplement were identified as positive determinants for cell growth. Their optimal concentrations were determined by the response surface method. The maximum viable cell concentration in the developed SFM was enhanced by more than 1.5-fold when compared to that in RPMI1640 supplemented with 10% fetal bovine serum (FBS. Furthermore, a cytotoxicity assay and an enzyme-linked immunospot assay revealed that the effector function of cytotoxic T lymphocytes generated from PBMCs grown in SFM, by stimulation of peptide-presenting dendritic cells, was retained or even better than that in SCM. Conclusions The use of a developed SFM with cholesterol and polyamine supplement for human lymphocyte culture resulted in better growth without loss of cellular function when compared to SCM.
Development of a serum-free medium for in vitro expansion of human cytotoxic T lymphocytes using a statistical design.
Jeon, Min Kyoung; Lim, Jong-Baeck; Lee, Gyun Min
Serum-containing medium (SCM), which has a number of poorly defined components with varying concentrations, hampers standardization of lymphocyte cultures. In order to develop a serum-free medium (SFM) for the expansion of human lymphocytes from peripheral blood mononuclear cells (PBMCs), a statistical optimization approach based on a fractional factorial method and a response surface method was adopted. A basal medium was prepared by supplementing RPMI1640 medium with insulin, albumin, ferric citrate, ethanolamine, fatty acids, glutamine, sodium pyruvate, 2-mercaptoethanol, 1-thioglycerol, nonessential amino acids, and vitamins. We identified additional positive determinants and their optimal concentrations for cell growth through a statistical analysis. From a statistical analysis using the fractional factorial method, cholesterol and polyamine supplement were identified as positive determinants for cell growth. Their optimal concentrations were determined by the response surface method. The maximum viable cell concentration in the developed SFM was enhanced by more than 1.5-fold when compared to that in RPMI1640 supplemented with 10% fetal bovine serum (FBS). Furthermore, a cytotoxicity assay and an enzyme-linked immunospot assay revealed that the effector function of cytotoxic T lymphocytes generated from PBMCs grown in SFM, by stimulation of peptide-presenting dendritic cells, was retained or even better than that in SCM. The use of a developed SFM with cholesterol and polyamine supplement for human lymphocyte culture resulted in better growth without loss of cellular function when compared to SCM.
Anaerobic thermophilic culture-system
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Ljungdahl, L G; Wiegel, J K.W.
A mixed culture system of Thermoanaerobacter ethanolicus and Clostridium thermocellum is employed for anaerobic, thermophilic ethanol fermentation of cellulose. By cellulase action, monosaccharides are formed which inhibit the growth of C. thermocellum, but are fermented by T. ethanolicus. Thus, at a regulated pH-value of 7.5, this mixed culture system of micro organisms results in a cellulose fermentation with a considerably higher ethanol yield.
Newly-derived neuroblastoma cell lines propagated in serum-free media recapitulate the genotype and phenotype of primary neuroblastoma tumours.
Bate-Eya, Laurel T; Ebus, Marli E; Koster, Jan; den Hartog, Ilona J M; Zwijnenburg, Danny A; Schild, Linda; van der Ploeg, Ida; Dolman, M Emmy M; Caron, Huib N; Versteeg, Rogier; Molenaar, Jan J
Recently protocols have been devised for the culturing of cell lines from fresh tumours under serum-free conditions in defined neural stem cell medium. These cells, frequently called tumour initiating cells (TICs) closely retained characteristics of the tumours of origin. We report the isolation of eight newly-derived neuroblastoma TICs from six primary neuroblastoma tumours and two bone marrow metastases. The primary tumours from which these TICs were generated have previously been fully typed by whole genome sequencing (WGS). Array comparative genomic hybridisation (aCGH) analysis showed that TIC lines retained essential characteristics of the primary tumours and exhibited typical neuroblastoma chromosomal aberrations such as MYCN amplification, gain of chromosome 17q and deletion of 1p36. Protein analysis showed expression for neuroblastoma markers MYCN, NCAM, CHGA, DBH and TH while haematopoietic markers CD19 and CD11b were absent. We analysed the growth characteristics and confirmed tumour-forming potential using sphere-forming assays, subcutaneous and orthotopic injection of these cells into immune-compromised mice. Affymetrix mRNA expression profiling of TIC line xenografts showed an expression pattern more closely mimicking primary tumours compared to xenografts from classical cell lines. This establishes that these neuroblastoma TICs cultured under serum-free conditions are relevant and useful neuroblastoma tumour models. Copyright © 2013 Elsevier Ltd. All rights reserved.
Two different serum-free media and osmolality effect upon human 293 cell growth and adenovirus production.
Ferreira, Tiago B; Ferreira, Ana L; Carrondo, Manuel J T; Alves, Paula M
Adenoviruses are promising vectors for gene therapy and vaccination protocols. Consequently, the market demands for adenovirus are increasing, driving the search for new methodologies for large-scale production of concentrated vectors with warranted purity and efficacy, in a cost-effective way. Nevertheless, the production of adenovirus is currently limited by the so-called 'cell density effect', i.e. a drop in cell specific productivity concomitant with increased cell concentration at infection. Of two different serum-free culture media (CD293 and EX-Cell), evaluated for their effect on human 293 cells growth and adenovirus production at cell densities higher than 1x10(6) cells/ml, EX-Cell proved the better medium for cell growth. Although adenovirus production was equivalent in both media when the infection was performed at 1x10(6) cells/ml, at 3x10(6) cells/ml CD293 was the better. This result related to the high ammonia content in EX-Cell medium at the highest cell concentration at infection. Besides this, the large-scale production of these vectors at high cell densities often requires re-feed strategies, which increase medium osmolality. While a negative effect on cell growth was observed with increasing osmolalities, adenovirus productivity was only affected for osmolalities higher than 430 mOsm.
Elevated serum free thyroxine by thyroxine analog radioimmunoassays in euthyroid patients with familial dysalbuminemic hyperthyroxinemia
International Nuclear Information System (INIS)
Rajatanavin, R.; Fournier, L.; DeCosimo, D.; Abreau, C.; Braverman, L.E.
A study was done to ascertain whether the serum free T4 measured by free T4 radioimmunoassay kits would, like equilibrium dialysis, be normal in patients with familial dysalbuminemic hyperthyroxinemia. Five free T4 radioimmunoassay kits were used to measure free T4 in serum samples from 19 patients with familial dysalbuminemic hyperthyroxinemia and 20 healthy volunteers. Values (mean +/- SE) for T4, free T4 index, and free T4 (equilibrium dialysis) in these normal subjects and patients with familial dysalbuminemic hyperthyroxinemia, respectively, were as follows: T4, 8.1 +/- 0.2 and 18.3 +/- 0.7 μg/dL; free T4 index, 3.1 +/- 0.1 and 7.3 +/- 0.3 μg/dL; free T4, 1.4 +/- 0.1 and 1.2 +/- 0.1 ng/dL. The following free T4 radioimmunoassay methods were used: antibody coated microfine silica, microencapsulated antibody, two-step antibody-coated tube, and one-step 125 I-T4 analog (2 kits). The present findings in patients with familial dysalbuminemic hyperthyroxinemia and previous observations in ill euthyroid patients suggest that serum free T4 measured by some radioimmunoassay methods must be interpreted with caution in these two clinical situations
Serum free-thyroxine modifications with age and in normal pregnancy
International Nuclear Information System (INIS)
Degrossi, O.J.; Altschuler, N.; Watanabe, T.; Pinkas, M.; Damilano, S.; Garcia del Rio, H.
The possibility of using radioimmunoassay techniques (RIA) in the assessment of circulating thyroid hormones, total thyroxine (T4) and triiodothyronine (T3) has particularly increases the diagnosis of thyroid diseases. Thyroidal hormones circulate bound to proteins; therefore, variations in the transport capacity of the latter will produce important modifications in the T4 and T3 figures. Only small fractions, less than 0.05% for T4 and than 0.5% for T3, circulate in the free form and are considered as metabolically active forms of both hormones. In order to attain a correct clinical valuation, the rates of the free fractions as well as the total rates of these hormones must be known. Recently, these studies of free hormones are carried out by means of RIA, with the consequent advantages. The variations in serum free thyroxine (FT4) under certain physiogical conditions, such as for different ages and during pregnancy, were particularly studied
Interaction of estradiol and high density lipoproteins on proliferation of the human breast cancer cell line MCF-7 adapted to grow in serum free conditions
International Nuclear Information System (INIS)
Jozan, S.; Faye, J.C.; Tournier, J.F.; Tauber, J.P.; David, J.F.; Bayard, F.
The responsiveness of the human mammary carcinoma cell line MCF-7 to estradiol and tamoxifen treatment has been studied in different culture conditions. Cells from exponentially growing cultures were compared with cells in their initial cycles after replating from confluent cultures (''confluent-log'' cells). It has been observed that estradiol stimulation of tritiated thymidine incorporation decreases with cell density and that ''confluent-log'' cells are estrogen unresponsive for a period of four cell cycles in serum-free medium conditions. On the other hand, growth of cells replated from exponentially growing, as well as from confluent cultures, can be inhibited by tamoxifen or a combined treatment with tamoxifen and the progestin levonorgestrel. This growth inhibitory effect can be rescued by estradiol when cells are replated from exponentially growing cultures. The growth inhibitory effect cannot be rescued by estradiol alone (10(-10) to 10(-8) M) when cells are replated from confluent cultures. In this condition, the addition of steroid depleted serum is necessary to reverse the state of estradiol unresponsiveness. Serum can be replaced by high density lipoproteins but not by low density lipoproteins or lipoprotein deficient serum. The present data show that estradiol and HDL interact in the control of MCF-7 cell proliferation
[The value of serum free light chain in differential diagnosis of monoclonal gammopathy of renal significance].
Li, C; Wen, Y B; Li, H; Su, W; Li, J; Cai, J F; Chen, L M; Li, X M; Li, X W
Objective: To investigate the value of serum free light chain (FLC) in differential diagnosis of monoclonal gammopathy of renal significance (MGRS). Methods: Forty-nine hospitalized patients who underwent renal biopsy in Peking Union Medical College Hospital between January 2013 and December 2015 were included. Monoclonal gammopathy was detected by serum protein electrophoresis (SPE), serum immunofixation electrophoresis (IFE), urine IFE and serum FLC. All patients were classified as MGRS ( n =32) and monoclonal gammopathy of undetermined significance (MGUS) ( n =17). Results: Renal lesions in MGRS subgroup included light chain amyloidosis ( n =24, 75.0%), light chain deposition disease ( n =7, 21.9%), and fibrillary glomerulopathy ( n =1, 3.1%). Renal diseases in MGUS subgroup included membranous nephropathy ( n =10), focal segmental glomerulosclerosi (FSGS) ( n =3), diabetic glomerulopathy ( n =1), Henoch-Schonlein purpura nephritis ( n =1), anti-GBM disease concurrent with membranous nephropathy ( n =1) and glomerulomegaly ( n =1). Positive number of SPE, serum IFE, urine IFE and abnormal number of serum FLC ratio in MGRS subgroup were 12, 16, 23 and 30, respectively. Positive number of SPE, serum IFE, urine IFE and abnormal number of serum FLC ratio in MGUS subgroup were 11, 17, 6 and 3, respectively. MGRS and MGUS subgroups differed significantly in positive rate of serum IFE ( P value for MGRS, which was helpful for differential diagnosis of patients who had contraindication to renal biopsy.
Differentiating Graves' disease from subacute thyroiditis using ratio of serum free triiodothyronine to free thyroxine.
Sriphrapradang, Chutintorn; Bhasipol, Adikan
The measurement of free thyroid hormone, instead of the total form, is more commonly used in current practice. We aimed to evaluate the usefulness of the ratio of serum free triiodothyronine (FT3, pg/mL) to free thyroxine (FT4, ng/dL) for differentiating Graves' disease from subacute thyroiditis. Medical records of thyrotoxic patients aged >15 years who had measurement of FT3, FT4 and thyrotropin on the first diagnosis of thyrotoxicosis before initiating treatment were retrospectively reviewed. Data were collected from all clinics, and were not limited to the endocrine clinic. Pregnant women were excluded. A total of 548 patients (468 with Graves' disease, 40 with subacute thyroiditis and 40 with toxic adenoma/multinodular goiter) were recruited. Mean age was 43.9 ± 15.4 years. Most were female 434 (79.2%), and goiter was present in 55.3%. Prevalence of T3-toxicosis and T4-toxicosis were 5.6% and 6.6%, respectively. Mean FT3/FT4 ratios were 4.62 ± 2 (10(-2) pg/ng) in patients with Graves' disease and 2.73 ± 0.5 in subacute thyroiditis. The area under the ROC curve of the FT3/FT4 ratio for diagnosis of Graves' disease was 0.83 (95%CI, 0.76-0.91). Cutoff level of this ratio >4.4 offered sensitivity of 47.2% and specificity of 92.8%. FT3/FT4 ratio of >4.4 (10(-2) pg/ng) may help in differentiating the cause of thyrotoxicosis.
Increased serum free tryptophan in patients with diarrhea-predominant irritable bowel syndrome.
Christmas, David M; Badawy, Abdulla A-B; Hince, Dana; Davies, Simon J C; Probert, Christopher; Creed, Tom; Smithson, John; Afzal, Muhammad; Nutt, David J; Potokar, John P
Irregularities of serotonin function in irritable bowel syndrome (IBS) may be due to changes in the metabolism of the serotonin precursor l-tryptophan. Dietary alteration of tryptophan intake may impact upon the mood and bowel symptoms of IBS. We hypothesized that diarrhea-predominant irritable bowel syndrome (d-IBS) patients would exhibit an increase in plasma tryptophan due to alterations in tryptophan metabolism. We also hypothesized that a diet low in tryptophan would reverse this change and reduce symptoms. Thirteen patients with d-IBS had fasting serum free and total tryptophan, large neutral amino acids, and 6 kynurenine metabolites measured before and after 2 weeks of a strict dairy-free diet. Baseline tryptophan parameters were compared with an age- and sex-matched control group. Changes in the specific tryptophan parameters before and after dairy-free diet were correlated with symptoms of IBS and mood. Compared with the control group, d-IBS patients at baseline exhibited significantly higher free serum tryptophan (10.5 ± 4.35 vs 4.75 ± 2.43 μmol/L [means ± standard deviation], P = .006) and significantly lower tryptophan dioxygenase and total tryptophan oxidation as measured by the kynurenine to free tryptophan and total kynurenines to free tryptophan ratios (23.37 ± 10.12 vs 55.33 ± 16.02, P < .001 and 49.34 ± 17.84 vs 258.46 ± 98.67, P < .001, respectively). Dairy-free diet did not modulate metabolites of the kynurenine pathway or symptoms. Tryptophan metabolism along the kynurenine pathway is inhibited in d-IBS, and a dairy-free diet does not alter this. Our findings are consistent with possible enhanced serotonin activity in d-IBS. Copyright © 2010 Elsevier Inc. All rights reserved.
Bone Marrow Mesenchymal Stem Cells Enhance the Differentiation of Human Switched Memory B Lymphocytes into Plasma Cells in Serum-Free Medium
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Guillaume Bonnaure
Full Text Available The differentiation of human B lymphocytes into plasma cells is one of the most stirring questions with regard to adaptive immunity. However, the terminal differentiation and survival of plasma cells are still topics with much to be discovered, especially when targeting switched memory B lymphocytes. Plasma cells can migrate to the bone marrow in response to a CXCL12 gradient and survive for several years while secreting antibodies. In this study, we aimed to get closer to niches favoring plasma cell survival. We tested low oxygen concentrations and coculture with mesenchymal stem cells (MSC from human bone marrow. Besides, all cultures were performed using an animal protein-free medium. Overall, our model enables the generation of high proportions of CD38+CD138+CD31+ plasma cells (≥50% when CD40-activated switched memory B lymphocytes were cultured in direct contact with mesenchymal stem cells. In these cultures, the secretion of CXCL12 and TGF-β, usually found in the bone marrow, was linked to the presence of MSC. The level of oxygen appeared less impactful than the contact with MSC. This study shows for the first time that expanded switched memory B lymphocytes can be differentiated into plasma cells using exclusively a serum-free medium.
Does National Culture Impact Capital Budgeting Systems?
Peter J. Graham; Milind Sathye
We examine how national culture impacts organisational selection of capital budgeting systems to develop our understanding of what influence a holistic formulation of national culture has on capital budgeting systems. Such an understanding is important as it would not only provide a clearer link between national culture and capital budgeting systems and advance extant literature but would also help multinational firms that have business relationships with Indonesian firms in suita...
[The prognostic value of baseline serum free light chain in cardiac amyloidosis].
Zhao, Lei; Tian, Zhuang; Fang, Quan
To analyze the prognostic value of baseline serum free light chain (sFLC) in light-chain (AL) cardiac amyloidosis. Twenty-seven patients with AL cardiac amyloidosis were retrospectively reviewed from January 2014 to January 2015. sFLC was measured by immuoturbidimetric assay. Baseline characteristics, echocardiographic parameters and electrocardiogram data were analyzed. According to the median baseline dFLC (involved sFLC minus uninvolved sFLC), patients were categorized into either the low dFLC(≤ 307 mg/L) or the high dFLC group (>307 mg/L). More subjects in the high dFLC group with early/late diastolic mitral velocity ratio (E/A ratio) over 2 (71.4% vs 30.8%, P=0.035), and subjects in this group had a shorter median survival time than those in the low dFLC group (3 months vs 17 months, P=0.004). A similar phenomenon for median survival time was observed when the subjects were redivided either by a new cut-off value of 180 mg/L for dFLC (low dFLC group: 17 months; high dFLC group: 4 months, P=0.014) or a κ/λ ratio, in which subjects with κ type sFLC-ratio ≤ 19.6 and λ type sFLC-ratio>0.065 were in the low sFLC-ratio group (17 months) and those with κ type sFLC-ratio > 19.6 and λ type sFLC-ratio ≤ 0.065 were in the high sFLC-ratio group (4 months, P=0.023). In multivariate analysis, dFLC and New York Heart Association (NYHA)classification of cardiac function were two risk factors associated with all-cause mortality in patients, among which the hazard ratio for higher dFLC was 4.28 (95%CI 1.55-11.8, P=0.005). The level of sFLC could be a marker for the prognosis of AL cardiac amyloidosis.
Simple suspension culture system of human iPS cells maintaining their pluripotency for cardiac cell sheet engineering.
Haraguchi, Yuji; Matsuura, Katsuhisa; Shimizu, Tatsuya; Yamato, Masayuki; Okano, Teruo
In this study, a simple three-dimensional (3D) suspension culture method for the expansion and cardiac differentiation of human induced pluripotent stem cells (hiPSCs) is reported. The culture methods were easily adapted from two-dimensional (2D) to 3D culture without any additional manipulations. When hiPSCs were directly applied to 3D culture from 2D in a single-cell suspension, only a few aggregated cells were observed. However, after 3 days, culture of the small hiPSC aggregates in a spinner flask at the optimal agitation rate created aggregates which were capable of cell passages from the single-cell suspension. Cell numbers increased to approximately 10-fold after 12 days of culture. The undifferentiated state of expanded hiPSCs was confirmed by flow cytometry, immunocytochemistry and quantitative RT-PCR, and the hiPSCs differentiated into three germ layers. When the hiPSCs were subsequently cultured in a flask using cardiac differentiation medium, expression of cardiac cell-specific genes and beating cardiomyocytes were observed. Furthermore, the culture of hiPSCs on Matrigel-coated dishes with serum-free medium containing activin A, BMP4 and FGF-2 enabled it to generate robust spontaneous beating cardiomyocytes and these cells expressed several cardiac cell-related genes, including HCN4, MLC-2a and MLC-2v. This suggests that the expanded hiPSCs might maintain the potential to differentiate into several types of cardiomyocytes, including pacemakers. Moreover, when cardiac cell sheets were fabricated using differentiated cardiomyocytes, they beat spontaneously and synchronously, indicating electrically communicative tissue. This simple culture system might enable the generation of sufficient amounts of beating cardiomyocytes for use in cardiac regenerative medicine and tissue engineering. Copyright © 2013 John Wiley & Sons, Ltd.
Cultural Systems and Land Use Decision Making.
Schaefer, Larry; Pressman, Rob
This material includes student guide sheets, reference material, and tape script for the audio-tutorial unit on Cultural Systems. An audio tape is used with the materials. The material is designed for use with Connecticut schools, but can be adapted to other localities. The materials in this unit consider components of cultural systems, land use…
SERPINA3K plays antioxidant roles in cultured pterygial epithelial cells through regulating ROS system.
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Chengpeng Zhu
Full Text Available We recently demonstrated that SERPINA3K, a serine proteinase inhibitor, has antioxidant activity in the cornea. Here we investigated the antioxidant effects of SERPINA3K on the pterygial, which is partially caused by oxidative stress in pathogenesis. The head part of primary pterygial tissue was dissected and then cultured in keratinocyte serum-free defined medium (KSFM. The cultured pterygial epithelial cells (PECs were treated with SERPINA3K. The cell proliferation and migration of PECs were measured and analyzed. Western blot and quantitative real-time polymerase chain reaction (PCR assay were performed. It showed that SERPINA3K significantly suppressed the cell proliferation of PECs in a concentration-dependent manner, compared with cultured human conjunctival epithelial cells. SERPINA3K also inhibited the cell migration of PECs. Towards its underlying mechanism, SERPINA3K had antioxidant activities on the PECs by significantly inhibiting NADPH oxidase 4 (NOX4, which is an important enzyme of ROS generation, and by elevating the levels of key antioxidant factors of ROS: such as NAD(PH dehydrogenase (quinone 1 (NQO1, NF-E2-related factor-2 (NRF2 and superoxide dismutases (SOD2. Meanwhile, SERPINA3K down-regulated the key effectors of Wnt signaling pathway: β-catenin, nonphospho-β-catenin, and low-density lipoprotein receptor-related protein 6 (LRP6. We provided novel evidence that SERPINA3K had inhibitory effects on pterygium and SERPINA3K played antioxidant role via regulating the ROS system and antioxidants.
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Marilena Maldonado
Full Text Available Problems may arise when organizational culture is not considered in the development of information systems, such as difficulties in system implementation, since users do not accept changes in their work cultures. However, current methodology designs do not contemplate cultural factors. The objective of this investigation was to identify the main cultural features shared by the users of information systems in an Argentinean university. As result of this work it was possible to identify the memes shared by the members of the community selected, and to categorize such memes according to their incidence grade. This work seeks to be an initial step towards the construction of systems that evolve along with the organizational culture they are an integral part of.
Microfluidic cell culture systems for drug research.
Wu, Min-Hsien; Huang, Song-Bin; Lee, Gwo-Bin
In pharmaceutical research, an adequate cell-based assay scheme to efficiently screen and to validate potential drug candidates in the initial stage of drug discovery is crucial. In order to better predict the clinical response to drug compounds, a cell culture model that is faithful to in vivo behavior is required. With the recent advances in microfluidic technology, the utilization of a microfluidic-based cell culture has several advantages, making it a promising alternative to the conventional cell culture methods. This review starts with a comprehensive discussion on the general process for drug discovery and development, the role of cell culture in drug research, and the characteristics of the cell culture formats commonly used in current microfluidic-based, cell-culture practices. Due to the significant differences in several physical phenomena between microscale and macroscale devices, microfluidic technology provides unique functionality, which is not previously possible by using traditional techniques. In a subsequent section, the niches for using microfluidic-based cell culture systems for drug research are discussed. Moreover, some critical issues such as cell immobilization, medium pumping or gradient generation in microfluidic-based, cell-culture systems are also reviewed. Finally, some practical applications of microfluidic-based, cell-culture systems in drug research particularly those pertaining to drug toxicity testing and those with a high-throughput capability are highlighted.
Economic Systems: A Modular Approach. Cultural Anthropology.
Kassebaum, Peter
Designed for use as supplementary instructional material in a cultural anthropology course, this learning module uses a systems approach to allow students to see the connections and similarities which most cultural groups share on the basis of the type of economic organization that they exhibit. The module begins with a general discussion of…
Stepwise Differentiation of Pluripotent Stem Cells into Osteoblasts Using Four Small Molecules under Serum-free and Feeder-free Conditions
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Kosuke Kanke
Full Text Available Pluripotent stem cells are a promising tool for mechanistic studies of tissue development, drug screening, and cell-based therapies. Here, we report an effective and mass-producing strategy for the stepwise differentiation of mouse embryonic stem cells (mESCs and mouse and human induced pluripotent stem cells (miPSCs and hiPSCs, respectively into osteoblasts using four small molecules (CHIR99021 [CHIR], cyclopamine [Cyc], smoothened agonist [SAG], and a helioxanthin-derivative 4-(4-methoxyphenylpyrido[4′,3′:4,5]thieno[2,3-b]pyridine-2-carboxamide [TH] under serum-free and feeder-free conditions. The strategy, which consists of mesoderm induction, osteoblast induction, and osteoblast maturation phases, significantly induced expressions of osteoblast-related genes and proteins in mESCs, miPSCs, and hiPSCs. In addition, when mESCs defective in runt-related transcription factor 2 (Runx2, a master regulator of osteogenesis, were cultured by the strategy, they molecularly recapitulated osteoblast phenotypes of Runx2 null mice. The present strategy will be a platform for biological and pathological studies of osteoblast development, screening of bone-augmentation drugs, and skeletal regeneration.
Does National Culture Impact Capital Budgeting Systems?
Directory of Open Access Journals (Sweden)
Peter J. Graham
Full Text Available We examine how national culture impacts organisational selection of capital budgeting systems to develop our understanding of what influence a holistic formulation of national culture has on capital budgeting systems. Such an understanding is important as it would not only provide a clearer link between national culture and capital budgeting systems and advance extant literature but would also help multinational firms that have business relationships with Indonesian firms in suitably designing strategies. We conducted semi-structured interviews of selected finance managers of listed firms in Indonesia and Australia. Consistent with the contingency theory, we found that economic, political, legal and social uncertainty impact on the use of capital budgeting systems. The levels of uncertainty were higher in Indonesia than Australia and need to be reckoned in the selection of capital budgeting systems used by firms. We also found that firms are influenced by project size and complexity, when selecting capital budgeting systems.
Towards Culturally-Aware Virtual Agent Systems
DEFF Research Database (Denmark)
Endrass, Birgit; André, Elisabeth; Rehm, Matthias
Globalization leads to an increase in intercultural encounters with a risk of misunderstandings due to different patterns of behavior and understanding. Learning applications have been proposed that employ virtual agents as their primary tool. Through their embodiment, learning can be done...... in a game-like environment in a more interesting way than for example learning with a textbook. The authors support the idea that virtual agents are a great opportunity for teaching cultural awareness. Realizing this, the concept of culture needs to be translated into computational models and the advantages...... of different systems using virtual agents need to be considered. Therefore, the authors reflect in this chapter on how virtual agents can help to learn about culture, scan definitions of culture from the social sciences, give an overview on how multiagent systems developed over time and classify the state...
Managing organizational culture within a management system
International Nuclear Information System (INIS)
Comeau, L.; Watts, G.
The Point Lepreau Generating Station (PLGS) is currently undergoing a major refurbishment of its nuclear reactor. At the same time, a small team is designing the organization that will operate the plant after refurbishment. This paper offers a high level overview of the Post-Refurbishment Organization (PRO) project and will focus primarily on the approach used to address organizational culture and human system dynamics. We will describe how various tools, used to assess organization culture, team performance, and individual self-understanding, are used collectively to place the right person in the right position. We will explain how the career system, Pathfinder, is used to integrate these tools to support a comprehensive model for organization design and development. Finally, we demonstrate how the management of organizational cultural and human system dynamics are integrated into the PLGS Integrated Management System. (author)
Tourism Attraction Systems. Exploring cultural behavior
Richards, G.W.
Attractions are vital sub-elements in all whole tourism systems, and yet their study suffers from lack of theoretical depth and empirical foundation. This paper presents an empirical exploration of the attraction system model, based on a survey of over 6,000 tourists to cultural attractions. The
Differentiation of Dental Pulp Stem Cells into Neuron-Like Cells in Serum-Free Medium
Directory of Open Access Journals (Sweden)
Shahrul Hisham Zainal Ariffin
Full Text Available Dental pulp tissue contains dental pulp stem cells (DPSCs. Dental pulp cells (also known as dental pulp-derived mesenchymal stem cells are capable of differentiating into multilineage cells including neuron-like cells. The aim of this study was to examine the capability of DPSCs to differentiate into neuron-like cells without using any reagents or growth factors. DPSCs were isolated from teeth extracted from 6- to 8-week-old mice and maintained in complete medium. The cells from the fourth passage were induced to differentiate by culturing in medium without serum or growth factors. RT-PCR molecular analysis showed characteristics of Cd146+, Cd166+, and Cd31− in DPSCs, indicating that these cells are mesenchymal stem cells rather than hematopoietic stem cells. After 5 days of neuronal differentiation, the cells showed neuron-like morphological changes and expressed MAP2 protein. The activation of Nestin was observed at low level prior to differentiation and increased after 5 days of culture in differentiation medium, whereas Tub3 was activated only after 5 days of neuronal differentiation. The proliferation of the differentiated cells decreased in comparison to that of the control cells. Dental pulp stem cells are induced to differentiate into neuron-like cells when cultured in serum- and growth factor-free medium.
ABT-888 enhances cytotoxic effects of temozolomide independent of MGMT status in serum free cultured glioma cells
R.K. Balvers (Rutger); M.L.M. Lamfers (Martine); J.J. Kloezeman (Jenneke); A. Kleijn (Anne); L.M.E. Berghauser Pont (Lotte); C.M.F. Dirven (Clemens); S. Leenstra (Sieger)
textabstractBackground: The current standard of care for Glioblastoma Multiforme (GBM) consists of fractionated focal irradiation with concomitant temozolomide (TMZ) chemotherapy. A promising strategy to increase the efficacy of TMZ is through interference with the DNA damage repair machinery, by
Clinical significance of two-step magnetic radioimmunoassay for determining serum free T3 and free T4
International Nuclear Information System (INIS)
Chen Bing
The concentrations of the serum free T 3 (FT 3 ), free T 4 (FT 4 ), total triodothyronine (TT 3 ), total thyroxine (TT 4 ) and thyrotropin (TSH) are determined for 355 cases of normal persons, pregnant women and various thyropathetic patients. The normal values of FT 3 and FT 4 are 2.0-8.5 pmol/l, and 9.5-26.5 pmol/l, respectively. Neither FT 3 nor FT 4 is affected by the thyroxine combined with globulin (TBG), which is of unique diagnostic value for those with variable TBG (such as pregnant women hyperthyroidism, hypothyroidism, etc.), FT 3 and FT 4 are the most sensitive indices for diagnosis of hyperthyroidism, and hypothyroidism, respectively. In addition, FT 3 and FT 4 can greatly contribute to the observation of curative effectiveness under treatment
Creating a culture for information systems success
Belkhamza, Zakariya
It has been widely reported that issues related to organizational context appear frequently in discussions of information systems success. The statement that the information system did not fit the behavioral context in an organization is often part of the explanation of why particular information system encountered unanticipated resistance and never met expectation. While this context has been intensively studied, we still lack evidence on how this organizational context is affecting the success of information system from a managerial action perspective. This type of managerial involvement is often neglected to the extent that it became an essential obstacle to organizational performance. The objective of Creating a Culture for Information Systems Success is to assist CIOs and IT managers on how to use their managerial actions to create a suitable cultural environment in the organization, which leads to a successful implementation of information systems. This book will also provide guidelines fo...
Cultural tourism innovation systems - the roskilde festival
DEFF Research Database (Denmark)
Hjalager, Anne Mette
It is only recently that the "innovation systems approach" has become a framework for micro-economic research in new institutional economics in tourism-related businesses and activities. There is still much to be explored. Cultural tourism phenomena constitute noteworthy objects for illustrative...... case studies, embedded as they are in business as well as maintaining relations with public governance structures and voluntary organizations. Since 1971, Roskilde Festival (Denmark) has developed its role as a leading element in an emerging cultural innovation system. Festival organizers maintain long...
Understanding Nuclear Safety Culture: A Systemic Approach
International Nuclear Information System (INIS)
Afghan, A.N.
The Fukushima accident was a systemic failure (Report by Director General IAEA on the Fukushima Daiichi Accident). Systemic failure is a failure at system level unlike the currently understood notion which regards it as the failure of component and equipment. Systemic failures are due to the interdependence, complexity and unpredictability within systems and that is why these systems are called complex adaptive systems (CAS), in which "attractors" play an important role. If we want to understand the systemic failures we need to understand CAS and the role of these attractors. The intent of this paper is to identify some typical attractors (including stakeholders) and their role within complex adaptive system. Attractors can be stakeholders, individuals, processes, rules and regulations, SOPs etc., towards which other agents and individuals are attracted. This paper will try to identify attractors in nuclear safety culture and influence of their assumptions on safety culture behavior by taking examples from nuclear industry in Pakistan. For example, if the nuclear regulator is an attractor within nuclear safety culture CAS then how basic assumptions of nuclear plant operators and shift in-charges about "regulator" affect their own safety behavior?
Taurine Biosynthesis in a Fish Liver Cell Line (ZFL) Adapted to a Serum-Free Medium.
Liu, Chieh-Lun; Watson, Aaron M; Place, Allen R; Jagus, Rosemary
Although taurine has been shown to play multiple important physiological roles in teleosts, little is known about the molecular mechanisms underlying dietary requirements. Cell lines can provide useful tools for deciphering biosynthetic pathways and their regulation. However, culture media and sera contain variable taurine levels. To provide a useful cell line for the investigation of taurine homeostasis, an adult zebrafish liver cell line (ZFL) has been adapted to a taurine-free medium by gradual accommodation to a commercially available synthetic medium, UltraMEM™-ITES. Here we show that ZFL cells are able to synthesize taurine and be maintained in medium without taurine. This has allowed for the investigation of the effects of taurine supplementation on cell growth, cellular amino acid pools, as well as the expression of the taurine biosynthetic pathway and taurine transporter genes in a defined fish cell type. After taurine supplementation, cellular taurine levels increase but hypotaurine levels stay constant, suggesting little suppression of taurine biosynthesis. Cellular methionine levels do not change after taurine addition, consistent with maintenance of taurine biosynthesis. The addition of taurine to cells grown in taurine-free medium has little effect on transcript levels of the biosynthetic pathway genes for cysteine dioxygenase (CDO), cysteine sulfinate decarboxylase (CSAD), or cysteamine dioxygenase (ADO). In contrast, supplementation with taurine causes a 30% reduction in transcript levels of the taurine transporter, TauT. This experimental approach can be tailored for the development of cell lines from aquaculture species for the elucidation of their taurine biosynthetic capacity.
Dynamic cell culture system (7-IML-1)
Cogoli, Augusto
This experiment is one of the Biorack experiments being flown on the International Microgravity Laboratory 1 (MIL-1) mission as part of an investigation studying cell proliferation and performance in space. One of the objectives of this investigation is to assess the potential benefits of bioprocessing in space with the ultimate goal of developing a bioreactor for continuous cell cultures in space. This experiment will test the operation of an automated culture chamber that was designed for use in a Bioreactor in space. The device to be tested is called the Dynamic Cell Culture System (DCCS). It is a simple device in which media are renewed or chemicals are injected automatically, by means of osmotic pumps. This experiment uses four Type I/O experiment containers. One DCCS unit, which contains a culture chamber with renewal of medium and a second chamber without a medium supply fits in each container. Two DCCS units are maintained under zero gravity conditions during the on-orbit period. The other two units are maintained under 1 gh conditions in a 1 g centrifuge. The schedule for incubator transfer is given.
IgD multiple myeloma: Clinical, biological features and prognostic value of the serum free light chain assay.
Djidjik, R; Lounici, Y; Chergeulaïne, K; Berkouk, Y; Mouhoub, S; Chaib, S; Belhani, M; Ghaffor, M
IgD multiple myeloma (MM) is a rare subtype of myeloma, it affects less than 2% of patients with MM. To evaluate the clinical and prognostic attributes of serum free light chains (sFLCs) analysis, we examined 17 cases of IgD MM. From 1998 to 2012, we obtained 1250 monoclonal gammapathies including 590 multiple myeloma and 17 patients had IgD MM. With preponderance of men patients with a mean age at diagnosis of: 59±12years. Patients with IgD MM have a short survival (Median survival=9months). The presenting features included: bone pain (75%), lymphadenopathy (16%), hepatomegaly (25%), splenomegaly (8%), associated AL amyloidosis (6%), renal impairment function (82%), infections (47%), hypercalcemia (37%) and anemia (93%). Serum electrophoresis showed a subtle M-spike (Mean=13.22±10g/L) in all patients associated to a hypogammaglobulinemia. There was an over-representation of Lambda light chain (65%); high serum β2-microglobulin in 91% and Bence Jones proteinuria was identified in 71%. The median rate of sFLCs κ was 19.05mg/L and 296.75mg/L for sFLCs λ. sFLCR was abnormal in 93% of patients and it showed concordance between baseline sFLCR and the survival (P=0.034). The contribution of FLC assay is crucial for the prognosis of patients with IgD MM. Copyright © 2015 Elsevier Masson SAS. All rights reserved.
Can cultural differences lead to accidents? Team cultural differences and sociotechnical system operations.
Strauch, Barry
I discuss cultural factors and how they may influence sociotechnical system operations. Investigations of several major transportation accidents suggest that cultural factors may have played a role in the causes of the accidents. However, research has not fully addressed how cultural factors can influence sociotechnical systems. I review literature on cultural differences in general and cultural factors in sociotechnical systems and discuss how these differences can affect team performance in sociotechnical systems. Cultural differences have been observed in social and interpersonal dimensions and in cognitive and perceptual styles; these differences can affect multioperator team performance. Cultural factors may account for team errors in sociotechnical systems, most likely during high-workload, high-stress operational phases. However, much of the research on cultural factors has methodological and interpretive shortcomings that limit their applicability to sociotechnical systems. Although some research has been conducted on the role of cultural differences on team performance in sociotechnical system operations, considerable work remains to be done before the effects of these differences can be fully understood. I propose a model that illustrates how culture can interact with sociotechnical system operations and suggest avenues of future research. Given methodological challenges in measuring cultural differences and team performance in sociotechnical system operations, research in these systems should use a variety of methodologies to better understand how culture can affect multioperator team performance in these systems.
... lab's website in order to provide you with background information about the test(s) you had performed. You will need to return ... the free light chains have a much shorter half-life (3-5 hours) than ... cell disorders, the test is also used for assessing response and minimal ...
An Integrated Approach Using Liquid Culture System Can it Make ...
African Journals Online (AJOL)
These automated liquid culture systems, when combined with commercial molecular ... Culture System. Can it Make an Impact for Clinical Diagnosis of ... affected by this disease. .... Tuberculosis in non‑UK‑born persons, England and Wales ...
Systems Biology for Organotypic Cell Cultures
Energy Technology Data Exchange (ETDEWEB)
Grego, Sonia [RTI International, Research Triangle Park, NC (United States); Dougherty, Edward R. [Texas A & M Univ., College Station, TX (United States); Alexander, Francis J. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Auerbach, Scott S. [National Inst. of Environmental Health Sciences, Research Triangle Park, NC (United States); Berridge, Brian R. [GlaxoSmithKline, Research Triangle Park, NC (United States); Bittner, Michael L. [Translational Genomics Research Inst., Phoenix, AZ (United States); Casey, Warren [National Inst. of Environmental Health Sciences, Research Triangle Park, NC (United States); Cooley, Philip C. [RTI International, Research Triangle Park, NC (United States); Dash, Ajit [HemoShear Therapeutics, Charlottesville, VA (United States); Ferguson, Stephen S. [National Inst. of Environmental Health Sciences, Research Triangle Park, NC (United States); Fennell, Timothy R. [RTI International, Research Triangle Park, NC (United States); Hawkins, Brian T. [RTI International, Research Triangle Park, NC (United States); Hickey, Anthony J. [RTI International, Research Triangle Park, NC (United States); Kleensang, Andre [Johns Hopkins Univ., Baltimore, MD (United States). Center for Alternatives to Animal Testing; Liebman, Michael N. [IPQ Analytics, Kennett Square, PA (United States); Martin, Florian [Phillip Morris International, Neuchatel (Switzerland); Maull, Elizabeth A. [National Inst. of Environmental Health Sciences, Research Triangle Park, NC (United States); Paragas, Jason [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Qiao, Guilin [Defense Threat Reduction Agency, Ft. Belvoir, VA (United States); Ramaiahgari, Sreenivasa [National Inst. of Environmental Health Sciences, Research Triangle Park, NC (United States); Sumner, Susan J. [RTI International, Research Triangle Park, NC (United States); Yoon, Miyoung [The Hamner Inst. for Health Sciences, Research Triangle Park, NC (United States); ScitoVation, Research Triangle Park, NC (United States)
Translating in vitro biological data into actionable information related to human health holds the potential to improve disease treatment and risk assessment of chemical exposures. While genomics has identified regulatory pathways at the cellular level, translation to the organism level requires a multiscale approach accounting for intra-cellular regulation, inter-cellular interaction, and tissue/organ-level effects. Tissue-level effects can now be probed in vitro thanks to recently developed systems of three-dimensional (3D), multicellular, "organotypic" cell cultures, which mimic functional responses of living tissue. However, there remains a knowledge gap regarding interactions across different biological scales, complicating accurate prediction of health outcomes from molecular/genomic data and tissue responses. Systems biology aims at mathematical modeling of complex, non-linear biological systems. We propose to apply a systems biology approach to achieve a computational representation of tissue-level physiological responses by integrating empirical data derived from organotypic culture systems with computational models of intracellular pathways to better predict human responses. Successful implementation of this integrated approach will provide a powerful tool for faster, more accurate and cost-effective screening of potential toxicants and therapeutics. On September 11, 2015, an interdisciplinary group of scientists, engineers, and clinicians gathered for a workshop in Research Triangle Park, North Carolina, to discuss this ambitious goal. Participants represented laboratory-based and computational modeling approaches to pharmacology and toxicology, as well as the pharmaceutical industry, government, non-profits, and academia. Discussions focused on identifying critical system perturbations to model, the computational tools required, and the experimental approaches best suited to generating key data. This consensus report summarizes the discussions held.
Clinical usefulness of serum free light chains measurement in patients with multiple myeloma: comparative analysis of two different tests
Directory of Open Access Journals (Sweden)
Tadeusz Kubicki
Full Text Available Introduction: There are two commercially available tests for measurement of serum free light chains (sFLC in multiple myeloma (MM patients – Freelite and N Latex FLC. The aim of this study was to perform an assessment and direct comparison of the usefulness of the methods in routine clinical practice.Methods: 40 refractory/relapsed MM patients underwent routine disease activity assessment studies, along with sFLC analysis using both assays. Correlation and concordance between the tests and sensitivity of studied methods of sFLC assessment were established. Special attention was focused on sFLC results in patients finally evaluated after completing the treatment. Results: A weak correlation for the measurement of both κ [Passing–Bablok slope (PB = 0.7681] and λ chains [(PB = 1.542] was found. Using Bland–Altman plots, a bias of 0.0467 (κ and -0.2133 (λ between the measurements was documented. The concordance coefficient equaled 0.87 for κ, 0.62 for λ and 0.52 for κ/λ ratio. Ten patients had an abnormal Freelite assay κ/λ ratio and normal N Latex FLC κ/λ ratio. Three of these patients had negative serum protein electrophoresis results and fulfilled diagnostic criteria of stringent complete remission (sCR according to N Latex FLC (but not according to Freelite. When the κ/λ ratio obtained by both methods was compared to patients' serum/urine protein electrophoresis and immunofixation results, sensitivity of Freelite and N Latex FLC was established to be 62.5% and 41%, respectively. Conclusions: There was no strong correlation or concordance between the two assays, and the sensitivity in terms of sFLC detection was different. This may cause problems when diagnosis of sCR is considered.
Determination of serum free thyroxine concentration (FT4) by means of fT4-fraction and total thyroxine concentration
International Nuclear Information System (INIS)
Passath, A.; Leb, G.
A new equilibrium assay for the determination of serum free thyroxine was evaluated in 514 patients. The assay comprises a two-vial-procedure to measure total thyroxine and free thyroxine fraction by use of monoclonal antibodies. Free thyroxine concentrations are calculated from fT 4 -fraction and total thyroxine concentration readings. In euthyroidism the average free thyroxine fraction (%fT 4 ) was 0.011%, in hyperthyroidism this fraction was elevated, in hypothyroidism it was below normal. In patients with TBG anomalies, TBG values were inversely correlated with fT 4 fraction readings. The 'euthyroid reference range' of FT 4 (SPAC ET) was between 0.70 to 1.78ng/dl. This euthyroid range of FT 4 was determined from TT 4 concentrations measured by T 4 -RIA (SPAC T 4 MONO) which were 30% above TT 4 values measured by conventional T 4 -RIA (SPAC T 4 POLY; polyclonal antibodies). However, a different euthyroid range of FT 4 between 0.55 to 1.30 ng/dl was observed as well as by other investigators when conventional T 4 -RIA measurements were used for calculation of FT 4 values. Our results indicate that calculated FT 4 concentration values are highly dependent on the methods used for determination of total thyroxine concentrations. Precision and reproducability of this two vial equilibrium assay did not meet the requirements mandatory for the application as a clinical routine diagnostic procedure, and its general use for this purpose can as yet not be recommended. (Author)
The comparison of free androgen index and serum free testosterone levels in women with hirsutism or polycystic ovary syndrome
Directory of Open Access Journals (Sweden)
M. Metin Yıldırımkaya
Full Text Available In many laboratories free testosterone can not be measured, so that free androgen index is suggested instead. The aim of this study was to compare free androgen index and serum free testosterone levels measured by radioimmunoassay in women with hirsutism or polycystic ovary syndrome.Materials and methods: Totally 94 women referred to the polyclinics of Ankara Numune Hospital were retrospectively included. Three patient groups were composed; 55 of hirsutism, 20 of polycystic ovary syndrome and 19 of both hirsutism and polycystic ovary syndrome. Total testosterone and sex hormone binding globuline levels were measured by chemiluminescence method and free testosterone levels were measured by radioimmunoassay. Free androgen index was calculated from total testosterone and sex hormone binding globuline.Results: There was a significant positive correlation between free testosterone and free androgen index in patients with hirsutism, in patients with polycystic ovary syndrome, in patients with hirsutism and polycystic ovary syndrome, and in total patient group [r(hirsutism=0,597, r(PCOS=0,617, r(hirsutism and PCOS=0,779, r(total patient group=0,649, P<0,01].Receiver operating characteristics curves were drawn to assess the diagnostic power of parameters for all patient groups [For hirsutism (n=55 auROC (FT=0,431 auROC (FAI=0,485] [For PCOS (n=20 auROC (FT=0,431 auROC (FAI=0,359] [For hirsutism and PCOS (n=19 auROC (FT=0,676 auROC (FAI=0,669]. In our study, free testosterone and free androgen index were found useful to diagnose 'hirsutism and polycystic ovary syndrome' but not others.Conclusion: Free androgen index can be used instead of free testosterone in hirsutism and polycystic ovary syndrome for diagnosis. J Clin Exp Invest 2011;2(2:152-6
Effects of culture systems on growth and economic performance of ...
African Journals Online (AJOL)
The effect of culture system on growth and economics performance of Orechromis niloticus (Nile tilapia) in concrete tanks was investigated. Four outdoor concrete tanks measuring 2.5 x 2 m was used for the study for 24 weeks culture period. The culture systems included the use of algae only at the stocking rates of 4 ...
Effects of age and health on the euthyroid reference ranges for serum free thyroxine and free triiodothyronine
International Nuclear Information System (INIS)
Midgley, J.E.M.
Age-related trends in serum free thyroxine (FT 4 ) and free triiodothyronine (FT 3 ) concentrations were measured in 7248 euthyroid subjects (age-range 3 months to 106 years). 5700 were patients referred to hospitals for investigation of suspected thyroid dysfunction, but who were diagnosed euthyroid. 1548 were healthy blood donors (age-range 18-63 years) with no indication of thyroid dysfunction. FT 4 concentrations were little affected by the age, the sex or the state of health of the subjects in either group. Serum FT 3 concentrations were significantly affected by both age and health factors. The upper limit of the euthyroid reference range for young subjects up to 15 years was about 20% higher (10.4 pmol/l) than for adult subjects older than 25 years (8.8 pmol/l). The change in the upper limits typical of young subjects to that typical of adults occurred steadily over the decade 15-25 years. After this age, little further change occurred, especially in healthy subjects. Additionally, the lower limit of the euthyroid range for FT 3 was extended by the inclusion in the reference group of patients referred to hospitals. Compared with the lower limit of the FT 3 range for healthy subjects (5 pmol/l), the corresponding limit for referred subjects (young or adult) was 3.5-3.8 pmol/l. Broadening of the FT 3 reference range was probably brought about by a significant number of patients in the hospital-referred group with the 'low-T 3 syndrome' of mild non-thyroidal illness. Accordingly, FT 3 was inferior to FT 4 in the discrimination of hypothyroidism, as FT 4 was unaffected by this phenomenon. Effects of age and non-thyroidal illness on serum FT 3 concentrations require great care when selecting subjects for a laboratory euthyroid reference range typical of the routine workload. Constraints on the choice of subjects for FT 4 reference ranges are less stringent. (orig.) [de
In vitro expansion and differentiation of rat pancreatic duct-derived stem cells into insulin secreting cells using a dynamicthree-dimensional cell culture system.
Chen, X C; Liu, H; Li, H; Cheng, Y; Yang, L; Liu, Y F
In this study, a dynamic three-dimensional cell culture technology was used to expand and differentiate rat pancreatic duct-derived stem cells (PDSCs) into islet-like cell clusters that can secrete insulin. PDSCs were isolated from rat pancreatic tissues by in situ collagenase digestion and density gradient centrifugation. Using a dynamic three-dimensional culture technique, the cells were expanded and differentiated into functional islet-like cell clusters, which were characterized by morphological and phenotype analyses. After maintaining 1 x 108 isolated rat PDSCs in a dynamic three-dimensional cell culture for 7 days, 1.5 x 109 cells could be harvested. Passaged PDSCs expressed markers of pancreatic endocrine progenitors, including CD29 (86.17%), CD73 (90.73%), CD90 (84.13%), CD105 (78.28%), and Pdx-1. Following 14 additional days of culture in serum-free medium with nicotinamide, keratinocyte growth factor (KGF), and b fibroblast growth factor (FGF), the cells were differentiated into islet-like cell clusters (ICCs). The ICC morphology reflected that of fused cell clusters. During the late stage of differentiation, representative clusters were non-adherent and expressed insulin indicated by dithizone (DTZ)-positive staining. Insulin was detected in the extracellular fluid and cytoplasm of ICCs after 14 days of differentiation. Additionally, insulin levels were significantly higher at this time compared with the levels exhibited by PDSCs before differentiation (P cell culture system, PDSCs can be expanded in vitro and can differentiate into functional islet-like cell clusters.
Modular Approach to Designing Computer Cultural Systems: Culture as a Thermodynamic Machine
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Leland Gilsen
Full Text Available Culture is a complex non-linear system. In order to design computer simulations of cultural systems, it is necessary to break the system down into sub-systems. Human culture is modular. It consists of sets of people that belong to economic units. Access to, and control over matter, energy and information is postulated as the key to development of cultural simulations. Because resources in the real world are patchy, access to and control over resources is expressed in two related arenas: economics (direct control and politics (non-direct control. The best way to create models for cultural ecology/economics lies in an energy-information-economic paradigm based on general systems theory and an understanding of the "thermodynamics" of ecology, or culture as a thermodynamic machine.
Cultural systems for growing potatoes in space
Tibbitts, T.; Bula, R.; Corey, R.; Morrow, R.
Higher plants are being evaluated for life support to provide needed food, oxygen and water as well as removal of carbon dioxide from the atmosphere. The successful utilization of plants in space will require the development of not only highly productive growing systems but also highly efficient bioregenerative systems. It will be necessary to recycle all inedible plant parts and all human wastes so that the entire complement of elemental compounds can be reused. Potatoes have been proposed as one of the desirable crops because they are 1) extremely productive, yielding more than 100 metric tons per hectare from field plantings, 2) the edible tubers are high in digestible starch (70%) and protein (10%) on a dry weight basis, 3) up to 80% of the total plant production is in tubers and thus edible, 4) the plants are easily propagated either from tubers or from tissue culture plantlets, 5) the tubers can be utilized with a minimum of processing, and 6) potatoes can be prepared in a variety of different forms for the human diet (Tibbitts et al., 1982). However potatoes have a growth pattern that complicates the development of growing the plants in controlled systems. Tubers are borne on underground stems that are botanically termed 'rhizomes', but in common usage termed 'stolons'. The stolons must be maintained in a dark, moist area with sufficient provision for enlargement of tubers. Stems rapidly terminate in flowers forcing extensive branching and spreading of plants so that individual plants will cover 0.2 m2 or more area. Thus the growing system must be developed to provide an area that is darkened for tuber and root growth and of sufficient size for plant spread. A system developed for growing potatoes, or any plants, in space will have certain requirements that must be met to make them a useful part of a life support system. The system must 1) be constructed of materials, and involve media, that can be reused for many successive cycles of plant growth, 2
Ratio of serum free triiodothyronine to free thyroxine in Graves' hyperthyroidism and thyrotoxicosis caused by painless thyroiditis.
Yoshimura Noh, Jaeduk; Momotani, Naoko; Fukada, Shuji; Ito, Koichi; Miyauchi, Akira; Amino, Nobuyuki
The serum T3 to T4 ratio is a useful indicator for differentiating destruction-induced thyrotoxicosis from Graves' thyrotoxicosis. However, the usefulness of the serum free T3 (FT3) to free T4 (FT4) ratio is controversial. We therefore systematically evaluated the usefulness of this ratio, based on measurements made using two widely available commercial kits in two hospitals. Eighty-two untreated patients with thyrotoxicosis (48 patients with Graves' disease and 34 patients with painless thyroiditis) were examined in Kuma Hospital, and 218 patients (126 with Graves' disease and 92 with painless thyroiditis) and 66 normal controls were examined in Ito Hospital. The FT3 and FT4 values, as well as the FT3/FT4 ratios, were significantly higher in the patients with Graves' disease than in those with painless thyroiditis in both hospitals, but considerable overlap between the two disorders was observed. Receiver operating characteristic (ROC) curves for the FT3 and FT4 values and the FT3/FT4 ratios of patients with Graves' disease and those with painless thyroiditis seen in both hospitals were prepared, and the area under the curves (AUC), the cut-off points for discriminating Graves' disease from painless thyroiditis, the sensitivity, and the specificity were calculated. AUC and sensitivity of the FT(3)/FT(4) ratio were smaller than those of FT(3) and FT(4) in both hospitals. The patients treated at Ito hospital were then divided into 4 groups according to their FT4 levels (A: 2.3 approximately 5.4 ng/dl), and the AUC, cut-off points, sensitivity, and specificity of the FT(3)/FT(4) ratios were calculated. The AUC and sensitivity of each group increased with the FT4 levels (AUC: 57.8%, 72.1%, 91.1%, and 93.4%, respectively; sensitivity: 62.6%, 50.0%, 77.8%, and 97.0%, respectively). The means +/- SE of the FT3/FT4 ratio in the Graves' disease groups were 3.1 +/- 0.22, 3.1 +/- 0.09, 3.2 +/- 0.06, and 3.1 +/- 0.07, respectively, versus 2.9 +/- 0.1, 2.6 +/- 0.07, 2.5 +/- 0
Reproducibility in Research: Systems, Infrastructure, Culture
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Tom Crick
Full Text Available The reproduction and replication of research results has become a major issue for a number of scientific disciplines. In computer science and related computational disciplines such as systems biology, the challenges closely revolve around the ability to implement (and exploit novel algorithms and models. Taking a new approach from the literature and applying it to a new codebase frequently requires local knowledge missing from the published manuscripts and transient project websites. Alongside this issue, benchmarking, and the lack of open, transparent and fair benchmark sets present another barrier to the verification and validation of claimed results. In this paper, we outline several recommendations to address these issues, driven by specific examples from a range of scientific domains. Based on these recommendations, we propose a high-level prototype open automated platform for scientific software development which effectively abstracts specific dependencies from the individual researcher and their workstation, allowing easy sharing and reproduction of results. This new e-infrastructure for reproducible computational science offers the potential to incentivise a culture change and drive the adoption of new techniques to improve the quality and efficiency – and thus reproducibility – of scientific exploration.
The Influence Of Organizational Culture On Management Information System
Directory of Open Access Journals (Sweden)
Arlis Dewi Kuraesin
Full Text Available This article aims to know the culture of the organization and management accounting information system based on existing theories. The management information system is a collection of sub-systems which are interconnected with each other to work together in harmony to achieve one goal of process data into information needed by management in decision making. An important factor influencing the use of information systems is Cultural Organization. Management Information system success is influenced by several factors one of which is the organizations culture. Organizational culture has a very strong influence on the overall organizational and individual behavior due to the information system is a major component of the organization are influenced substantially by organizational culture.
Aseptic Plant Culture System (APCS), Phase II
National Aeronautics and Space Administration — Aseptic plant culture plays a significant role in biotechnology and plant physiology research, and in vegetative propagation of many plant species. The development...
Aseptic Plant Culture System (APCS), Phase I
National Aeronautics and Space Administration — Aseptic plant culture plays a significant role in biotechnology and plant physiology research and in vegetative propagation of many plant species. The development of...
Usability and Applicability of Microfluidic Cell Culture Systems
DEFF Research Database (Denmark)
Hemmingsen, Mette
possibilities for, for example, precise control of the chemical environment, 3D cultures, controlled co-culture of different cell types or automated, individual control of up to 96 cell culture chambers in one integrated system. Despite the great new opportunities to perform novel experimental designs......Microfluidic cell culture has been a research area with great attention the last decade due to its potential to mimic the in vivo cellular environment more closely compared to what is possible by conventional cell culture methods. Many exciting and complex devices have been presented providing......, these devices still lack general implementation into biological research laboratories. In this project, the usability and applicability of microfluidic cell culture systems have been investigated. The tested systems display good properties regarding optics and compatibility with standard laboratory equipment...
Plant assessment system and safety culture
International Nuclear Information System (INIS)
Chun, Chuyoung
The government, upon these events, keenly felt the necessity for developing the safety culture which was already forwarded in nuclear industries and started taking actions to propagate it to all parts of society. The government established a social safety director position under the Prime Minister's jurisdiction and also established a Safety Culture Promotion Headquarters in which 7 ministries and other organizations, such as Korea Economic Council, Federation of Korea Trade Union and Women's Federation Council were participating. In accordance with the government's strong will to enhance the safety consciousness of people, safety campaigns are being developed voluntarily in the private sector. The formation of non-governmental organizations, such as People's Central Council of Safety Culture Promotion, shows a good example of such movement
Tumour-cytolytic human monocyte-derived macrophages: a simple and efficient method for the generation and long-term cultivation as non-adherent cells in a serum-free medium.
Streck, R J; Hurley, E L; Epstein, D A; Pauly, J L
We report a simple and efficient culture procedure for the generation of tumour-cytolytic human monocyte-derived macrophages (MAC). In this method, normal human peripheral blood mononuclear cells, isolated using a conventional Ficoll-Hypaque density gradient procedure, are cultured as a heterogenous leukocyte population in Teflon or other hydrophobic cultureware, in a commercially available serum-free culture medium (M-SFM) that has been formulated specifically for the cultivation and ex vivo stimulation of human monocytes and MAC, and in the absence of exogenous mitogens, antigens, cytokines or other stimulants. This procedure features a negative-selection technique that takes advantage of the differential survival of blood leukocytes. Using the prescribed in vitro conditions, lymphocytes survived relatively poorly, whereas monocytes differentiated in the absence of exogenous stimulants into mature tumour-cytolytic MAC. The MAC were present as non-adherent, single cells that expressed good viability (greater than 95%) for a prolonged period (greater than 60 days). When compared to conventional procedures for generating MAC, the prescribed technique is thought to offer several important advantages in that it: (a) eliminates the tedious and cumbersome monocyte isolation procedures, thus providing a significant savings not only in time and money but also in eliminating repetitive cell manipulations that have often been associated with damage to monocyte morphology and/or function; (b) reduces the loss of monocyte subsets that are not recovered during specific isolation procedures; (c) facilitates harvesting a single cell, non-adherent suspension of immunocompetent MAC suitable for various examinations including analyses defining MAC morphology, cytochemistry, phenotype and function; and (d) eliminates variability and artifacts associated with different sera that are utilised frequently as medium supplements. The utility of the prescribed method is illustrated by the
Biona-C Cell Culture pH Monitoring System
Friedericks, C.
Sensors 2000! is developing a system to demonstrate the ability to perform accurate, real-time measurements of pH and CO2 in a cell culture media in Space. The BIONA-C Cell Culture pH Monitoring System consists of S2K! developed ion selective sensors and control electronics integrated with the fluidics of a cell culture system. The integrated system comprises a "rail" in the Cell Culture Module (CCM) of WRAIR (Space Biosciences of Walter Read Army Institute of Research). The CCM is a Space Shuttle mid-deck locker experiment payload. The BIONA-C is displayed along with associated graphics and text explanations. The presentation will stimulate interest in development of sensor technology for real-time cell culture measurements. The transfer of this technology to other applications will also be of interest. Additional information is contained in the original document.
Performance of Gram staining on blood cultures flagged negative by an automated blood culture system.
Peretz, A; Isakovich, N; Pastukh, N; Koifman, A; Glyatman, T; Brodsky, D
Blood is one of the most important specimens sent to a microbiology laboratory for culture. Most blood cultures are incubated for 5-7 days, except in cases where there is a suspicion of infection caused by microorganisms that proliferate slowly, or infections expressed by a small number of bacteria in the bloodstream. Therefore, at the end of incubation, misidentification of positive cultures and false-negative results are a real possibility. The aim of this work was to perform a confirmation by Gram staining of the lack of any microorganisms in blood cultures that were identified as negative by the BACTEC™ FX system at the end of incubation. All bottles defined as negative by the BACTEC FX system were Gram-stained using an automatic device and inoculated on solid growth media. In our work, 15 cultures that were defined as negative by the BACTEC FX system at the end of the incubation were found to contain microorganisms when Gram-stained. The main characteristic of most bacteria and fungi growing in the culture bottles that were defined as negative was slow growth. This finding raises a problematic issue concerning the need to perform Gram staining of all blood cultures, which could overload the routine laboratory work, especially laboratories serving large medical centers and receiving a large number of blood cultures.
Culture and cognition in health systems change.
Evans, Jenna M; Baker, G Ross; Berta, Whitney; Barnsley, Jan
Large-scale change involves modifying not only the structures and functions of multiple organizations, but also the mindsets and behaviours of diverse stakeholders. This paper focuses on the latter: the informal, less visible, and often neglected psychological and social factors implicated in change efforts. The purpose of this paper is to differentiate between the concepts of organizational culture and mental models, to argue for the value of applying a shared mental models (SMM) framework to large-scale change, and to suggest directions for future research. The authors provide an overview of SMM theory and use it to explore the dynamic relationship between culture and cognition. The contributions and limitations of the theory to change efforts are also discussed. Culture and cognition are complementary perspectives, providing insight into two different levels of the change process. SMM theory draws attention to important questions that add value to existing perspectives on large-scale change. The authors outline these questions for future research and argue that research and practice in this domain may be best served by focusing less on the potentially narrow goal of "achieving consensus" and more on identifying, understanding, and managing cognitive convergences and divergences as part of broader research and change management programmes. Drawing from both cultural and cognitive paradigms can provide researchers with a more complete picture of the processes by which coordinated action are achieved in complex change initiatives in the healthcare domain.
Sustaining organizational culture change in health systems.
Willis, Cameron David; Saul, Jessie; Bevan, Helen; Scheirer, Mary Ann; Best, Allan; Greenhalgh, Trisha; Mannion, Russell; Cornelissen, Evelyn; Howland, David; Jenkins, Emily; Bitz, Jennifer
The questions addressed by this review are: first, what are the guiding principles underlying efforts to stimulate sustained cultural change; second, what are the mechanisms by which these principles operate; and, finally, what are the contextual factors that influence the likelihood of these principles being effective? The paper aims to discuss these issues. The authors conducted a literature review informed by rapid realist review methodology that examined how interventions interact with contexts and mechanisms to influence the sustainability of cultural change. Reference and expert panelists assisted in refining the research questions, systematically searching published and grey literature, and helping to identify interactions between interventions, mechanisms and contexts. Six guiding principles were identified: align vision and action; make incremental changes within a comprehensive transformation strategy; foster distributed leadership; promote staff engagement; create collaborative relationships; and continuously assess and learn from change. These principles interact with contextual elements such as local power distributions, pre-existing values and beliefs and readiness to engage. Mechanisms influencing how these principles sustain cultural change include activation of a shared sense of urgency and fostering flexible levels of engagement. The principles identified in this review, along with the contexts and mechanisms that influence their effectiveness, are useful domains for policy and practice leaders to explore when grappling with cultural change. These principles are sufficiently broad to allow local flexibilities in adoption and application. This is the first study to adopt a realist approach for understanding how changes in organizational culture may be sustained. Through doing so, this review highlights the broad principles by which organizational action may be organized within enabling contextual settings.
Growing B Lymphocytes in a Three-Dimensional Culture System
Wu, J. H. David; Bottaro, Andrea
A three-dimensional (3D) culture system for growing long-lived B lymphocytes has been invented. The capabilities afforded by the system can be expected to expand the range of options for immunological research and related activities, including testing of immunogenicity of vaccine candidates in vitro, generation of human monoclonal antibodies, and immunotherapy. Mature lymphocytes, which are the effectors of adaptive immune responses in vertebrates, are extremely susceptible to apoptotic death, and depend on continuous reception of survival-inducing stimulation (in the forms of cytokines, cell-to-cell contacts, and antigen receptor signaling) from the microenvironment. For this reason, efforts to develop systems for long-term culture of functional, non-transformed and non-activated mature lymphocytes have been unsuccessful until now. The bone-marrow microenvironment supports the growth and differentiation of many hematopoietic lineages, in addition to B-lymphocytes. Primary bone-marrow cell cultures designed to promote the development of specific cell types in vitro are highly desirable experimental systems, amenable to manipulation under controlled conditions. However, the dynamic and complex network of stromal cells and insoluble matrix proteins is disrupted in prior plate- and flask-based culture systems, wherein the microenvironments have a predominantly two-dimensional (2D) character. In 2D bone-marrow cultures, normal B-lymphoid cells become progressively skewed toward precursor B-cell populations that do not retain a normal immunophenotype, and such mature B-lymphocytes as those harvested from the spleen or lymph nodes do not survive beyond several days ex vivo in the absence of mitogenic stimulation. The present 3D culture system is a bioreactor that contains highly porous artificial scaffolding that supports the long-term culture of bone marrow, spleen, and lymph-node samples. In this system, unlike in 2D culture systems, B-cell subpopulations developing
Cytoarchitecture in cultured rat neocortex explants
de Jong, B. M.; Ruijter, J. M.; Romijn, H. J.
Neocortex explants obtained from 6-day-old rat pups and cultured in a serum-free medium from 5 hr to 13 days in vitro (DIV) show preservation of cytoarchitectural characteristics. Major changes in the size of the explants and their layers occur during the first 2 DIV. A radial arrangement of neurons
Development of a microfluidic perfusion 3D cell culture system
Park, D. H.; Jeon, H. J.; Kim, M. J.; Nguyen, X. D.; Morten, K.; Go, J. S.
Recently, 3-dimensional in vitro cell cultures have gained much attention in biomedical sciences because of the closer relevance between in vitro cell cultures and in vivo environments. This paper presents a microfluidic perfusion 3D cell culture system with consistent control of long-term culture conditions to mimic an in vivo microenvironment. It consists of two sudden expansion reservoirs to trap incoming air bubbles, gradient generators to provide a linear concentration, and microchannel mixers. Specifically, the air bubbles disturb a flow in the microfluidic channel resulting in the instability of the perfusion cell culture conditions. For long-term stable operation, the sudden expansion reservoir is designed to trap air bubbles by using buoyancy before they enter the culture system. The performance of the developed microfluidic perfusion 3D cell culture system was examined experimentally and compared with analytical results. Finally, it was applied to test the cytotoxicity of cells infected with Ewing's sarcoma. Cell death was observed for different concentrations of H2O2. For future work, the developed microfluidic perfusion 3D cell culture system can be used to examine the behavior of cells treated with various drugs and concentrations for high-throughput drug screening.
An Integrated Approach Using Liquid Culture System Can it Make ...
African Journals Online (AJOL)
Diagnostic dilemma is a common problem faced as culture and polymerase chain reaction results vary in their sensitivity and specificity. A thorough knowledge of epidemiology, immunopathogenesis, and spectrum of the disease and importance of including liquid culture system for the diagnosis of this disease are ...
Culturally grounded indicators of resilience in social-ecological systems
Eleanor Sterling; Tamara Ticktin; Tē Kipa Kepa Morgan; Georgina Cullman; Diana Alvira; Pelika Andrade; Nadia Bergamini; Erin Betley; Kate Burrows; Sophie Caillon; Joachim Claudet; Rachel Dacks; Pablo Eyzaguirre; Chris Filardi; Nadav Gazit; Christian Giardina; Stacy Jupiter; Kealohanuiopuna Kinney; Joe McCarter; Manuel Mejia; Kanoe Morishige; Jennifer Newell; Lihla Noori; John Parks; Pua'ala Pascua; Ashwin Ravikumar; Jamie Tanguay; Amanda Sigouin; Tina Stege; Mark Stege; Alaka Wali
Measuring progress toward sustainability goals is a multifaceted task. International, regional, and national organizations and agencies seek to promote resilience and capacity for adaptation at local levels. However, their measurement systems may be poorly aligned with local contexts, cultures, and needs. Understanding how to build effective, culturally grounded...
a Cultural Landscape Information System Developed with Open Source Tools
Chudyk, C.; Müller, H.; Uhler, M.; Würriehausen, F.
Since 2010, the state of Rhineland-Palatinate in Germany has developed a cultural landscape information system as a process to secure and further enrich aggregate data about its cultural assets. In an open dialogue between governing authorities and citizens, the intention of the project is an active cooperation of public and private actors. A cultural landscape information system called KuLIS was designed as a web platform, combining semantic wiki software with a geographic information system. Based on data sets from public administrations, the information about cultural assets can be extended and enhanced by interested participants. The developed infrastructure facilitates local information accumulation through a crowdsourcing approach. This capability offers new possibilities for e-governance and open data developments. The collaborative approach allows governing authorities to manage and supervise official data, while public participation enables affordable information acquisition. Gathered cultural heritage information can provide incentives for touristic valorisation of communities or concepts for strengthening regional identification. It can also influence political decisions in defining significant cultural regions worth of protecting from industrial influences. The presented cultural landscape information allows citizens to influence the statewide development of cultural landscapes in a democratic way.
A Managerial Approach to NASA's Cultural Changes: Open System Model
National Research Council Canada - National Science Library
Aytekin, Yasin; Long, Nicholas
This project describes NASA's culture during two important time periods (1958-1972) and (1996-2004) and explains its relative fit with its system components -- task, people, resources, and structure...
Simple Impeller Systems for Maintenance of Oil Palm Culture Aggregates
International Nuclear Information System (INIS)
Tarmizi, A.H.; Zaiton, R.; Rosli, M.Y.
Scaling up of liquid culture systems generally involves moving from the use of simple shake flasks to bioreactors or specialised vessels; this is costly. A new innovation called the Two-in-One MPOB Simple Impeller (2-in-1 MoSLIM) was developed using commonly available Schott bottles in the laboratory. This system provided simultaneous aeration and agitation (two-in-one) in a single device for tissue propagation in liquid culture. The 2-in-1 MoSLIM produced cell aggregates with fresh weight increments of two- to six-fold over 30-40 days. This system was a convenient alternative compared to the conventional shake flask system. Multiplication of cultures in the 2-in-1 MoSLIM did not require any shaker or a big space area. This system with a working volume of 300 - 700 ml used a simple impeller and a pump for agitation and aeration purposes. However, with the 2-in-1 MoSLIM, media replenishment remained a tedious task. To overcome this, modifications were made to the system to enable media replenishment on-site without the need of a sterile hood. The adaptation of 2-in-1 MoSLIM with an earlier innovation, Fast Transfer Technique (MoFaTT) in Liquid Culture System, resulted in the development of the Simple Impeller with Fast Transfer Technique (SLIM-FaTT) system. This new system can be applied to the liquid culture system of any crop with a potential towards automation. (author)
Miniature Bioreactor System for Long-Term Cell Culture
Gonda, Steve R.; Kleis, Stanley J.; Geffert, Sandara K.
A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays.
Cultural relativism: maintenance of genomic imprints in pluripotent stem cell culture systems.
Greenberg, Maxim Vc; Bourc'his, Déborah
Pluripotent stem cells (PSCs) in culture have become a widely used model for studying events occurring during mammalian development; they also present an exciting avenue for therapeutics. However, compared to their in vivo counterparts, cultured PSC derivatives have unique properties, and it is well established that their epigenome is sensitive to medium composition. Here we review the specific effects on genomic imprints in various PSC types and culture systems. Imprinted gene regulation is developmentally important, and imprinting defects have been associated with several human diseases. Therefore, imprint abnormalities in PSCs may have considerable consequences for downstream applications. Copyright © 2015 Elsevier Ltd. All rights reserved.
Differentiation of mammalian skeletal muscle cells cultured on microcarrier beads in a rotating cell culture system
Torgan, C. E.; Burge, S. S.; Collinsworth, A. M.; Truskey, G. A.; Kraus, W. E.
The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.
Efficiency of liquid culture systems over conventional ...
African Journals Online (AJOL)
The most common methods of micropropagation involve the proliferation of shoots via a semi solid system. While such semi solid systems have been moderately to highly successful in terms of multiplication yields, it has become increasingly important to improve productivity and reduce the time taken to multiply ...
IAEA Safety Standards on Management Systems and Safety Culture
International Nuclear Information System (INIS)
Persson, Kerstin Dahlgren
The IAEA has developed a new set of Safety Standard for applying an integrated Management System for facilities and activities. The objective of the new Safety Standards is to define requirements and provide guidance for establishing, implementing, assessing and continually improving a Management System that integrates safety, health, environmental, security, quality and economic related elements to ensure that safety is properly taken into account in all the activities of an organization. With an integrated approach to management system it is also necessary to include the aspect of culture, where the organizational culture and safety culture is seen as crucial elements of the successful implementation of this management system and the attainment of all the goals and particularly the safety goals of the organization. The IAEA has developed a set of service aimed at assisting it's Member States in establishing. Implementing, assessing and continually improving an integrated management system. (author)
Geographic Names Information System (GNIS) Cultural Features
Department of Homeland Security — The Geographic Names Information System (GNIS) is the Federal standard for geographic nomenclature. The U.S. Geological Survey developed the GNIS for the U.S. Board...
Influence of serum extraction from the culture medium and of sublethal X-ray irradiation upon microvilli and invaginations of the membrane of Ehrlich ascites tumor cells in monolayer culture
International Nuclear Information System (INIS)
Laudenbach, G.; Pfab, R.; Hess, F.; Schachtschabel, D.O.
In order to find out modifications of microvilli and invaginations, the cellular surfaces of Ehrlich ascites tumor cells in monolayer culture (basal medium of Eagle + 10% fetal calf serum) were investigated with the aid of electron-microscopic cross-sections. The tumor cells had been cultured without serum 24 hours prior to investigation or irradiated with 2 Gy. Morphometric evaluation after cell culture in a serum-free medium showed a reduced number of microvilli and a diminution of sections of microvilli. As already described before, a reduction of cell proliferation, of the microtubule-microfilament system, and of the endocytosis activity occurs under these serum-free conditions. The number of invaginations (related to a constant membrane part) was reduced by nearly 50% after serum extraction. Similarly to serum extraction, sublethal X-ray irradiation reduced the sections of microvilli, whereas the number of microvilli increased slightly. Contrary to the effect of serum extraction, the irradiated cells showed twice as many invaginations as the non-irradiated control cells. These differences in the surface structures are interpreted as a result of modified growth stimulations (+- serum) and radiogenic reparation processes. (orig.) [de
Cultural Factors in Systems Design Decision Making and Action
Proctor, Robert W; Yih, Yuehwern
This book brings together an interdisciplinary group of experts to provide increased understanding of the ways in which cultural differences may influence decision making and action. It brings together current knowledge about decision processes, culture and cognition, design of products and interfaces for human interaction with machines and organizational processes culled from a wide variety of sources and puts them into one comprehensive resource. It examines how to design systems used by individuals from different cultures and accommodate the varied backgrounds that affect the users' decisio
Cultural selection drives the evolution of human communication systems.
Tamariz, Monica; Ellison, T Mark; Barr, Dale J; Fay, Nicolas
Human communication systems evolve culturally, but the evolutionary mechanisms that drive this evolution are not well understood. Against a baseline that communication variants spread in a population following neutral evolutionary dynamics (also known as drift models), we tested the role of two cultural selection models: coordination- and content-biased. We constructed a parametrized mixed probabilistic model of the spread of communicative variants in four 8-person laboratory micro-societies engaged in a simple communication game. We found that selectionist models, working in combination, explain the majority of the empirical data. The best-fitting parameter setting includes an egocentric bias and a content bias, suggesting that participants retained their own previously used communicative variants unless they encountered a superior (content-biased) variant, in which case it was adopted. This novel pattern of results suggests that (i) a theory of the cultural evolution of human communication systems must integrate selectionist models and (ii) human communication systems are functionally adaptive complex systems.
Three-dimensional alginate spheroid culture system of murine osteosarcoma.
Akeda, Koji; Nishimura, Akinobu; Satonaka, Haruhiko; Shintani, Ken; Kusuzaki, Katsuyuki; Matsumine, Akihiko; Kasai, Yuichi; Masuda, Koichi; Uchida, Atsumasa
Osteosarcoma (OS) is the most common primary malignant tumor of the bone and often forms pulmonary metastases, which are the most important prognostic factor. For further elucidation of the mechanism underlying the progression and metastasis of human OS, a culture system mimicking the microenvironment of the tumor in vivo is needed. We report a novel three-dimensional (3D) alginate spheroid culture system of murine osteosarcoma. Two different metastatic clones, the parental Dunn and its derivative line LM8, which has a higher metastatic potential to the lungs, were encapsulated in alginate beads to develop the 3D culture system. The beads containing murine OS cells were also transplanted into mice to determine their metastatic potential in vivo. In this culture system, murine OS cells encapsulated in alginate beads were able to grow in a 3D structure with cells detaching from the alginate environment. The number of detaching cells was higher in the LM8 cell line than the Dunn cell line. In the in vivo alginate bead transplantation model, the rate of pulmonary metastasis was higher with LM8 cells compared with that of Dunn cells. The cell characteristics and kinetics in this culture system closely reflect the original malignant potential of the cells in vivo.
A second inheritance system: the extension of biology through culture.
Whiten, Andrew
By the mid-twentieth century (thus following the 'Modern Synthesis' in evolutionary biology), the behavioural sciences offered only the sketchy beginnings of a scientific literature documenting evidence for cultural inheritance in animals-the transmission of traditional behaviours via learning from others (social learning). By contrast, recent decades have seen a massive growth in the documentation of such cultural phenomena, driven by long-term field studies and complementary laboratory experiments. Here, I review the burgeoning scope of discoveries in this field, which increasingly suggest that this 'second inheritance system', built on the shoulders of the primary genetic inheritance system, occurs widely among vertebrates and possibly in invertebrates too. Its novel characteristics suggest significant implications for our understanding of evolutionary biology. I assess the extent to which this second system extends the scope of evolution, both by echoing principal properties of the primary, organic evolutionary system, and going beyond it in significant ways. This is well established in human cultural evolution; here, I address animal cultures more generally. The further major, and related, question concerns the extent to which the consequences of widespread animal cultural transmission interact with the primary, genetically based inheritance systems, shaping organic evolution.
A Situated Cultural Festival Learning System Based on Motion Sensing
Chang, Yi-Hsing; Lin, Yu-Kai; Fang, Rong-Jyue; Lu, You-Te
A situated Chinese cultural festival learning system based on motion sensing is developed in this study. The primary design principle is to create a highly interactive learning environment, allowing learners to interact with Kinect through natural gestures in the designed learning situation to achieve efficient learning. The system has the…
Culture temperature affects human chondrocyte messenger RNA expression in monolayer and pellet culture systems.
Directory of Open Access Journals (Sweden)
Akira Ito
Full Text Available Cell-based therapy has been explored for articular cartilage regeneration. Autologous chondrocyte implantation is a promising cell-based technique for repairing articular cartilage defects. However, there are several issues such as chondrocyte de-differentiation. While numerous studies have been designed to overcome some of these issues, only a few have focused on the thermal environment that can affect chondrocyte metabolism and phenotype. In this study, the effects of different culture temperatures on human chondrocyte metabolism- and phenotype-related gene expression were investigated in 2D and 3D environments. Human chondrocytes were cultured in a monolayer or in a pellet culture system at three different culture temperatures (32°C, 37°C, and 41°C for 3 days. The results showed that the total RNA level, normalized to the threshold cycle value of internal reference genes, was higher at lower temperatures in both culture systems. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH and citrate synthase (CS, which are involved in glycolysis and the citric acid cycle, respectively, were expressed at similar levels at 32°C and 37°C in pellet cultures, but the levels were significantly lower at 41°C. Expression of the chondrogenic markers, collagen type IIA1 (COL2A1 and aggrecan (ACAN, was higher at 37°C than at 32°C and 41°C in both culture systems. However, this phenomenon did not coincide with SRY (sex-determining region Y-box 9 (SOX9, which is a fundamental transcription factor for chondrogenesis, indicating that a SOX9-independent pathway might be involved in this phenomenon. In conclusion, the expression of chondrocyte metabolism-related genes at 32°C was maintained or enhanced compared to that at 37°C. However, chondrogenesis-related genes were further induced at 37°C in both culture systems. Therefore, manipulating the culture temperature may be an advantageous approach for regulating human chondrocyte metabolic activity and
Culture temperature affects human chondrocyte messenger RNA expression in monolayer and pellet culture systems.
Ito, Akira; Nagai, Momoko; Tajino, Junichi; Yamaguchi, Shoki; Iijima, Hirotaka; Zhang, Xiangkai; Aoyama, Tomoki; Kuroki, Hiroshi
Cell-based therapy has been explored for articular cartilage regeneration. Autologous chondrocyte implantation is a promising cell-based technique for repairing articular cartilage defects. However, there are several issues such as chondrocyte de-differentiation. While numerous studies have been designed to overcome some of these issues, only a few have focused on the thermal environment that can affect chondrocyte metabolism and phenotype. In this study, the effects of different culture temperatures on human chondrocyte metabolism- and phenotype-related gene expression were investigated in 2D and 3D environments. Human chondrocytes were cultured in a monolayer or in a pellet culture system at three different culture temperatures (32°C, 37°C, and 41°C) for 3 days. The results showed that the total RNA level, normalized to the threshold cycle value of internal reference genes, was higher at lower temperatures in both culture systems. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and citrate synthase (CS), which are involved in glycolysis and the citric acid cycle, respectively, were expressed at similar levels at 32°C and 37°C in pellet cultures, but the levels were significantly lower at 41°C. Expression of the chondrogenic markers, collagen type IIA1 (COL2A1) and aggrecan (ACAN), was higher at 37°C than at 32°C and 41°C in both culture systems. However, this phenomenon did not coincide with SRY (sex-determining region Y)-box 9 (SOX9), which is a fundamental transcription factor for chondrogenesis, indicating that a SOX9-independent pathway might be involved in this phenomenon. In conclusion, the expression of chondrocyte metabolism-related genes at 32°C was maintained or enhanced compared to that at 37°C. However, chondrogenesis-related genes were further induced at 37°C in both culture systems. Therefore, manipulating the culture temperature may be an advantageous approach for regulating human chondrocyte metabolic activity and chondrogenesis.
Cross-cultural Human-Machine-Systems: selected aspects of a cross-cultural system engineering; Interkulturelle Mensch-Maschine-Systeme: ausgewaehlte Aspekte einer interkulturellen Systemgestaltung
Energy Technology Data Exchange (ETDEWEB)
Roese, K. [Technische Univ. Kaiserslautern (Germany). AG Nutzergerechte Produktentwicklung
Cross-cultural Human-Machine-Systems are one key factor for success in the global market era. Nowadays the machine producer have to offer their products worldwide. With the export to other nations they have to consider on the user behaviour in these other cultures. The analysis of cross-cultural user requirements and their integration into the product development process is a real chance to cape with these challenge. This paper describe two aspects of cross-cultural user aspects. It gives an impression of the complex and sometimes unknown cultural influencing factors and their impact on Human-Machine-System-Engineering. (orig.)
A Hybrid Robotic Control System Using Neuroblastoma Cultures
Ferrández, J. M.; Lorente, V.; Cuadra, J. M.; Delapaz, F.; Álvarez-Sánchez, José Ramón; Fernández, E.
The main objective of this work is to analyze the computing capabilities of human neuroblastoma cultured cells and to define connection schemes for controlling a robot behavior. Multielectrode Array (MEA) setups have been designed for direct culturing neural cells over silicon or glass substrates, providing the capability to stimulate and record simultaneously populations of neural cells. This paper describes the process of growing human neuroblastoma cells over MEA substrates and tries to modulate the natural physiologic responses of these cells by tetanic stimulation of the culture. We show that the large neuroblastoma networks developed in cultured MEAs are capable of learning: establishing numerous and dynamic connections, with modifiability induced by external stimuli and we propose an hybrid system for controlling a robot to avoid obstacles.
Smart management of sample dilution using an artificial neural network to achieve streamlined processes and saving resources: the automated nephelometric testing of serum free light chain as case study.
Ialongo, Cristiano; Pieri, Massimo; Bernardini, Sergio
Saving resources is a paramount issue for the modern laboratory, and new trainable as well as smart technologies can be used to allow the automated instrumentation to manage samples more efficiently in order to achieve streamlined processes. In this regard the serum free light chain (sFLC) testing represents an interesting challenge, as it usually causes using a number of assays before achieving an acceptable result within the analytical range. An artificial neural network based on the multi-layer perceptron (MLP-ANN) was used to infer the starting dilution status of sFLC samples based on the information available through the laboratory information system (LIS). After the learning phase, the MLP-ANN simulation was applied to the nephelometric testing routinely performed in our laboratory on a BN ProSpec® System analyzer (Siemens Helathcare) using the N Latex FLC kit. The MLP-ANN reduced the serum kappa free light chain (κ-FLC) and serum lambda free light chain (λ-FLC) wasted tests by 69.4% and 70.8% with respect to the naïve stepwise dilution scheme used by the automated analyzer, and by 64.9% and 66.9% compared to a "rational" dilution scheme based on a 4-step dilution. Although it was restricted to follow-up samples, the MLP-ANN showed good predictive performance, which alongside the possibility to implement it in any automated system, made it a suitable solution for achieving streamlined laboratory processes and saving resources.
Deciphering defective amelogenesis using in vitro culture systems.
Arinawati, Dian Yosi; Miyoshi, Keiko; Tanimura, Ayako; Horiguchi, Taigo; Hagita, Hiroko; Noma, Takafumi
The conventional two-dimensional (2D) in vitro culture system is frequently used to analyze the gene expression with or without extracellular signals. However, the cells derived from primary culture and cell lines frequently deviate the gene expression profile compared to the corresponding in vivo samples, which sometimes misleads the actual gene regulation in vivo. To overcome this gap, we developed the comparative 2D and 3D in vitro culture systems and applied them to the genetic study of amelogenesis imperfecta (AI) as a model. Recently, we found specificity protein 6 (Sp6) mutation in an autosomal-recessive AI rat that was previously named AMI. We constructed 3D structure of ARE-B30 cells (AMI-derived rat dental epithelial cells) or G5 (control wild type cells) combined with RPC-C2A cells (rat pulp cell line) separated by the collagen membrane, while in 2D structure, ARE-B30 or G5 was cultured with or without the collagen membrane. Comparative analysis of amelogenesis-related gene expression in ARE-B30 and G5 using our 2D and 3D in vitro systems revealed distinct expression profiles, showing the causative outcomes. Bone morphogenetic protein 2 and follistatin were reciprocally expressed in G5, but not in ARE-B30 cells. All-or-none expression of amelotin, kallikrein-related peptidase 4, and nerve growth factor receptor was observed in both cell types. In conclusion, our in vitro culture systems detected the phenotypical differences in the expression of the stage-specific amelogenesis-related genes. Parallel analysis with 2D and 3D culture systems may provide a platform to understand the molecular basis for defective amelogenesis caused by Sp6 mutation. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.
[Comparison of two types of cell cultures for preparation of sTNFRII-gAD fusion protein].
Huang, Shigao; Yin, Yuting; Xiong, Chunhui; Wang, Caihong; Lü, Jianxin; Gao, Jimin
In this study we used two types of cell cultures, i.e., anchorage-dependent basket and full suspension batch cultures of sTNFRII-gAD-expressing CHO cells in the CelliGen 310 bioreactor (7.5 L) to compare their yields in order to optimize the culturing conditions for efficient expression of sTNFRII-gAD fusion protein consisting of soluble tumor necrosis factor receptor II and globular domain of adiponectin. The anchorage-dependent basket culture was performed in 4L 10% serum-containing medium with the final inoculating concentration of 3 x 10(5) to 4 x 10(5) cells/mL of sTNFRII-gAD-expressing CHO cells for 3 days, and then switched to 4 L serum-free LK021 medium to continue the culture for 4 days. The full suspension batch culture was carried out in the 4 L serum-free LK021 medium with the final inoculating concentration of 3 x 10(5) to 4 x 10(5) cells/mL of sTNFRII-gAD-expressing CHO cells for 7 days. The culturing conditions were monitored in real-time to maintain pH and dissolved oxygen stability through the whole process. The supernatants were collected by centrifuge, and the protein was concentrated through Pellicon flow ultrafiltration system and then purified by DEAE anion exchange. The results showed that the yields of sTNFRII-gAD fusion protein were 8.0 mg/L with 95% purity and 7.5 mg/L with 98% purity in the anchorage-dependent basket and the full suspension batch cultures, respectively. The study provided the framework for the pilot production of sTNFRII-gAD fusion protein.
The effect of three culture methods on intensive culture system of pacific white shrimp ( Litopenaeus vannamei)
Ma, Zhen; Wan, Rong; Song, Xiefa; Gao, Lei
Different culture methods may affect the intensive culture system of Pacific white shrimp ( Litopenaeus vannamei) regarding water quality and growth and economic performance. This study evaluated the potential effects of three culture methods through cultivation of juvenile shrimps under consistent tank management conditions for 84 d. The three methods involved shrimp cultivation in different tanks, i.e., outdoor tanks with cement bottom (mode-C), greenhouse tanks with cement bottom (mode-G) and outdoor tanks with mud-substrate (mode-M). Results showed that water temperature was significantly higher in mode-G than that in mode-C ( P shrimps. In the mid-late period, the average concentrations of TAN, NO2-N, DIP and COD were significantly lower in mode-M and mode-G compared with those in mode-C ( P shrimp weight among different treatments ( P > 0.05), mode-M had significantly higher shrimp yield, survival rate and feed conversion rate ( P < 0.05) than other modes. There were significant differences in revenue and net return among different treatments ( P < 0.05). These demonstrated that the treatments of mode-G and mode-M were conductive to the intensive culture system of L. vannamei.
The Cultural Analysis of Soft Systems Methodology and the Configuration Model of Organizational Culture
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Jürgen Staadt
Full Text Available Organizations that find themselves within a problematic situation connected with cultural issues such as politics and power require adaptable research and corresponding modeling approaches so as to grasp the arrangements of that situation and their impact on the organizational development. This article originates from an insider-ethnographic intervention into the problematic situation of the leading public housing provider in Luxembourg. Its aim is to describe how the more action-oriented cultural analysis of soft systems methodology and the theory-driven configuration model of organizational culture are mutually beneficial rather than contradictory. The data collected between 2007 and 2013 were analyzed manually as well as by means of ATLAS.ti. Results demonstrate that the cultural analysis enables an in-depth understanding of the power-laden environment within the organization bringing about the so-called "socio-political system" and that the configuration model makes it possible to depict the influence of that system on the whole organization. The overall research approach thus contributes toward a better understanding of the influence and the impact of oppressive social environments and evolving power relations on the development of an organization.
Moving Towards Culturally Competent Health Systems: Organizational and Market Factors
Weech-Maldonado, Robert; Elliott, Marc; Pradhan, Rohit; Schiller, Cameron; Dreachslin, Janice; Hays, Ron D.
Cultural competency has been proposed as an organizational strategy to address racial/ethnic disparities in the health care system; disparities are a long-standing policy challenge whose relevance is only increasing with the increasing population diversity of the US and across the world. Using an integrative conceptual framework based on the resource dependency and institutional theories, we examine the relationship between organizational and market factors and hospitals' degree of cultural competency. Our sample consists of 119 hospitals located in the state of California (US) and is constructed using the following datasets for the year 2006: Cultural Competency Assessment Tool of Hospitals (CCATH) Survey, California's Office of Statewide Health Planning & Development's Hospital Inpatient Discharges and Annual Hospital Financial Data, American Hospital Association's Annual Survey, and the Area Resource File. The dependent variable consists of the degree of hospital cultural competency, as assessed by the CCATH overall score. Organizational variables include ownership status, teaching hospital, payer mix, size, system membership, financial performance, and the proportion of inpatient racial/ethnic minorities. Market characteristics included hospital competition, the proportion of racial/ethnic minorities in the area, metropolitan area, and per capita income. Regression analyses were conducted to assess the relationship between the CCATH overall score and organizational and market variables. Our results show that hospitals which are not-for-profit, serve a more diverse inpatient population, and are located in more competitive and affluent markets exhibit a higher degree of cultural competency. Our results underscore the importance of both institutional and competitive market pressures in guiding hospital behavior. For instance, while not-for-profit may adopt innovative/progressive policies like cultural competency simply as a function of their organizational
Moving towards culturally competent health systems: organizational and market factors.
Weech-Maldonado, Robert; Elliott, Marc N; Pradhan, Rohit; Schiller, Cameron; Dreachslin, Janice; Hays, Ron D
Cultural competency has been proposed as an organizational strategy to address racial/ethnic disparities in the healthcare system; disparities are a long-standing policy challenge whose relevance is only increasing with the increasing population diversity of the US and across the world. Using an integrative conceptual framework based on the resource dependency and institutional theories, we examine the relationship between organizational and market factors and hospitals' degree of cultural competency. Our sample consists of 119 hospitals located in the state of California (US) and is constructed using the following datasets for the year 2006: Cultural Competency Assessment Tool of Hospitals (CCATH) Survey, California's Office of Statewide Health Planning & Development's Hospital Inpatient Discharges and Annual Hospital Financial Data, American Hospital Association's Annual Survey, and the Area Resource File. The dependent variable consists of the degree of hospital cultural competency, as assessed by the CCATH overall score. Organizational variables include ownership status, teaching hospital, payer mix, size, system membership, financial performance, and the proportion of inpatient racial/ethnic minorities. Market characteristics included hospital competition, the proportion of racial/ethnic minorities in the area, metropolitan area, and per capita income. Regression analyses were conducted to assess the relationship between the CCATH overall score and organizational and market variables. Our results show that hospitals which are not-for-profit, serve a more diverse inpatient population, and are located in more competitive and affluent markets exhibit a higher degree of cultural competency. Our results underscore the importance of both institutional and competitive market pressures in guiding hospital behavior. For instance, while not-for-profit may adopt innovative/progressive policies like cultural competency simply as a function of their organizational goals
Analytical Criticalities Associated to Different Immunological Methods for Serum Free Light Chain Detection in Plasma Cell Dyscrasias: A Description of Particular Clinical Cases
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Rocco Sabatino
Full Text Available Current criteria for differential diagnosis of multiple myeloma (MM, Monoclonal gammopathy of undetermined significance (MGUS, and smoldering multiple myeloma (SMM are included in the 2003 guidelines by the International Myeloma Working Group (IMWG. An updated version was then published in 2014, highlighting the importance of serum free light chain (sFLC detection, as well as the κ/λ ratio as excellent indicators of clonality. At present, two commercial assays for sFLC quantification are available: the Freelite™ assay and the N-Latex assay. The first was developed by The Binding Site based on a mixture of polyclonal antibodies directed against a variety of FLC epitopes. It may be run on a wide range of nephelometers, as well as on turbidimeters. The second method was developed by Siemens and runs exclusively on Siemens instruments. It employs a probe mixture of mouse monoclonal antibodies. The aim of our study was to evaluate sFLC measurement and calculated κ/λ ratio in 85 patients with monoclonal gammopathies (MGs in order to compare methods. We demonstrated that there is only a moderate concordance between the two FLC assays. In particular, in one case, we observed no qualitative alterations of the serum protein pattern, and in the absence of a Freelite™ assay, sFLC measurement would not have been possible to highlight the increase of λ FLC.
Transient gene expression in serum-free suspension-growing mammalian cells for the production of foot-and-mouth disease virus empty capsids.
Directory of Open Access Journals (Sweden)
Ana Clara Mignaqui
Full Text Available Foot-and-mouth disease (FMD is a highly contagious disease of cloven-hoofed animals. It produces severe economic losses in the livestock industry. Currently available vaccines are based on inactivated FMD virus (FMDV. The use of empty capsids as a subunit vaccine has been reported to be a promising candidate because it avoids the use of virus in the vaccine production and conserves the conformational epitopes of the virus. In this report, we explored transient gene expression (TGE in serum-free suspension-growing mammalian cells for the production of FMDV recombinant empty capsids as a subunit vaccine. The recombinant proteins produced, assembled into empty capsids and induced protective immune response against viral challenge in mice. Furthermore, they were recognized by anti-FMDV bovine sera. By using this technology, we were able to achieve expression levels that are compatible with the development of a vaccine. Thus, TGE of mammalian cells is an easy to perform, scalable and cost-effective technology for the production of a recombinant subunit vaccine against FMDV.
Organoid culture systems to study host-pathogen interactions
Dutta, Devanjali; Clevers, Hans
Recent advances in host-microbe interaction studies in organoid cultures have shown great promise and have laid the foundation for much more refined future studies using these systems. Modeling of Zika virus (ZIKV) infection in cerebral organoids have helped us understand its association with
Pedagogical System of Future Teachers' Professional Thinking Culture Formation
Abildina, Saltanat K.; Sarsekeyeva, Zhanar Y.; Aidarbekova, Kulzhan A.; Asetova, Zhannur B.; Adanov, Kuanysbek B.
Research objective is to theoretically justify and to develop a pedagogical system of development of future teachers' professional thinking culture. In the research there are used a set of theoretical methods: systematic analysis of the philosophical, psychological and pedagogical literature on the researched topic; compilation and classification…
Clinical comparison of two commercial blood culture systems
Spanjaard, L.; Kuijper, E. J.; Dankert, J.
A prospective, volume-controlled comparison of the BacT/Alert FAN (Organon Teknika, USA) and Vital (bioMérieux, France) blood culture systems was performed in a university hospital during a period of 11 months. Twenty to 40 ml of blood drawn from an adult patient was distributed equally between a
World Culture in the Capitalist World-System in Transition
Griffiths, Tom G.; Arnove, Robert F.
World culture theory (WCT) offers an explanatory framework for macro-level comparative analyses of systems of mass education, including their structures, accompanying policies and their curricular and pedagogical practices. WCT has contributed to broader efforts to overcome methodological nationalism in comparative research. In this paper, we…
Effects of culture systems on growth and economic performance of ...
African Journals Online (AJOL)
Jul 3, 2013 ... The effect of culture system on growth and economics performance of Orechromis niloticus ( ... from the Food and Agriculture Organization (FAO) stated ... in the reduction of the availability of natural fish food ..... lowest profit, while algae only had the lowest cost and ... Also, maximizing production in terms of.
Culture and basic psychological processes--toward a system view of culture: comment on Oyserman et al. (2002).
Kitayama, Shinobu
D. Oyserman, H. M. Coon, and M. Kemmelmeier (2002) provide a most comprehensive review of empirical studies that used attitudinal surveys to capture cultural variations in individualism and collectivism. In the present article, the author suggests that the cross-cultural validity of attitudinal surveys can no longer be taken for granted. Moreover, the meta-theory underlying this literature (called the entity view of culture) is called into question. The author presents an alternative meta-theory (called the system view of culture) and discusses its implications for future work in cultural and cross-cultural psychology.
Application of cultural algorithm to generation scheduling of hydrothermal systems
International Nuclear Information System (INIS)
Yuan Xiaohui; Yuan Yanbin
The daily generation scheduling of hydrothermal power systems plays an important role in the operation of electric power systems for economics and security, which is a large scale dynamic non-linear constrained optimization problem. It is difficult to solve using traditional optimization methods. This paper proposes a new cultural algorithm to solve the optimal daily generation scheduling of hydrothermal power systems. The approach takes the water transport delay time between connected reservoirs into consideration and can conveniently deal with the complicated hydraulic coupling simultaneously. An example is used to verify the correctness and effectiveness of the proposed cultural algorithm, comparing with both the Lagrange method and the genetic algorithm method. The simulation results demonstrate that the proposed algorithm has rapid convergence speed and higher solution precision. Thus, an effective method is provided to solve the optimal daily generation scheduling of hydrothermal systems
Gender and cultural issues in psychiatric nosological classification systems.
van de Water, Tanya; Suliman, Sharain; Seedat, Soraya
Much has changed since the two dominant mental health nosological systems, the International Classification of Diseases (ICD) and the Diagnostic and Statistical Manual of Mental Disorders (DSM), were first published in 1900 and 1952, respectively. Despite numerous modifications to stay up to date with scientific and cultural changes (eg, exclusion of homosexuality as a disorder) and to improve the cultural sensitivity of psychiatric diagnoses, the ICD and DSM have only recently renewed attempts at harmonization. Previous nosological iterations demonstrate the oscillation in the importance placed on the biological focus, highlighting the tension between a gender- and culture-free nosology (solely biological) and a contextually relevant understanding of mental illness. In light of the release of the DSM 5, future nosological systems, such as the ICD 11, scheduled for release in 2017, and the Research Development Criteria (RDoC), can learn from history and apply critiques. This article aims to critically consider gender and culture in previous editions of the ICD and DSM to inform forthcoming classifications.
Thymic epithelial cells. I. Expression of strong suppressive (veto) activity in mouse thymic epithelial cell cultures
DEFF Research Database (Denmark)
Claesson, Mogens Helweg; Ropke, C
We show that thymic epithelial cells grown under serum-free conditions in a chemically defined culture medium can act as veto cells in vitro. The veto activity of thymic epithelial cells results in inactivation of specific alloreactive cytotoxic T-cell precursors at the clonal level. It is conclu......We show that thymic epithelial cells grown under serum-free conditions in a chemically defined culture medium can act as veto cells in vitro. The veto activity of thymic epithelial cells results in inactivation of specific alloreactive cytotoxic T-cell precursors at the clonal level...
Smith, Timothy B; Rodríguez, Melanie Domenech; Bernal, Guillermo
This article summarizes the definitions, means, and research of adapting psychotherapy to clients' cultural backgrounds. We begin by reviewing the prevailing definitions of cultural adaptation and providing a clinical example. We present an original meta-analysis of 65 experimental and quasi-experimental studies involving 8,620 participants. The omnibus effect size of d = .46 indicates that treatments specifically adapted for clients of color were moderately more effective with that clientele than traditional treatments. The most effective treatments tended to be those with greater numbers of cultural adaptations. Mental health services targeted to a specific cultural group were several times more effective than those provided to clients from a variety of cultural backgrounds. We recommend a series of research-supported therapeutic practices that account for clients' culture, with culture-specific treatments being more effective than generally culture-sensitive treatments. © 2010 Wiley Periodicals, Inc.
Information support systems for cultural heritage protection against flooding
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K. Nedvedova
Full Text Available The goal of this paper is to present use of different kind of software applications to create complex support system for protection of cultural heritage against flooding. The project is very complex and it tries to cover the whole area of the problem from prevention to liquidation of aftermath effects. We used GIS for mapping the risk areas, ontology systems for vulnerability assessment application and the BORM method (Business Object Relation Modelling for flood protection system planning guide. Those modern technologies helped us to gather a lot of information in one place and provide the knowledge to the broad audience.
Culture and the Immune System: Cultural Consonance in Social Support and C-reactive Protein in Urban Brazil.
Dressler, William W; Balieiro, Mauro C; Ribeiro, Rosane P; Dos Santos, José Ernesto
In this article, we examine the distribution of a marker of immune system stimulation-C-reactive protein-in urban Brazil. Social relationships are associated with immunostimulation, and we argue that cultural dimensions of social support, assessed by cultural consonance, are important in this process. Cultural consonance is the degree to which individuals, in their own beliefs and behaviors, approximate shared cultural models. A measure of cultural consonance in social support, based on a cultural consensus analysis regarding sources and patterns of social support in Brazil, was developed. In a survey of 258 persons, the association of cultural consonance in social support and C-reactive protein was examined, controlling for age, sex, body mass index, low-density lipoprotein cholesterol, depressive symptoms, and a social network index. Lower cultural consonance in social support was associated with higher C-reactive protein. Implications of these results for future research are discussed. © 2016 by the American Anthropological Association.
Impact of initial FDG-PET/CT and serum-free light chain on transformation of conventionally defined solitary plasmacytoma to multiple myeloma.
Fouquet, Guillemette; Guidez, Stéphanie; Herbaux, Charles; Van de Wyngaert, Zoé; Bonnet, Sarah; Beauvais, David; Demarquette, Hélène; Adib, Salim; Hivert, Bénédicte; Wemeau, Mathieu; Berthon, Céline; Terriou, Louis; Coiteux, Valérie; Macro, Margaret; Decaux, Olivier; Facon, Thierry; Huglo, Damien; Leleu, Xavier
Solitary plasmacytoma (SP) is a localized proliferation of monoclonal plasma cells in either bone or soft tissue, without evidence of multiple myeloma (MM), and whose prognosis is marked by a high risk of transformation to MM. We studied the impact of FDG-PET/CT (2[18F]fluoro-2-deoxy-D-glucose positron emission tomography-computed tomography) on the risk of transformation of SP to overt MM among other markers in a series of 43 patients diagnosed with SP. Median age was 57.5 years; 48% of patients had an abnormal involved serum-free light chain (sFLC) value, and 64% had an abnormal sFLC ratio at diagnosis. Thirty-three percent had two or more hypermetabolic lesions on initial PET/CT, and 20% had two or more focal lesions on initial MRI. With a median follow-up of 50 months, 14 patients transformed to MM with a median time (TTMM) of 71 months. The risk factors that significantly shortened TTMM at diagnosis were two or more hypermetabolic lesions on PET/CT, abnormal sFLC ratio and involved sFLC, and to a lesser extent at completion of treatment, absence of normalized involved sFLC and PET/CT or MRI. In a multivariate analysis, abnormal initial involved sFLC [OR = 10; 95% confidence interval (CI), 1-87; P = 0.008] and PET/CT (OR = 5; 95% CI, 0-9; P = 0.032) independently shortened TTMM. An abnormal involved sFLC value and the presence of at least two hypermetabolic lesions on PET/CT at diagnosis of SP were the two predictors of early evolution to myeloma in our series. This data analysis will need confirmation in a larger study, and the study of these two risk factors may lead to a different management of patients with SP in the future. . ©2014 American Association for Cancer Research.
Cultural Requirements of Policy Making System for Hijab and Dignity
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Shahla Bagheri
Full Text Available Policy making and policy measures is important in the social system. occurs. Policy maker aimed to achieve cultural requirements of policy making system by interaction stale and society. After the Islamic Revolution of Iran. the strengths and weaknesses of the different levels of the system politically has been accompanied in the field of moral and sexual dignity and chastity, aside from the basic necessity of building systems - Iranian, coordination and harmony of the system was not relevant. That is in the realm of theoretical ideas and goals are expressed in practice, the relationship between logical and measurable programs are executed with the goals and policies have been developed. measures to improve processes, motivate and educate individuals and groups, and to monitor the development of information systems.
In vitro culture of human osteosarcoma cell lines: a comparison of functional characteristics for cell lines cultured in medium without and with fetal calf serum.
Bruserud, Oystein; Tronstad, Karl Johan; Berge, Rolf
Experimental in vitro models including well-characterised cell lines can be used to identify possible new therapeutic targets for the treatment of osteosarcoma. Culture media including inactivated serum is often recommended for in vitro culture of osteosarcoma cells, but the serum component then represents a nonstandardised parameter including a wide range of unidentified mediators. To improve the standardisation we have investigated whether serum-free culture media can be used in experimental in vitro studies of osteosarcoma cell lines. The seven osteosarcoma cell lines Cal72, SJSA-1, Saos-2, SK-ES-1, U2OS, 143.98.2, and KHOS-32IH were cultured in vitro in various serum-free media and media supplemented with 10% heat-inactivated fetal calf serum (FCS). Although proliferation often was relatively low in serum-free media (X-vivo 10, X-vivo 15, X-vivo 20, Stem Span SFEM), some cell lines (Cal72, KHOS-32IH, Saos-2) showed proliferation comparable with the recommended FCS-containing media even when using serum-free conditions. The optimal serum-free medium then varied between cell lines. We also compared 6 different FCS-containing media (including Stem Span with 10% FCS) and the optimal FCS-containing medium varied between cell lines. However, all cell lines proliferated well in Stem Span with FCS, and this medium was regarded as optimal for four of the lines. FCS could not be replaced by fatty acids or low density lipoprotein when testing the Stem Span medium. The release of a wide range of soluble mediators showed only minor differences when using serum-free and FCS-containing media (including Stem Span with and without FCS), and serum-free Stem Span could also be used for in vitro studies of mitogen-stimulated T cell activation in the presence of accessory osteosarcoma cells. The use of Stem Span with 10% FCS allowed the release of a wide range of chemokines by osteosarcoma cell lines (Cal72, SJSA-1), and the chemokine release profile was very similar to the
Safety climate and culture: Integrating psychological and systems perspectives.
Casey, Tristan; Griffin, Mark A; Flatau Harrison, Huw; Neal, Andrew
Safety climate research has reached a mature stage of development, with a number of meta-analyses demonstrating the link between safety climate and safety outcomes. More recently, there has been interest from systems theorists in integrating the concept of safety culture and to a lesser extent, safety climate into systems-based models of organizational safety. Such models represent a theoretical and practical development of the safety climate concept by positioning climate as part of a dynamic work system in which perceptions of safety act to constrain and shape employee behavior. We propose safety climate and safety culture constitute part of the enabling capitals through which organizations build safety capability. We discuss how organizations can deploy different configurations of enabling capital to exert control over work systems and maintain safe and productive performance. We outline 4 key strategies through which organizations to reconcile the system control problems of promotion versus prevention, and stability versus flexibility. (PsycINFO Database Record (c) 2017 APA, all rights reserved).
Advanced imaging systems for diagnostic investigations applied to Cultural Heritage
International Nuclear Information System (INIS)
Peccenini, E; Bettuzzi, M; Brancaccio, R; Casali, F; Morigi, M P; Albertin, F; Petrucci, F
The diagnostic investigations are an important resource in the studies on Cultural Heritage to enhance the knowledge on execution techniques, materials and conservation status of a work of art. In this field, due to the great historical and artistic value of the objects, preservation is the main concern; for this reason, new technological equipment has been designed and developed in the Physics Departments of the Universities of Ferrara and Bologna to enhance the non-invasive approach to the study of pictorial artworks and other objects of cultural interest. Infrared (IR) reflectography, X-ray radiography and computed tomography (CT), applied to works of art, are joined by the same goal: to get hidden information on execution techniques and inner structure pursuing the non-invasiveness of the methods, although using different setup and physical principles. In this work transportable imaging systems to investigate large objects in museums and galleries are presented. In particular, 2D scanning devices for IR reflectography and X-ray radiography, CT systems and some applications to the Cultural Heritage are described
Advanced imaging systems for diagnostic investigations applied to Cultural Heritage
Peccenini, E.; Albertin, F.; Bettuzzi, M.; Brancaccio, R.; Casali, F.; Morigi, M. P.; Petrucci, F.
The diagnostic investigations are an important resource in the studies on Cultural Heritage to enhance the knowledge on execution techniques, materials and conservation status of a work of art. In this field, due to the great historical and artistic value of the objects, preservation is the main concern; for this reason, new technological equipment has been designed and developed in the Physics Departments of the Universities of Ferrara and Bologna to enhance the non-invasive approach to the study of pictorial artworks and other objects of cultural interest. Infrared (IR) reflectography, X-ray radiography and computed tomography (CT), applied to works of art, are joined by the same goal: to get hidden information on execution techniques and inner structure pursuing the non-invasiveness of the methods, although using different setup and physical principles. In this work transportable imaging systems to investigate large objects in museums and galleries are presented. In particular, 2D scanning devices for IR reflectography and X-ray radiography, CT systems and some applications to the Cultural Heritage are described.
Serum Free Light Chains in Neoplastic Monoclonal Gammopathies: Relative Under-Detection of Lambda Dominant Kappa/Lambda Ratio, and Underproduction of Free Lambda Light Chains, as Compared to Kappa Light Chains, in Patients With Neoplastic Monoclonal Gammopathies.
Lee, Won Sok; Singh, Gurmukh
Quantitative evaluation of serum free light chains is recommended for the work up of monoclonal gammopathies. Immunoglobulin light chains are generally produced in excess of heavy chains. In patients with monoclonal gammopathy, κ/λ ratio is abnormal less frequently with lambda chain lesions. This study was undertaken to ascertain if the levels of overproduction of the two light chain types and their detection rates are different in patients with neoplastic monoclonal gammopathies. Results of serum protein electrophoresis (SPEP), serum protein immunofixation electrophoresis (SIFE), urine protein electrophoresis (UPEP), urine protein immunofixation electrophoresis (UIFE), and serum free light chain assay (SFLCA) in patients with monoclonal gammopathies were examined retrospectively. The κ/λ ratios were appropriately abnormal more often in kappa chain lesions. Ratios of κ/λ were normal in about 25% of patients with lambda chain lesions in whom free homogenous lambda light chains were detectable in urine. An illustrative case suggests underproduction of free lambda light chains, in some instances. The lower prevalence of lambda dominant κ/λ ratio in lesions with lambda light chains is estimated to be due to relative under-detection of lambda dominant κ/λ ratio in about 25% of the patients and because lambda chains are not produced in as much excess of heavy chains as are kappa chains, in about 5% of the patients. The results question the medical necessity and clinical usefulness of the serum free light chain assay. UPEP/UIFE is under-utilized.
Development of the Continued Improvement System for Nuclear Safety Culture
International Nuclear Information System (INIS)
Park, H. C.; Park, H. G.; Park, Y. W.; Park, J. Y.
It has been found that almost 80 % of the incidents and accidents occurred recently, such as the Fukushima Daiichi disaster and Domestic SBO accident etc. were analyzed to be caused from human errors. (IAEA NES NG-G-2.1) Which strongly claims the importance of the safety culture system. Accordingly, it should be away from a cursory approach like one-off field survey or Snap shop which were being conducted at present for the continued improvement of safety culture. This study introduces an analytical methodology which approaches the generic form of the safety both consciously and unconsciously expressed with behavior, thoughts, and attitude etc. This study was implemented only for open materials such as Inspection report, incidents and accidents reports, QA documents because of the limitation in accessibility to data. More effective use with securing operational data will be possible in future
Development of the Continued Improvement System for Nuclear Safety Culture
Energy Technology Data Exchange (ETDEWEB)
Park, H. C.; Park, H. G.; Park, Y. W.; Park, J. Y. [KAIST, Daejeon (Korea, Republic of)
It has been found that almost 80 % of the incidents and accidents occurred recently, such as the Fukushima Daiichi disaster and Domestic SBO accident etc. were analyzed to be caused from human errors. (IAEA NES NG-G-2.1) Which strongly claims the importance of the safety culture system. Accordingly, it should be away from a cursory approach like one-off field survey or Snap shop which were being conducted at present for the continued improvement of safety culture. This study introduces an analytical methodology which approaches the generic form of the safety both consciously and unconsciously expressed with behavior, thoughts, and attitude etc. This study was implemented only for open materials such as Inspection report, incidents and accidents reports, QA documents because of the limitation in accessibility to data. More effective use with securing operational data will be possible in future.
An Information System for European culture collections: the way forward.
Casaregola, Serge; Vasilenko, Alexander; Romano, Paolo; Robert, Vincent; Ozerskaya, Svetlana; Kopf, Anna; Glöckner, Frank O; Smith, David
Culture collections contain indispensable information about the microorganisms preserved in their repositories, such as taxonomical descriptions, origins, physiological and biochemical characteristics, bibliographic references, etc. However, information currently accessible in databases rarely adheres to common standard protocols. The resultant heterogeneity between culture collections, in terms of both content and format, notably hampers microorganism-based research and development (R&D). The optimized exploitation of these resources thus requires standardized, and simplified, access to the associated information. To this end, and in the interest of supporting R&D in the fields of agriculture, health and biotechnology, a pan-European distributed research infrastructure, MIRRI, including over 40 public culture collections and research institutes from 19 European countries, was established. A prime objective of MIRRI is to unite and provide universal access to the fragmented, and untapped, resources, information and expertise available in European public collections of microorganisms; a key component of which is to develop a dynamic Information System. For the first time, both culture collection curators as well as their users have been consulted and their feedback, concerning the needs and requirements for collection databases and data accessibility, utilised. Users primarily noted that databases were not interoperable, thus rendering a global search of multiple databases impossible. Unreliable or out-of-date and, in particular, non-homogenous, taxonomic information was also considered to be a major obstacle to searching microbial data efficiently. Moreover, complex searches are rarely possible in online databases thus limiting the extent of search queries. Curators also consider that overall harmonization-including Standard Operating Procedures, data structure, and software tools-is necessary to facilitate their work and to make high-quality data easily accessible
Management and supervisory training: Changing the culture/system
International Nuclear Information System (INIS)
Martin, L.F.
Professional development training courses, and a unique promoting appraisal system have been combined in the form of a training matrix and are being used as a culture change agent. This training and system modification is currently being implemented within the Nuclear Materials Processing Division of the Savannah River Site. It is designed to help solve the systems problems of managers and supervisors while enabling subordinates to develop themselves professionally, and starts with a promoting appraisal between a manager and a subordinate. These easy to apply appraisals are becoming an integral part of the business system, and result in giving a manager greater control over business change and greater influence over the work each employee accomplishes
Micro fluidic System for Culturing and Monitoring of Neuronal Cells and Tissue
DEFF Research Database (Denmark)
Bakmand, Tanya; Waagepetersen, Helle S.
The aim of this Ph.D. project was to combine experience within cell and tissue culturing, electrochemistry and microfabrication in order to develop an in vivo-like fluidic culturing platform, challenging the traditional culturing methods. The first goal was to develope a fluidic system for cultur...... with mass production. The last part of this thesis also includes perspectives on how to expand the latest designed device to facilitate culturing of tissue and co-culturing of cells....
Development of bovine embryos cultured in CR1aa and IVD101 media using different oxygen tensions and culture systems.
Somfai, Tamás; Inaba, Yasushi; Aikawa, Yoshio; Ohtake, Masaki; Kobayashi, Shuji; Konishi, Kazuyuki; Nagai, Takashi; Imai, Kei
The aim of the present study was to optimise the culture conditions for the in vitro production of bovine embryos. The development of in vitro fertilised bovine oocytes in CR1aa supplemented with 5% calf serum and IVD101 culture media were compared using traditional microdrops and Well of the Well (WOW) culture systems either under 5% or 20% oxygen tension. After 7 days of culture, a significantly higher blastocyst formation rate was obtained for embryos cultured in CR1aa medium compared to those cultured in IVD101, irrespective of O2 tensions and culture systems. The blastocyst formation in IVD101 was suppressed under 20% O2 compared to 5% O2 . Despite their similar total cell numbers, higher rates of inner cell mass (ICM) cells were observed in blastocysts developed in IVD101 medium than in those developed in CR1aa, irrespective of O2 tensions. There was no significant difference in blastocyst formation, total, ICM and trophectoderm (TE) cell numbers between embryos obtained by microdrop and WOW culture systems irrespective of the culture media and O2 tensions used. In conclusion, CR1aa resulted in higher blastocyst formation rates irrespective of O2 tension, whereas IVD101 supported blastocyst formation only under low O2 levels but enhanced the proliferation of ICM cells.
Discussion on establishment and improvement of the nuclear safety culture system
International Nuclear Information System (INIS)
Lu Weiqiang; Na Fuli
By discussion of the problems in the manufacture process of nuclear power equipment enterprisers, puts forwards the tentative idea of establishment the nuclear safety culture system, meanwhile, gives some suggestions in order to improving the nuclear safety culture system. (authors)
Culture, Pedagogy and Equity in a Meritocratic Education System: Teachers' Work and the Politics of Culture in Singapore
Lim, Leonel; Tan, Michael
Drawing upon insights gained from the extant work on culture and pedagogy, this paper explores the ways in which, in an ostensibly meritocratic education system, ideas about students' cultural backgrounds and its relevance for teaching are interpreted, negotiated, and ultimately drawn upon to engage students in the low-progress academic tracks.…
Modeling human gastrointestinal inflammatory diseases using microphysiological culture systems.
Hartman, Kira G; Bortner, James D; Falk, Gary W; Ginsberg, Gregory G; Jhala, Nirag; Yu, Jian; Martín, Martín G; Rustgi, Anil K; Lynch, John P
Gastrointestinal illnesses are a significant health burden for the US population, with 40 million office visits each year for gastrointestinal complaints and nearly 250,000 deaths. Acute and chronic inflammations are a common element of many gastrointestinal diseases. Inflammatory processes may be initiated by a chemical injury (acid reflux in the esophagus), an infectious agent (Helicobacter pylori infection in the stomach), autoimmune processes (graft versus host disease after bone marrow transplantation), or idiopathic (as in the case of inflammatory bowel diseases). Inflammation in these settings can contribute to acute complaints (pain, bleeding, obstruction, and diarrhea) as well as chronic sequelae including strictures and cancer. Research into the pathophysiology of these conditions has been limited by the availability of primary human tissues or appropriate animal models that attempt to physiologically model the human disease. With the many recent advances in tissue engineering and primary human cell culture systems, it is conceivable that these approaches can be adapted to develop novel human ex vivo systems that incorporate many human cell types to recapitulate in vivo growth and differentiation in inflammatory microphysiological environments. Such an advance in technology would improve our understanding of human disease progression and enhance our ability to test for disease prevention strategies and novel therapeutics. We will review current models for the inflammatory and immunological aspects of Barrett's esophagus, acute graft versus host disease, and inflammatory bowel disease and explore recent advances in culture methodologies that make these novel microphysiological research systems possible. © 2014 by the Society for Experimental Biology and Medicine.
Information and Communication Technologies – and Culturally Sensitive Systems
Directory of Open Access Journals (Sweden)
Nancy Michail
Full Text Available This paper discusses the perceptions of Egyptian minority groups in relation to internet information technology with which they feel empowered to protect, affirm and communicate their oppressed existence, on local and global dimensions. The research employs qualitative methods and interpretive analysis, to focus on the use of Internet information technology tools by Egyptian minority groups, in particular, their online platforms and chat rooms, and the related issues associated with these practices and usages. The paper argues that cyberspace is used by specific minority groups in Egypt as a "gateway to freedom" in which it constitutes an ally to establish newly founded cyber identities that aide them to exercise their basic human rights of freedom of thought, speech and expression. The paper thus examines cyberspace a medium or tool for the carrying out of information exchange without the traditional fear of politics and power that is deeply engraved in the roots of the Egyptian culture. In this way, these minority groups are analysed as the newly conceived human information systems (HIS residing on Internet information technology and infrastructure. The paper proposes an adaptive and culturally sensitive model of human information systems as well as human information systems development life cycle (HISDLC to aid in establishing effective processes of information exchange and creation, hence assisting in the emancipation of conflicting parties residing in Egypt, elsewhere in the Middle East and globally.
Promoting a Culture of Tailoring for Systems Engineering Policy Expectations
Blankenship, Van A.
NASA's Marshall Space Flight Center (MSFC) has developed an integrated systems engineering approach to promote a culture of tailoring for program and project policy requirements. MSFC's culture encourages and supports tailoring, with an emphasis on risk-based decision making, for enhanced affordability and efficiency. MSFC's policy structure integrates the various Agency requirements into a single, streamlined implementation approach which serves as a "one-stop-shop" for our programs and projects to follow. The engineers gain an enhanced understanding of policy and technical expectations, as well as lesson's learned from MSFC's history of spaceflight and science missions, to enable them to make appropriate, risk-based tailoring recommendations. The tailoring approach utilizes a standard methodology to classify projects into predefined levels using selected mission and programmatic scaling factors related to risk tolerance. Policy requirements are then selectively applied and tailored, with appropriate rationale, and approved by the governing authorities, to support risk-informed decisions to achieve the desired cost and schedule efficiencies. The policy is further augmented by implementation tools and lifecycle planning aids which help promote and support the cultural shift toward more tailoring. The MSFC Customization Tool is an integrated spreadsheet that ties together everything that projects need to understand, navigate, and tailor the policy. It helps them classify their project, understand the intent of the requirements, determine their tailoring approach, and document the necessary governance approvals. It also helps them plan for and conduct technical reviews throughout the lifecycle. Policy tailoring is thus established as a normal part of project execution, with the tools provided to facilitate and enable the tailoring process. MSFC's approach to changing the culture emphasizes risk-based tailoring of policy to achieve increased flexibility, efficiency
Effect of different culture media and deswelling agents on survival of human corneal endothelial and epithelial cells in vitro.
Valtink, Monika; Donath, Patricia; Engelmann, Katrin; Knels, Lilla
To examine the effects of media and deswelling agents on human corneal endothelial and epithelial cell viability using a previously developed screening system. The human corneal endothelial cell line HCEC-12 and the human corneal epithelial cell line HCE-T were cultured in four different corneal organ culture media (serum-supplemented: MEM +2 % FCS, CorneaMax®/CorneaJet®, serum-free: Human Endothelial-SFM, Stemalpha-2 and -3) with and without 6 % dextran T500 or 7 % HES 130/0.4. Standard growth media F99HCEC and DMEM/F12HCE-T served as controls. In additional controls, the stress inducers staurosporine or hydrogen peroxide were added. After 5 days in the test media, cell viability was assessed by flow cytometrically quantifying apoptotic and necrotic cells (sub-G1 DNA content, vital staining with YO-PRO-1® and propidium iodide) and intracellular reactive oxygen species (ROS). The MEM-based media were unable to support HCEC-12 and HCE-T survival under stress conditions, resulting in significantly increased numbers of apoptotic and necrotic cells. HCEC-12 survival was markedly improved in SFM-based media even under staurosporine or hydrogen peroxide. Likewise, HCE-T survival was improved in SFM with or without dextran. The media CorneaMax®, CorneaJet®, and CorneaMax® with HES supported HCEC-12 survival better than MEM-based media, but less well than SFM-based media. HCE-T viability was also supported by CorneaJet®, but not by CorneaMax® with or without HES. Stemalpha-based media were not suitable for maintaining viability of HCEC-12 or HCE-T in the applied cell culture system. The use of serum-supplemented MEM-based media for corneal organ culture should be discontinued in favour of serum-free media like SFM.
Lean management systems: creating a culture of continuous quality improvement.
Clark, David M; Silvester, Kate; Knowles, Simon
This is the first in a series of articles describing the application of Lean management systems to Laboratory Medicine. Lean is the term used to describe a principle-based continuous quality improvement (CQI) management system based on the Toyota production system (TPS) that has been evolving for over 70 years. Its origins go back much further and are heavily influenced by the work of W Edwards Deming and the scientific method that forms the basis of most quality management systems. Lean has two fundamental elements--a systematic approach to process improvement by removing waste in order to maximise value for the end-user of the service and a commitment to respect, challenge and develop the people who work within the service to create a culture of continuous improvement. Lean principles have been applied to a growing number of Healthcare systems throughout the world to improve the quality and cost-effectiveness of services for patients and a number of laboratories from all the pathology disciplines have used Lean to shorten turnaround times, improve quality (reduce errors) and improve productivity. Increasingly, models used to plan and implement large scale change in healthcare systems, including the National Health Service (NHS) change model, have evidence-based improvement methodologies (such as Lean CQI) as a core component. Consequently, a working knowledge of improvement methodology will be a core skill for Pathologists involved in leadership and management.
Organisational Culture Matters for System Integration in Health Care
Munir, Samina K.; Kay, Stephen
This paper illustrates the importance of organisational culture for Clinical Information Systems (CIS) integration. The study is based on data collected in intensive care units in the UK and Denmark. Data were collected using qualitative methods, i.e., observations, interviews and shadowing of health care providers, together with a questionnaire at each site. The data are analysed to extract salient variables for CIS integration, and it is shown that these variables can be separated into two categories that describe the 'Actual Usefulness' of the system and the 'Organisational Culture'. This model is then extended to show that CIS integration directly affects the work processes of the organisation, forming an iterative process of change as a CIS is introduced and integrated. PMID:14728220
Organisational Culture as a Dominant in Enterprise Activity: System Approach
Directory of Open Access Journals (Sweden)
Serikov Anatoliy V.
Full Text Available The article offers a "conceptual carcass" of the enterprise model, which is based on known results of studies in sociology, biology, system theory and mathematics. The article lists main features of growth of main indicators of economic activity and development of an enterprise. Dynamics of changes at an enterprise is described with a system of non-linear differential equations. One of the global and dominating factors in it is entrepreneurship of personnel, which is an integral part of its labour mentality or organisational culture. The article proves for the first time ever, using mathematical modelling, that namely entrepreneurship, innovation capability, is a comprehensive and dominant factor of enterprise growth and development.
A Cross-Cultural Study of How Usability Professionals Experience the Usability of Everyday Systems
DEFF Research Database (Denmark)
Jiang, Y.; Sun, X.; Li, H.
in the study, their understanding of fun systems differs across cultural backgrounds. Also, easy-to-use and useful systems are perceived as being similar or different depending on the usability professional's cultural background. Most other cross-cultural differences relate to categories of construct......Culture influences many aspects of the design and use of computer systems; understanding better this influence on their own thinking may benefit usability professionals who do cross-cultural usability work. Using Kelly's notion of personal constructs, we focus on one mediator of culture: how...
Development and evaluation of the quick anaero-system-a new disposable anaerobic culture system.
Yang, Nam Woong; Kim, Jin Man; Choi, Gwang Ju; Jang, Sook Jin
We developed a new disposable anaerobic culture system, namely, the Quick anaero-system, for easy culturing of obligate anaerobes. Our system consists of 3 components: 1) new disposable anaerobic gas pack, 2) disposable culture-envelope and sealer, and 3) reusable stainless plate rack with mesh containing 10 g of palladium catalyst pellets. To evaluate the efficiency of our system, we used 12 anaerobic bacteria. We prepared 2 sets of ten-fold serial dilutions of the 12 anaerobes, and inoculated these samples on Luria-Bertani (LB) broth and LB blood agar plate (LB-BAP) (BD Diagnostic Systems, USA). Each set was incubated in the Quick anaero-system (DAS Tech, Korea) and BBL GasPak jar with BD GasPak EZ Anaerobe Container System (BD Diagnostic Systems) at 35-37 degrees C for 48 hr. The minimal inoculum size showing visible growth of 12 anaerobes when incubated in both the systems was compared. The minimal inoculum size showing visible growth for 2 out of the 12 anaerobes in the LB broth and 9 out of the 12 anaerobes on LB-BAP was lower for the Quick anaero-system than in the BD GasPak EZ Anaerobe Container System. The mean time (+/-SD) required to achieve absolute anaerobic conditions of the Quick anaero-system was 17 min and 56 sec (+/-3 min and 25 sec). The Quick anaero-system is a simple and effective method of culturing obligate anaerobes, and its performance is superior to that of the BD GasPak EZ Anaerobe Container System.
Adhesion of cultured human endothelial cells onto methacrylate polymers with varying surface wettability and charge
van Wachem, P.B.; Hogt, A.H.; Beugeling, T.; Feijen, Jan; Bantjes, A.; Detmers, J.P.; van Aken, W.G.
The adhesion of human endothelial cells (HEC) onto a series of well-characterized methacrylate polymer surfaces with varying wettabilities and surface charges was studied either in serum-containing (CMS) or in serum-free (CM) culture medium. HEC adhesion in CMS onto (co)polymers * of hydroxyethyl
A Neuronal Culture System to Detect Prion Synaptotoxicity.
Directory of Open Access Journals (Sweden)
Cheng Fang
Full Text Available Synaptic pathology is an early feature of prion as well as other neurodegenerative diseases. Although the self-templating process by which prions propagate is well established, the mechanisms by which prions cause synaptotoxicity are poorly understood, due largely to the absence of experimentally tractable cell culture models. Here, we report that exposure of cultured hippocampal neurons to PrPSc, the infectious isoform of the prion protein, results in rapid retraction of dendritic spines. This effect is entirely dependent on expression of the cellular prion protein, PrPC, by target neurons, and on the presence of a nine-amino acid, polybasic region at the N-terminus of the PrPC molecule. Both protease-resistant and protease-sensitive forms of PrPSc cause dendritic loss. This system provides new insights into the mechanisms responsible for prion neurotoxicity, and it provides a platform for characterizing different pathogenic forms of PrPSc and testing potential therapeutic agents.
Directory of Open Access Journals (Sweden)
Oleksandr A. Ratsul
Full Text Available The article deals with the analysis of foreign experience of systemic development of future social pedagogists' informational culture. A number of cultural universals are identified, each of them is treated as the core of culture. A list of components of future social pedagogists' information culture is given. Personality traits that enable future social pedagogists to participate effectively in all kinds of work with information are characterized. Two structural levels (contents and functions in future social pedagogists' information culture are singled out. Main functions of future social pedagogists' information culture are defined. The structural organization of future social pedagogists' information culture is analyzed.
Mobile system for in-situ imaging of cultural objects
International Nuclear Information System (INIS)
Zemlicka, J; Jakubek, J; Krejci, F; Hradil, D; Hradilova, J; Mislerova, H
Non-invasive analytical techniques recently developed with the Timepix pixel detector have shown great potential for the inspection of objects of cultural heritage. We have developed new instrumentation and methodology for in-situ X-ray transmission radiography and X-ray fluorescence imaging and successfully tested and evaluated a mobile system for remote terrain tasks. The prototype portable imaging device comprises the radiation source tube and the spectral sensitive X-ray camera. Both components can be moreover mounted on independent motorized positioning systems allowing adaptation of irradiation geometry to the object shape. Both parts are placed onto a pair of universal portable holders (tripods). The detector is placed in a shielded box with exchangeable entrance window (beam filters and pinhole collimator). This adjustable setup allows performing in-situ measurements for both transmission and emission (XRF) radiography. The assembled system has been successfully tested in our laboratory with phantoms and real samples. The obtained and evaluated results are presented in this paper. Future work will include successive adaptation of the current system for real in-situ utilization and preparation of software allowing semi-automatic remote control of measurements.
National Innovation System And Culture A Cross-Country Analysis
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Joseph Gogodze
Full Text Available This study assesses the relationship between Hofstedes cultural dimensions and the constituents of a National Innovation System NIS. We consider an NIS as a special kind of intangible latent asset and identify its two constituents input and output capital. These are extracted through a modern NIS measurement model based on the Global Innovation Index. Using structural equation models we show that power distance and uncertainty avoidance and long-term orientation and indulgence vs. restraint act through the latent constructs PDUA and LTIV respectively. Moreover individualism IDV and NIS constituents are directly and negatively affected by PDUA. IDV and LTIV directly and positively affect the NIS constituents. Further the results show that masculinity vs. femininity significantly and negatively affects the NIS input constituent and significantly affects the NIS output constituent but its impact is negative for high-income countries and positive for non-high income countries.
Safety Culture in Modern Aviation Systems – Civil and Military
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Valentin-Marian IORDACHE
Full Text Available Understanding important aspects of the safety culture should be the main objective for identifying hazards, mitigate and manage risk and find solutions to problems before accidents and incidents occur. The two defining elements of aeronautical decision-making are hazard and risk; risk management is an important component of decisional process and by understanding some issues regarding risk and safety, we will be able to realize the feasible solutions that we may have to apply in flight or ground operations. As aviation is in continous development and worldwide expansion, in order to better understand the associated risks and mitigate them, proper control methods which can give a thoroughly comprehension of the aeronautical system must be used.
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Marija Merdzhanovska
Full Text Available The paper explores the need of connecting the technology, political and cultural system in the right way to achieve the companies' goals in terms of globalization and changes. The leaders have the main role in realizing the company's strategy. They must be strategy focused and they are responsible for connecting the employees, processes and technologies. In the beginning of the paper, the characteristics of strategy focused leadership are analyzed. The available tools of strategy focused leadership through the model are presented. The matrix of strategy management can be used like a manual which helps the management in connecting systems with managerial instruments: mission and strategy, organization structure and management of human resources. The goal of the paper is to show that with professional approach, the strategy focused leaders play the main role in connecting and changing the existing systems in companies. This is the way of reaching the best performance of companies and surviving at the global world market.
Directory of Open Access Journals (Sweden)
Irene Kreutz
Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.
Comparative studies of types 1 and 2 herpes simplex virus infection of cultured normal keratinocytes.
Su, S J; Wu, H H; Lin, Y H; Lin, H Y
AIMS--To investigate the differences in biological properties, multiplication patterns, and cytopathic effects between type 1 and type 2 herpes simplex virus (HSV) through the replication of HSV in cultured normal human keratinocytes. METHODS--Keratinocytes were obtained from surgical specimens of normal gingiva, cervix, trunk skin, and newborn foreskin. They were cultured in serum free, chemically defined, culture medium and infected with a pool of HSV collected from clinical specimens. RESU...
Culturing of PC12 Cells, Neuronal Cells, Astrocytes Cultures and Brain Slices in an Open Microfluidic System
DEFF Research Database (Denmark)
Al Atraktchi, Fatima Al-Zahraa; Bakmand, Tanya; Rømer Sørensen, Ane
The brain is the center of the nervous system, where serious neurodegenerative diseases such as Parkinson's, Alzheimer's and Huntington's are products of functional loss in the neural cells (1). Typical techniques used to investigate these diseases lack precise control of the cellular surroundings......, in addition to isolating the neural tissue from nutrient delivery and to creating unwanted gradients (2). This means that typical techniques used to investigate neurodegenerative diseases cannot mimic in vivo conditions, as closely as desired. We have developed a novel microfluidic system for culturing PC12...... cells, neuronal cells, astrocytes cultures and brain slices. The microfluidic system provides efficient nutrient delivery, waste removal, access to oxygen, fine control over the neurochemical environment and access to modern microscopy. Additionally, the setup consists of an in vitro culturing...
Arab management accounting systems under the influence of their culture
Dik, Roula
Due to globalization, business opportunities can be established in new countries more than ever before. The Arab countries are gaining more attention as emerging markets in global business. Applying managerial accounting on a global basis requires an understanding of the cultural aspects of the countries. Arabic culture has a long history on the one hand and contradicts the Western cultures sharply on the other hand. Research on comparative management accounting (CMA) emerged in the early ...
Planning Smalltalk Behavior with Cultural Influences for Multiagent Systems
DEFF Research Database (Denmark)
Endrass, Birgit; Rehm, Matthias; André, Elisabeth
There are several factors that influence communicative behavior, such as gender, personality or culture. As virtual agents interact in a more and more human-like manner, their behavior should be dependent on social factors as well. Culture is a phenomenon that affects one's behavior without one...... realizing it. Behavior is thus sometimes perceived as inappropriate because there is no awareness of the cultural gap. Thus, we think cultural background should also influence the communication behavior of virtual agents. Behavioral differences are sometimes easy to recognize by humans but still hard...
A managerial approach to NASA's cultural changes open-system model
Long, Nicholas.
This project describes NASA's culture during two important time periods (1958-1972) and (1996-2004) and explains its relative fit with its system components-task, people, resources, and structure. The open-system model is used to explain how system components affect culture and how culture affects them. During the first period (1958- 1972), NASA was established and it landed the first man on the moon, a remarkable accomplishment given the advances in science and technology required to com...
Cell-type-specific and differentiation-status-dependent variations in cytotoxicity of tributyltin in cultured rat cerebral neurons and astrocytes.
Oyanagi, Koshi; Tashiro, Tomoko; Negishi, Takayuki
Tributyltin (TBT) is an organotin used as an anti-fouling agent for fishing nets and ships and it is a widespread environmental contaminant at present. There is an increasing concern about imperceptible but serious adverse effect(s) of exposure to chemicals existing in the environment on various organs and their physiological functions, e.g. brain and mental function. Here, so as to contribute to improvement of and/or advances in in vitro cell-based assay systems for evaluating brain-targeted adverse effect of chemicals, we tried to evaluate cell-type-specific and differentiation-status-dependent variations in the cytotoxicity of TBT towards neurons and astrocytes using the four culture systems differing in the relative abundance of these two types of cells; primary neuron culture (> 95% neurons), primary neuron-astrocyte (2 : 1) mix culture, primary astrocyte culture (> 95% astrocytes), and passaged astrocyte culture (100% proliferative astrocytes). Cell viability was measured at 48 hr after exposure to TBT in serum-free medium. IC50's of TBT were 198 nM in primary neuron culture, 288 nM in primary neuron-astrocyte mix culture, 2001 nM in primary astrocyte culture, and 1989 nM in passaged astrocyte culture. Furthermore, in primary neuron-astrocyte mix culture, vulnerability of neurons cultured along with astrocytes to TBT toxicity was lower than that of neurons cultured purely in primary neuron culture. On the other hand, astrocytes in primary neuron-astrocyte mix culture were considered to be more vulnerable to TBT than those in primary or passaged astrocyte culture. The present study demonstrated variable cytotoxicity of TBT in neural cells depending on the culture condition.
Further characterization of the adhesive-tumor-cell culture system for measuring the radiosensitivity of human tumor primary cultures
International Nuclear Information System (INIS)
Brock, W.A.; Bock, S.P.; Williams, M.; Baker, F.L.
This study extends the use of the adhesive-tumor-cell culture system to include: over 100 sensitivity measurements at 2.0 Gy; tumorgenicity determinations in nude mice; and flow cytometry of the cells grown in the system. The malignant nature of the growing cells was proved by injecting cells into nude mice. Tumors resulted in 60% of the cases and the histology of each xenograft was similar to that of the human tumor. Flow cytometry was used to obtain DNA histograms of the original cell suspension and of cultures during the two week culture period in order to obtain quantitative information about the growth of aneuploid versus diploid populations. The results thus far demonstrate that 95% of aneuploid populations yield aneuploid growth; of the first 20 cases studied, only one suspension with an aneuploid peak resulted in diploid growth. Of further interest was the observation that it is not unusual for a minor aneuploid population to become the predominate growth fraction after two weeks in culture. These results demonstrate that the adhesive-tumor-cell culture system supports the growth of malignant cells, that multiple cell populations exist in cell suspensions derived from solid tumors, and that differences exist between the radiosensitivity of cells at 2.0 Gy in different histology types
Indigenous culture as a knowledge system | du Plessis | Tydskrif vir ...
African Journals Online (AJOL)
This article makes a contribution to the debate by addressing cultural issues encountered during a cross-cultural research project based in India and South Africa. ... The combined approach provided a method that allowed researchers to develop interventions that capitalised on existing skills, practices and social ...
Human cultures as niche constructions within the solar system
Van de Vliert, Evert
This commentary seeks to refine Kashima's (2016) timely and topical but too-general call for embedding culture within the planetary ecosystem. My starting point is that cultures are to an underestimated extent ongoing niche constructions within the merry-go-round of the Sun's radiation, the Earth's
Flowering of Woody Bamboo in Tissue Culture Systems
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Jin-Ling Yuan
Full Text Available Flowering and subsequent seed set are not only normal activities in the life of most plants, but constitute the very reason for their existence. Woody bamboos can take a long time to flower, even over 100 years. This makes it difficult to breed bamboo, since flowering time cannot be predicted and passing through each generation takes too long. Another unique characteristic of woody bamboo is that a bamboo stand will often flower synchronously, both disrupting the supply chain within the bamboo industry and affecting local ecology. Therefore, an understanding of the mechanism that initiates bamboo flowering is important not only for biology research, but also for the bamboo industry. Induction of flowering in vitro is an effective way to both shorten the flowering period and control the flowering time, and has been shown for several species of bamboo. The use of controlled tissue culture systems allows investigation into the mechanism of bamboo flowering and facilitates selective breeding. Here, after a brief introduction of flowering in bamboo, we review the research on in vitro flowering of bamboo, including our current understanding of the effects of plant growth regulators and medium components on flower induction and how in vitro bamboo flowers can be used in research.
Cultural adaptation of visual attention: calibration of the oculomotor control system in accordance with cultural scenes.
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Yoshiyuki Ueda
Full Text Available Previous studies have found that Westerners are more likely than East Asians to attend to central objects (i.e., analytic attention, whereas East Asians are more likely than Westerners to focus on background objects or context (i.e., holistic attention. Recently, it has been proposed that the physical environment of a given culture influences the cultural form of scene cognition, although the underlying mechanism is yet unclear. This study examined whether the physical environment influences oculomotor control. Participants saw culturally neutral stimuli (e.g., a dog in a park as a baseline, followed by Japanese or United States scenes, and finally culturally neutral stimuli again. The results showed that participants primed with Japanese scenes were more likely to move their eyes within a broader area and they were less likely to fixate on central objects compared with the baseline, whereas there were no significant differences in the eye movements of participants primed with American scenes. These results suggest that culturally specific patterns in eye movements are partly caused by the physical environment.
Does the concept of safety culture help or hinder systems thinking in safety?
Reiman, Teemu; Rollenhagen, Carl
The concept of safety culture has become established in safety management applications in all major safety-critical domains. The idea that safety culture somehow represents a "systemic view" on safety is seldom explicitly spoken out, but nevertheless seem to linger behind many safety culture discourses. However, in this paper we argue that the "new" contribution to safety management from safety culture never really became integrated with classical engineering principles and concepts. This integration would have been necessary for the development of a more genuine systems-oriented view on safety; e.g. a conception of safety in which human, technological, organisational and cultural factors are understood as mutually interacting elements. Without of this integration, researchers and the users of the various tools and methods associated with safety culture have sometimes fostered a belief that "safety culture" in fact represents such a systemic view about safety. This belief is, however, not backed up by theoretical or empirical evidence. It is true that safety culture, at least in some sense, represents a holistic term-a totality of factors that include human, organisational and technological aspects. However, the departure for such safety culture models is still human and organisational factors rather than technology (or safety) itself. The aim of this paper is to critically review the various uses of the concept of safety culture as representing a systemic view on safety. The article will take a look at the concepts of culture and safety culture based on previous studies, and outlines in more detail the theoretical challenges in safety culture as a systems concept. The paper also presents recommendations on how to make safety culture more systemic. Copyright © 2013 Elsevier Ltd. All rights reserved.
Application of cell co-culture system to study fat and muscle cells.
Pandurangan, Muthuraman; Hwang, Inho
Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.
Biljana Bogićević Milikić
The paper intends to investigate how companies can efficiently manage their organisational cultures through changes in the reward system. The paper is based on a research which has taken place in one Serbian company which decided to change its organisational culture, as a prerequisite for further organisational changes. As the main instrument for changing organisational culture, the top management used changes in the reward system. The findings suggest that in the short run only narrow change...
Establishment of sorghum cell suspension culture system for ...
African Journals Online (AJOL)
Mar 18, 2008 ... Additionally, sorghum cell suspension cultures have been initiated from the friable ... proteomics technologies. The field of proteomics is .... air dried at room temperature and resuspended in 2 ml of urea buffer [9 M urea, 2 M ...
A Cultural Framework for the Interoperability of C2 Systems
National Research Council Canada - National Science Library
Slay, Jill
In considering some of the difficulties experienced in coalition operations, it becomes apparent that attention is needed, is in establishing a cultural framework for the interoperability of personnel (the human agents...
Impact of Implementation of an Automated Liquid Culture System on Diagnosis of Tuberculous Pleurisy.
Lee, Byung Hee; Yoon, Seong Hoon; Yeo, Hye Ju; Kim, Dong Wan; Lee, Seung Eun; Cho, Woo Hyun; Lee, Su Jin; Kim, Yun Seong; Jeon, Doosoo
This study was conducted to evaluate the impact of implementation of an automated liquid culture system on the diagnosis of tuberculous pleurisy in an HIV-uninfected patient population. We retrospectively compared the culture yield, time to positivity, and contamination rate of pleural effusion samples in the BACTEC Mycobacteria Growth Indicator Tube 960 (MGIT) and Ogawa media among patients with tuberculous pleurisy. Out of 104 effusion samples, 43 (41.3%) were culture positive on either the MGIT or the Ogawa media. The culture yield of MGIT was higher (40.4%, 42/104) than that of Ogawa media (18.3%, 19/104) (Pliquid culture system could provide approximately twice as high yields and fast results in effusion culture, compared to solid media. Supplemental solid media may have a limited impact on maximizing sensitivity in effusion culture; however, further studies are required.
Standards in chestnut coppice system: cultural heritage or coltural requirement?
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Manetti MC
Full Text Available Standards in chestnut coppice system: cultural heritage or coltural requirement? This paper aims at evaluating the role of standards in chestnut coppices from a biological and functional perspective. In addition to a detailed analysis of Italian regulations on the issue, the technical definition of the term is analysed: (i as for the functional role of standards; (ii to assess whether the required functions are technically necessary and are being actually performed. In this contex, the results of an experimental trial are reported. The goal of the trial were to assess the shoots' parameters, the stand productivity, the dynamics of canopy cover in coppices with or without standards. In 2001, at harvesting operations in a coppice aged 30 with standards managed by the local community, two experimental plots 2500 m2 each were established. The two theses being compared were: simple coppice and coppixce with standards (100 standards per hectare. The released standards were qualified immediately after final harvesting. Sprouting ability, growth pattern and stool vitality were surveyed in March 2004 (at age 2, in May 2008 (at age 6 and in April 2010 (at age 8. First results highlighted the evidence of statistically significant differences between the two thesis. The high number of standards effected negatively both vitality and growth pattern of the stools. Simple coppice recorded a lower shoot mortality, a higher diametrical growth and canopy cover degree as well; the heigth growth was, on the opposite, significantly lower. These results, although referred to a limited lifespan (1/3 of the rotation time and to one site only, underline productive, ecological and environmental benefits and as a consequence suggest the widening of the experimental network and the development of new, more relevant and consistent rules, making acceptable the simple coppice as a possible silvicultural choice to be applied to chestnut coppices.
Rat brain sagittal organotypic slice cultures as an ex vivo dopamine cell loss system.
McCaughey-Chapman, Amy; Connor, Bronwen
Organotypic brain slice cultures are a useful tool to study neurological function as they provide a more complex, 3-dimensional system than standard 2-dimensional in vitro cell cultures. Building on a previously developed mouse brain slice culture protocol, we have developed a rat sagittal brain slice culture system as an ex vivo model of dopamine cell loss. We show that rat brain organotypic slice cultures remain viable for up to 6 weeks in culture. Using Fluoro-Gold axonal tracing, we demonstrate that the slice 3-dimensional cytoarchitecture is maintained over a 4 week culturing period, with particular focus on the nigrostriatal pathway. Treatment of the cultures with 6-hydroxydopamine and desipramine induces a progressive loss of Fluoro-Gold-positive nigral cells with a sustained loss of tyrosine hydroxylase-positive nigral cells. This recapitulates the pattern of dopaminergic degeneration observed in the rat partial 6-hydroxydopamine lesion model and, most importantly, the progressive pathology of Parkinson's disease. Our slice culture platform provides an advance over other systems, as we demonstrate for the first time 3-dimensional cytoarchitecture maintenance of rat nigrostriatal sagittal slices for up to 6 weeks. Our ex vivo organotypic slice culture system provides a long term cellular platform to model Parkinson's disease, allowing for the elucidation of mechanisms involved in dopaminergic neuron degeneration and the capability to study cellular integration and plasticity ex vivo. Copyright © 2017 Elsevier B.V. All rights reserved.
The socio-cultural significance of the diagnostic label "neurasthenia" in Japan's mental health care system.
Munakata, T
This paper is an attempt to explore the socio-cultural significance of deliberately disguising schizophrenia as neurasthenia, neurosis or malfunction of autonomic nervous system. To understand its significance, the socio-cultural background of Japanese attitudes toward mental illness and Japan's mental health care system is also examined from a non-Western standpoint.
Revisiting the Role of Cultural Capital in East Asian Educational Systems: The Case of South Korea
Byun, Soo-yong; Schofer, Evan; Kim, Kyung-keun
The concept of cultural capital has proved invaluable in understanding educational systems in Western countries, and recent work seeks to extend those insights to the diverse educational systems of other geographic regions. Using data from the 2000 Programme for International Student Assessment, the authors explored cultural capital in South Korea…
On China's Social Security System and Traditional Chinese Culture
Institute of Scientific and Technical Information of China (English)
This paper explores the interrelation between China's social security system and traditional Chinese culture, pointing out the meaning of the study, and that China's social security system is a carrier and representation of traditional Chinese culture and
Military Force and Culture Change: Systems, Narratives, and the Social Transmission of Behavior in Counter-Terrorism Strategy
National Research Council Canada - National Science Library
Casebeer, William D
.... Operationalizing culture as socially transmitted behavior, and treating culture systematically using open systems theory, best allows us to understand the perils and prospects of acting upon culture with force...
Synthetic surface for expansion of human mesenchymal stem cells in xeno-free, chemically defined culture conditions.
Directory of Open Access Journals (Sweden)
Paula J Dolley-Sonneville
Full Text Available Human mesenchymal stem cells (HMSCS possess three properties of great interest for the development of cell therapies and tissue engineering: multilineage differentiation, immunomodulation, and production of trophic factors. Efficient ex vivo expansion of hMSCs is a challenging requirement for large scale production of clinical grade cells. Low-cost, robust, scalable culture methods using chemically defined materials need to be developed to address this need. This study describes the use of a xeno-free synthetic peptide acrylate surface, the Corning® Synthemax® Surface, for culture of hMSCs in serum-free, defined medium. Cell performance on the Corning Synthemax Surface was compared to cells cultured on biological extracellular matrix (ECM coatings in xeno-free defined medium and in traditional conditions on tissue culture treated (TCT plastic in fetal bovine serum (FBS supplemented medium. Our results show successful maintenance of hMSCs on Corning Synthemax Surface for eight passages, with cell expansion rate comparable to cells cultured on ECM and significantly higher than for cells in TCT/FBS condition. Importantly, on the Corning Synthemax Surface, cells maintained elongated, spindle-like morphology, typical hMSC marker profile and in vitro multilineage differentiation potential. We believe the Corning Synthemax Surface, in combination with defined media, provides a complete synthetic, xeno-free, cell culture system for scalable production of hMSCs.
Formation and importance of corporate culture in the system of management
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A. V. Bogomolov
Full Text Available The article considers corporate culture as a management tool in the economic model of the activity underlying the organization of management of all factors of production (labor, materials, capital and information. Companies with well developed corporate culture are developing successfully. Weak corporate culture can become a source of deep crisis of the entrepreneurial structure. It is emphasized that the set of factors influencing the development of entrepreneurial structures must be supplemented with a factor of effective corporate culture. Reforms aimed at changes in property relations require the creation of an adequate organizational and economic mechanism for the management of corporate enterprises, taking into account current trends in the concentration and specialization of agro-industrial production. A corporate culture is presented that includes the strategic objective of the firm; Standards of personnel behavior; Structural characteristics of personnel, nature, content, working conditions and methods of its organization; Incentive system; System of personnel training. The types of corporate culture and their features are singled out, namely the culture of power, the culture of roles, the culture of tasks and the culture of individuals. The unique essence of the corporate culture, the complexity of its assessment, create certain problems in the management of corporate culture. Corporate culture provides an opportunity to competently and effectively manage the organization. A strong and solid corporate culture is a necessary attribute of a successful company, as it unites employees who identify with their organization and strive to achieve a common goal by joint efforts. In such a company, key corporate values are shared by all members of the organization, the dominant culture strongly influences the behavior of employees, the need for high structuring and formalization of the company's activity disappears, and the turnover of staff
A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands.
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Takako Noguchi
Full Text Available In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR, respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that
Of Models and Meanings: Cultural Resilience in Social-Ecological Systems
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Todd A. Crane
Full Text Available Modeling has emerged as a key technology in analysis of social-ecological systems. However, the tendency for modeling to focus on the mechanistic materiality of biophysical systems obscures the diversity of performative social behaviors and normative cultural positions of actors within the modeled system. The fact that changes in the biophysical system can be culturally constructed in different ways means that the perception and pursuit of adaptive pathways can be highly variable. Furthermore, the adoption of biophysically resilient livelihoods can occur under conditions that are subjectively experienced as the radical transformation of cultural systems. The objectives of this work are to: (1 highlight the importance of understanding the place of culture within social-ecological systems, (2 explore the tensions between empirical and normative positions in the analysis of social-ecological resilience, and (3 suggest how empirical modeling of social-ecological systems can synergistically interact with normative aspects of livelihoods and lifeways.
Efficient generation of patient-matched malignant and normal primary cell cultures from clear cell renal cell carcinoma patients: clinically relevant models for research and personalized medicine
International Nuclear Information System (INIS)
Lobo, Nazleen C.; Gedye, Craig; Apostoli, Anthony J.; Brown, Kevin R.; Paterson, Joshua; Stickle, Natalie; Robinette, Michael; Fleshner, Neil; Hamilton, Robert J.; Kulkarni, Girish; Zlotta, Alexandre; Evans, Andrew; Finelli, Antonio; Moffat, Jason; Jewett, Michael A. S.; Ailles, Laurie
Patients with clear cell renal cell carcinoma (ccRCC) have few therapeutic options, as ccRCC is unresponsive to chemotherapy and is highly resistant to radiation. Recently targeted therapies have extended progression-free survival, but responses are variable and no significant overall survival benefit has been achieved. Commercial ccRCC cell lines are often used as model systems to develop novel therapeutic approaches, but these do not accurately recapitulate primary ccRCC tumors at the genomic and transcriptional levels. Furthermore, ccRCC exhibits significant intertumor genetic heterogeneity, and the limited cell lines available fail to represent this aspect of ccRCC. Our objective was to generate accurate preclinical in vitro models of ccRCC using tumor tissues from ccRCC patients. ccRCC primary single cell suspensions were cultured in fetal bovine serum (FBS)-containing media or defined serum-free media. Established cultures were characterized by genomic verification of mutations present in the primary tumors, expression of renal epithelial markers, and transcriptional profiling. The apparent efficiency of primary cell culture establishment was high in both culture conditions, but genotyping revealed that the majority of cultures contained normal, not cancer cells. ccRCC characteristically shows biallelic loss of the von Hippel Lindau (VHL) gene, leading to accumulation of hypoxia-inducible factor (HIF) and expression of HIF target genes. Purification of cells based on expression of carbonic anhydrase IX (CA9), a cell surface HIF target, followed by culture in FBS enabled establishment of ccRCC cell cultures with an efficiency of >80 %. Culture in serum-free conditions selected for growth of normal renal proximal tubule epithelial cells. Transcriptional profiling of ccRCC and matched normal cell cultures identified up- and down-regulated networks in ccRCC and comparison to The Cancer Genome Atlas confirmed the clinical validity of our cell cultures. The ability
An empirical investigation to analysis the dimensions of cultural security: A case study of educational system
Directory of Open Access Journals (Sweden)
Elyas Nouraei
Full Text Available The purpose of this study is to explore the concept of cultural security and its practical implications in a case study of educational system. There are several definitions of culture, which describe different aspects of cultural security. In fact, cultural security tends to show various behaviors, thinking, beliefs and other people's perspectives in a society, which represents the overall circumstances. Cultural security and its issues make it possible to have a clear image of a society or community. This paper aims to identify the important dimensions of cultural security in terms of basic components of cultural security identified in a framework. Therefore, after identifying the components, using a questionnaire, required data are gathered from the universities of the city of Ilam, Iran and they are analyzed by SPSS18.0 software and its appropriate statistical tests.
Establishment of automated culture system for murine induced pluripotent stem cells
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Koike Hiroyuki
Full Text Available Abstract Background Induced pluripotent stem (iPS cells can differentiate into any cell type, which makes them an attractive resource in fields such as regenerative medicine, drug screening, or in vitro toxicology. The most important prerequisite for these industrial applications is stable supply and uniform quality of iPS cells. Variation in quality largely results from differences in handling skills between operators in laboratories. To minimize these differences, establishment of an automated iPS cell culture system is necessary. Results We developed a standardized mouse iPS cell maintenance culture, using an automated cell culture system housed in a CO2 incubator commonly used in many laboratories. The iPS cells propagated in a chamber uniquely designed for automated culture and showed specific colony morphology, as for manual culture. A cell detachment device in the system passaged iPS cells automatically by dispersing colonies to single cells. In addition, iPS cells were passaged without any change in colony morphology or expression of undifferentiated stem cell markers during the 4 weeks of automated culture. Conclusions Our results show that use of this compact, automated cell culture system facilitates stable iPS cell culture without obvious effects on iPS cell pluripotency or colony-forming ability. The feasibility of iPS cell culture automation may greatly facilitate the use of this versatile cell source for a variety of biomedical applications.
Long-term maintenance of human induced pluripotent stem cells by automated cell culture system.
Konagaya, Shuhei; Ando, Takeshi; Yamauchi, Toshiaki; Suemori, Hirofumi; Iwata, Hiroo
Pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem (iPS) cells, are regarded as new sources for cell replacement therapy. These cells can unlimitedly expand under undifferentiated conditions and be differentiated into multiple cell types. Automated culture systems enable the large-scale production of cells. In addition to reducing the time and effort of researchers, an automated culture system improves the reproducibility of cell cultures. In the present study, we newly designed a fully automated cell culture system for human iPS maintenance. Using an automated culture system, hiPS cells maintained their undifferentiated state for 60 days. Automatically prepared hiPS cells had a potency of differentiation into three germ layer cells including dopaminergic neurons and pancreatic cells.
Biomaterials in co-culture systems: towards optimizing tissue integration and cell signaling within scaffolds.
Battiston, Kyle G; Cheung, Jane W C; Jain, Devika; Santerre, J Paul
Most natural tissues consist of multi-cellular systems made up of two or more cell types. However, some of these tissues may not regenerate themselves following tissue injury or disease without some form of intervention, such as from the use of tissue engineered constructs. Recent studies have increasingly used co-cultures in tissue engineering applications as these systems better model the natural tissues, both physically and biologically. This review aims to identify the challenges of using co-culture systems and to highlight different approaches with respect to the use of biomaterials in the use of such systems. The application of co-culture systems to stimulate a desired biological response and examples of studies within particular tissue engineering disciplines are summarized. A description of different analytical co-culture systems is also discussed and the role of biomaterials in the future of co-culture research are elaborated on. Understanding the complex cell-cell and cell-biomaterial interactions involved in co-culture systems will ultimately lead the field towards biomaterial concepts and designs with specific biochemical, electrical, and mechanical characteristics that are tailored towards the needs of distinct co-culture systems. Copyright © 2014 Elsevier Ltd. All rights reserved.
A systems biology framework for pathway level culture media engineering: pplication to Pichia pastoris cultures
Ferreira, Ana Raquel Santos
Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica Culture media (CM) formulations contain hundreds of ingredients in aqueous solutions that may be involved in complex interactions in the same or competing pathways within the cell. This thesis proposes a new methodology for determining the optimal composition of CM that migrates from an empirical to a mechanistic or hybrid mechanistic CM development approach. A framework consisting in the execution of an a...
Assessing cultural competence at a local hospital system in the United States.
Polacek, Georgia N L J; Martinez, Rubén
Cultural competence in health care has come to the forefront with the changing demographics in the United States. Standards have been created by the Office of Minority Health for culturally appropriate health care. This article presents the findings of one hospital system's cultural competency assessment. Employee surveys and patient and physician focus groups were conducted to gain insight into cultural differences and challenges encountered in this system. Statistically significant effects of ethnicity and gender on language skills and awareness, as well as differences in awareness and knowledge by the respondent's employment position, were found. Patient concerns included access to care and respect from staff. The need for cross-cultural education and training for all health care delivery personnel was reinforced. Cultural competency will not be achieved if education, attention to diversity, trained interpreters, and the understanding that social factors have a profound influence on health and health outcomes are not considered.
A Novel Greeting Selection System for a Culture-Adaptive Humanoid Robot
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Gabriele Trovato
Full Text Available Robots, especially humanoids, are expected to perform human-like actions and adapt to our ways of communication in order to facilitate their acceptance in human society. Among humans, rules of communication change depending on background culture: greetings are a part of communication in which cultural differences are strong. Robots should adapt to these specific differences in order to communicate effectively, being able to select the appropriate manner of greeting for different cultures depending on the social context. In this paper, we present the modelling of social factors that influence greeting choice, and the resulting novel culture-dependent greeting gesture and words selection system. An experiment with German participants was run using the humanoid robot ARMAR-IIIb. Thanks to this system, the robot, after interacting with Germans, can perform greeting gestures appropriate to German culture in addition to a repertoire of greetings appropriate to Japanese culture.
Educational Policy vs. Culturally Sensitive Programs in Turkish Educational System
Arslan, Hasan
The purpose of this study is to examine the perceptions of elementary school teachers about the sensitiveness of principals, teachers, and curriculum on multicultural education. Education provides the transmission and the advancement of its culture while it is developing and enhancing the common values, the integrity and the progress of…
Isolation and Characterization of Poliovirus in Cell Culture Systems.
Thorley, Bruce R; Roberts, Jason A
The isolation and characterization of enteroviruses by cell culture was accepted as the "gold standard" by clinical virology laboratories. Methods for the direct detection of all enteroviruses by reverse transcription polymerase chain reaction, targeting a conserved region of the genome, have largely supplanted cell culture as the principal diagnostic procedure. However, the World Health Organization's Global Polio Eradication Initiative continues to rely upon cell culture to isolate poliovirus due to the lack of a reliable sensitive genetic test for direct typing of enteroviruses from clinical specimens. Poliovirus is able to infect a wide range of mammalian cell lines, with CD155 identified as the primary human receptor for all three seroytpes, and virus replication leads to an observable cytopathic effect. Inoculation of cell lines with extracts of clinical specimens and subsequent passaging of the cells leads to an increased virus titre. Cultured isolates of poliovirus are suitable for testing by a variety of methods and remain viable for years when stored at low temperature.This chapter describes general procedures for establishing a cell bank and routine passaging of cell lines. While the sections on specimen preparation and virus isolation focus on poliovirus, the protocols are suitable for other enteroviruses.
Radiation-induced PKC signaling system in cultured rat hepatocytes
International Nuclear Information System (INIS)
Nakajima, Tetsuo; Yukawa, Osami
Radiation effects on living organisms are mainly caused through reactive oxygen species (ROS) on living cells. It is known that ROS damages various membranes and the bio membranes play an important role in cellular signal transduction pathways. The effects of radiation on cellular signal transduction pathways in cultured rat hepatocytes have been studied
Animal-cell culture in aqueous two-phase systems
Zijlstra, G.M.
In current industrial biotechnology, animal-cell culture is an important source of therapeutic protein products. The conventional animal-cell production processes, however, include many unit operations as part of the fermentation and downstream processing strategy. The research described in
Organoid culture systems for prostate epithelial and cancer tissue
Drost, Jarno; Karthaus, Wouter R; Gao, Dong; Driehuis, Else; Sawyers, Charles L; Chen, Yu; Clevers, Hans
This protocol describes a strategy for the generation of 3D prostate organoid cultures from healthy mouse and human prostate cells (either bulk or FACS-sorted single luminal and basal cells), metastatic prostate cancer lesions and circulating tumor cells. Organoids derived from healthy material
Design and Evaluation of a Cross-Cultural Training System
Santarelli, Thomas; Stagl, Kevin C.
Cross-cultural competency, and the underlying communication and affective skills required to develop such expertise, is becoming increasingly important for a wide variety of domains. To address this need, we developed a blended learning platform which combines virtual role-play with tutorials, assessment and feedback. A Middle-Eastern Curriculum (MEC) exemplar for cross-cultural training U.S. military personnel was developed to guide the refinement of an existing game-based training platform. To complement this curriculum, we developed scenario authoring tools to enable end-users to define training objectives, link performance measures and feedback/remediation to these objectives, and deploy experiential scenarios within a game-based virtual environment (VE). Lessons learned from the design and development of this exemplar cross-cultural competency curriculum, as well as formative evaluation results, are discussed. Initial findings suggest that the underlying training technology promotes deep levels of semantic processing of the key information of relevant cultural and communication skills.
Bridging the Two Cultures: Disciplinary Divides and Educational Reward Systems
Schiferl, E. I.
In 1959 C.P. Snow believed that communication and education could span the cultural gap between the sciences and the humanities. In the twenty-first century, language, research models, and academic structures hinder intellectual communication between art history, cognitive neuroscience and perceptual psychology--three disciplines dedicated to…
Influence of mixed culture system on the growth performance of ...
African Journals Online (AJOL)
This study describes a novel strategy to improve the α-galactosidase and invertase production of Saccharomyces cerevisiae by co-cultivating it with Aspergillus oryzae. In the mixed culture, the growth of the both strains was repressed, and the protein synthesis for the yeast cell wall was promoted significantly. As a result ...
A Systemic Approach to Culturally Responsive Assessment Practices and Evaluation
Slee, June
In an earlier paper, Slee and Keenan demonstrated that it was possible for tertiary education institutions to design culturally responsive assessment procedures that complied with standardised assessment policy. The authors' paper described "Growing Our Own," an initiative between Charles Darwin University and Northern Territory Catholic…
How Theaters Remember: Cultures of Memory in Institutionalized Systems
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Milena Dragićević Šešić
Full Text Available The aim of this study is to explore organizational policies and strategies regarding the institutional memory of Belgrade's repertoire theaters. The concept of institutional (organizational memory has not been developed within the culture of memory theory. The role of theater in the culture of memory has been researched mostly through studies of its repertoire, corresponding to how theaters deal with issues of glory, guilt, or shame. This study explores how theaters rethink their own past and organizational culture, how they use their capacity for re-imagining themselves, for clarifying their role and function in different historical moments. The objective of this research is to identify the main institutional policies and types of strategies used for preserving institutional memory through key narratives of remembering, and key methods of inter-generational transfer. The sample comprises of four Belgrade-based public repertoire theaters: the Yugoslav Dramatic Theater (JDP, the Belgrade Dramatic Theater (BDP, Atelje 212, and Bitef Theater. Specific attention is given to the means of transmission, of individual (episodic memories into the collective consciousness, influencing organizational cultures and shaping a theater's identity (semantic memory. Research has shown that there are important differences in active policies of preserving institutional memory among Belgrade's theaters. Different organizational and programming strategies were implemented in order to safeguard institutional identity and memory, particularly in theaters with a permanent ensemble. The major difference is between theaters whose culture of memory might be called "non-existent" (Bitef, or "in storage" (BDP, and those succeeding in creating a functional memory (JDP, Atelje 212.
Examining the Relationship Between Safety Management System Implementation and Safety Culture in Collegiate Flight Schools
Robertson, Michael F
Safety management systems (SMS) are becoming the industry standard for safety management throughout the aviation industry. As the Federal Aviation Administration continues to mandate SMS for different segments, the assessment of an organization's safety culture becomes more important. An SMS can facilitate the development of a strong aviation safety culture. This study describes how safety culture and SMS are integrated. The purpose of this study was to examine the relationship between an ...
Scientific culture in Colombia. A proposal of an indicator system for science technology and innovation
Energy Technology Data Exchange (ETDEWEB)
Pardo Martinez, C.I.; Alfonso, W.H
In last decades, scientific culture has become a key element of Governance of Science, Technology and Innovation in the countries where it is important to determine measurement to analysis trends on scientific culture. Research questions that guide this paper are the following: i. What are information needs on scientific culture in Colombia?; ii. How can be measured scientific culture?; iii. What is the adequate structure for indicators of scientific culture?. In order to answer these questions, a mix of methodologies is used. First, we review the literature on scientific culture and indicators related to this topic. Second, we made a series of interviews with staff members of Colciencias to determine requirements of measurement on scientific culture. Third, with this information, we built an information matrix to prioritise information and determine indicators with respective metrics, and sources according to relevance and cost-effectiveness of estimation. Fourth, from indicators formulated and an indicator system is proposed determining for every dimension of scientific culture indicators related to inputs, process, and outputs designed indicator sheets that includes definition, objective, sources aggregation levels, time series, and calculation methods for indicators proposed. This study achieves formulate an indicator system from the definition of scientific culture a and its dimension proposing around 60 indicators through a multidimensional model that integrates different elements of scientific culture such as the individual and society establishing indicators to measure inputs, process and outputs in general form and specific initiatives for Colciencias. (Author)
Methodology for considering environments and culture in developing information security systems
Mwakalinga, G Jeffy; Kowalski, Stewart; Yngström, Louise
In this paper, we describe a methodology for considering culture of users and environments when developing information security systems. We discuss the problem of how researchers and developers of security for information systems have had difficulties in considering culture of users and environments when they develop information security systems. This has created environments where people serve technology instead of technology serving people. Users have been considered just as any other compo...
Effects of all-trans retinol and cigarette smoke condensate on hamster tracheal epithelium in organ culture. I. A cell proliferation study
Rutten, A.A.J.J.L.; Wilmer, J.W.G.M.; Beems, R.B.
The effects of cigarette smoke condensate (CSC) and all-trans retinol on the cell proliferative activity of vitamin A-deprived hamster tracheal epithelium have been studied in vitamin A-deficient, serum-free, hormone-supplemented medium in organ culture. In the absence of retinol, CSC induced a
Highly efficient full-length hepatitis C virus genotype 1 (strain TN) infectious culture system
DEFF Research Database (Denmark)
Li, Yi-Ping; Ramirez, Santseharay; Jensen, Sanne B
Chronic infection with hepatitis C virus (HCV) is an important cause of end stage liver disease worldwide. In the United States, most HCV-related disease is associated with genotype 1 infection, which remains difficult to treat. Drug and vaccine development was hampered by inability to culture...... full-length TN infection dose-dependently. Given the unique importance of genotype 1 for pathogenesis, this infectious 1a culture system represents an important advance in HCV research. The approach used and the mutations identified might permit culture development for other HCV isolates, thus......) culture systems in Huh7.5 cells. Here, we developed a highly efficient genotype 1a (strain TN) full-length culture system. We initially found that the LSG substitutions conferred viability to an intergenotypic recombinant composed of TN 5' untranslated region (5'UTR)-NS5A and JFH1 NS5B-3'UTR; recovered...
Visual Estimation of Bacterial Growth Level in Microfluidic Culture Systems
Directory of Open Access Journals (Sweden)
Kyukwang Kim
Full Text Available Microfluidic devices are an emerging platform for a variety of experiments involving bacterial cell culture, and has advantages including cost and convenience. One inevitable step during bacterial cell culture is the measurement of cell concentration in the channel. The optical density measurement technique is generally used for bacterial growth estimation, but it is not applicable to microfluidic devices due to the small sample volumes in microfluidics. Alternately, cell counting or colony-forming unit methods may be applied, but these do not work in situ; nor do these methods show measurement results immediately. To this end, we present a new vision-based method to estimate the growth level of the bacteria in microfluidic channels. We use Fast Fourier transform (FFT to detect the frequency level change of the microscopic image, focusing on the fact that the microscopic image becomes rough as the number of cells in the field of view increases, adding high frequencies to the spectrum of the image. Two types of microfluidic devices are used to culture bacteria in liquid and agar gel medium, and time-lapsed images are captured. The images obtained are analyzed using FFT, resulting in an increase in high-frequency noise proportional to the time passed. Furthermore, we apply the developed method in the microfluidic antibiotics susceptibility test by recognizing the regional concentration change of the bacteria that are cultured in the antibiotics gradient. Finally, a deep learning-based data regression is performed on the data obtained by the proposed vision-based method for robust reporting of data.
Sociocultural Systems: The Next Step in Army Cultural Capability
chapters in her conclusion of the anthology, pulling together overarching themes for the reader. She discusses multiple perspectives for defining and...driver pulling up to a gas pump is likely to be low on gas) or knowing the implication of a particular set of activities (we know that shopping in...relevance to civilian and military helpers. These are referred to by anthropologists as gatekeepers, culture brokers, and (my term) goalies . They are
Visual Estimation of Bacterial Growth Level in Microfluidic Culture Systems.
Kim, Kyukwang; Kim, Seunggyu; Jeon, Jessie S
Microfluidic devices are an emerging platform for a variety of experiments involving bacterial cell culture, and has advantages including cost and convenience. One inevitable step during bacterial cell culture is the measurement of cell concentration in the channel. The optical density measurement technique is generally used for bacterial growth estimation, but it is not applicable to microfluidic devices due to the small sample volumes in microfluidics. Alternately, cell counting or colony-forming unit methods may be applied, but these do not work in situ; nor do these methods show measurement results immediately. To this end, we present a new vision-based method to estimate the growth level of the bacteria in microfluidic channels. We use Fast Fourier transform (FFT) to detect the frequency level change of the microscopic image, focusing on the fact that the microscopic image becomes rough as the number of cells in the field of view increases, adding high frequencies to the spectrum of the image. Two types of microfluidic devices are used to culture bacteria in liquid and agar gel medium, and time-lapsed images are captured. The images obtained are analyzed using FFT, resulting in an increase in high-frequency noise proportional to the time passed. Furthermore, we apply the developed method in the microfluidic antibiotics susceptibility test by recognizing the regional concentration change of the bacteria that are cultured in the antibiotics gradient. Finally, a deep learning-based data regression is performed on the data obtained by the proposed vision-based method for robust reporting of data.
Traveller: An Interactive Cultural Training System Controlled by User-Defined Body Gestures
Kistler, F.; André, E.; Mascarenhas, S.; Silva, A.; Paiva, A.; Degens, D.M.; Hofstede, G.J.; Krumhuber, E.; Kappas, A.; Aylett, R.
In this paper, we describe a cultural training system based on an interactive storytelling approach and a culturally-adaptive agent architecture, for which a user-defined gesture set was created. 251 full body gestures by 22 users were analyzed to find intuitive gestures for the in-game actions in
The Influence of Academic Culture on Quality Management System ISO 9001 Maintenance within Malaysian Universities
Basir, Siti Arni; Davies, John; Douglas, Jacqueline; Douglas, Alexander
This study investigates the influence of the elements of academic culture on quality management system ISO 9001 maintenance within Malaysian universities. There is a dearth of empirical studies on maintaining ISO 9001, particularly in the higher education context. From the literature review, academic culture was classified according to four…
Enhanced production and isotope enrichment of recombinant glycoproteins produced in cultured mammalian cells
International Nuclear Information System (INIS)
Skelton, David; Goodyear, Abbey; Ni, DaQun; Walton, Wendy J.; Rolle, Myron; Hare, Joan T.; Logan, Timothy M.
NMR studies of post-translationally modified proteins are complicated by the lack of an efficient method to produce isotope enriched recombinant proteins in cultured mammalian cells. We show that reducing the glucose concentration and substituting glutamate for glutamine in serum-free medium increased cell viability while simultaneously increasing recombinant protein yield and the enrichment of non-essential amino acids compared to culture in unmodified, serum-free medium. Adding dichloroacetate, a pyruvate dehydrogenase kinase inhibitor, further improves cell viability, recombinant protein yield, and isotope enrichment. We demonstrate the method by producing partially enriched recombinant Thy1 glycoprotein from Lec1 Chinese hamster ovary (CHO) cells using U- 13 C-glucose and 15 N-glutamate as labeled precursors. This study suggests that uniformly 15 N, 13 C-labeled recombinant proteins may be produced in cultured mammalian cells starting from a mixture of labeled essential amino acids, glucose, and glutamate.
Enhanced production and isotope enrichment of recombinant glycoproteins produced in cultured mammalian cells
Energy Technology Data Exchange (ETDEWEB)
Skelton, David; Goodyear, Abbey [Florida State University, Department of Chemistry and Biochemistry (United States); Ni, DaQun; Walton, Wendy J.; Rolle, Myron; Hare, Joan T. [Florida State University, Institute of Molecular Biophysics (United States); Logan, Timothy M., E-mail: email@example.com [Florida State University, Department of Chemistry and Biochemistry (United States)
NMR studies of post-translationally modified proteins are complicated by the lack of an efficient method to produce isotope enriched recombinant proteins in cultured mammalian cells. We show that reducing the glucose concentration and substituting glutamate for glutamine in serum-free medium increased cell viability while simultaneously increasing recombinant protein yield and the enrichment of non-essential amino acids compared to culture in unmodified, serum-free medium. Adding dichloroacetate, a pyruvate dehydrogenase kinase inhibitor, further improves cell viability, recombinant protein yield, and isotope enrichment. We demonstrate the method by producing partially enriched recombinant Thy1 glycoprotein from Lec1 Chinese hamster ovary (CHO) cells using U-{sup 13}C-glucose and {sup 15}N-glutamate as labeled precursors. This study suggests that uniformly {sup 15}N,{sup 13}C-labeled recombinant proteins may be produced in cultured mammalian cells starting from a mixture of labeled essential amino acids, glucose, and glutamate.
Multi-Organ toxicity demonstration in a functional human in vitro system composed of four organs.
Oleaga, Carlota; Bernabini, Catia; Smith, Alec S T; Srinivasan, Balaji; Jackson, Max; McLamb, William; Platt, Vivien; Bridges, Richard; Cai, Yunqing; Santhanam, Navaneetha; Berry, Bonnie; Najjar, Sarah; Akanda, Nesar; Guo, Xiufang; Martin, Candace; Ekman, Gail; Esch, Mandy B; Langer, Jessica; Ouedraogo, Gladys; Cotovio, Jose; Breton, Lionel; Shuler, Michael L; Hickman, James J
We report on a functional human model to evaluate multi-organ toxicity in a 4-organ system under continuous flow conditions in a serum-free defined medium utilizing a pumpless platform for 14 days. Computer simulations of the platform established flow rates and resultant shear stress within accepted ranges. Viability of the system was demonstrated for 14 days as well as functional activity of cardiac, muscle, neuronal and liver modules. The pharmacological relevance of the integrated modules were evaluated for their response at 7 days to 5 drugs with known side effects after a 48 hour drug treatment regime. The results of all drug treatments were in general agreement with published toxicity results from human and animal data. The presented phenotypic culture model exhibits a multi-organ toxicity response, representing the next generation of in vitro systems, and constitutes a step towards an in vitro "human-on-a-chip" assay for systemic toxicity screening.
Revisiting the Role of Cultural Capital in East Asian Educational Systems: The Case of South Korea.
Byun, Soo-Yong; Schofer, Evan; Kim, Kyung-Keun
The concept of cultural capital has proved invaluable in understanding educational systems in Western countries, and recent work seeks to extend those insights to the diverse educational systems of other geographic regions. We explored cultural capital in South Korea by investigating the relationships among family socioeconomic status (SES), cultural capital, and children's academic achievement using data from the 2000 Programme for International Student Assessment. South Korea was compared with Japan, France, and the United States to understand how institutional features of South Korean education shape the role of cultural capital in academic success. Results showed that family SES had a positive effect on both parental objectified cultural capital and children's embodied cultural capital in South Korea, consistent with evidence from the other countries. Moreover, parental objectified cultural capital had a positive effect on children's academic achievement in South Korea. In contrast to other countries, however, children's embodied cultural capital had a negative effect on academic achievement in South Korea controlling for the other variables. We highlighted several institutional features of South Korean education including a standardized curriculum, extreme focus on test preparation, and extensive shadow education, which may combine to suppress the effect of children's embodied cultural capital on academic achievement.
Regulatory Oversight of Safety Culture in Finland: A Systemic Approach to Safety
International Nuclear Information System (INIS)
Oedewald, P.; Väisäsvaara, J.
In Finland the Radiation and Nuclear Safety Authority STUK specifies detailed regulatory requirements for good safety culture. Both the requirements and the practical safety culture oversight activities reflect a systemic approach to safety: the interconnections between the technical, human and organizational factors receive special attention. The conference paper aims to show how the oversight of safety culture can be integrated into everyday oversight activities. The paper also emphasises that the scope of the safety culture oversight is not specific safety culture activities of the licencees, but rather the overall functioning of the licence holder or the new build project organization from safety point of view. The regulatory approach towards human and organizational factors and safety culture has evolved throughout the years of nuclear energy production in Finland. Especially the recent new build projects have highlighted the need to systematically pay attention to the non-technical aspects of safety as it has become obvious how the HOF issues can affect the design processes and quality of construction work. Current regulatory guides include a set of safety culture related requirements. The requirements are binding to the licence holders and they set both generic and specific demands on the licencee to understand, monitor and to develop safety culture of their own organization but also that of their supplier network. The requirements set for the licence holders has facilitated the need to develop the regulator's safety culture oversight practices towards a proactive and systemic approach.
Enhanced Chondrocyte Proliferation in a Prototyped Culture System with Wave-Induced Agitation
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Pilarek Maciej
Full Text Available One of the actual challenges in tissue engineering applications is to efficiently produce as high of number of cells as it is only possible, in the shortest time. In static cultures, the production of animal cell biomass in integrated forms (i.e. aggregates, inoculated scaffolds is limited due to inefficient diffusion of culture medium components observed in such non-mixed culture systems, especially in the case of cell-inoculated fiber-based dense 3D scaffolds, inside which the intensification of mass transfer is particularly important. The applicability of a prototyped, small-scale, continuously wave-induced agitated system for intensification of anchorage-dependent CP5 chondrocytes proliferation outside and inside three-dimensional poly(lactic acid (PLA scaffolds has been discussed. Fibrous PLA-based constructs have been inoculated with CP5 cells and then maintained in two independent incubation systems: (i non-agitated conditions and (ii culture with wave-induced agitation. Significantly higher values of the volumetric glucose consumption rate have been noted for the system with the wave-induced agitation. The advantage of the presented wave-induced agitation culture system has been confirmed by lower activity of lactate dehydrogenase (LDH released from the cells in the samples of culture medium harvested from the agitated cultures, in contrast to rather high values of LDH activity measured for static conditions. Results of the proceeded experiments and their analysis clearly exhibited the feasibility of the culture system supported with continuously wave-induced agitation for robust proliferation of the CP5 chondrocytes on PLA-based structures. Aside from the practicability of the prototyped system, we believe that it could also be applied as a standard method offering advantages for all types of the daily routine laboratory-scale animal cell cultures utilizing various fiber-based biomaterials, with the use of only regular laboratory
Crafting decision options and alternatives for designing cultural observation system using general morphological modelling
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Jaber Moghaddampour
Full Text Available According to connoisseurs, cultural system is encountering a fully new space in future decades and cultural indicators will be exposed by some dangers; for the same reason, cultural observation activities in management dialogue of Iran are emphasized; the concept of "observation" in Iran is facing with challenges including being far from the concepts of futures studies and the focus on "cultural indicators monitoring" while cultural engineering needs foresight and identification of affecting progressive and deterring factors on the culture. The present study aims at providing alternatives to design an observation system by considering the monitoring and scanning simultaneously to bring out strategic and futurist vision for cultural organizations. To this end, the solution space and morphological field of observation (parameters and values by using MA/Carma Viewer software package is designed based on the literature review and forming a five-member expert group and specialized conversations. Upon Internal Consistency Assessment of parameters and by considering some values as drivers, Parameters Activity Check was conducted to study the reactions by other parameters and values. After sense-making and proper understanding of the model behavior, an Inference and "What-If" model were devised; some configurations were studied and provided in order to aware a part of proper alternatives for designing a cultural observation system (two scenarios. Scenario selection is different due to contingencies and conditions of executing the process as well as the users' needs and goals in cultural observation system and cultural organizations can make decisions and design detailed observation systems by using morphological models, solution space and alternatives provided in the present study and depended on their goals and needs.
[Research progress of co-culture system for constructing vascularized tissue engineered bone].
Fu, Weili; Xiang, Zhou
To review the research progress of the co-culture system for constructing vascularized tissue engineered bone. The recent literature concerning the co-culture system for constructing vascularized tissue engineered bone was reviewed, including the selection of osteogenic and endothelial lineages, the design and surface modification of scaffolds, the models and dimensions of the co-culture system, the mechanism, the culture conditions, and their application progress. The construction of vascularized tissue engineered bone is the prerequisite for their survival and further clinical application in vivo. Mesenchymal stem cells (owning the excellent osteogenic potential) and endothelial progenitor cells (capable of directional differentiation into endothelial cell) are considered as attractive cell types for the co-culture system to construct vascularized tissue engineered bone. The culture conditions need to be further optimized. Furthermore, how to achieve the clinical goals of minimal invasion and autologous transplantation also need to be further studied. The strategy of the co-culture system for constructing vascularized tissue engineered bone would have a very broad prospects for clinical application in future.
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Theerasak Thanasankit
Full Text Available This paper explores the impact of Thai culture on managing the decision making process in requirements engineering and contribution a better understand of its influence on the management of requirements engineering process. The paper illustrates the interaction of technology and culture and shows that rather than technology changing culture, culture can change the way technology is used. Thai culture is naturally inherent in Thai daily life and Thais bring that into their work practices. The concepts of power and uncertainty in Thai culture contribute toward hierarchical forms of communication and decision making process in Thailand, especially during requirements engineering, where information systems requirements need to be established for further development. The research shows that the decision making process in Thailand tends to take a much longer time, as every stage during requirements engineering needs to be reported to management for final decisions. The tall structure of Thai organisations also contributes to a bureaucratic, elongated decision-making process during information systems development. Understanding the influence of Thai culture on requirements engineering and information systems development will assist multinational information systems consulting organisations to select, adapt, better manage, or change requirements engineering process and information systems developments methodologies to work best with Thai organisations.
SCRMS: An RFID and Sensor Web-Enabled Smart Cultural Relics Management System.
Xiao, Changjiang; Chen, Nengcheng; Li, Dandan; Lv, You; Gong, Jianya
Cultural relics represent national or even global resources of inestimable value. How to efficiently manage and preserve these cultural relics is a vitally important issue. To achieve this goal, this study proposed, designed, and implemented an RFID and Sensor Web-enabled smart cultural relics management system (SCRMS). In this system, active photovoltaic subtle energy-powered Radio Frequency Identification (RFID) is used for long-range contactless identification and lifecycle management of cultural relics during their storage and circulation. In addition, different types of ambient sensors are integrated with the RFID tags and deployed around cultural relics to monitor their environmental parameters, helping to ensure that they remain in good condition. An Android-based smart mobile application, as middleware, is used in collaboration with RFID readers to collect information and provide convenient management for the circulation of cultural relics. Moreover, multiple sensing techniques are taken advantage of simultaneously for preservation of cultural relics. The proposed system was successfully applied to a museum in the Yongding District, Fujian Province, China, demonstrating its feasibility and advantages for smart and efficient management and preservation of cultural relics.
SCRMS: An RFID and Sensor Web-Enabled Smart Cultural Relics Management System
Directory of Open Access Journals (Sweden)
Changjiang Xiao
Full Text Available Cultural relics represent national or even global resources of inestimable value. How to efficiently manage and preserve these cultural relics is a vitally important issue. To achieve this goal, this study proposed, designed, and implemented an RFID and Sensor Web–enabled smart cultural relics management system (SCRMS. In this system, active photovoltaic subtle energy-powered Radio Frequency Identification (RFID is used for long-range contactless identification and lifecycle management of cultural relics during their storage and circulation. In addition, different types of ambient sensors are integrated with the RFID tags and deployed around cultural relics to monitor their environmental parameters, helping to ensure that they remain in good condition. An Android-based smart mobile application, as middleware, is used in collaboration with RFID readers to collect information and provide convenient management for the circulation of cultural relics. Moreover, multiple sensing techniques are taken advantage of simultaneously for preservation of cultural relics. The proposed system was successfully applied to a museum in the Yongding District, Fujian Province, China, demonstrating its feasibility and advantages for smart and efficient management and preservation of cultural relics.
Concerns and Opportunities around Cultural Heritage in East Asian Globally Important Agricultural Heritage Systems (GIAHS
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Hiroyuki Kajihara
Full Text Available Fifteen years have passed since Food and Agriculture Organization (FAO launched the Globally Important Agricultural Heritage Systems (GIAHS project in 2002. In this time, participation from East Asian countries has been increasing rapidly with interest flowing over into several related subjects and disciplines. Culture is one of the selection criteria that has to be satisfied to become a GIAHS site, and equally culture plays an important role in the development of tourism to a destination. However, few scientists or GIAHS members have discussed directly how to apply cultural features in GIAHS. Therefore, the purposes of this paper are firstly to recognize the importance and contribution of culture in GIAHS. Then, through detailing the current forms of cultural management in the GIAHS located in Japan, Korea, and China, we identify some of the key cultural problems and prospects in those sites. Two social surveys conducted in Japan show that culture is a prime motivation for tourist visitation, as well as being a core GIAHS selection criteria. These surveys further highlight that GIAHS needs to incorporate culture more effectively into their management strategies. Detailed descriptions of the three countries analyzed in this paper outline each has to engage with particular cultural management challenges: Japan has a well-arranged list of cultural assets, but is unclear how to move forward with that information and data. Korea has just begun to generate a strategy on how to manage cultural heritage features in GIAHS with the use of approaches such as Agrostories or Gil tourism, in recognition of the gradual changes that are occurring in local identity. China has the longest history of engagement with GIAHS in the East Asia region. However, the utilization of the model here has recognized further issues of change in cultural identity not least through commercialization. This paper therefore identifies, discusses and arranges eight problems and
Fluidic system for long-term in vitro culturing and monitoring of organotypic brain slices
DEFF Research Database (Denmark)
Bakmand, Tanya; Troels-Smith, Ane R.; Dimaki, Maria
Brain slice preparations cultured in vitro have long been used as a simplified model for studying brain development, electrophysiology, neurodegeneration and neuroprotection. In this paper an open fluidic system developed for improved long term culturing of organotypic brain slices is presented....... The positive effect of continuous flow of growth medium, and thus stability of the glucose concentration and waste removal, is simulated and compared to the effect of stagnant medium that is most often used in tissue culturing. Furthermore, placement of the tissue slices in the developed device was studied...... by numerical simulations in order to optimize the nutrient distribution. The device was tested by culturing transverse hippocampal slices from 7 days old NMRI mice for a duration of 14 days. The slices were inspected visually and the slices cultured in the fluidic system appeared to have preserved...
Mycobacterium tuberculosis bacteremia detected by the Isolator lysis-centrifugation blood culture system.
Kiehn, T E; Gold, J W; Brannon, P; Timberger, R J; Armstrong, D
Mycobacterium tuberculosis was detected by the Isolator lysis-centrifugation blood culture system from the blood of a patient with tuberculosis of the breast. The organism also grew on conventional laboratory media inoculated with pleural fluid from the patient.
Behaviour of marine oil-degrading bacterial populations in a continuous culture system
Digital Repository Service at National Institute of Oceanography (India)
Mohandass, C.; David, J.J.; Nair, S.; LokaBharathi, P.A.; Chandramohan, D.
In pursuit of developing an oil-degrading microbial consortium, we used the principle of "plasmid assisted molecular breeding" (PAMB) in a continuous culture system. Three marine bacteria, Pseudomonas putida, Brevibacterium epidermidis...
PDMS/glass microfluidic cell culture system for cytotoxicity tests and cells passage
DEFF Research Database (Denmark)
Ziolkowska, K.; Jedrych, E.; Kwapiszewski, R.
In this paper, hybrid (PDMS/glass) microfluidic cell culture system (MCCS) integrated with the concentration gradient generator (CGG) is presented. PDMS gas permeability enabled cells' respiration in the fabricated microdevices and excellent glass hydrophilicity allowed successful cells' seeding...
Monitoring System of Sustainable Development in Cultural and Mountain Tourism Destinations
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Jurigova Zuzana
Full Text Available Tourism destinations are vulnerable to negative impacts of tourism development and thus require a sustainable approach. It is significant mainly in destinations with fragile environments such as cultural destinations with their historical value and mountain destinations with specific natural conditions. The aim of this paper is to propose the monitoring system for sustainable development of cultural and mountain destinations based on the critical scientific literature review. The added value of this work resides in defining specific indicators (creating monitoring system for measurement of sustainability in cultural routes and mountain destinations.
Tolerance as a factor of value system formation within process of cross-cultural communication
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J. Y. Hanas
Full Text Available Crosscultural communication relates to particular phenomenon in two or more cultures and has an additional value for communicative competence comparison of different cultures representatives. The realization of communicative competence capacity is culturally conditioned, in addition, it also caused by unique individual experience of person. Intercultural communication became one of the most urgent issues of humanity in modern society. Study of intercultural communication becomes increasingly important in recent years due to globalization. Features of intercultural communication are studied within the sciences such as philosophy, linguistics, cultural studies, psychology, sociology, anthropology, ethnology, cybernetics, and an interdisciplinary process. Intercultural communication as a social phenomenon was called to the practical needs of the postwar world, reinforced by ideological interest, which of the early twentieth century was formed in academia and in the public mind for the different cultures and languages. The study of intercultural communication is a result of rapid economic development of many countries and regions, revolutionary changes in technology associated with this globalization of economic activity. On the level of historical evolutionary approach to the development of complex systems tolerance phenomenon could not be reduced to everyday perspective of tolerance. Tolerance is works as cultural norm and as a civilization principle. A key feature of tolerance as long as multiculturalism is support of complex systems diversity. Tolerance also provides a right of each individual to be a different personality. The concept of tolerance is understood as a norm that provides a balance opposing sides and the possibility of dialogue of various world views, religions and cultures. Initial thesis that each person is a unique individual and unlike the others, is characterized by different manifestations of their own individuality, is the
Analysis of GMO Plum Plant Culture in System Operations Failure
Mercado, Dianne
GMO plum trees are being evaluated at the Kennedy Space Center as a possible candidate for future space crops. Previously conducted horticultural testing compared the performance of several plum genotypes in controlled environment chambers, resulting in a down-selection to the NASA-11 genotype. Precursory studies determined the water use requirements to sustain the plants as well as the feasibility of grafting non-GMO plum scions onto GMO plum rootstocks of NASA-5, NASA-10, and NASA-11 genotypes. This study follows the growth and horticultural progress of plum trees and in-vitro cultures from August 2017 to November 2017, and provides supplemental support for future GMO plum studies. The presence of Hurricane Irma in early September 2017 resulted in the plants undergoing material deterioration from major changes to their overall horticultural progress.
Model-aided optimization of delta-endotoxin-formation in continuous culture systems
Energy Technology Data Exchange (ETDEWEB)
Schulz, V; Schorcht, R; Ignatenko, Yu N; Sakharova, Z V; Khovrychev, M P
A mathematical model of growth, sporulation and delta-endotoxin-formation of bac. thuringiensis is given. The results of model-aided optimization of steady-state continuous culture systems indicate that the productivity in the one-stage system is 1.9% higher and in the two-stage system is 18.5% higher than in the batch process.
Three-dimensional spheroid culture of human umbilical cord mesenchymal stem cells promotes cell yield and stemness maintenance.
Li, Yi; Guo, Gang; Li, Li; Chen, Fei; Bao, Ji; Shi, Yu-Jun; Bu, Hong
Mesenchymal stem cell (MSC) transplantation is a promising treatment of many diseases. However, conventional techniques with cells being cultured as a monolayer result in slow cell proliferation and insufficient yield to meet clinical demands. Three-dimensional (3D) culture systems are gaining attention with regard to recreating a complex microenvironment and to understanding the conditions experienced by cells. Our aim is to establish a novel 3D system for the culture of human umbilical cord MSCs (hUC-MSCs) within a real 3D microenvironment but with no digestion or passaging. Primary hUC-MSCs were isolated and grown in serum-free medium (SFM) on a suspension Rocker system. Cell characteristics including proliferation, phenotype and multipotency were recorded. The therapeutic effects of 3D-cultured hUC-MSCs on carbon tetrachloride (CCl4)-induced acute liver failure in mouse models were examined. In the 3D Rocker system, hUC-MSCs formed spheroids in SFM and maintained high viability and active proliferation. Compared with monolayer culture, the 3D-culture system yielded more hUC-MSCs cells within the same volume. The spheroids expressed higher levels of stem cell markers and displayed stronger multipotency. After transplantation into mouse, 3D hUC-MSCs significantly promoted the secretion of interferon-γ and interleukin-6 but inhibited that of tumor necrosis factor-α, thereby alleviating liver necrosis and promoting regeneration following CCl4 injury. The 3D culture of hUC-MSCs thus promotes cell yield and stemness maintenance and represents a promising strategy for hUC-MSCs expansion on an industrial scale with great potential for cell therapy and biotechnology.
Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production.
Ferraz, Marcia A M M; Henning, Heiko H W; Stout, Tom A E; Vos, Peter L A M; Gadella, Bart M
The oviduct was long considered a largely passive conduit for gametes and embryos. However, an increasing number of studies into oviduct physiology have demonstrated that it specifically and significantly influences gamete interaction, fertilization and early embryo development. While oviduct epithelial cell (OEC) function has been examined during maintenance in conventional tissue culture dishes, cells seeded into these two-dimensional (2-D) conditions suffer a rapid loss of differentiated OEC characteristics, such as ciliation and secretory activity. Recently, three-dimensional (3-D) cell culture systems have been developed that make use of cell inserts to create basolateral and apical medium compartments with a confluent epithelial cell layer at the interface. Using such 3-D culture systems, OECs can be triggered to redevelop typical differentiated cell properties and levels of tissue organization can be developed that are not possible in a 2-D culture. 3-D culture systems can be further refined using new micro-engineering techniques (including microfluidics and 3-D printing) which can be used to produce 'organs-on-chips', i.e. live 3-D cultures that bio-mimic the oviduct. In this review, concepts for designing bio-mimic 3-D oviduct cultures are presented. The increased possibilities and concomitant challenges when trying to more closely investigate oviduct physiology, gamete activation, fertilization and embryo production are discussed.
A multiplexed chip-based assay system for investigating the functional development of human skeletal myotubes in vitro.
Smith, A S T; Long, C J; Pirozzi, K; Najjar, S; McAleer, C; Vandenburgh, H H; Hickman, J J
This report details the development of a non-invasive in vitro assay system for investigating the functional maturation and performance of human skeletal myotubes. Data is presented demonstrating the survival and differentiation of human myotubes on microscale silicon cantilevers in a defined, serum-free system. These cultures can be stimulated electrically and the resulting contraction quantified using modified atomic force microscopy technology. This system provides a higher degree of sensitivity for investigating contractile waveforms than video-based analysis, and represents the first system capable of measuring the contractile activity of individual human muscle myotubes in a reliable, high-throughput and non-invasive manner. The development of such a technique is critical for the advancement of body-on-a-chip platforms toward application in pre-clinical drug development screens. Copyright © 2014 Elsevier B.V. All rights reserved.
[Biological characteristics of mesenchymal stem cell and hematopoietic stem cell in the co-culture system].
Wei, Wei; Xu, Chao; Ye, Zhi-Yong; Huang, Xiao-Jun; Yuan, Jia-En; Ma, Tian-Bao; Lin, Han-Biao; Chen, Xiu-Qiong
The aim of the present study was to obtain the qualified hematopoietic stem/progenitor cells (HSC/HPC) and human umbilical cord-mesenchymal stem cells (MSC) in vitro in the co-culture system. Cord blood mononuclear cells were separated from umbilical cord blood by Ficoll lymphocyte separation medium, and then CD34 + HSC was collected by MACS immunomagnetic beads. The selected CD34 + HSC/HPC and MSC were transferred into culture flask. IMDM culture medium with 15% AB-type cord plasma supplemented with interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand (Flt-3L) factors were used as the co-culture system for the amplification of HSC/HPC and MSC. The cellular growth status and proliferation on day 6 and 10 after co-culture were observed by using inverted microscope. The percentage of positive expression of CD34 in HSC/HPC, as well as the percentages of positive expressions of CD105, CD90, CD73, CD45, CD34 and HLA-DR in the 4 th generation MSC, was tested by flow cytometry. Semisolid colony culture was used to test the HSC/HPC colony forming ability. The osteogenic, chondrogenesis and adipogenic ability of the 4 th generation MSC were assessed. The karyotype analysis of MSC was conducted by colchicines. The results demonstrated that the HSC/HPC of co-culture group showed higher ability of amplification, CFU-GM and higher CD34 + percentage compared with the control group. The co-cultured MSC maintained the ability to differentiate into bone cells, fat cells and chondrocytes. And the karyotype stability of MSC remained normal. These results reveal that the appropriate co-culture system for MSC and HSC is developed, and via this co-culture system we could gain both two kinds of these cells. The MSCs under the co-culture system maintain the biological characteristics. The CFU-GM ability, cell counting and the flow cytometry results of HSC/HPC under the co-culture system are conform to the criterion, showing that
Spatially monitoring oxygen level in 3D microfabricated cell culture systems using optical oxygen sensing beads.
Wang, Lin; Acosta, Miguel A; Leach, Jennie B; Carrier, Rebecca L
Capability of measuring and monitoring local oxygen concentration at the single cell level (tens of microns scale) is often desirable but difficult to achieve in cell culture. In this study, biocompatible oxygen sensing beads were prepared and tested for their potential for real-time monitoring and mapping of local oxygen concentration in 3D micro-patterned cell culture systems. Each oxygen sensing bead is composed of a silica core loaded with both an oxygen sensitive Ru(Ph2phen3)Cl2 dye and oxygen insensitive Nile blue reference dye, and a poly-dimethylsiloxane (PDMS) shell rendering biocompatibility. Human intestinal epithelial Caco-2 cells were cultivated on a series of PDMS and type I collagen based substrates patterned with micro-well arrays for 3 or 7 days, and then brought into contact with oxygen sensing beads. Using an image analysis algorithm to convert florescence intensity of beads to partial oxygen pressure in the culture system, tens of microns-size oxygen sensing beads enabled the spatial measurement of local oxygen concentration in the microfabricated system. Results generally indicated lower oxygen level inside wells than on top of wells, and local oxygen level dependence on structural features of cell culture surfaces. Interestingly, chemical composition of cell culture substrates also appeared to affect oxygen level, with type-I collagen based cell culture systems having lower oxygen concentration compared to PDMS based cell culture systems. In general, results suggest that oxygen sensing beads can be utilized to achieve real-time and local monitoring of micro-environment oxygen level in 3D microfabricated cell culture systems.
Investigating Educational Systems, Leadership, and School Culture: A Holistic Approach
Pratt, Jill Elizabeth
Most populous school districts operate using a bureaucratic hierarchical organizational structure developed primarily for industry, a system structure that has remained intact for a century despite evolving from a manufacturing to a knowledge-based economy. Although strong for efficiency, this system structure is resistant to change and promotes…
Comparison of the lysis-centrifugation and agitated biphasic blood culture systems for detection of fungemia.
Murray, P R
Although the detection of fungemia has been improved by the use of vented or biphasic blood culture bottles, the best recovery and earliest detection have been reported in the Isolator lysis-centrifugation system. It was recently demonstrated that improved detection of both bacteria and fungi was accomplished by mechanically agitating blood culture bottles for the first 24 h of incubation. In this study the detection of fungemia by use of the Isolator system was compared with that of an agitated biphasic system. A total of 182 fungi were isolated from blood specimens inoculated into both culture systems. No difference in the overall recovery of fungi or individual species of yeasts was observed between the two systems. However, all seven isolates of Histoplasma capsulatum were recovered in the Isolator system only. The time required to detect fungemia with each of the two systems was also compared. No statistically significant difference was observed. From the data collected during this 18-month study, it can be concluded that the overall recovery and time of detection of yeasts are equivalent in the lysis-centrifugation system and the agitated biphasic blood culture system. The lysis-centrifugation system is still superior for the detection of filamentous fungi such as H. capsulatum. PMID:1993772
Development of an automated chip culture system with integrated on-line monitoring for maturation culture of retinal pigment epithelial cells
Directory of Open Access Journals (Sweden)
Mee-Hae Kim
Full Text Available In cell manufacturing, the establishment of a fully automated, microfluidic, cell culture system that can be used for long-term cell cultures, as well as for process optimization is highly desirable. This study reports the development of a novel chip bioreactor system that can be used for automated long-term maturation cultures of retinal pigment epithelial (RPE cells. The system consists of an incubation unit, a medium supply unit, a culture observation unit, and a control unit. In the incubation unit, the chip contains a closed culture vessel (2.5 mm diameter, working volume 9.1 μL, which can be set to 37 °C and 5% CO2, and uses a gas-permeable resin (poly- dimethylsiloxane as the vessel wall. RPE cells were seeded at 5.0 × 104 cells/cm2 and the medium was changed every day by introducing fresh medium using the medium supply unit. Culture solutions were stored either in the refrigerator or the freezer, and fresh medium was prepared before any medium change by warming to 37 °C and mixing. Automated culture was allowed to continue for 30 days to allow maturation of the RPE cells. This chip culture system allows for the long-term, bubble-free, culture of RPE cells, while also being able to observe cells in order to elucidate their cell morphology or show the presence of tight junctions. This culture system, along with an integrated on-line monitoring system, can therefore be applied to long-term cultures of RPE cells, and should contribute to process control in RPE cell manufacturing.
Mixed culture polyhydroxyalkanoates production from sugar molasses: the use of a 2-stage CSTR system for culture selection.
Albuquerque, M G E; Concas, S; Bengtsson, S; Reis, M A M
Polyhydroxyalkanoates (PHAs) are promising biodegradable polymers. The use of mixed microbial cultures (MMC) and low cost feedstocks have a positive impact on the cost-effectiveness of the process. It has typically been carried out in Sequencing Batch Reactors (SBR). In this study, a 2-stage CSTR system (under Feast and Famine conditions) was used to effectively select for PHA-storing organisms using fermented molasses as feedstock. The effect of influent substrate concentration (60-120 Cmmol VFA/L) and HRT ratio between the reactors (0.2-0.5h/h) on the system's selection efficiency was assessed. It was shown that Feast reactor residual substrate concentration impacted on the selective pressure for PHA storage (due to substrate-dependent kinetic limitation). Moreover, a residual substrate concentration coming from the Feast to the Famine reactor did not jeopardize the physiological adaptation required for enhanced PHA storage. The culture reached a maximum PHA content of 61%. This success opens new perspectives to the use of wastewater treatment infrastructure for PHA production, thus valorizing either excess sludge or wastewaters. Copyright 2010 Elsevier Ltd. All rights reserved.
Evaluating Safety Culture Under the Socio-Technical Complex Systems Perspective
International Nuclear Information System (INIS)
Lemos, F. L. de
Since the term "safety culture" was coined, it has gained more and more attention as an effort to achieve higher levels of system safety. A good deal of effort has been done in order to better define, evaluate and implement safety culture programs in organizations throughout all industries, and especially in the Nuclear Industry. Unfortunately, despite all those efforts, we continue to witness accidents that are, in great part, attributed to flaws in the safety culture of the organization. Fukushima nuclear accident is one example of a serious accident in which flaws in the safety culture has been pointed to as one of the main contributors. In general, the definitions of safety culture emphasise the social aspect of the system. While the definitions also include the relations with the technical aspects, it does so in a general sense. For example, the International Nuclear Safety Advisory Group (INSAG) defines safety culture as: "The assembly of characteristics and attitudes in organizations and individuals which establishes that, as an overriding priority, nuclear plant safety issues receives the attention warranted by their significance." By the way safety culture is defined we can infer that it represents a property of a social system, or a property of the social aspect of the system. In this sense, the social system is a component of the whole system. Where, "system" is understood to be comprised of a social (humans) and technical (equipment) aspects, as a Nuclear Power Plant, for example. Therefore, treating safety culture as an identity on its own right, finding and fixing flaws in the safety culture may not be enough to improve safety of the system. We also needed to evaluate all the interactions between the components that comprise all the aspects of the system. In some cases a flaw in the safety culture can easily be detected, such as an employee not wearing appropriate individual protection equipment, e.g., dosimeter, or when basic safety
Design of biomimetic cellular scaffolds for co-culture system and their application
Kook, Yun-Min; Jeong, Yoon; Lee, Kangwon; Koh, Won-Gun
The extracellular matrix of most natural tissues comprises various types of cells, including fibroblasts, stem cells, and endothelial cells, which communicate with each other directly or indirectly to regulate matrix production and cell functionality. To engineer multicellular interactions in vitro, co-culture systems have achieved tremendous success achieving a more realistic microenvironment of in vivo metabolism than monoculture system in the past several decades. Recently, the fields of tissue engineering and regenerative medicine have primarily focused on three-dimensional co-culture systems using cellular scaffolds, because of their physical and biological relevance to the extracellular matrix of actual tissues. This review discusses several materials and methods to create co-culture systems, including hydrogels, electrospun fibers, microfluidic devices, and patterning for biomimetic co-culture system and their applications for specific tissue regeneration. Consequently, we believe that culture systems with appropriate physical and biochemical properties should be developed, and direct or indirect cell–cell interactions in the remodeled tissue must be considered to obtain an optimal tissue-specific microenvironment. PMID:29081966
Dynamic cell culture system: a new cell cultivation instrument for biological experiments in space
Gmunder, F. K.; Nordau, C. G.; Tschopp, A.; Huber, B.; Cogoli, A.
The prototype of a miniaturized cell cultivation instrument for animal cell culture experiments aboard Spacelab is presented (Dynamic cell culture system: DCCS). The cell chamber is completely filled and has a working volume of 200 microliters. Medium exchange is achieved with a self-powered osmotic pump (flowrate 1 microliter h-1). The reservoir volume of culture medium is 230 microliters. The system is neither mechanically stirred nor equipped with sensors. Hamster kidney (Hak) cells growing on Cytodex 3 microcarriers were used to test the biological performance of the DCCS. Growth characteristics in the DCCS, as judged by maximal cell density, glucose consumption, lactic acid secretion and pH, were similar to those in cell culture tubes.
Organ Culture as a Model System for Studies on Enterotoxin Interactions with the Intestinal Epithelium
DEFF Research Database (Denmark)
Lorenzen, Ulver Spangsberg; Hansen, Gert H; Danielsen, E Michael
Studies on bacterial enterotoxin-epithelium interactions require model systems capable of mimicking the events occurring at the molecular and cellular levels during intoxication. In this chapter, we describe organ culture as an often neglected alternative to whole-animal experiments or enterocyte......-like cell lines. Like cell culture, organ culture is versatile and suitable for studying rapidly occurring events, such as enterotoxin binding and uptake. In addition, it is advantageous in offering an epithelium with more authentic permeability/barrier properties than any cell line, as well...
System-level modeling and simulation of the cell culture microfluidic biochip ProCell
DEFF Research Database (Denmark)
Minhass, Wajid Hassan; Pop, Paul; Madsen, Jan
Microfluidic biochips offer a promising alternative to a conventional biochemical laboratory. There are two technologies for the microfluidic biochips: droplet-based and flow-based. In this paper we are interested in flow-based microfluidic biochips, where the liquid flows continuously through pre......-defined micro-channels using valves and pumps. We present an approach to the system-level modeling and simulation of a cell culture microfluidic biochip called ProCell, Programmable Cell Culture Chip. ProCell contains a cell culture chamber, which is envisioned to run 256 simultaneous experiments (viewed...
Non-Mulberry and Mulberry Silk Protein Sericins as Potential Media Supplement for Animal Cell Culture
Directory of Open Access Journals (Sweden)
Neety Sahu
Full Text Available Silk protein sericins, in the recent years, find application in cosmetics and pharmaceuticals and as biomaterials. We investigate the potential of sericin, extracted from both mulberry Bombyx mori and different non-mulberry sources, namely, tropical tasar, Antheraea mylitta; muga, Antheraea assama; and eri, Samia ricini, as growth supplement in serum-free culture medium. Sericin supplemented media containing different concentrations of sericins from the different species are examined for attachment, growth, proliferation, and morphology of fibrosarcoma cells. The optimum sericin supplementation seems to vary with the source of sericins. The results indicate that all the sericins promote the growth of L929 cells in serum-free culture media; however, S. ricini sericin seems to promote better growth of cells amongst other non-mulberry sericins.
Employing Data Fusion in Cultural Analysis and Counterinsurgency in Tribal Social Systems
Merten, Steffen
This article was published in Culture and Conflict Review (Fall 2009), v.3 no.3 "The point of this essay has been to outline ways that data fusion may be achieved, and how it can dramatically enhance the analytical capabilities of cultural analysts, especially in tribal social systems. By using Visual Analytics theory and technology to conduct the labor intensive aspects of data fusion, and accepting the theoretical justification of fusion between the geospatial, relational, and temporal d...
Using Cultural Modeling to Inform a NEDSS-Compatible System Functionality Evaluation
Anderson, Olympia; Torres-Urquidy, Miguel
Objective The culture by which public health professionals work defines their organizational objectives, expectations, policies, and values. These aspects of culture are often intangible and difficult to qualify. The introduction of an information system could further complicate the culture of a jurisdiction if the intangibles of a culture are not clearly understood. This report describes how cultural modeling can be used to capture intangible elements or factors that may affect NEDSS-compatible (NC) system functionalities within the culture of public health jurisdictions. Introduction The National Notifiable Disease Surveillance System (NNDSS) comprises many activities including collaborations, processes, standards, and systems which support gathering data from US states and territories. As part of NNDSS, the National Electronic Disease Surveillance System (NEDSS) provides the standards, tools, and resources to support reporting public health jurisdictions (jurisdictions). The NEDSS Base System (NBS) is a CDC-developed, software application available to jurisdictions to collect, manage, analyze and report national notifiable disease (NND) data. An evaluation of NEDSS with the objective of identifying the functionalities of NC systems and the impact of these features on the user's culture is underway. Methods We used cultural models to capture additional NC system functionality gaps within the culture of the user. Cultural modeling is a process of graphically depicting people and organizations referred to as influencers and the intangible factors that affect the user's operations or work as influences. Influencers are denoted as bubbles while influences are depicted as arrows penetrating the bubbles. In the cultural model, influence can be seen by the size and proximity (or lack of) in the model. We restricted the models to secondary data sources and interviews of CDC programs (data users) and public health jurisdictions (data reporters). Results Three cultural
Plant regeneration system from cotyledons-derived calluses cultures ...
African Journals Online (AJOL)
Sep 26, 2011 ... The objective of this study was to successfully establish plant regeneration system with cotyledons of. Stylosanthes guianensis Sw. cv. 'Reyan 2' as explants. In this study, the following results were obtained; (1) the highest rates of callus induction on medium MS with 3.0 mg L-1 2, 4-D with cotyledons.
Mobile system for in-situ imaging of cultural objects
Czech Academy of Sciences Publication Activity Database
Žemlička, J.; Jakůbek, J.; Krejčí, F.; Hradil, David; Hradilová, J.; Mislerová, H.
Roč. 7, January (2012), C01108/1-C01108/8 ISSN 1748-0221 R&D Projects: GA MŠk(CZ) LC06041 Institutional research plan: CEZ:AV0Z40320502 Keywords : inspection with x-rays * X-ray fluorescence (XRF) systems Subject RIV: CA - Inorganic Chemistry Impact factor: 1.869, year: 2011
Production systems, diversity and richness of cavy culture in ...
African Journals Online (AJOL)
In order to assess existing cavy (Cavia porcellus) production systems of Cameroon western highlands and rain forest agro-ecological zones, a household ... for a well designed and comprehensive national breeding program for cavies, and increased capacity building of farmers to address mortality rates and health issues.
Development of an in vitro culture system adapted to banana ...
African Journals Online (AJOL)
The beneficial impact of arbuscular mycorrhizal (AM) fungi on banana nutrition and resistance against abiotic and biotic stresses is well documented. However, most studies were conducted under greenhouse or field conditions and none reported the life cycle of the AM fungi on banana roots. It is obvious that any system ...
A thin-layer liquid culture technique for the growth of Helicobacter pylori.
Joo, Jung-Soo; Park, Kyung-Chul; Song, Jae-Young; Kim, Dong-Hyun; Lee, Kyung-Ja; Kwon, Young-Cheol; Kim, Jung-Min; Kim, Kyung-Mi; Youn, Hee-Shang; Kang, Hyung-Lyun; Baik, Seung-Chul; Lee, Woo-Kon; Cho, Myung-Je; Rhee, Kwang-Ho
Several attempts have been successful in liquid cultivation of Helicobaccter pylori. However, there is a need to improve the growth of H. pylori in liquid media in order to get affluent growth and a simple approach for examining bacterial properties. We introduce here a thin-layer liquid culture technique for the growth of H. pylori. A thin-layer liquid culture system was established by adding liquid media to a 90-mm diameter Petri dish. Optimal conditions for bacterial growth were investigated and then viability, growth curve, and released proteins were examined. Maximal growth of H. pylori was obtained by adding 3 mL of brucella broth supplemented with 10% horse to a Petri dish. H. pylori grew in both DMEM and RPMI-1640 supplemented with 10% fetal bovine serum and 0.5% yeast extract. Serum-free RPMI-1640 supported the growth of H. pylori when supplemented with dimethyl-beta-cyclodextrin (200 microg/mL) and 1% yeast extract. Under optimal growth, H. pylori grew exponentially for 28 hours, reaching a density of 3.4 OD(600) with a generation time of 3.3 hours. After 24 hours, cultures at a cell density of 1.0 OD(600) contained 1.3 +/- 0.1 x 10(9 )CFU/mL. gamma-Glutamyl transpeptidase, nuclease, superoxide dismutase, and urease were not detected in culture supernatants at 24 hours in thin-layer liquid culture, but were present at 48 hours, whereas alcohol dehydrogenase, alkylhydroperoxide reductase, catalase, and vacuolating cytotoxin were detected at 24 hours. Thin-layer liquid culture technique is feasible, and can serve as a versatile liquid culture technique for investigating bacterial properties of H. pylori.
Efficient expansion of mesenchymal stromal cells in a disposable fixed bed culture system.
Mizukami, Amanda; Orellana, Maristela D; Caruso, Sâmia R; de Lima Prata, Karen; Covas, Dimas T; Swiech, Kamilla
The need for efficient and reliable technologies for clinical-scale expansion of mesenchymal stromal cells (MSC) has led to the use of disposable bioreactors and culture systems. Here, we evaluate the expansion of cord blood-derived MSC in a disposable fixed bed culture system. Starting from an initial cell density of 6.0 × 10(7) cells, after 7 days of culture, it was possible to produce of 4.2(±0.8) × 10(8) cells, which represents a fold increase of 7.0 (±1.4). After enzymatic retrieval from Fibra-Cell disks, the cells were able to maintain their potential for differentiation into adipocytes and osteocytes and were positive for many markers common to MSC (CD73, CD90, and CD105). The results obtained in this study demonstrate that MSC can be efficiently expanded in the culture system. This novel approach presents several advantages over the current expansion systems, based on culture flasks or microcarrier-based spinner flasks and represents a key element for MSC cellular therapy according to GMP compliant clinical-scale production system. Copyright © 2013 American Institute of Chemical Engineers.
Effectiveness of a Novel Specimen Collection System in Reducing Blood Culture Contamination Rates.
Bell, Mary; Bogar, Catherine; Plante, Jessica; Rasmussen, Kristen; Winters, Sharon
False-positive blood-culture results due to skin contamination of samples remain a persistent problem for health care providers. Our health system recognized that our rates of contamination across the 4 emergency department campuses were above the national average. A unique specimen collection system was implemented throughout the 4 emergency departments and became the mandatory way to collect adult blood cultures. The microbiology laboratory reported contamination rates weekly to manage potential problems; 7 months of data are presented here. There was an 82.8% reduction in false positives with the unique specimen collection system compared with the standard method (chi-squared test with Yates correction, 2-tailed, P = 0.0001). Based on the historical 3.52% rate of blood-culture contamination for our health facilities, 2.92 false positives were prevented for every 100 blood cultures drawn, resulting from adoption of the unique specimen collection system as the standard of care. This unique collection system can reduce the risk of blood culture contamination significantly and is designed to augment, rather than replace, the standard phlebotomy protocol already in use in most health care settings. Copyright © 2018 Elsevier Inc. All rights reserved.
Formation of Pedagogical System for Individual Self-Development by Means of Physical Culture and Sport
Panachev, Valery D.; Zelenin, Leonid A.; Opletin, Anatoly A.; Verbytskyi, Sergei A.
Problems of formation, development and introduction of the modern pedagogical selfdevelopment system in university educational process by means of physical culture and sport have been considered in this article. Such generated pedagogical system reflects practical implementation of social order on the modern educational paradigm aimed at creation…
Of Models and Meanings: Cultural Resilience in Social–Ecological Systems
Crane, T.A.
Modeling has emerged as a key technology in analysis of social–ecological systems. However, the tendency for modeling to focus on the mechanistic materiality of biophysical systems obscures the diversity of performative social behaviors and normative cultural positions of actors within the modeled
The Transformation of Ergonomic Affordances into Cultural Affordances: The Case of the Alnuset System
Chiappini, Giampaolo
Is it possible to study the ergonomic affordances offered by a system designed for educational aims and their transformation into cultural affordances? To this purpose, what references can we adopt? This work describes the theoretical framework used to realise this study referring to AlNuSet, a system realised within the EC ReMath project to…
Revisiting Bourdieu: Alternative Educational Systems in the Light of the Theory of Social and Cultural Reproduction
Azaola, Marta Cristina
The paper reflects upon the principles and practice of an alternative educational system operating in rural Mexico in the light of Bourdieu's theory of cultural and social reproduction. Bourdieu's theory seeks to explain processes of reproduction of power relations within schools and society; whereas alternative educational systems seek to expand…
Influence of cell type and cell culture media on the propagation of foot-and-mouth disease virus with regard to vaccine quality.
Dill, Veronika; Hoffmann, Bernd; Zimmer, Aline; Beer, Martin; Eschbaumer, Michael
Suspension culture of BHK cells allows large-scale virus propagation and cost-efficient vaccine production, while the shift to animal-component-free cell culture media without serum is beneficial for the quality and downstream processing of the product. Foot-and-mouth disease virus is still endemic in many parts of the world and high-quality vaccines are essential for the eradication of this highly contagious and economically devastating disease. Changes to the viral genome sequence during passaging in an adherent and a suspension cell culture system were compared and the impact of amino acid substitutions on receptor tropism, antigenicity and particle stability was examined. Virus production in suspension cells in animal-component-free media and in serum-containing media as well as in adherent cells in serum-containing media was compared. Infection kinetics were determined and the yield of intact viral particles was estimated in all systems using sucrose density gradient centrifugation. Capsid protein sequence alterations were serotype-specific, but varied between cell lines. But The A 24 -2P virus variant had expanded its receptor tropism, but virus neutralization tests found no changes in the antigenic profile in comparison to the original viruses. There were no differences in viral titer between a suspension and an adherent cell culture system, independent of the type of media used. Also, the usage of a serum-free suspension culture system promoted viral growth and allowed an earlier harvest. For serotype O isolates, no differences were seen in the yield of 146S particles. Serotype A preparations revealed a decreased yield of 146S particles in suspension cells independent of the culture media. The selective pressure of the available surface receptors in different cell culture systems may be responsible for alterations in the capsid coding sequence of culture-grown virus. Important vaccine potency characteristics such as viral titer and the neutralization
Cultural impacts on e-learning systems' success
Aparicio, M.; Bação, F.; Oliveira, T.
WOS:000383295100007 (Nº de Acesso Web of Science) E-learning systems are enablers in the learning process, strengthening their importance as part of the educational strategy. Understanding the determinants of e-learning success is crucial for defining instructional strategies. Several authors have studied e-learning implementation and adoption, and various studies have addressed e-learning success from different perspectives. However, none of these studies have verified whether students' c...
Impact of culture on the application of quality management system
M.Ing. The management of quality in companies has become more and more strategically important over recent years. The emphasis on quality is vital to managing projects and achieving excellence in today's global economy. Modern petrochemical construction project management has incorporated quality management principles and initiatives in their activities. The Quality Management System is used to ensure that the project will satisfy the requirements for which it was undertaken. Improving pro...
Development of a definition, classification system, and model for cultural geology
Mitchell, Lloyd W., III
The concept for this study is based upon a personal interest by the author, an American Indian, in promoting cultural perspectives in undergraduate college teaching and learning environments. Most academicians recognize that merged fields can enhance undergraduate curricula. However, conflict may occur when instructors attempt to merge social science fields such as history or philosophy with geoscience fields such as mining and geomorphology. For example, ideologies of Earth structures derived from scientific methodologies may conflict with historical and spiritual understandings of Earth structures held by American Indians. Specifically, this study addresses the problem of how to combine cultural studies with the geosciences into a new merged academic discipline called cultural geology. This study further attempts to develop the merged field of cultural geology using an approach consisting of three research foci: a definition, a classification system, and a model. Literature reviews were conducted for all three foci. Additionally, to better understand merged fields, a literature review was conducted specifically for academic fields that merged social and physical sciences. Methodologies concentrated on the three research foci: definition, classification system, and model. The definition was derived via a two-step process. The first step, developing keyword hierarchical ranking structures, was followed by creating and analyzing semantic word meaning lists. The classification system was developed by reviewing 102 classification systems and incorporating selected components into a system framework. The cultural geology model was created also utilizing a two-step process. A literature review of scientific models was conducted. Then, the definition and classification system were incorporated into a model felt to reflect the realm of cultural geology. A course syllabus was then developed that incorporated the resulting definition, classification system, and model. This
The rat whole embryo culture assay using the Dysmorphology Score system.
Zhang, Cindy; Panzica-Kelly, Julie; Augustine-Rauch, Karen
The rat whole embryo culture (WEC) system has been used extensively for characterizing teratogenic properties of test chemicals. In this chapter, we describe the methodology for culturing rat embryos as well as a new morphological score system, the Dysmorphology Score (DMS) system for assessing morphology of mid gestation (gestational day 11) rat embryos. In contrast to the developmental stage focused scoring associated with the Brown and Fabro score system, this new score system assesses the respective degree of severity of dysmorphology, which delineates normal from abnormal morphology of specific embryonic structures and organ systems. This score system generates an approach that allows rapid identification and quantification of adverse developmental findings, making it conducive for characterization of compounds for teratogenic properties and screening activities.
Evaluation of a rainbow trout (Oncorhynchus mikyss culture water recirculating system
Directory of Open Access Journals (Sweden)
Iván Sánchez O.
Full Text Available Objective. To evaluate a water recirculation system for rainbow trout fish cultures at the recirculating laboratory of the Aquaculture Engineering Production Program of University of Nariño. Materials and Methods. 324 rainbow trout (Oncorhynchus mikyss fries were cultured in 12 plastic tanks with a capacity of 250 L in an aquaculture recirculating system the treatment system of which was made up by a conventional sedimentation tank, a fixed stand upflow biofilter with recycled PVC tube pieces and a natural degassing system; the sedimentation unit effluent was pumped up to a reservoir tank using a 2 HP centrifugal pump after being subject to gravity through the biofilter and to be then distributed to the 12 culture units to which a constant amount of air from a blower was injected. Results. The water treatment system removed 31% of total suspended solids, 9.5% of total ammonia nitrogen, and increased dissolved oxygen to the final effluent in 6.5%. An increase of 305% in biomass was calculated during 75 days, the mortality percentage registered throughout the study period was 4.9%. Conclusions. The water treatment system maintained the physicochemical water quality parameters within the values recommended for the species. The increase in weight and size, food conversion, mortality and biomass production reported normal values for rainbow trout fish culture in recirculating systems.
Garlic exerts allelopathic effects on pepper physiology in a hydroponic co-culture system
Directory of Open Access Journals (Sweden)
Haiyan Ding
Full Text Available A hydroponic co-culture system was adopted to determine the allelopathic potential of garlic on the growth of pepper plants. Different numbers of garlic plants (0, 2, 4, 8 and 12 were hydroponically co-cultured with two pepper plants to investigate allelopathic effects on the growth attributes and antioxidative defense system of the test pepper plants. The responses of the pepper plants depended on the number of garlic plants included in the co-culture system, indicating an association of pepper growth with the garlic root exudate concentration. When grown at a pepper/garlic ratio of 1:1 or 1:2, the pepper plant height, chlorophyll content, and peroxidase (POD, catalase (CAT and phenylalanine ammonia-lyase (PAL activities were significantly increased after 30 days of co-culture; in contrast, reduction in methane dicarboxylic aldehyde (MDA content was observed. However, when the pepper/garlic ratio was 1:4 or higher, these morphological indices and protective enzyme activities were significantly inhibited, whereas MDA levels in the pepper leaves were significantly increased due to severe membrane lipid peroxidation. The results indicate that although low concentrations of garlic root exudates appear to induce protective enzyme systems and promote pepper growth, high concentrations have deleterious effects. These findings suggest that further investigations should optimize the co-culture pepper/garlic ratio to reduce continuous cropping obstacles in pepper production.
Large-scale clinical comparison of the lysis-centrifugation and radiometric systems for blood culture
International Nuclear Information System (INIS)
Brannon, P.; Kiehn, T.E.
The Isolator 10 lysis-centrifugation blood culture system (E. I. du Pont de Nemours and Co., Inc., Wilmington, Del.) was compared with the BACTEC radiometric method (Johnston Laboratories, Inc., Towson, Md.) with 6B and 7D broth media for the recovery of bacteria and yeasts. From 11,000 blood cultures, 1,174 clinically significant organisms were isolated. The Isolator system recovered significantly more total organisms, members of the family Enterobacteriaceae, Staphylococcus spp., and yeasts. The BACTEC system recovered significantly more Pseudomonas spp., Streptococcus spp., and anaerobes. Of the Isolator colony counts, 87% measured less than 11 CFU/ml of blood. Organisms, on an average, were detected the same day from each of the two culture systems. Only 13 of the 975 BACTEC isolates (0.01%) were recovered by subculture of growth-index-negative bottles, and 12 of the 13 were detected in another broth blood culture taken within 24 h. Contaminants were recovered from 4.8% of the Isolator 10 and 2.3% of the BACTEC cultures
Development of a mechanical testing and loading system for trabecular bone studies for long term culture
Directory of Open Access Journals (Sweden)
DB Jones
Full Text Available A highly accurate (�3% mechanical loading and measurement system combined with a trabecular bone diffusion culture-loading chamber has been developed, which provides the ability to study trabecular bone (and possibly cartilage under controlled culture and loading conditions over long periods of time. The loading device has been designed to work in two main modes, either to apply a specific compressive strain to a trabecular bone cylinder or to apply a specific force and measure the resulting deformation. Presently, precisely machined bone cylinders can be loaded at frequencies between 0.1 Hz to 50 Hz and amplitudes over 7,000�e. The system allows accurate measurement of many mechanical properties of the tissue in real time, including visco-elastic properties. This paper describes the technical components, reproducibility, precision, and the calibration procedures of the loading system. Data on long term culture and mechanical responses to different loading patterns will be published separately.
Hand-made cloned buffalo (Bubalus bubalis) embryos: comparison of different media and culture systems.
Shah, Riaz A; George, Aman; Singh, Manoj K; Kumar, Dharmendra; Chauhan, Manmohan S; Manik, Radhaysham; Palta, Prabhat; Singla, Suresh K
Hand-made cloning (HMC) has proved to be an efficient alternative to the conventional micromanipulator-based technique in some domestic animal species. This study reports the development of an effective culture system for in vitro culture of zona-free cloned buffalo (Bubalus bubalis) embryos reconstructed using adult skin fibroblast cells as nucleus donor. Cleavage and blastocyst rates observed were 52 and 0% in modified Charles Rosenkrans 2 (mCR2), 61 and 4.6% in modified Synthetic Oviductal Fluid (mSOF), and 82 and 40.3% in Research Vitro Cleave (RVCL; Cook, Australia) medium, respectively. Similarly, higher blastocyst rates (24.5 +/- 4.1%) were observed when zona-free parthenotes were cultured in RVCL medium. Culturing zona-free cloned buffalo embryos on flat surfaces (FS) yielded significantly higher (p WOW) or microdrops (MD). Furthermore, development in WOW was found to be significantly better than MD culture. The quality of HMC blastocysts was examined using differential staining. This study establishes the application of zona-free nuclear transfer procedures for the production of hand-made cloned buffalo embryos and the development of efficient culture system and appropriate media requirements for enhancing their preimplantation development.
Culturing Security System of Chemical Laboratory in Indonesia
Directory of Open Access Journals (Sweden)
Eka Dian Pusfitasari
Full Text Available Indonesia has experiences on the lack of chemical security such as: a number of bombing terrors and hazardous chemicals found in food. Bomb used in terror is a homemade bomb made from chemicals which are widely spread in the research laboratories such as a mixture of pottasium chlorate, sulphur, and alumunium. Therefore, security of chemicals should be implemented to avoid the misused of the chemicals. Although it has experienced many cases of the misuse of chemicals, and many regulations and seminars related to chemical security have been held, but the implementation of chemical security is still a new thing for Indonesian citizens. The evident is coming from the interviews conducted in this study. Questions asked in this interview/survey included: the implementation of chemical safety and chemical security in laboratory; chemical inventory system and its regulation; and training needed for chemical security implementation. Respondents were basically a researcher from Government Research Institutes, University laboratories, senior high school laboratories, and service laboratories were still ambiguous in distinguishing chemical safety and chemical security. Because of this condition, most Indonesia chemical laboratories did not totally apply chemical security system. Education is very important step to raise people awareness and address this problem. Law and regulations should be sustained by all laboratory personnel activities to avoid chemical diversion to be used for harming people and environment. The Indonesia Government could also develop practical guidelines and standards to be applied to all chemical laboratories in Indonesia. These acts can help Government's efforts to promote chemical security best practices which usually conducted by doing seminars and workshop.
Arts and cultural activity: A vital part of the health and care system.
Cann, Paul L
This article discusses how the arts and cultural activities are a vital part of a health and care system and have potential to fulfil the theme of active ageing. The changing nature of care provision in response to demographic change, fiscal pressure and increasingly consumerist attitudes on the part of care users, is considered. Selected examples of how participation in arts and cultural activities increases not only well-being but also health outcomes are then outlined. The article highlights the potential of 'cultural commissioning' and within that 'arts on prescription' - public funding of arts-related activities for people with care needs - and advocates investment in arts and cultural activities to better meet the demands of health, social care and aged care. Concluding remarks are made, and a way forward is suggested. © 2017 AJA Inc.
The use of plant tissue culture system in the mutagenesis of Secale cereale L
International Nuclear Information System (INIS)
Rybczynski, J.J.; KozIowska, W.; Turzynski, D.
Full text: Among cereals, Secale cereale L. is the worst species for 'in vitro' mutagenesis. In the case of seed mutagenesis of rye each seed is expected to be a different genotype and only somatic embryogenesis assures propagation towards numerous individuals possessing the same genotype. Therefore, another system of in-vitro mutagenesis is explored. Immature embryos were isolated from spikes of field growing plants. The established cultures were irradiated with 0.5; 1.0 and 1.5 kR gamma rays on the first day of the culture and after 6 weeks in culture. After irradiation all cultures were subcultured. For mutagenesis in general uniformity of the original material is very important. Therefore, in rye, irradiation of regenerated somatic embryos may be a good approach. (author)
Effect Of The Use Of Information Technology And Organization Cultural Of The Quality Accounting Information System
Directory of Open Access Journals (Sweden)
Full Text Available The result of the application of effective accounting information system and provide quality and effective accounting information quality. Fundamental rule accounting information systems in an organization is generating accounting information quality through the process of collecting raw data and then processed and then presented in the form of accounting information useful for user information. The purpose of this study was to know how the effect of Use of information technology on the quality of accounting information systems organizational culture on the quality of accounting information systems and the quality of accounting information system on the quality of accounting information. Based on the literature of some previous researchers proved that a technology affects the quality of accounting information systems using information system AIS Effectively requires an understanding of the organization management and information technology shaping the system. the use of information technology within an organization intended to provide information to the user. B. Organizational culture affects the quality of AIS at the stage of design and implementation of the system required careful consideration of the information attitude is the main component of the organization information systems can be substantially influenced by the culture of the organization. C accounting information quality influence on information accounting quality is built with the main purpose to process accounting data from various sources into the accounting information needed by a wide range of users to reduce risk when making decisions.
Evaluation of the antimicrobial removal device when used with the BACTEC blood culture system
International Nuclear Information System (INIS)
Strand, C.L.
A study to determine the value of the Antimicrobial Removal Device (ARD) used in conjunction with radiometric detection of bacteremia using three media was conducted. During a 12-month period, 622 duplicate ARD/BACTEC blood-culture sets were collected. There were 88 positive cultures that yielded 68 pathogenic isolates and 28 probable contaminant isolates. When all patients were considered, 31 pathogenic isolates were detected by both systems, 25 pathogenic isolates were detected faster or only by the BACTEC system, and 12 pathogenic isolates were detected faster or only when the ARD was employed. This difference is statistically significant (P less than 0.05). Thus, the standard BACTEC blood-culture system using three different media was superior to the same BACTEC system using ARD-processed blood specimens. When only patients receiving antimicrobial therapy were considered, there were more pathogenic isolates detected from unprocessed blood than from blood processed in the ARD; however, this difference was not statistically significant. In conclusion, there appears to be no advantage to using the ARD system in conjunction with the three-bottle BACTEC blood-culture system
Nencki Affective Picture System: Cross-Cultural Study in Europe and Iran
Riegel, Monika; Moslehi, Abnoos; Michałowski, Jarosław M.; Żurawski, Łukasz; Horvat, Marko; Wypych, Marek; Jednoróg, Katarzyna; Marchewka, Artur
Although emotions have been assumed conventionally to be universal, recent studies have suggested that various aspects of emotions may be mediated by cultural background. The purpose of our research was to test these contradictory views, in the case of the subjective evaluation of visual affective stimuli. We also sought to validate the recently introduced Nencki Affective Picture System (NAPS) database on a different cultural group. Since there has been, to date, no attempt to compare the emotions of a culturally distinct sample of Iranians with those of Europeans, subjective ratings were collected from 40 Iranians and 39 Europeans. Each cultural group was asked separately to provide normative affective ratings and classify pictures according to discrete emotions. The results were analyzed to identify cultural differences in the ratings of individual images. One hundred and seventy NAPS pictures were rated with regard to the intensity of the basic emotions (happiness, sadness, fear, surprise, anger, and disgust) they elicited, as well as in terms of affective dimensions (valence and arousal). Contrary to previous studies using the International Affective Picture System, our results for Europeans and Iranians show that neither the ratings for affective dimensions nor for basic emotions differed across cultural groups. In both cultural groups, the relationship between valence and arousal ratings could be best described by a classical boomerang-shaped function. However, the content of the pictures (animals, faces, landscapes, objects, or people) had a significant effect on the ratings for valence and arousal. These findings indicate that further studies in cross-cultural affective research should control for the content of stimuli. PMID:28316576
Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
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Maria Mrakovcic
Full Text Available Nano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic cytotoxicity must therefore be evaluated. In order to perform chronic cytotoxicity testing of plain polystyrene nanoparticles on the endothelial cell line EAhy 926, we established a microcarrier cell culture system for anchorage-dependent cells (BioLevitator(TM. Cells were cultured for four weeks and exposed to doses, which were not cytotoxic upon 24 hours of exposure. For comparison, these particles were also studied in regularly sub-cultured cells, a method that has traditionally been used to assess chronic cellular effects. Culturing on basal membrane coated microcarriers produced very high cell densities. Fluorescent particles were mainly localized in the lysosomes of the exposed cells. After four weeks of exposure, the number of cells exposed to 20 nm polystyrene particles decreased by 60% as compared to untreated controls. When tested in sub-cultured cells, the same particles decreased cell numbers to 80% of the untreated controls. Dose-dependent decreases in cell numbers were also noted after exposure of microcarrier cultured cells to 50 nm short multi-walled carbon nanotubes. Our findings support that necrosis, but not apoptosis, contributed to cell death of the exposed cells in the microcarrier culture system. In conclusion, the established microcarrier model appears to be more sensitive for the identification of cellular effects upon prolonged and repeated exposure to nanoparticles than traditional sub-culturing.
Nencki Affective Picture System: Cross-Cultural Study in Europe and Iran.
Riegel, Monika; Moslehi, Abnoos; Michałowski, Jarosław M; Żurawski, Łukasz; Horvat, Marko; Wypych, Marek; Jednoróg, Katarzyna; Marchewka, Artur
Although emotions have been assumed conventionally to be universal, recent studies have suggested that various aspects of emotions may be mediated by cultural background. The purpose of our research was to test these contradictory views, in the case of the subjective evaluation of visual affective stimuli. We also sought to validate the recently introduced Nencki Affective Picture System (NAPS) database on a different cultural group. Since there has been, to date, no attempt to compare the emotions of a culturally distinct sample of Iranians with those of Europeans, subjective ratings were collected from 40 Iranians and 39 Europeans. Each cultural group was asked separately to provide normative affective ratings and classify pictures according to discrete emotions. The results were analyzed to identify cultural differences in the ratings of individual images. One hundred and seventy NAPS pictures were rated with regard to the intensity of the basic emotions (happiness, sadness, fear, surprise, anger, and disgust) they elicited, as well as in terms of affective dimensions (valence and arousal). Contrary to previous studies using the International Affective Picture System, our results for Europeans and Iranians show that neither the ratings for affective dimensions nor for basic emotions differed across cultural groups. In both cultural groups, the relationship between valence and arousal ratings could be best described by a classical boomerang-shaped function. However, the content of the pictures (animals, faces, landscapes, objects, or people) had a significant effect on the ratings for valence and arousal. These findings indicate that further studies in cross-cultural affective research should control for the content of stimuli.
Socio-cultural barriers to the development of a sustainable energy system - the case of hydrogen
DEFF Research Database (Denmark)
Petersen, Lars Kjerulf; Andersen, Anne Holst
Any transition to a more sustainable energy system, radically reducing greenhouse gas emissions, is bound to run in to a host of different barriers - technological and economic, but also socio-cultural. This will also be the case for any large-scale application of hydrogen as energy carrier......, especially if the system is going to be based on renewable energy sources. The aim of these research notes is to review and discuss major socio-cultural barriers to new forms of energy supply in general and to hydrogen specifically. Reaching sufficient reductions in greenhouse gas emissions may require more...
[Progress in microalgae culture system for biodiesel combined with reducing carbon dioxide emission].
Su, Hongyang; Zhou, Xuefei; Xia, Xuefen; Sun, Zhen; Zhang, Yalei
Wastewater resources, CO2 emission reduction and microalgae biodiesel are considered as current frontier fields of energy and environmental researches. In this paper, we reviewed the progress in system of microalgae culture for biodiesel production by wastewater and stack gas. Multiple factors including microalgal species, nutrition, culture methods and photobioreactor, which were crucial to the cultivation of microalgae for biodiesel production, were discussed in detail. A valuable culture system of microalgae for biodiesel production or other high value products combined with the treatment of wastewater by microalgae was put forward through the optimizations of algal species and culture technology. The culture system coupled with the treatment of wastewater, the reduction of CO2 emission with the cultivation of microalgae for biodiesel production will reduce the production cost of microalgal biofuel production and the treatment cost of wastewater simultaneously. Therefore, it would be a promising technology with important environmental value, social value and economic value to combine the treatment of wastewater with the cultivation of microalgae for biodiesel production.
The potential use of constructed wetlands in a recirculating aquaculture system for shrimp culture
International Nuclear Information System (INIS)
Lin, Y.-F.; Jing, S.-R.; Lee, D.-Y.
Constructed wetlands improved water qualities and consequently increased the shrimp growth and survival in a recirculating system. - A pilot-scale constructed wetland unit, consisting of free water surface (FWS) and subsurface flow (SF) constructed wetlands arranged in series, was integrated into an outdoor recirculating aquaculture system (RAS) for culturing Pacific white shrimp (Litopenaeus vannamei). This study evaluated the performance of the wetland unit in treating the recirculating wastewater and examined the effect of improvement in water quality of the culture tank on the growth and survival of shrimp postlarvae. During an 80-day culture period, the wetland unit operated at a mean hydraulic loading rate of 0.3 m/day and effectively reduced the influent concentrations of 5-day biochemical oxygen demand (BOD 5 , 24%), suspended solids (SS, 71%), chlorophyll a (chl-a, 88%), total ammonium (TAN, 57%), nitrite nitrogen (NO 2 -N, 90%) and nitrate nitrogen (NO 3 -N, 68%). Phosphate (PO 4 -P) reduction was the least efficient (5.4%). The concentrations of SS, Chl-a, turbidity and NO 3 -N in the culture tank water in RAS were significantly (P≤0.05) lower than those in a control aquaculture system (CAS) that simulated static pond culture without wetland treatment. However, no significant difference (P≤0.05) in BOD 5 , TAN and NO 2 -N was found between the two systems. At the end of the study, the harvest results showed that shrimp weight and survival rate in the RAS (3.8±1.8 g/shrimp and 90%) significantly (P≤0.01) exceeded those in the CAS (2.3±1.5 g/shrimp and 71%). This study concludes that constructed wetlands can improve the water quality and provide a good culture environment, consequently increasing the shrimp growth and survival without water exchange, in a recirculating system
The potential use of constructed wetlands in a recirculating aquaculture system for shrimp culture
Energy Technology Data Exchange (ETDEWEB)
Lin, Y.-F.; Jing, S.-R.; Lee, D.-Y
Constructed wetlands improved water qualities and consequently increased the shrimp growth and survival in a recirculating system. - A pilot-scale constructed wetland unit, consisting of free water surface (FWS) and subsurface flow (SF) constructed wetlands arranged in series, was integrated into an outdoor recirculating aquaculture system (RAS) for culturing Pacific white shrimp (Litopenaeus vannamei). This study evaluated the performance of the wetland unit in treating the recirculating wastewater and examined the effect of improvement in water quality of the culture tank on the growth and survival of shrimp postlarvae. During an 80-day culture period, the wetland unit operated at a mean hydraulic loading rate of 0.3 m/day and effectively reduced the influent concentrations of 5-day biochemical oxygen demand (BOD{sub 5}, 24%), suspended solids (SS, 71%), chlorophyll a (chl-a, 88%), total ammonium (TAN, 57%), nitrite nitrogen (NO{sub 2}-N, 90%) and nitrate nitrogen (NO{sub 3}-N, 68%). Phosphate (PO{sub 4}-P) reduction was the least efficient (5.4%). The concentrations of SS, Chl-a, turbidity and NO{sub 3}-N in the culture tank water in RAS were significantly (P{<=}0.05) lower than those in a control aquaculture system (CAS) that simulated static pond culture without wetland treatment. However, no significant difference (P{<=}0.05) in BOD{sub 5}, TAN and NO{sub 2}-N was found between the two systems. At the end of the study, the harvest results showed that shrimp weight and survival rate in the RAS (3.8{+-}1.8 g/shrimp and 90%) significantly (P{<=}0.01) exceeded those in the CAS (2.3{+-}1.5 g/shrimp and 71%). This study concludes that constructed wetlands can improve the water quality and provide a good culture environment, consequently increasing the shrimp growth and survival without water exchange, in a recirculating system.
The reduced serum free triiodothyronine and increased dorsal hippocampal SNAP-25 and Munc18-1 had existed in middle-aged CD-1 mice with mild spatial cognitive impairment.
Cao, Lei; Jiang, Wei; Wang, Fang; Yang, Qi-Gang; Wang, Chao; Chen, Yong-Ping; Chen, Gui-Hai
Changes of synaptic proteins in highlighted brain regions and decreased serum thyroid hormones (THs) have been implied in age-related learning and memory decline. Previously, we showed significant pairwise correlations among markedly impaired spatial learning and memory ability, decreased serum free triiodothyronine (FT3) and increased hippocampal SNAP-25 and Munc18-1 in old Kunming mice. However, whether these changes and the correlations occur in middle-age mice remains unclear. Since this age is one of the best stages to study age-related cognitive decline, we explored the spatial learning and memory ability, serum THs, cerebral SNAP-25 and Munc18-1 levels and their relationships of middle-aged mice in this study. The learning and memory abilities of 35 CD-1 mice (19 mice aged 6 months and 16 mice aged 12 months) were measured with a radial six-arm water maze (RAWM). The SNAP-25 and Munc18-1 levels were semi-quantified by Western blotting and the serum THs were detected by radioimmunoassay. The results showed the middle-aged mice had decreased serum FT3, increased dorsal hippocampal (DH) SNAP-25 and Munc18-1, and many or long number of errors and latency in both learning and memory phases of the RAWM. The Pearson's correlation test showed that the DH SANP-25 and Munc18-1 levels were positively correlated with the number of errors and latency in learning phases of the RAWM. Meanwhile, the DH SANP-25 and Munc18-1 levels negatively correlated with the serum FT3 level. These results suggested that reduced FT3 with increased DH SNAP-25 and Munc18-1 levels might be involved in the spatial learning ability decline in the middle-aged mice. © 2013 Elsevier B.V. All rights reserved.
Nonlinear impulsive system of fed-batch culture in fermentative production and its properties
International Nuclear Information System (INIS)
Gao Caixia; Li Kezan; Feng Enmin; Xiu Zhilong
In this study, the nonlinear dynamical system of fed-batch fermentation is investigated in the process of bio-dissimilation of glycerol to 1,3-propanediol by Klebsiella pneumoniae. Considering the abrupt increase of glycerol in fed-batch culture, this paper proposes a nonlinear impulsive system of the culture process, which is fit for formulating the factual fermentation better than the continuous models in being. We study the questions of existence and properties of mild solutions for the system and the continuous dependence of solutions on initial values and the controllable variable. Finally, the numerical simulations show that the errors between experimental and computational values using the impulsive system are less than those using the previous continuous system
A self-contained, programmable microfluidic cell culture system with real-time microscopy access
DEFF Research Database (Denmark)
Skafte-Pedersen, Peder; Hemmingsen, Mette; Sabourin, David
Utilizing microfluidics is a promising way for increasing the throughput and automation of cell biology research. We present a complete self-contained system for automated cell culture and experiments with real-time optical read-out. The system offers a high degree of user-friendliness, stability...... enables the system to perform parallel, programmable and multiconditional assays on a single chip. A modular approach provides system versatility and allows many different chips to be used dependent upon application. We validate the system's performance by demonstrating on-chip passive switching...... and mixing by peristaltically driven flows. Applicability for biological assays is demonstrated by on-chip cell culture including on-chip transfection and temporally programmable gene expression....
Cutting the gordian knot-development and biological relevance of hepatitis C virus cell culture systems
DEFF Research Database (Denmark)
Gottwein, Judith Margarete; Bukh, Jens
described. Research on the viral life cycle, efficient therapeutics, and a vaccine has been hampered by the absence of suitable cell culture systems. The first system permitting studies of the full viral life cycle was intrahepatic transfection of RNA transcripts of HCV consensus complementary DNA (c...... studies of the function of viral proteins, their interaction with each other and host proteins, new antivirals, and neutralizing antibodies in the context of the full viral life cycle. However, several challenges remain, including development of cell culture systems for all major HCV genotypes...... isolate JFH1, which for unknown reasons showed an exceptional replication capability and resulted in formation of infectious viral particles in the human hepatoma cell line Huh7, led in 2005 to the development of the first full viral life cycle in vitro systems. JFH1-based systems now enable in vitro...
Animal cell cultures: recent achievements and perspectives in the production of biopharmaceuticals.
Butler, Michael
There has been a rapid increase in the number and demand for approved biopharmaceuticals produced from animal cell culture processes over the last few years. In part, this has been due to the efficacy of several humanized monoclonal antibodies that are required at large doses for therapeutic use. There have also been several identifiable advances in animal cell technology that has enabled efficient biomanufacture of these products. Gene vector systems allow high specific protein expression and some minimize the undesirable process of gene silencing that may occur in prolonged culture. Characterization of cellular metabolism and physiology has enabled the design of fed-batch and perfusion bioreactor processes that has allowed a significant improvement in product yield, some of which are now approaching 5 g/L. Many of these processes are now being designed in serum-free and animal-component-free media to ensure that products are not contaminated with the adventitious agents found in bovine serum. There are several areas that can be identified that could lead to further improvement in cell culture systems. This includes the down-regulation of apoptosis to enable prolonged cell survival under potentially adverse conditions. The characterization of the critical parameters of glycosylation should enable process control to reduce the heterogeneity of glycoforms so that production processes are consistent. Further improvement may also be made by the identification of glycoforms with enhanced biological activity to enhance clinical efficacy. The ability to produce the ever-increasing number of biopharmaceuticals by animal cell culture is dependent on sufficient bioreactor capacity in the industry. A recent shortfall in available worldwide culture capacity has encouraged commercial activity in contract manufacturing operations. However, some analysts indicate that this still may not be enough and that future manufacturing demand may exceed production capacity as the number
Numerical calculations for diffusion effects in the well-of-the-well culture system for mammalian embryos.
Matsuura, Koji
Recent studies suggest that the microenvironment and embryo density used during embryo culture considerably affect development to the blastocyst stage. High embryo density allows for autocrine secretions to diffuse to neighbouring embryos during group culture, with a positive effect on further development. A variation of group culture is the well-of-the-well (WOW) culture system, allowing for individual identification of embryos cultured in small holes in a microdroplet. Bovine blastocyst development is higher in the WOW culture system than in conventional group culture. To compare the concentration of chemical factors between conventional and WOW culture, a model was constructed to calculate the concentration of secreted factors based on Fick's second law of diffusion using spreadsheet software. Furthermore, model was used to determine the concentration of growth factors and waste materials adjacent to the embryo periphery. The results of these calculations suggest that the highest difference in the concentration of secreted small molecules and macromolecules was at the most two- to threefold, with the concentrations reduced more and diffusion kinetics facilitated to a greater extent in the WOW culture system. The average ratio of the concentration of secreted macromolecules (10nm diameter) around the embryos was also compared between systems with well widths of 0.1 and 0.3mm. The concentration of secreted materials surrounding embryos increased in a narrow tapered well. The findings suggest that the WOW culture system is better than conventional group culture because of the increased final concentration of autocrine factors and higher diffusion kinetics of waste materials.
Three-dimensional hydrogel cell culture systems for modeling neural tissue
Frampton, John
Two-dimensional (2-D) neural cell culture systems have served as physiological models for understanding the cellular and molecular events that underlie responses to physical and chemical stimuli, control sensory and motor function, and lead to the development of neurological diseases. However, the development of three-dimensional (3-D) cell culture systems will be essential for the advancement of experimental research in a variety of fields including tissue engineering, chemical transport and delivery, cell growth, and cell-cell communication. In 3-D cell culture, cells are provided with an environment similar to tissue, in which they are surrounded on all sides by other cells, structural molecules and adhesion ligands. Cells grown in 3-D culture systems display morphologies and functions more similar to those observed in vivo, and can be cultured in such a way as to recapitulate the structural organization and biological properties of tissue. This thesis describes a hydrogel-based culture system, capable of supporting the growth and function of several neural cell types in 3-D. Alginate hydrogels were characterized in terms of their biomechanical and biochemical properties and were functionalized by covalent attachment of whole proteins and peptide epitopes. Methods were developed for rapid cross-linking of alginate hydrogels, thus permitting the incorporation of cells into 3-D scaffolds without adversely affecting cell viability or function. A variety of neural cell types were tested including astrocytes, microglia, and neurons. Cells remained viable and functional for longer than two weeks in culture and displayed process outgrowth in 3-D. Cell constructs were created that varied in cell density, type and organization, providing experimental flexibility for studying cell interactions and behavior. In one set of experiments, 3-D glial-endothelial cell co-cultures were used to model blood-brain barrier (BBB) structure and function. This co-culture system was
Three-dimensional Huh7 cell culture system for the study of Hepatitis C virus infection
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Uprichard Susan L
Full Text Available Abstract Background In order to elucidate how Hepatitis C Virus (HCV interacts with polarized hepatocytes in vivo and how HCV-induced alterations in cellular function contribute to HCV-associated liver disease, a more physiologically relevant hepatocyte culture model is needed. As such, NASA-engineered three-dimensional (3-D rotating wall vessel (RWV bioreactors were used in effort to promote differentiation of HCV-permissive Huh7 hepatoma cells. Results When cultured in the RWV, Huh7 cells became morphologically and transcriptionally distinct from more standard Huh7 two-dimensional (2-D monolayers. Specifically, RWV-cultured Huh7 cells formed complex, multilayered 3-D aggregates in which Phase I and Phase II xenobiotic drug metabolism genes, as well as hepatocyte-specific transcripts (HNF4α, Albumin, TTR and α1AT, were upregulated compared to 2-D cultured Huh7 cells. Immunofluorescence analysis revealed that these HCV-permissive 3-D cultured Huh7 cells were more polarized than their 2D counterparts with the expression of HCV receptors, cell adhesion and tight junction markers (CD81, scavenger receptor class B member 1, claudin-1, occludin, ZO-1, β-Catenin and E-Cadherin significantly increased and exhibiting apical, lateral and/or basolateral localization. Conclusion These findings show that when cultured in 3-D, Huh7 cells acquire a more differentiated hepatocyte-like phenotype. Importantly, we show that these 3D cultures are highly permissive for HCV infection, thus providing an opportunity to study HCV entry and the effects of HCV infection on host cell function in a more physiologically relevant cell culture system.
Neuroprotective effect of the endogenous neural peptide apelin in cultured mouse cortical neurons
International Nuclear Information System (INIS)
Zeng, Xiang Jun; Yu, Shan Ping; Zhang, Like; Wei, Ling
The adipocytokine apelin and its G protein-coupled APJ receptor were initially isolated from a bovine stomach and have been detected in the brain and cardiovascular system. Recent studies suggest that apelin can protect cardiomyocytes from ischemic injury. Here, we investigated the effect of apelin on apoptosis in mouse primary cultures of cortical neurons. Exposure of the cortical cultures to a serum-free medium for 24 h induced nuclear fragmentation and apoptotic death; apelin-13 (1.0-5.0 nM) markedly prevented the neuronal apoptosis. Apelin neuroprotective effects were mediated by multiple mechanisms. Apelin-13 reduced serum deprivation (SD)-induced ROS generation, mitochondria depolarization, cytochrome c release and activation of caspase-3. Apelin-13 prevented SD-induced changes in phosphorylation status of Akt and ERK1/2. In addition, apelin-13 attenuated NMDA-induced intracellular Ca 2+ accumulation. These results indicate that apelin is an endogenous neuroprotective adipocytokine that may block apoptosis and excitotoxic death via cellular and molecular mechanisms. It is suggested that apelins may be further explored as a potential neuroprotective reagent for ischemia-induced brain damage.
Toward an integrated system concept for monitoring and evaluation of safety culture
International Nuclear Information System (INIS)
Makino, Maomi; Sakaue, Takeharu
The concept of ''nuclear safety culture'' has been advocated and has been much discussed internationally by INSAG (The International Nuclear Safety Advisory Group) under IAEA (the International Atomic Energy Agency) and other institutions since Chernobyl accident. On the safety front, Japan had maintained an excellent track record in nuclear power operations throughout the 1990s. However, there have been a series of new type of problems strongly implying degradation of safety culture, e.g., Monju accident, fire and explosion accident at an Asphalt Solidification Process Facility at Tokai, falsification of annealing data at nuclear power plants (NPP), another data falsification for transport cask of spent fuel and JCO criticality accident. Then the TEPCO (Tokyo Electric Power Company) issue was revealed in 2002. Triggered by this issue, the Nuclear and Industrial Safety Agency (NISA) has been implementing a variety of improvements, one of which was the establishment of a study group in 2003, which invited experts from other fields as well as from nuclear-related industries, to study on how to implement safety culture sufficiently and possible recommendations. Subjects such as the followings piled in the study report will indicate leading keys in case it is going to realize such efforts: ''Foundation of safety culture is a quality management'' and ''Realistic and scientific technique is necessary for the evaluation of safety culture''. In order to respond to these requests, JNES have been advancing the development toward an Integrated System Concept for Monitoring and Evaluation of Safety Culture. This paper describes the outline of the study results reported by the study group and then introduces one of subsystems, SCEST, structuring the integrated system concept for Monitoring and Evaluation of Safety Culture. (author)
Optimization of the Static Human Osteoblast/Osteoclast Co-culture System
Directory of Open Access Journals (Sweden)
James Jam Jolly
Full Text Available Osteoblasts (OBs and osteoclasts (OCs are 2 major groups of bone cells. Their cell-to-cell interactions are important to ensure the continuity of the bone-remodeling process. Therefore, the present study was carried out to optimize an OB/OC co-culture system utilizing the human OB cell line hFOB 1.19 and OCs extracted from peripheral blood mononuclear cells (PBMNCs. It was a 2-step procedure, involving the optimization of the OB culture and the co-culture of the OBs with PBMNCs at an optimum ratio. Firstly, pre-OBs were cultured to 90% confluency and the time required for differentiation was determined. OB differentiation was determined using the van Gieson staining to detect the presence of collagen and Alizarin Red for calcium. Secondly, OBs and OCs were co-cultured at the ratios of 1 OC: 1 OB, 1 OC: 4 OBs, 2 OCs: 1 OB, and 1 OC: 2 OBs. Tartrate-resistant acid phosphatase (TRAP staining was used to detect the differentiation of the OCs. The results showed that collagen was present on day 1, whereas calcium was detected as early as day 3. Based on the result of TRAP staining, 1 OC: 2 OBs was taken as the most appropriate ratio. No macrophage colony-stimulating factor and receptor activator of the nuclear factor-κB ligand were added because they were provided by the OBs. In conclusion, these optimization processes are vital as they ensure the exact time point and ratio of the OB/OC co-culture in order to produce a reliable and reproducible co-culture system.
Co-culture systems and technologies: taking synthetic biology to the next level.
Goers, Lisa; Freemont, Paul; Polizzi, Karen M
Co-culture techniques find myriad applications in biology for studying natural or synthetic interactions between cell populations. Such techniques are of great importance in synthetic biology, as multi-species cell consortia and other natural or synthetic ecology systems are widely seen to hold enormous potential for foundational research as well as novel industrial, medical and environmental applications with many proof-of-principle studies in recent years. What is needed for co-cultures to fulfil their potential? Cell-cell interactions in co-cultures are strongly influenced by the extracellular environment, which is determined by the experimental set-up, which therefore needs to be given careful consideration. An overview of existing experimental and theoretical co-culture set-ups in synthetic biology and adjacent fields is given here, and challenges and opportunities involved in such experiments are discussed. Greater focus on foundational technology developments for co-cultures is needed for many synthetic biology systems to realize their potential in both applications and answering biological questions. © 2014 The Author(s) Published by the Royal Society. All rights reserved.
A dense cell retention culture system using stirred ceramic membrane reactor.
Suzuki, T; Sato, T; Kominami, M
A novel reactor design incorporating porous ceramic tubes into a stirred jar fermentor was developed. The stirred ceramic membrane reactor has two ceramic tubular membrane units inside the vessel and maintains high filtration flux by alternating use for filtering and recovering from clogging. Each filter unit was linked for both extraction of culture broth and gas sparging. High permeability was maintained for long periods by applying the periodical control between filtering and air sparging during the stirred retention culture of Saccharomyces cerevisiae. The ceramic filter aeration system increased the k(L)a to about five times that of ordinary gas sparing. Using the automatic feeding and filtering system, cell mass concentration reached 207 g/L in a short time, while it was 64 g/L in a fed-batch culture. More than 99% of the growing cells were retained in the fermentor by the filtering culture. Both yield and productivity of cells were also increased by controlling the feeding of fresh medium and filtering the supernatant of the dense cells culture. (c) 1994 John Wiley & Sons, Inc.
Raising Cultural Self-Efficacy among Faculty and Staff of a Private Native Hawaiian School System
Fong, Randie Kamuela
The Hawaiian cultural revitalization movement in Hawai`i is an important driver for many Hawaiian organizations as well as educational institutions that serve Native Hawaiians. One such organization is Kamehameha Schools, a private school system founded and endowed by Princess Bernice Pauahi Bishop in 1887 to educate Native Hawaiian children. From…
Improving Hospital Quality and Patient Safety an Examination of Organizational Culture and Information Systems
Gardner, John Wallace
This dissertation examines the effects of safety culture, including operational climate and practices, as well as the adoption and use of information systems for delivering high quality healthcare and improved patient experience. Chapter 2 studies the influence of both general and outcome-specific hospital climate and quality practices on process…
National Contexts Influencing Principals' Time Use and Allocation: Economic Development, Societal Culture, and Educational System
Lee, Moosung; Hallinger, Philip
This study examines the impact of macro-context factors on the behavior of school principals. More specifically, the article illuminates how a nation's level of economic development, societal culture, and educational system influence the amount of time principals devote to their job role and shape their allocation of time to instructional…
Examining the Relationship between Safety Management System Implementation and Safety Culture in Collegiate Flight Schools
Robertson, Mike Fuller
Safety Management Systems (SMS) are becoming the industry standard for safety management throughout the aviation industry. As the Federal Aviation Administration (FAA) continues to mandate SMS for different segments, the assessment of an organization's safety culture becomes more important. An SMS can facilitate the development of a strong…
Ethno-cultural Component Implementation in the System of Higher Education of Kazakhstan
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Gulnaz K. Tleuzhanova
Full Text Available The article considers the ethno-cultural aspect of the educational process of the higher school on the example of one of the leading institutes of higher education of Kazakhstan. Priorities and tendencies of language education and language policy of Kazakhstan, opportunities of its realization through the prism of the system of higher education are presented
Planning Cultures and Histories: Influences on the Evolution of Planning Systems and Spatial Development Patterns
Stead, D.; de Vries, J.; Tasan-Kok, T.
This special issue addresses the influences of planning cultures and histories on the evolution of planning systems and spatial development. As well as providing an international comparative perspective on these issues, the collection of articles also engages in a search for new conceptual
In vitro culture systems and acclimatization of Aechmea setigera Mart. ex Schult. & Schult. f. (Bromeliaceae
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Janaína Medeiros Vasconcelos
Full Text Available Aechmea setigera is an endemic bromeliad from Amazon with ornamental potential. Bromeliads have been propagated by tissue culture. The consistency of the culture medium in vitro multiplication influences the rate of propagation. In this sense, the objective of this study was to evaluate different culture systems with the use of 6-benzylaminopurine (BAP on in vitro propagation and the effect of different substrates in acclimatization of plantlets Aechmea setigera. In vitro germinated seedlings were inoculated in MS medium in liquid stationary, semisolid, double-phase systems, plus 6-benzylaminopurine (BAP in different concentrations (0, 2.2, 4.4, 8.8 and 17.7 μM. The ex vitro rooting and acclimatization were performed on substrate Plantmax Forest ®, vermiculite and sawdust eucalyptus. After three successive subcultures, the double-phase system showed a higher number of regenerated shoots in comparison to other systems. Acclimatization using the combination of commercial substrate Plantmax Forest ® and vermiculite favored the growth of micropropagated plants. The use of a culture medium double-phase without growth regulator, and the rooting in acclimatization are feasible strategy for the micropropagation of A. setigera. Indexação
Policies on and Practices of Cultural Inclusivity in Learning Management Systems: Perspectives of Indigenous Holistic Pedagogies
Dreamson, Neal; Thomas, Gary; Lee Hong, Anita; Kim, Soyoung
Online learning has become a conventional term and practice in Australian higher education, yet cultural inclusivity for Indigenous (Indigenous for the purposes of this paper refers to Australian Aboriginal and Torres Strait Islander peoples) students is insufficiently reflected in learning management system (LMS) policies and design. This study…
Development of Quality Assurance System in Culture and Nation Character Education in Primary Education in Indonesia
Susilana, Rudi; Asra
The purpose of national education is to develop skills and build dignified national character and civilization in educating nation life (Act No. 20, 2003). The paper describes a system of quality assurance in culture and character education in primary education. This study employs the six sigma model which consists of the formula DMAIC (Define,…
Improving Achievement for Linguistically and Culturally Diverse Learners through an Inquiry-Based Earth Systems Curriculum
Lambert, Julie; Ariza, Eileen N. Whelan
This report describes an inquiry-based Earth systems curriculum and strategies for teaching diverse students, which were embedded in the curriculum. The curriculum was implemented with 5th-grade students with varied linguistic, cultural, and socioeconomic backgrounds in five schools in a large, southeastern U.S., urban school district. At the end…
Effects of carbohydrate addition on production in extensive shrimp culture systems
Hari, B.; Kurup, B.M.; Varghese, J.T.; Schrama, J.W.; Verdegem, M.C.J.
One indoor and one on-farm trial were conducted to evaluate the effect of control of carbon/ nitrogen ratio (C/N ratio) by addition of carbohydrate to the water column in extensive types of shrimp culture systems. In the indoor experiment, 25% and 40% dietary protein ('P25' and 'P40') with or
Culture in embryonic kidney serum and xeno-free media as renal cell carcinoma and renal cell carcinoma cancer stem cells research model.
Krawczyk, Krzysztof M; Matak, Damian; Szymanski, Lukasz; Szczylik, Cezary; Porta, Camillo; Czarnecka, Anna M
The use of fetal bovine serum hinders obtaining reproducible experimental results and should also be removed in hormone and growth factor studies. In particular hormones found in FBS act globally on cancer cell physiology and influence transcriptome and metabolome. The aim of our study was to develop a renal carcinoma serum free culture model optimized for (embryonal) renal cells in order to select the best study model for downstream auto-, para- or endocrine research. Secondary aim was to verify renal carcinoma stem cell culture for this application. In the study, we have cultured renal cell carcinoma primary tumour cell line (786-0) as well as human kidney cancer stem cells in standard 2D monolayer cultures in Roswell Park Memorial Institute Medium or Dulbecco's Modified Eagle's Medium and Complete Human Kidney Cancer Stem Cell Medium, respectively. Serum-free, animal-component free Human Embryonic Kidney 293 media were tested. Our results revealed that xeno-free embryonal renal cells optimized culture media provide a useful tool in RCC cancer biology research and at the same time enable effective growth of RCC. We propose bio-mimic RCC cell culture model with specific serum-free and xeno-free medium that promote RCC cell viability.
An aeroponic culture system for the study of root herbivory on Arabidopsis thaliana
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Vaughan Martha M
Full Text Available Abstract Background Plant defense against herbivory has been studied primarily in aerial tissues. However, complex defense mechanisms have evolved in all parts of the plant to combat herbivore attack and these mechanisms are likely to differ in the aerial and subterranean environment. Research investigating defense responses belowground has been hindered by experimental difficulties associated with the accessibility and quality of root tissue and the lack of bioassays using model plants with altered defense profiles. Results We have developed an aeroponic culture system based on a calcined clay substrate that allows insect herbivores to feed on plant roots while providing easy recovery of the root tissue. The culture method was validated by a root-herbivore system developed for Arabidopsis thaliana and the herbivore Bradysia spp. (fungus gnat. Arabidopsis root mass obtained from aeroponically grown plants was comparable to that from other culture systems, and the plants were morphologically normal. Bradysia larvae caused considerable root damage resulting in reduced root biomass and water absorption. After feeding on the aeroponically grown root tissue, the larvae pupated and emerged as adults. Root damage of mature plants cultivated in aeroponic substrate was compared to that of Arabidopsis seedlings grown in potting mix. Seedlings were notably more susceptible to Bradysia feeding than mature plants and showed decreased overall growth and survival rates. Conclusions A root-herbivore system consisting of Arabidopsis thaliana and larvae of the opportunistic herbivore Bradysia spp. has been established that mimics herbivory in the rhizosphere. Bradysia infestation of Arabidopsis grown in this culture system significantly affects plant performance. The culture method will allow simple profiling and in vivo functional analysis of root defenses such as chemical defense metabolites that are released in response to belowground insect attack.
The culture of Chlorella vulgaris with human urine in multibiological life support system experiments
Li, Ming; Liu, Hong; Tong, Ling; Fu, Yuming; He, Wenting; Hu, Enzhu; Hu, Dawei
The Integrative Experimental System (IES) was established as a tool to evaluate the rela-tionship of the subsystems in Bioregenerative Life Support System, and Multibiological Life Support System Experiments (MLSSE) have been conducted in the IES. The IES consists of a higher plant chamber, an animal chamber and a plate photo bioreactor (PPB) which cultivated lettuce (Lactuca sativa L.), silkworm (Bombyx Mori L.) and microalgae (Chlorella vulgaris), respectively. In MLSSE, four volunteers took turns breathing the system air through a tube connected with the animal chamber periodically. According to the CO2 concentration in the IES, the automotive control system of the PPB changed the light intensity regulating the photosynthesis of Chlorella vulgaris to make CO2 /O2 in the system maintain at stable levels. Chlorella vulgaris grew with human urine by carrying certain amount of alga liquid out of the bioreactor every day with synthetic urine replenished into the system, and O2 was regenerated, at the same time human urine was purified. Results showed that this IES worked stably and Chlorella vulgaris grew well; The culture of Chlorella vulgaris could be used to keep the balance of CO2 and O2 , and the change of light intensity could control the gas composition in the IES; Microalgae culture could be used in emergency in the system, the culture of Chlorella vulgaris could recover to original state in 5 days; 15.6 ml of condensation water was obtained every day by the culture of Chlorella vulgaris; The removal efficiencies of N, P in human urine could reach to 98.2% and 99.5%.
The Effect of Primary Cancer Cell Culture Models on the Results of Drug Chemosensitivity Assays: The Application of Perfusion Microbioreactor System as Cell Culture Vessel
Chen, Yi-Dao; Huang, Shiang-Fu; Wang, Hung-Ming
To precisely and faithfully perform cell-based drug chemosensitivity assays, a well-defined and biologically relevant culture condition is required. For the former, a perfusion microbioreactor system capable of providing a stable culture condition was adopted. For the latter, however, little is known about the impact of culture models on the physiology and chemosensitivity assay results of primary oral cavity cancer cells. To address the issues, experiments were performed. Results showed that minor environmental pH change could significantly affect the metabolic activity of cells, demonstrating the importance of stable culture condition for such assays. Moreover, the culture models could also significantly influence the metabolic activity and proliferation of cells. Furthermore, the choice of culture models might lead to different outcomes of chemosensitivity assays. Compared with the similar test based on tumor-level assays, the spheroid model could overestimate the drug resistance of cells to cisplatin, whereas the 2D and 3D culture models might overestimate the chemosensitivity of cells to such anticancer drug. In this study, the 3D culture models with same cell density as that in tumor samples showed comparable chemosensitivity assay results as the tumor-level assays. Overall, this study has provided some fundamental information for establishing a precise and faithful drug chemosensitivity assay. PMID:25654105
Enhanced Geothermal System Development of the AmeriCulture Leasehold in the Animas Valley; FINAL
International Nuclear Information System (INIS)
Duchane, David V; Seawright, Gary L; Sewright, Damon E; Brown, Don; Witcher, James c.; Nichols, Kenneth E.
Working under the grant with AmeriCulture, Inc., and its team of geothermal experts, assembled a plan to apply enhanced geothermal systems (EGS) techniques to increase both the temperature and flow rate of the geothermal waters on its leasehold. AmeriCulture operates a commercial aquaculture facility that will benefit from the larger quantities of thermal energy and low cost electric power that EGS technology can provide. The project brought together a team of specialists that, as a group, provided the full range of expertise required to successfully develop and implement the project
Effect of national cultural values on safety climate, and safety management system
International Nuclear Information System (INIS)
Ali, T.H.; Memon, N.A.
This paper investigates the critical role played by the national culture in influencing how workers safely or otherwise behave (mainly in risky situations) on construction sites, and how site managers implement safety management processes and practices. The paper presents the findings of an empirical research study based on a questionnaire survey, administered in Pakistan, targeting construction site managers and workers to gauge the effect national culture has on managers preferences for and perceptions of safety management systems (policies and practices) and than linking this effect to predict workers attitudes and intentional behaviors. (author)
Spatially monitoring oxygen level in 3D microfabricated cell culture systems using optical oxygen sensing beads
Wang, Lin; Acosta, Miguel A.; Leach, Jennie B.; Carrier, Rebecca L.
Capability of measuring and monitoring local oxygen concentration at the single cell level (tens of microns scale) is often desirable but difficult to achieve in cell culture. In this study, biocompatible oxygen sensing beads were prepared and tested for their potential for real-time monitoring and mapping of local oxygen concentration in 3D micro-patterned cell culture systems. Each oxygen sensing bead is composed of a silica core loaded with both an oxygen sensitive Ru(Ph2phen3)Cl2 dye and ...
Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy.
Kim, Jinho; Henley, Beverley M; Kim, Charlene H; Lester, Henry A; Yang, Changhuei
Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period.
An All-Recombinant Protein-Based Culture System Specifically Identifies Hematopoietic Stem Cell Maintenance Factors
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Aki Ieyasu
Full Text Available Hematopoietic stem cells (HSCs are considered one of the most promising therapeutic targets for the treatment of various blood disorders. However, due to difficulties in establishing stable maintenance and expansion of HSCs in vitro, their insufficient supply is a major constraint to transplantation studies. To solve these problems we have developed a fully defined, all-recombinant protein-based culture system. Through this system, we have identified hemopexin (HPX and interleukin-1α as responsible for HSC maintenance in vitro. Subsequent molecular analysis revealed that HPX reduces intracellular reactive oxygen species levels within cultured HSCs. Furthermore, bone marrow immunostaining and 3D immunohistochemistry revealed that HPX is expressed in non-myelinating Schwann cells, known HSC niche constituents. These results highlight the utility of this fully defined all-recombinant protein-based culture system for reproducible in vitro HSC culture and its potential to contribute to the identification of factors responsible for in vitro maintenance, expansion, and differentiation of stem cell populations.
A continuous culture system of direct somatic embryogenesis in microspore-derived embryos of Brassica juncea.
Prabhudesai, V; Bhaskaran, S
An efficient culture system has been developed for repeated cycles of somatic embryogenesis in microspore-derived embryos of Brassica juncea without a callus phase. Haploid embryos produced through anther culture showed a high propensity for direct production of somatic embryos in response to 2 mgL(-1) BA and 0.1 mgL(-1) NAA. The embryogenic cultures which comprised the elongated embryonal axis of microspore-derived embryos when explanted and grown on the medium of same composition produced a large number of secondary embryos. These somatic embryos in turn underwent axis elongation and produced more somatic embryos when explanted and cultured. This cycle of repetitive somatic embryogenesis continued with undiminished vigour passage after passage and was monitored for more than a year. Somatic embryos from any passage when isolated at cotyledonary stage and grown on auxin-free medium for 5 days and then on a medium containing NAA (0.1 mgL(-1)), developed into complete plants with a profuse root system and were easily established in the soil. The cytology of the root tips of these plants confirmed their haploid nature. The total absence of callus phase makes the system ideal for continuous cloning of androgenic lines, Agrobacterium-mediated transformation and mutation induction studies.
Systemic Analysis of the Cultural Production of a Virtual Learning Community
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Germán Alejandro Miranda Díaz
Full Text Available This paper describes a systemic analysis from the standpoint of activity theory of the cultural emergence of a virtual learning community as a complex system. Three levels of analysis were employed: data mining, visualization of complex systems and analysis of discursive interactions, with the aim of understanding the emerging phenomena of online learning and in order to have the necessary elements to assist in planning the formation of virtual communities in formal settings. This was carried out using six years of activity from 3,324 people from different documentary sources: 9,871,531 CMS records; 1,371,907 from LMS; 67,828 IRC statements; and 27,798 online forum comments. In the process, we observed how action aimed at socialization and discussion of its object evolve into historical-cultural milestones such as the culture of merit as opposed to certification, the division of labor and the process of transition from free software to free culture.
Assessment of patient safety culture in clinical laboratories in the Spanish National Health System.
Giménez-Marín, Angeles; Rivas-Ruiz, Francisco; García-Raja, Ana M; Venta-Obaya, Rafael; Fusté-Ventosa, Margarita; Caballé-Martín, Inmaculada; Benítez-Estevez, Alfonso; Quinteiro-García, Ana I; Bedini, José Luis; León-Justel, Antonio; Torra-Puig, Montserrat
There is increasing awareness of the importance of transforming organisational culture in order to raise safety standards. This paper describes the results obtained from an evaluation of patient safety culture in a sample of clinical laboratories in public hospitals in the Spanish National Health System. A descriptive cross-sectional study was conducted among health workers employed in the clinical laboratories of 27 public hospitals in 2012. The participants were recruited by the heads of service at each of the participating centers. Stratified analyses were performed to assess the mean score, standardized to a base of 100, of the six survey factors, together with the overall patient safety score. 740 completed questionnaires were received (88% of the 840 issued). The highest standardized scores were obtained in Area 1 (individual, social and cultural) with a mean value of 77 (95%CI: 76-78), and the lowest ones, in Area 3 (equipment and resources), with a mean value of 58 (95%CI: 57-59). In all areas, a greater perception of patient safety was reported by the heads of service than by other staff. We present the first multicentre study to evaluate the culture of clinical safety in public hospital laboratories in Spain. The results obtained evidence a culture in which high regard is paid to safety, probably due to the pattern of continuous quality improvement. Nevertheless, much remains to be done, as reflected by the weaknesses detected, which identify areas and strategies for improvement.
Global Incorporation and Cultural Survival: The Surinamese Maroons at the Margins of the World-System
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Aonghas St.-Hilaire
Full Text Available The Maroons of Suriname enter the twenty-? rst century as the only surviving, culturally and politically autonomous Maroon communities in the Americas. The paper adopts a world-systems approach to explain the evolution of the Surinamese Maroon nations vis-à-vis the ever expanding Surinamese state and global political economy, with particular attention paid to the cultural survival of the Maroon nations. Prior to emancipation, limited colonial integration and mutual fear between the Maroons and Surinamese coastal society facilitated the development of the Maroon communities as relatively independent nations. Despite the best efforts of colonial authorities after emancipation, the weak economy and infrastructure of the Surinamese colony as well as continued mutual fear and distrust enabled the Maroons to guard their autonomy. However, after the Second World War, a period of global economic expansion, the rapid development of the Suriname as an integrated political, economic and cultural unit, and the depletion of and strain on natural resources in traditional Maroon territory dealt serious blows to Maroon autonomy. Maroon cultural survival depends on the ability of the Maroon nations to navigate the rapid changes currently affecting Maroon society, guarding political autonomy and cultivating the most treasured aspects of their cultural heritage, while participating, albeit peripherally, in modern global capitalism.
A Refined Culture System for Human Induced Pluripotent Stem Cell-Derived Intestinal Epithelial Organoids
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Yu Takahashi
Full Text Available Gut epithelial organoids are routinely used to investigate intestinal biology; however, current culture methods are not amenable to genetic manipulation, and it is difficult to generate sufficient numbers for high-throughput studies. Here, we present an improved culture system of human induced pluripotent stem cell (iPSC-derived intestinal organoids involving four methodological advances. (1 We adopted a lentiviral vector to readily establish and optimize conditioned medium for human intestinal organoid culture. (2 We obtained intestinal organoids from human iPSCs more efficiently by supplementing WNT3A and fibroblast growth factor 2 to induce differentiation into definitive endoderm. (3 Using 2D culture, followed by re-establishment of organoids, we achieved an efficient transduction of exogenous genes in organoids. (4 We investigated suspension organoid culture without scaffolds for easier harvesting and assays. These techniques enable us to develop, maintain, and expand intestinal organoids readily and quickly at low cost, facilitating high-throughput screening of pathogenic factors and candidate treatments for gastrointestinal diseases.
Supply Chain Collaboration Roles of Interorganizational Systems, Trust, and Collaborative Culture
Cao, Mei
To survive and thrive in the competition, firms have strived to achieve greater supply chain collaboration to leverage the resources and knowledge of suppliers and customers. Internet based technologies, particularly interorganizational systems, further extend the firms' opportunities to strengthen their supply chain partnerships and share real-time information to optimize their operations. Supply Chain Collaboration: Roles of Interorganizational Systems, Trust, and Collaborative Culture explores the nature and characteristics, antecedents, and consequences of supply chain collaboration from multiple theoretical perspectives. Supply Chain Collaboration: Roles of Interorganizational Systems, Trust, and Collaborative Culture conceptualizes supply chain collaboration as seven interconnecting elements including information sharing, incentive alignment, goal congruence, decision synchronization, resource sharing, as well as communication and joint knowledge creation. These seven components define the occur...
Photo irradiation Systems for In-Vitro Cultured Cells Phototherapy and Photobiology Experiments
International Nuclear Information System (INIS)
Serrano Navarro, Joel; Morales Lopez, Orestes M.; Hernandez Quintanas, Luis F.; Lopez Silva, Y.; Fabila Bustos, Diego A.; De la Rosa Vazquez, Jose M.; Valor Reed, Alma; Stolik Isakina, Suren; Brodin, Patrik N.; Guha, Chandan; Tome, Wolfgang A.
The increase in research and application of various phototherapy methods, especially photodynamic therapy (PDT) has created the need to study in depth the mechanisms of interaction of light with biological tissue using a photosensitizing drug in order to increase the therapeutic effectiveness. In this issue, two systems for controlled irradiation of in-vitro cell culture and temperature monitoring of the culture are presented. The first system was designed to irradiate 24 wells in a 96-well microplate. The second one was constructed for the irradiation and control of a 24-well microplate using larger volumes of cultured cells. Both systems can independently irradiate and control the temperature of each well. The systems include a module for contactless measurement of the temperature in each well. Light sources are located in an interchangeable module, so that it can be replaced to irradiate with different wavelengths. These prototypes count with various operation modes, controlled by a computer, which permits establishing specific settings in accordance with the desired experiment. The systems allow the automated experiment execution with precise control of dosimetry, irradiation and temperature, which reduces the sample-handling while, saves time. (Author)
Ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride as a novel matrix for rapid quantitative and qualitative determination of serum free fatty acids by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry
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Zhang, Yaping [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China); Wang, Yanmin [Department of Clinical Laboratory, Heze Municipal Hospital, Shandong (China); Guo, Shuai; Guo, Yumei; Liu, Hui [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China); Li, Zhili, E-mail: firstname.lastname@example.com [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China)
Graphical abstract: -- Highlights: •A novel MALDI matrix for the detection of serum free fatty acids is ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride. •Multiple point internal standard calibration curves were constructed for nine FFAs, respectively, with excellent correlation coefficients between 0.991 and 0.999. •The MALDI-MS approach was used to rapidly differentiate the patients with and without hyperglycemia and healthy controls. -- Abstract: The blood free fatty acids (FFAs), which provide energy to the cell and act as substrates in the synthesis of fats, lipoproteins, liposaccharides, and eicosanoids, involve in a number of important physiological processes. In the present study, matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS) with ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride (ATNEDC) as a novel MALDI matrix in a negative ion mode was employed to directly quantify serum FFAs. Multiple point internal standard calibration curves between the concentration ratios of individual fatty acids to internal standard (IS, C{sub 17:0}) versus their corresponding intensity ratios were constructed for C{sub 14:0}, C{sub 16:1}, C{sub 16:0}, C{sub 18:0}, C{sub 18:1}, C{sub 18:2}, C{sub 18:3}, C{sub 20:4}, and C{sub 22:6}, respectively, in their mixture, with correlation coefficients between 0.991 and 0.999 and limits of detection (LODs) between 0.2 and 5.4 μM, along with the linear dynamic range of more than two orders of magnitude. The results indicate that the multiple point internal standard calibration could reduce the impact of ion suppression and improve quantification accuracy in the MALDI mode. The quantitative results of nine FFAs from 339 serum samples, including 161 healthy controls, 118 patients with hyperglycemia and 60 patients without hyperglycemia show that FFAs levels in hyperglycemic patient sera are significantly higher than those in healthy
Ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride as a novel matrix for rapid quantitative and qualitative determination of serum free fatty acids by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry
International Nuclear Information System (INIS)
Zhang, Yaping; Wang, Yanmin; Guo, Shuai; Guo, Yumei; Liu, Hui; Li, Zhili
Graphical abstract: -- Highlights: •A novel MALDI matrix for the detection of serum free fatty acids is ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride. •Multiple point internal standard calibration curves were constructed for nine FFAs, respectively, with excellent correlation coefficients between 0.991 and 0.999. •The MALDI-MS approach was used to rapidly differentiate the patients with and without hyperglycemia and healthy controls. -- Abstract: The blood free fatty acids (FFAs), which provide energy to the cell and act as substrates in the synthesis of fats, lipoproteins, liposaccharides, and eicosanoids, involve in a number of important physiological processes. In the present study, matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS) with ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride (ATNEDC) as a novel MALDI matrix in a negative ion mode was employed to directly quantify serum FFAs. Multiple point internal standard calibration curves between the concentration ratios of individual fatty acids to internal standard (IS, C 17:0 ) versus their corresponding intensity ratios were constructed for C 14:0 , C 16:1 , C 16:0 , C 18:0 , C 18:1 , C 18:2 , C 18:3 , C 20:4 , and C 22:6 , respectively, in their mixture, with correlation coefficients between 0.991 and 0.999 and limits of detection (LODs) between 0.2 and 5.4 μM, along with the linear dynamic range of more than two orders of magnitude. The results indicate that the multiple point internal standard calibration could reduce the impact of ion suppression and improve quantification accuracy in the MALDI mode. The quantitative results of nine FFAs from 339 serum samples, including 161 healthy controls, 118 patients with hyperglycemia and 60 patients without hyperglycemia show that FFAs levels in hyperglycemic patient sera are significantly higher than those in healthy controls and patients without
A filter paper-based liquid culture system for citrus shoot organogenesis - a mixture-amount experiment
The objective of this study was to determine the effects of a static liquid culture system on shoot regeneration from citrus epicotyl explants. Two citrus types were used, Carrizo citrange and Ridge Pineapple sweet orange. A liquid culture system comprised of a Petri dish, cellulose filter paper dis...
The Impact of Corporate Culture, the Reward System, and Perceived Moral Intensity on Marketing Students' Ethical Decision Making
Nill, Alexander; Schibrowsky, John A.
An experiment was conducted to study how marketing students' ethical decision making was influenced by their perceived moral intensity (PMI), corporate culture, and the reward system. The findings indicate that levels of awareness of the ethical consequences of a decision, the corporate culture, and the reward system all significantly affect…
Effect of Changing Governance System: Result of Western Style Management Adoption to Japanese Culture of Ambiguity
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Kohichiro Hotta
Full Text Available This paper considers the difficulty of management style change through observation of the management style of Company-A, one of the biggest Japanese IT companies. Japanese economy grew after World War II until the early 1990's. During that era, Ba or SECI process worked in Japanese organizations very well. Further, there was an ambiguous culture in the background of such characteristics. Some kinds of ambiguity or adhocracy made positive effects for Japanese organizational activity, or ambiguity played an important role for Ba activity. There were nested Ba's in each organization with ambiguity. Ambiguous descriptions of roles for each organizational unit activated nested Ba's and generated hot groups. After the economic crisis, Company-A changed its governance and gave clear targets for each organizational unit and for each employee. This change gave new difficulty and diminishes its competence. The change denied the ambiguity in the organization but it was the basis of the competence. Adopting a new system of governance is not a simple activity. Systems must be adjusted to the culture of the organization. Company-A should study competitors in different cultures and adjust the methodology for its culture.
Enhanced clinical-scale manufacturing of TCR transduced T-cells using closed culture system modules.
Jin, Jianjian; Gkitsas, Nikolaos; Fellowes, Vicki S; Ren, Jiaqiang; Feldman, Steven A; Hinrichs, Christian S; Stroncek, David F; Highfill, Steven L
Genetic engineering of T-cells to express specific T cell receptors (TCR) has emerged as a novel strategy to treat various malignancies. More widespread utilization of these types of therapies has been somewhat constrained by the lack of closed culture processes capable of expanding sufficient numbers of T-cells for clinical application. Here, we evaluate a process for robust clinical grade manufacturing of TCR gene engineered T-cells. TCRs that target human papillomavirus E6 and E7 were independently tested. A 21 day process was divided into a transduction phase (7 days) and a rapid expansion phase (14 days). This process was evaluated using two healthy donor samples and four samples obtained from patients with epithelial cancers. The process resulted in ~ 2000-fold increase in viable nucleated cells and high transduction efficiencies (64-92%). At the end of culture, functional assays demonstrated that these cells were potent and specific in their ability to kill tumor cells bearing target and secrete large quantities of interferon and tumor necrosis factor. Both phases of culture were contained within closed or semi-closed modules, which include automated density gradient separation and cell culture bags for the first phase and closed GREX culture devices and wash/concentrate systems for the second phase. Large-scale manufacturing using modular systems and semi-automated devices resulted in highly functional clinical-grade TCR transduced T-cells. This process is now in use in actively accruing clinical trials and the NIH Clinical Center and can be utilized at other cell therapy manufacturing sites that wish to scale-up and optimize their processing using closed systems.
HB&L System: rapid determination of antibiotic sensitivity of bacteria isolated from blood cultures.
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Simone Barocci
Full Text Available Introduction. Blood culture is an important method to detect microbial pathogens on blood, very useful for diagnosing bacterial infections. Unfortunately, classical diagnostic protocols cannot directly identify bacteria responsible for sepsis and accordingly their antimicrobial profiles. This problem causes a delay of almost two days in the availability of a specific antimicrobial profile. Objective. Among the main causes of death, sepsis have a relevant importance. For this reason it is important both to identify pathogens and to perform an antimicrobial susceptibility test in the shortest time as possible. For this purpose, the main aim of this study is the evaluation of the performances of an antimicrobial susceptibility determination directly performed on positive blood cultures. Materials and methods. This study has been performed on 70 positive blood cultures, during the period from January to July 2009. A number of 35 blood cultures were positive for Gram negative bacteria, and 35 were positive for Gram positive bacteria. From these positive blood cultures, after a short sample preparation, it has been possible to directly determine antimicrobial susceptibility profiles by using the HB&L (formerly URO-QUICK instrument. Results. The HB&L system results showed a very good correlation with both the classical disk diffusion method and VITEK 2 automatic system.The performances between the methods carried out in this study were equivalent. Conclusions. From data reported, thanks to the rapidity and simplicity of the method used, we can assert that the direct susceptibility test available with the HB&L system, is useful for a rapid and early choice of the antibiotic treatment.
Organizational culture focused on quality management and benefits derived from an ERP system implementation
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Oscar F. Bustinza
Full Text Available Purpose: Organizational culture focused on quality management aims to meet customer needs and enhance teamwork, being oriented toward a dynamic process of continuous improvement. The purpose of this paper is to analyze whether, indeed, the quality-oriented culture has an effect on the management of business processes. In doing so, we analyze their relationship with the benefits of a resource management system or ERP.Design/methodology/approach: A survey is used to collect data, with valid questionnaires obtained for 200 Spain based respondents. Empirical analysis utilises Structural Equation Modelling (SEM.Findings: The results confirm that firm's commitment with quality management, customer focus, and confidence of workers has a positive effect on the results of operational, strategic and managerial benefits derived from an ERP system implementation. However, there is not relationship between customer focus and organizational benefits, neither to increase system capacity.Originality/value: The present study analyzes the relationship between quality-oriented culture and the resource management systems of the firm clarifying their strengths and limitations. In this sense, the customer orientation may limit the flexibility of business as require a lot of resources, and generate dissatisfaction among workers resulting from the attention to customer complaints.
A Novel Miniature Culture System to Screen CO2-Sequestering Microalgae
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Xiaoling Miao
Full Text Available In this study, a novel 96-well microplate swivel system (M96SS was built for high-throughput screening of microalgal strains for CO2 fixation. Cell growth under different CO2 supply conditions (0.2, 0.4, 0.8, and 1.2 g L−1 d−1, residual nitrate, and pH value of Chlorella sp. SJTU-3, Chlorella pyrenoidosa SJTU-2, and Scenedesmus obliquus SJTU-3 were examined in the M96SS and traditional flask cultures. The dynamic data showed there was a good agreement between the systems. Two critical problems in miniature culture systems (intra-well mixing and evaporation loss were improved by sealed vertical mixing of the M96SS. A sample screen of six microalgal species (Chlorella sp. SJTU-3, Chlorella pyrenoidosa SJTU-2, Selenastrum capricornutum, Scenedesmus obliquus SJTU-3, Chlamydomonas sajao, Dunaliella primolecta was carried out in flasks and the M96SS. Chlamydomonas sajao appeared to be a robust performer (highest cell density: 1.437 g L−1 in anaerobic pond water with 0.8, and 1.2 g L−1 d−1 CO2. The reliability and efficiency of the M96SS were verified through a comparison of traditional flask culture, M96SS, Lukavský's system, and a microplate shaker.
In vitro Differentiation of Functional Human Skeletal Myotubes in a Defined System.
Guo, Xiufang; Greene, Keshel; Akanda, Nesar; Smith, Alec; Stancescu, Maria; Lambert, Stephen; Vandenburgh, Herman; Hickman, James
In vitro human skeletal muscle systems are valuable tools for the study of human muscular development, disease and treatment. However, published in vitro human muscle systems have so far only demonstrated limited differentiation capacities. Advanced differentiation features such as cross-striations and contractility have only been observed in co-cultures with motoneurons. Furthermore, it is commonly regarded that cultured human myotubes do not spontaneously contract, and any contraction has been considered to originate from innervation. This study developed a serum-free culture system in which human skeletal myotubes demonstrated advanced differentiation. Characterization by immunocytochemistry, electrophysiology and analysis of contractile function revealed these major features: A) well defined sarcomeric development, as demonstrated by the presence of cross-striations. B) finely developed excitation-contraction coupling apparatus characterized by the close apposition of dihydropyridine receptors on T-tubules and Ryanodine receptors on sarcoplasmic reticulum membranes. C) spontaneous and electrically controlled contractility. This report not only demonstrates an improved level of differentiation of cultured human skeletal myotubes, but also provides the first published evidence that such myotubes are capable of spontaneous contraction. Use of this functional in vitro human skeletal muscle system would advance studies concerning human skeletal muscle development and physiology, as well as muscle-related disease and therapy.
New method for culture of zona-included or zona-free embryos: the Well of the Well (WOW) system.
Vajta, G; Peura, T T; Holm, P; Páldi, A; Greve, T; Trounson, A O; Callesen, H
Culture of mammalian zygotes individually and in small groups results in lower developmental rates than culture of large groups. Zona-free zygotes also have impaired developmental potential in current culture systems. This paper describes a new approach to resolve the problems, the Well of the Well (WOW) system. Small wells (WOWs) were formed in four-well dishes by melting the bottom with heated steel rods. The WOWs were then rinsed, the wells were filled with medium, and the embryos were placed into the WOWs. To test the value of the WOW system a 3 x 3 factorial experiment was performed. Bovine presumptive zygotes were cultured from day 1 to day 7 (day 0: day of insemination) using three modules (single embryos, embryo groups of five, or single zona-digested embryos) and three different culture systems (400 microl medium, 200 microl drops, or WOWs). An additional control group consisted of 40 to 50 embryos cultured in 400 microl medium. The WOW system resulted in higher blastocyst/oocyte rates for all three modules (single: 59%; group of five: 61%; single zona-digested: 53%) than the culture in drops or in wells (P WOWs per well. The cell number of blastocysts cultured in the WOW system did not differ from that of the controls. Apart from its theoretical value in revealing the role of different factors influencing embryo development in vitro, the WOW system may have immediate practical consequences in certain areas of mammalian embryo production. Copyright 2000 Wiley-Liss, Inc.
Effect of agitation and terminal subcultures on yield and speed of detection of the Oxoid Signal blood culture system versus the BACTEC radiometric system
International Nuclear Information System (INIS)
Weinstein, M.P.; Mirrett, S.; Reimer, L.G.; Reller, L.B.
In an initial evaluation, we found the Oxoid Signal blood culture system inferior to the BACTEC radiometric system for detection of some microorganisms causing septicemia. To determine whether modified processing of the Oxoid Signal blood culture system could improve its yield and speed of detecting positive cultures relative to the BACTEC radiometric system, we agitated all Oxoid bottles during the first 24 to 48 h of incubation and performed aerobic and anaerobic subcultures of all Oxoid bottles negative after 7 days of incubation. These modifications improved the overall performance of the Oxoid system, particularly with regard to the yield of streptococci, members of the family Enterobacteriaceae, and Haemophilus, Neisseria, and Acinetobacter spp. The speed of detecting positive cultures also was improved, especially within the first 24 h of incubation. However, the BACTEC system still detected more positive cultures (P less than 0.005), especially of obligate aerobes such as Pseudomonas aeruginosa (P less than 0.05) and yeasts (P less than 0.005). The BACTEC system also detected positive cultures earlier than the Oxoid system (e.g., at 24 h of incubation, 70.5% of BACTEC positive cultures detected versus 62.1% of Oxoid positive cultures detected). Further modifications of the Oxoid system which might include a revised medium, additional processing modifications, altered headspace atmosphere, or a complementary second broth medium should be considered, since the system is attractive in concept and is easy to use in the clinical laboratory
Intangible Knowledge. The Culture of Knowledge within Organisations from the Perspective of the Sociological Systems Theory
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Tilia Stingl de Vasconcelos
Full Text Available Knowledge can get lost when workers leave the company, or it may be missed when new challenges emerge. Specific knowledge may be important for the value-added chain of an organization, and its inaccessibility could be a problem. The work on this paper seeks to juxtapose this problem with the concept of intangible knowledge. This concept is developed as an observation model for particular situations within organisations, in which specific, useful, knowledge is no longer available and is being missed. This paper considers a potentially useful way to deal with absence of such knowledge by using the social science approach. In addition to social systems theory, the communication and cultural science view was selected here to propose a new understanding of the function of knowledge as a communicational or cultural parameter within structures and meanings of a social system. This should facilitate a better perception of the actions and dynamics inside organizations regarding knowledge or the lack thereof.
LIIS: A web-based system for culture collections and sample annotation
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Matthew S Forster
Full Text Available The Lab Information Indexing System (LIIS is a web-driven database application for laboratories looking to store their sample or culture metadata on a central server. The design was driven by a need to replace traditional paper storage with an easier to search format, and extend current spreadsheet storage methods. The system supports the import and export of CSV spreadsheets, and stores general metadata designed to complement the environmental packages provided by the Genomic Standards Consortium. The goals of the LIIS are to simplify the storage and archival processes and to provide an easy to access library of laboratory annotations. The program will find utility in microbial ecology laboratories or any lab that needs to annotate samples/cultures.
Cell culture system of a hepatitis C genotype 3a and 2a chimera
DEFF Research Database (Denmark)
A robust and genetically stable cell culture system for Hepatitis C Virus (HCV) genotype 3a is provided. A genotype 3a/2a (S52/JFH1) recombinant containing the structural genes (Core, E1, E2), p7 and NS2 of strain S52 was constructed and characterized in Huh7.5 cells. S52/JFH1 and J6/JFH viruses ...
In vitro culture of pre-implanted mouse embryos. A model system for studying combined effects
International Nuclear Information System (INIS)
Streffer, C.; Beuningen, D. van; Molls, M.; Pon, A.; Schulz, S.; Zamboglou, N.
Studies on combined effects, e.g. interaction between chemical toxicants and ionizing radiation, are difficult to perform, as they are dependent on many factors (substance concentration, radiation dose, sequence of treatments, etc.). In order to obtain data from such studies it is necessary to establish a comparatively simple experimental model system. We have established such a model system by studying combined effects on pre-implanted mouse embryos cultured in vitro. This system has the following advantages: (1) The embryos can be cultivated for several days in vitro; (2) Their physiological intactness can be tested; and (3) Cell proliferation, cell killing and chromosomal damage can be investigated comparatively easily. The embryos are isolated at the 2-cell stage and incubated in a culture medium in vitro. The development of the embryos is followed under the microscope until the development of blastocysts or the hatching of blastocysts is observed. These blastocysts can be transplanted to fostered mice and the development of normal animals determined. The proliferation kinetics can be studied easily, and the methods are described. A method has also been developed to measure the DNA content of individual cells by microscope fluorometry. After treatment of the embryos with ionizing radiation or drugs the release of micronuclei has been observed from the cell nuclei, which is an expression for chromosomal damage. Substances or radionuclides can be added to the culture medium or external irradiation can be performed during the culture period. Also the combined effects of radiation and heating can be studied. The effects of X-rays and tritiated compounds have also been investigated. The combined effects of radiation with antibiotics such as actinomycin D, and environmental toxicants such as lead, have been determined. The system described has been useful to evaluate cytological, teratogenic and cytogenetic effects
Bioreactors to influence stem cell fate: augmentation of mesenchymal stem cell signaling pathways via dynamic culture systems.
Yeatts, Andrew B; Choquette, Daniel T; Fisher, John P
Mesenchymal stem cells (MSCs) are a promising cell source for bone and cartilage tissue engineering as they can be easily isolated from the body and differentiated into osteoblasts and chondrocytes. A cell based tissue engineering strategy using MSCs often involves the culture of these cells on three-dimensional scaffolds; however the size of these scaffolds and the cell population they can support can be restricted in traditional static culture. Thus dynamic culture in bioreactor systems provides a promising means to culture and differentiate MSCs in vitro. This review seeks to characterize key MSC differentiation signaling pathways and provides evidence as to how dynamic culture is augmenting these pathways. Following an overview of dynamic culture systems, discussion will be provided on how these systems can effectively modify and maintain important culture parameters including oxygen content and shear stress. Literature is reviewed for both a highlight of key signaling pathways and evidence for regulation of these signaling pathways via dynamic culture systems. The ability to understand how these culture systems are affecting MSC signaling pathways could lead to a shear or oxygen regime to direct stem cell differentiation. In this way the efficacy of in vitro culture and differentiation of MSCs on three-dimensional scaffolds could be greatly increased. Bioreactor systems have the ability to control many key differentiation stimuli including mechanical stress and oxygen content. The further integration of cell signaling investigations within dynamic culture systems will lead to a quicker realization of the promise of tissue engineering and regenerative medicine. This article is part of a Special Issue entitled Biochemistry of Stem Cells. Copyright © 2012 Elsevier B.V. All rights reserved.
Intimal permeability evaluated in a short-term organ culture of diabetic guinea pig aorta
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Schlosser, M.J.; Verlangieri, A.J.
A novel short-term organ culture system was used to evaluate intimal permeability changes by measuring aortic [ 14 C]methylated albumin accumulation. Aortic plugs were removed from the upper thoracic aorta of male guinea pigs and maintained in serum-free media. The accumulation of [ 14 C]albumin in the intimal-medial layer was determined after a 5 h incubation. In preliminary studies, albumin recovered from intimal-injured aortic plugs was significantly greater than those from non-injured plugs. Aortic plugs from streptozotocin-treated guinea pigs, diabetic for 3 weeks, also accumulated significantly more [ 14 C]albumin than plugs from nondiabetic controls. Histological changes were not observed in the aorta of either the diabetic or control group. A strong significant inverse correlation was found between plasma ascorbic acid levels and [ 14 C]-activity recovered from aortic plugs. This study demonstrates a simple and rapid method for assessing aortic permeability changes under a well-defined in vitro system, and suggests that vascular permeability changes in the streptozotocin-diabetic guinea pig may be associated with an ascorbic acid deficit
Soft material-based microculture system having air permeable cover sheet for the protoplast culture of Nicotiana tabacum.
Ju, Jong Il; Ko, Jung-Moon; Kim, So Hyeon; Baek, Ju Yeoul; Cha, Hyeon-Cheol; Lee, Sang Hoon
In plant cell culture, the delivery of nutrition and gas (mainly oxygen) to the cells is the most important factor for viability. In this paper, we propose a polydimethylsiloxane (PDMS)-based microculture system that is designed to have good aeration. PDMS is known to have excellent air permeability, and through the experimental method, we investigated the relation between the degree of air delivery and the thickness of the PDMS sheet covering the culture chamber. We determined the proper thickness of the cover sheet, and cultured protoplasts of Nicotiana tabacum in a culture chamber covered with a PDMS sheet having thickness of 400 microm. The cells were successfully divided, and lived well inside the culture chamber for 10 days. In addition, protoplasts were cultured inside the culture chambers covered with the cover glass and the PDMS sheet, respectively, and the microcolonies were formed well inside the PDMS covered chamber after 10 days.
Effect of explant density and medium culture volumes on cassava micropropagation in Temporal Immersion System
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Milagros Basail
Full Text Available Due to the need of producing high quality planting material available to cassava growers, it has been necessary to look for alternatives in order to increase the efficiancy of in vitro propagation methods and their automation, such as the use of the Temporal Immersion Systems (RITA®. This work was carried out to increase the multiplication coefficient for cassava mass propagation through out Temporal Immersion Systems. The clone 'CMC-40' was used. Different medium volumes per explant, and material density per unit at a given Immersion frequency were tested. The highest results were obtained in the 2.8 multiplication coefficient with 20 ml culture medium volume and 3.2 using a density of 40 explants/flask. When the Temporal Immersion System is used with these results, a more efficient method for cassava micropropagation is established and also higher quality vitroplants for the rooting stage and further acclimatization in field conditions are produced. Key Words: Tissue Culture, liquid culture medium, Manihot esculenta Crantz
Brain Aggregates: An Effective In Vitro Cell Culture System Modeling Neurodegenerative Diseases.
Ahn, Misol; Kalume, Franck; Pitstick, Rose; Oehler, Abby; Carlson, George; DeArmond, Stephen J
Drug discovery for neurodegenerative diseases is particularly challenging because of the discrepancies in drug effects between in vitro and in vivo studies. These discrepancies occur in part because current cell culture systems used for drug screening have many limitations. First, few cell culture systems accurately model human aging or neurodegenerative diseases. Second, drug efficacy may differ between dividing and stationary cells, the latter resembling nondividing neurons in the CNS. Brain aggregates (BrnAggs) derived from embryonic day 15 gestation mouse embryos may represent neuropathogenic processes in prion disease and reflect in vivo drug efficacy. Here, we report a new method for the production of BrnAggs suitable for drug screening and suggest that BrnAggs can model additional neurological diseases such as tauopathies. We also report a functional assay with BrnAggs by measuring electrophysiological activities. Our data suggest that BrnAggs could serve as an effective in vitro cell culture system for drug discovery for neurodegenerative diseases. © 2016 American Association of Neuropathologists, Inc. All rights reserved.
Implementing oxygen control in chip-based cell and tissue culture systems.
Oomen, Pieter E; Skolimowski, Maciej D; Verpoorte, Elisabeth
Oxygen is essential in the energy metabolism of cells, as well as being an important regulatory parameter influencing cell differentiation and function. Interest in precise oxygen control for in vitro cultures of tissues and cells continues to grow, especially with the emergence of the organ-on-a-chip and the desire to emulate in vivo conditions. This was recently discussed in this journal in a Critical Review by Brennan et al. (Lab Chip (2014). DOI: ). Microfluidics can be used to introduce flow to facilitate nutrient supply to and waste removal from in vitro culture systems. Well-defined oxygen gradients can also be established. However, cells can quickly alter the oxygen balance in their vicinity. In this Tutorial Review, we expand on the Brennan paper to focus on the implementation of oxygen analysis in these systems to achieve continuous monitoring. Both electrochemical and optical approaches for the integration of oxygen monitoring in microfluidic tissue and cell culture systems will be discussed. Differences in oxygen requirements from one organ to the next are a challenging problem, as oxygen delivery is limited by its uptake into medium. Hence, we discuss the factors determining oxygen concentrations in solutions and consider the possible use of artificial oxygen carriers to increase dissolved oxygen concentrations. The selection of device material for applications requiring precise oxygen control is discussed in detail, focusing on oxygen permeability. Lastly, a variety of devices is presented, showing the diversity of approaches that can be employed to control and monitor oxygen concentrations in in vitro experiments.
Formation of pedagogical system for individual selfdevelopment by means of physical culture and sport
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Valery Panachev
Full Text Available Problems of formation, development and introduction of the modern pedagogical selfdevelopment system in university educational process by means of physical culture and sport have been considered in this article. Such generated pedagogical system reflects practical implementation of social order on the modern educational paradigm aimed at creation of competitive, physically and morally strong individuals. This system promotes selfrealization of students' individuality in formation of physical culture and sport competencies as well as competencies of selfdevelopment. Contemporary conditions of society's development and analysis of the world social cultural and educational tendencies show that recently the object of scrupulous society's attention and the subject of interdisciplinary research have become different aspects of person's behavior in respect of his health in many highly developed countries. The slogan of such relation is: "Health is not everything but everything without health is nothing". And this very principle specifies the problems of students' selfdevelopment during intensive preparation for professional activity in the course of university training. These problems are aimed at maintenance of their health and improvement of physical preparation considering formation of professional motor competences for effective and qualitative mastering of future profession and career after graduation from the university.
Floating rice-culture system for nutrient remediation and feed production in a eutrophic lake.
Srivastava, Ankita; Chun, Seong-Jun; Ko, So-Ra; Kim, Junhwan; Ahn, Chi-Yong; Oh, Hee-Mock
The increased inputs of nutrients have been demonstrated to be a major contributing factor to the eutrophication of lakes and reservoirs which can lead to the production of harmful algal/cyanobacterial blooms and deleteriously affect the aesthetics of water-bodies. Floating plant-culture systems have been widely used for the ecological remediation of eutrophic water in a cost-effective manner. We investigated the applicability of Korean japonica rice variety 'Nampyeong' in a floating-culture system in a eutrophic lake for nutrient uptake and biomass production. Chemical and organic compound compositions were analyzed two times during the growth stages of the rice plant: 98 DAT (days after transplanting) and 165 DAT. Total nitrogen and phosphorus contributed around 1.36 and 0.15 (% dry weight), respectively, in rice plant components at 165 DAT. Crude protein, lipids, fiber and ash were 4.35, 1.91, 23.66 and 5.55 (% dry weight), respectively. In addition, microcystin levels in the rice plant components ranged from 0.0008 to 0.002 μg/g and did not exceed the recommended tolerable limits. These results suggested that the developed floating rice-culture system showed a good potential as a holistic management approach in terms of nutrient reduction, rice production for further use as feed and for bloom control. Copyright © 2017 Elsevier Ltd. All rights reserved.
Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell-based immunotherapies.
Fekete, Natalie; Béland, Ariane V; Campbell, Katie; Clark, Sarah L; Hoesli, Corinne A
In recent years, cell-based therapies targeting the immune system have emerged as promising strategies for cancer treatment. This review summarizes manufacturing challenges related to production of antigen presenting cells as a patient-tailored cancer therapy. Understanding cell-material interactions is essential because in vitro cell culture manipulations to obtain mature antigen-producing cells can significantly alter their in vivo performance. Traditional antigen-producing cell culture protocols often rely on cell adhesion to surface-treated hydrophilic polystyrene flasks. More recent commercial and investigational cancer immunotherapy products were manufactured using suspension cell culture in closed hydrophobic fluoropolymer bags. The shift to closed cell culture systems can decrease risks of contamination by individual operators, as well as facilitate scale-up and automation. Selecting closed cell culture bags over traditional open culture systems entails different handling procedures and processing controls, which can affect product quality. Changes in culture vessels also entail changes in vessel materials and geometry, which may alter the cell microenvironment and resulting cell fate decisions. Strategically designed culture systems will pave the way for the generation of more sophisticated and highly potent cell-based cancer vaccines. As an increasing number of cell-based therapies enter the clinic, the selection of appropriate cell culture vessels and materials becomes a critical consideration that can impact the therapeutic efficacy of the product, and hence clinical outcomes and patient quality of life. © 2018 The Authors Transfusion published by Wiley Periodicals, Inc. on behalf of AABB.
Culturing Toxic Benthic Blooms: The Fate of Natural Biofilms in a Microcosm System
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Francesca Di Pippo
Full Text Available A microcosm designed for culturing aquatic phototrophic biofilms on artificial substrata was used to perform experiments with microphytobenthos sampled during summer toxic outbreaks of Ostreopsis cf. ovata along the Middle Tyrrhenian coast. This dynamic approach aimed at exploring the unique and complex nature of O. cf. ovata bloom development in the benthic system. Epibenthic assemblages were used as inocula for co-cultures of bloom organisms on polycarbonate slides at controlled environmental conditions. Biofilm surface adhesion, growth, and spatial structure were evaluated along with shifts in composition and matrix production in a low disturbance regime, simulating source habitat. Initial adhesion and substratum colonisation appeared as stochastic processes, then community structure and physiognomy markedly changed with time. Dominance of filamentous cyanobacteria and diatoms, and dense clusters of Amphidinium cf. carterae at the mature biofilm phases, were recorded by light and confocal microscopy, whilst O. cf. ovata growth was visibly limited in the late culture phases. Life-form strategies, competitiveness for resources, and possibly allelopathic interactions shaped biofilm structure during culture growth. HPLC (High Performance Liquid Chromatography analysis of exopolysaccharidic matrix revealed variations in sugar total amounts and composition. No toxic compounds were detected in the final communities tested by LC-MS (Liquid Chromatography- Mass Spectrometry and MALDI-TOF MS (Matrix Assisted Laser Desorption Ionization Time OF Flight Mass Spectroscopy techniques.
Triple co-culture cell model as an in vitro model for oral particulate vaccine systems
DEFF Research Database (Denmark)
Nielsen, Line Hagner; De Rossi, C.; Lehr, C-M.
; this was not observed with ovalbumin and blank solution. An example of the results is shown in Figure 2 for IL-17A. An established co-culture of Caco-2, THP-1 and MUTZ-3 cells showed promise as an in vitro model for testing of oral vaccine formulations. Mobility of co-culture immune cells as well as cytokine production......A triple co-culture cell model of Caco-2 cells, dendritic cells and macrophages (Figure 1) has previously been developed for studying intestinal permeability in a state of inflammation [1],[2]. The aim of this study was to investigate the applicability of this cell model for testing...... the model antigen ovalbumin was spray dried to obtain a particulate vaccine model system for testing in the cell model. The precursors were shown to form cubosomes when dispersed in aqueous medium, and was therefore used as the vaccine formulation for testing on the co-cultures. After 11 days, the TEER...
Engineering systems for the generation of patterned co-cultures for controlling cell-cell interactions.
Kaji, Hirokazu; Camci-Unal, Gulden; Langer, Robert; Khademhosseini, Ali
Inside the body, cells lie in direct contact or in close proximity to other cell types in a tightly controlled architecture that often regulates the resulting tissue function. Therefore, tissue engineering constructs that aim to reproduce the architecture and the geometry of tissues will benefit from methods of controlling cell-cell interactions with microscale resolution. We discuss the use of microfabrication technologies for generating patterned co-cultures. In addition, we categorize patterned co-culture systems by cell type and discuss the implications of regulating cell-cell interactions in the resulting biological function of the tissues. Patterned co-cultures are a useful tool for fabricating tissue engineered constructs and for studying cell-cell interactions in vitro, because they can be used to control the degree of homotypic and heterotypic cell-cell contact. In addition, this approach can be manipulated to elucidate important factors involved in cell-matrix interactions. Patterned co-culture strategies hold significant potential to develop biomimetic structures for tissue engineering. It is expected that they would create opportunities to develop artificial tissues in the future. This article is part of a Special Issue entitled Nanotechnologies - Emerging Applications in Biomedicine. 2010 Elsevier B.V. All rights reserved.
Co-culture systems-based strategies for articular cartilage tissue engineering.
Zhang, Yu; Guo, Weimin; Wang, Mingjie; Hao, Chunxiang; Lu, Liang; Gao, Shuang; Zhang, Xueliang; Li, Xu; Chen, Mingxue; Li, Penghao; Jiang, Peng; Lu, Shibi; Liu, Shuyun; Guo, Quanyi
Cartilage engineering facilitates repair and regeneration of damaged cartilage using engineered tissue that restores the functional properties of the impaired joint. The seed cells used most frequently in tissue engineering, are chondrocytes and mesenchymal stem cells. Seed cells activity plays a key role in the regeneration of functional cartilage tissue. However, seed cells undergo undesirable changes after in vitro processing procedures, such as degeneration of cartilage cells and induced hypertrophy of mesenchymal stem cells, which hinder cartilage tissue engineering. Compared to monoculture, which does not mimic the in vivo cellular environment, co-culture technology provides a more realistic microenvironment in terms of various physical, chemical, and biological factors. Co-culture technology is used in cartilage tissue engineering to overcome obstacles related to the degeneration of seed cells, and shows promise for cartilage regeneration and repair. In this review, we focus first on existing co-culture systems for cartilage tissue engineering and related fields, and discuss the conditions and mechanisms thereof. This is followed by methods for optimizing seed cell co-culture conditions to generate functional neo-cartilage tissue, which will lead to a new era in cartilage tissue engineering. © 2017 Wiley Periodicals, Inc.
Manifestation of radiaton injury of human lymphocytes using PHA mitogenic stimulation in different culture systems
International Nuclear Information System (INIS)
Horky, J.
The proliferative response of human lymphocytes to phytohemagglutinin in vitro is affected by X-irradiation. Dose-related changes in mitogenic stimulation of irradiated lymphocytes were compared for two culture systems - the cultivation of separated lymphocytes and the cultivation of whole blood. In the whole blood cultures, the proliferative activity of stimulated lyphocytes was markedly and reproducibly depressed by irradiation. An exponential curve could be fitted to the values of mitogenic response within a dose range from 0 to 2.5 Gy with high correlation. In a modified test where the mitogenic stimulus was given after a 24 h delay, depression in the response was even more pronounced. Radiosensitivity of human lymphocytes as determined by means of mitogenic stimulation in the whole blood cultures appears to be a characteristic individual feature. The mean D 37 value of the radiation-induced depression in mitogenic response in a group of 20 healthy donors was 2.5 Gy in the standard test and 2.0 Gy in the test with a delayed mitogenic stimulus. In contrast, the data obtained from separated lymphocyte cultures were characterized by a high degree of test-to-test variability and by much lower radiosensitivity. The possible mechanisms of these distinctive manifestations of the same primary radiation injury are discussed. (author) 3 tabs., 2 figs., 12 refs
A cultural trauma: Outcomes of the system change in post-socialist Poland
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Kabzinska Iwona
Full Text Available This paper discusses the outcomes of the system change in post-socialist Poland. The author discusses various important and inter-related issues in the Polish societal sphere: shock therapy accompanied by the changes since the 1990's, unemployment, uncontrolled privatization, cultural trauma and cultural plaint. Theoretically, the paper belongs to anthropology of transformation, and it is based on sociological literature for the most part. The paper also discusses 'societal diagnosis', its creators, crisis in confidence as a consequence of social and cultural traumas, the weakness of political elite and criteria used to measure poverty levels. Lately, there has been a change in mythical representation about easy life in Poland related to the state' affiliation with EU. The change includes a lack of global crisis influence, resistance of the Polish society toward media influence, a rise in optimism and decrease of cultural plaint. Is this change in attitude due to cyclical alteration between phases of depression and euphoria? What will happen if depression returns? Did the Polish handle the trauma of transformation exceptionally well? Possible answers to these and other relevant questions are sought by the author in this paper, who uses, as additional sources for research, a world of local communities and individual accounts.
Effect of Culture System on Developmental Competence, Cryosurvival and DNA-Fragmentation of In Vitro Bovine Blastocysts
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Mahdi Hajian
Full Text Available Background: This study investigated the effect of two in vitro embryo culture systems (co-culturesystem versus cell-free sequential-media on developmental competence, cryosurvival and DNAfragmentationof in vitro developed bovine blastocysts.Materials and Methods: Bovine presumptive zygotes were cultured in Ménézo's B2 (B2 plusvero-cells or sequential synthetic oviductal fluid (SOF for eight days. Subsequently, half of theexpanded blastocysts developed in both groups were vitrified, warmed within 30 minutes and postwarmingembryos along with their corresponding non-vitrified embryos were cultured for twoadditional days in the same medium used before vitrification. Embryo development, cryosurvivaland apoptosis were compared between the groups.Results: For non-vitrified embryos, culture in SOF significantly promoted the potency of embryosto develop into blastocysts compared with the co-culture system. The difference in post vitrificationsurvival rate of SOF blastocysts (83.3% was insignificant compared with co-culture (84.3%.However, while total cell number of warmed blastocysts in the co-culture system was significantlyhigher in the co-culture versus the sequential system (215.4 vs. 170.4, the quality of survived embryosin terms of hatching ability and apoptosis was adversely affected by co-culture compared with SOF(65.0% vs. 74.3%, and 13.5% vs. 10.0%, respectively; p<0.05.Conclusion: Although co-culture system may increase the viability of embryos followingcryopreservation, the potency and dynamics of blastocyst formation significantly increased withsequential media compared to the co-culture system which can compensate for the lower efficiency ofsequential media for vitrification/warming purposes.
Development and characterization of cell culture systems from Puntius (Tor) chelynoides (McClelland).
Goswami, M; Sharma, B S; Tripathi, A K; Yadav, Kamalendra; Bahuguna, S N; Nagpure, N S; Lakra, W S; Jena, J K
Puntius (Tor) chelynoides, commonly known as dark mahseer, is a commercially important coldwater fish species which inhabits fast-flowing hill-streams of India and Nepal. Cell culture systems were developed from eye, fin, heart and swim bladder tissues of P. chelynoides using explant method. The cell culture system developed from eye has been maintained towards a continuous cell line designated as PCE. The cells were grown in 25cm(2) tissue culture flasks with Leibovitz' L-15 media supplemented with 20 % fetal bovine serum (FBS) at 24°C. The PCE cell line consists of predominantly fibroblast-like cells and showed high plating efficiency. The monolayer formed from the fin and heart explants were comprised of epithelial as well as fibroblast-like cells, a prominent and rhythmic heartbeat was also observed in heart explants. Monolayer formed from swim bladder explants showed the morphology of fibroblast-like cells. All the cells from different tissues are able to grow at an optimum temperature of 24°C and growth rate increased as the FBS concentration increased. The PCE cell line was characterized using amplification of mitochondrial cytochrome oxidase subunit I (COI) & 16S rRNA genes which confirmed that the cell line originated from P. chelynoides. Cytogenetic analysis of PCE cell line and cells from fin revealed a diploid count of 100 chromosomes. Upon transfection with pEGFP-C1 plasmid, bright fluorescent signals were observed, suggesting that this cell line can be used for transgenic and genetic manipulation studies. Further, genotoxicity assessment of PCE cells illustrated the utility of this cell line as an in vitro model for aquatic toxicological studies. The PCE cell line was successfully cryopreserved and revived at different passage levels. The cell line and culture systems are being maintained to develop continuous cell lines for further studies. Copyright © 2012 Elsevier B.V. All rights reserved.
Cell Monitoring and Manipulation Systems (CMMSs based on Glass Cell-Culture Chips (GC3s
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Sebastian M. Buehler
Full Text Available We developed different types of glass cell-culture chips (GC3s for culturing cells for microscopic observation in open media-containing troughs or in microfluidic structures. Platinum sensor and manipulation structures were used to monitor physiological parameters and to allocate and permeabilize cells. Electro-thermal micro pumps distributed chemical compounds in the microfluidic systems. The integrated temperature sensors showed a linear, Pt1000-like behavior. Cell adhesion and proliferation were monitored using interdigitated electrode structures (IDESs. The cell-doubling times of primary murine embryonic neuronal cells (PNCs were determined based on the IDES capacitance-peak shifts. The electrical activity of PNC networks was detected using multi-electrode arrays (MEAs. During seeding, the cells were dielectrophoretically allocated to individual MEAs to improve network structures. MEA pads with diameters of 15, 20, 25, and 35 µm were tested. After 3 weeks, the magnitudes of the determined action potentials were highest for pads of 25 µm in diameter and did not differ when the inter-pad distances were 100 or 170 µm. Using 25-µm diameter circular oxygen electrodes, the signal currents in the cell-culture media were found to range from approximately −0.08 nA (0% O2 to −2.35 nA (21% O2. It was observed that 60-nm thick silicon nitride-sensor layers were stable potentiometric pH sensors under cell-culture conditions for periods of days. Their sensitivity between pH 5 and 9 was as high as 45 mV per pH step. We concluded that sensorized GC3s are potential animal replacement systems for purposes such as toxicity pre-screening. For example, the effect of mefloquine, a medication used to treat malaria, on the electrical activity of neuronal cells was determined in this study using a GC3 system.
Comparing the Cultural Dimensions and Learner's Perceived Effectiveness of Online Learning Systems (OLS) among American and Malaysian Learners
Seng C. Keng
With the rapid and exponential growth of Internet use worldwide, online learning has become one of the most widely used learning paradigms in the education environment. Yet despite the rapidly increasing cultural diversity of online learners, few studies have investigated the effectiveness of cross-cultural Online Learning Systems (OLS) using a suitable measurement to answer the question, "Do culturally different learners perceive OLS effectiveness differently?" The aim of this co...
Psychosocial and Cultural Modeling in Human Computation Systems: A Gamification Approach
Energy Technology Data Exchange (ETDEWEB)
Sanfilippo, Antonio P.; Riensche, Roderick M.; Haack, Jereme N.; Butner, R. Scott
"Gamification", the application of gameplay to real-world problems, enables the development of human computation systems that support decision-making through the integration of social and machine intelligence. One of gamification's major benefits includes the creation of a problem solving environment where the influence of cognitive and cultural biases on human judgment can be curtailed through collaborative and competitive reasoning. By reducing biases on human judgment, gamification allows human computation systems to exploit human creativity relatively unhindered by human error. Operationally, gamification uses simulation to harvest human behavioral data that provide valuable insights for the solution of real-world problems.
Copyright for Interactive Systems: Stratagems for Tourism and Cultural Heritage Promotion
Cipolla-Ficarra, Francisco V.; Cipolla-Ficarra, Miguel; Ficarra, Valeria M.
We present a series of strategies followed from the interactive design for the realization of a hypermedia system aimed at promoting in an original, simple and universal way the cultural and tourism heritage of a wide rural area in two Italian regions: Emilia Romagna and Lombardy. Besides, the main stratagems followed are disclosed to overcome the existing hurdles when it comes to copyright for the free diffusion of the tourism view of the area, such as can be photography or video, for instance. Finally, we present the first vademecum to be considered before making on-line and off-line interactive systems in Italy.
Use of an automated blood culture system (BD BACTEC™) for diagnosis of prosthetic joint infections: easy and fast.
Minassian, Angela M; Newnham, Robert; Kalimeris, Elizabeth; Bejon, Philip; Atkins, Bridget L; Bowler, Ian C J W
For the diagnosis of prosthetic joint infection (PJI) automated BACTEC™ blood culture bottle methods have comparable sensitivity, specificity and a shorter time to positivity than traditional cooked meat enrichment broth methods. We evaluate the culture incubation period required to maximise sensitivity and specificity of microbiological diagnosis, and the ability of BACTEC™ to detect slow growing Propionibacteria spp. Multiple periprosthetic tissue samples taken by a standardised method from 332 patients undergoing prosthetic joint revision arthroplasty were cultured for 14 days, using a BD BACTEC™ instrumented blood culture system, in a prospective study from 1st January to 31st August 2012. The "gold standard" definition for PJI was the presence of at least one histological criterion, the presence of a sinus tract or purulence around the device. Cases where > =2 samples yielded indistinguishable isolates were considered culture-positive. 1000 BACTEC™ bottle cultures which were negative after 14 days incubation were sub-cultured for Propionibacteria spp. 79 patients fulfilled the definition for PJI, and 66 of these were culture-positive. All but 1 of these 66 culture-positive cases of PJI were detected within 3 days of incubation. Only one additional (clinically-insignificant) Propionibacterium spp. was identified on terminal subculture of 1000 bottles. Prolonged microbiological culture for 2 weeks is unnecessary when using BACTEC™ culture methods. The majority of clinically significant organisms grow within 3 days, and Propionibacteria spp. are identified without the need for terminal subculture. These findings should facilitate earlier decisions on final antimicrobial prescribing.
Using Remote Sensing Technology on the Delimitation of the Conservation Area for the Jianan Irrigation System Cultural Landsccape
Wang, C. H.
In recent years the cultural landscape has become an important issue for cultural heritages throughout the world. It represents the "combined works of nature and of man" designated in Article 1 of the World Heritage Convention. When a landscape has a cultural heritage value, important features should be marked and mapped through the delimitation of a conservation area, which may be essential for further conservation work. However, a cultural landscape's spatial area is usually wider than the ordinary architectural type of cultural heritage, since various elements and impact factors, forming the cultural landscape's character, lie within a wide geographic area. It is argued that the conservation of a cultural landscape may be influenced by the delimitation of the conservation area, the corresponding land management measures, the limits and encouragements. The Jianan Irrigation System, an historical cultural landscape in southern Taiwan, was registered as a living cultural heritage site in 2009. However, the system's conservation should not be limited to just only the reservoir or canals, but expanded to irrigated areas where farmland may be the most relevant. Through the analysis process, only approximately 42,000 hectares was defined as a conservation area, but closely related to agricultural plantations and irrigated by the system. This is only half of the 1977 irrigated area due to urban sprawl and continuous industrial expansion.
Understanding the earth systems of Malawi: Ecological sustainability, culture, and place-based education
Glasson, George E.; Frykholm, Jeffrey A.; Mhango, Ndalapa A.; Phiri, Absalom D.
The purpose of this 2-year study was to investigate Malawian teacher educators' perspectives and dispositions toward teaching about ecological sustainability issues in Malawi, a developing country in sub-Sahara Africa. This study was embedded in a larger theoretical framework of investigating earth systems science through the understanding of nature-knowledge-culture systems from local, place-based perspectives. Specifically, we were interested in learning more about eco-justice issues that are related to environmental degradation in Malawi and the potential role of inquiry-oriented pedagogies in addressing these issues. In a science methods course, the African educators' views on deforestation and teaching about ecological sustainability were explored within the context of the local environment and culture. Teachers participated in inquiry pedagogies designed to promote the sharing of perspectives related to the connections between culture and ecological degradation. Strategies encouraging dialogue and reflection included role-playing, class discussions, curriculum development activities, teaching experiences with children, and field trips to a nature preserve. Data were analyzed from postcolonial and critical pedagogy of place theoretical perspectives to better understand the hybridization of viewpoints influenced by both Western and indigenous science and the political hegemonies that impact sustainable living in Malawi. Findings suggested that the colonial legacy of Malawi continues to impact the ecological sustainability issue of deforestation. Inquiry-oriented pedagogies and connections to indigenous science were embraced by the Malawian educators as a means to involve children in investigation, decision making, and ownership of critical environmental issues.
A two dimensional clinostat experiment for microalgae cultures - basic work for bio- regenerativ life support systems
Harting, Benjamin; Slenzka, Klaus
To investigate the influence of microgravity environments on photosynthetic organisms we designed a 2 dimensional clinostatexperiment for a suspended cell culture of Chlamydomonas reinhardtii. A novel approach of online measurments concerning relevant parameters important for the clasification of photosynthesis was obtained. To adress the photosynthesis rate we installed and validated an optical mesurement system to monitor the evolution and consumption of dissolved oxygen. Simultaneously a PAM sensor to analyse the flourescence quantum yield of the photochemical reaction was integarted. Thus it was possible to directly classify important parameters of the phototrophic metabolism during clinorotation. The experiment design including well suited light conditions and further biochemical analysis were directly performed for microalgal cell cultures. Changes in the photosynthetic efficiancy of phototrophic cyanobacteria has been observed during parabolic flight campaign but the cause is already not understood. Explenations could be the dependency of gravitaxis by intracellular ionconcentartion or the existance of mechanosensitive ionchannels for example associated in chloroplasts of Chlamydomonas reinhardtii. The purpuse of the microalgal clinostat are studies in a qasi microgravity environment for the process design of future bioregenerative life suport systems in spaceflight missions. First results has indicated the need for special nourishment of the cell culture during microgravity experiments. Further data will be presented during the assembly.
Development of a Continuous Phytoplankton Culture System for Ocean Acidification Experiments
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Cathryn Wynn-Edwards
Full Text Available Around one third of all anthropogenic CO2 emissions have been absorbed by the oceans, causing changes in seawater pH and carbonate chemistry. These changes have the potential to affect phytoplankton, which are critically important for marine food webs and the global carbon cycle. However, our current knowledge of how phytoplankton will respond to these changes is limited to a few laboratory and mesocosm experiments. Long-term experiments are needed to determine the vulnerability of phytoplankton to enhanced pCO2. Maintaining phytoplankton cultures in exponential growth for extended periods of time is logistically difficult and labour intensive. Here we describe a continuous culture system that greatly reduces the time required to maintain phytoplankton cultures, and minimises variation in experimental pCO2 treatments over time. This system is simple, relatively cheap, flexible, and allows long-term experiments to be performed to further our understanding of chronic responses and adaptation by phytoplankton species to future ocean acidification.
Socio-cultural barriers to the development of a sustainable energy system - the case of hydrogen
Energy Technology Data Exchange (ETDEWEB)
Kjerulf Petersen, L.; Holst Andersen, A.
Any transition to a more sustainable energy system, radically reducing greenhouse gas emissions, is bound to run in to a host of different barriers - technological and economic, but also socio-cultural. This will also be the case for any large-scale application of hydrogen as energy carrier, especially if the system is going to be based on renewable energy sources. The aim of these research notes is to review and discuss major socio-cultural barriers to new forms of energy supply in general and to hydrogen specifically. Reaching sufficient reductions in greenhouse gas emissions may require more than large-scale dissemination of renewable energy sources. Also reductions or moderations in energy demand may be necessary. Hence, a central point in the research note is to consider not only socio-cultural obstacles for changing technologies in energy production, distribution and consumption but also obstacles for changing the scale of energy consumption, i.e. moderating the growth in how much energy is consumed or even reducing consumption volumes. (au)
Application of Biomaterials and Inkjet Printing to Develop Bacterial Culture System
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Tithimanan Srimongkon
Full Text Available We created an automated bioassay system based on inkjet printing. Compared to conventional manual bacterial culture systems our printing approach improves the quality as well as the processing speed. A hydrophobic/hydrophilic pattern as a container supporting a culture medium was built on filter paper using a toluene solution of polystyrene for hydrophobization, followed by toluene printing to create several hydrophilic areas. As culture media we used a novel poly(vinyl alcohol based hydrogel and a standard calcium alginate hydrogel. The poly(vinyl alcohol hydrogel was formed by physical crosslinking poly(vinyl alcohol with adipic acid dihydrazide solutions. The conditions of poly(vinyl alcohol gelation were optimized for inkjet printability and the optimum mixture ratio was determined. The calcium alginate hydrogel was formed by chemical reaction between sodium alginate and CaCl2 solutions. Together with nutrients both hydrogel solutions were successfully printed on paper by means of the modified inkjet printer. The amount of each solution was demanded simply by outputting CMYK values. In the last step bacterial cells were printed on both hydrogel media. For both media we achieved a stable bacteria growth which was confirmed by microscopical imaging of the developed bacterial colonies.
[Clinical governance and patient safety culture in clinical laboratories in the Spanish National Health System].
Giménez-Marín, Á; Rivas-Ruiz, F
To conduct a situational analysis of patient safety culture in public laboratories in the Spanish National Health System and to determine the clinical governance variables that most strongly influence patient safety. A descriptive cross-sectional study was carried out, in which a Survey of Patient Safety in Clinical Laboratories was addressed to workers in 26 participating laboratories. In this survey, which consisted of 45 items grouped into 6 areas, scores were assigned on a scale from 0 to 100 (where 0 is the lowest perception of patient safety). Laboratory managers were asked specific questions about quality management systems and technology. The mean scores for the 26 participating hospitals were evaluated, and the following results observed: in 4of the 6areas, the mean score was higher than 70 points. In the third area (equipment and resources) and the fourth area (working conditions), the scores were lower than 60 points. Every hospital had a digital medical record system. This 100% level of provision was followed by that of an electronic request management system, which was implemented in 82.6% of the hospitals. The results obtained show that the culture of security is homogeneous and of high quality in health service laboratories, probably due to the steady improvement observed. However, in terms of clinical governance, there is still some way to go, as shown by the presence of weaknesses in crucial dimensions of safety culture, together with variable levels of implementation of fail-safe technologies and quality management systems. Copyright © 2017 SECA. Publicado por Elsevier España, S.L.U. All rights reserved.
Increased Arginine and Ornithine Flux in Islets of Langerhans Cultured in a Microgravity Model System
Tobin, B. W.; Sams, C. F.; Smith, S. M.
Microgravity is associated with alterations in protein metabolism of both muscle and bone. That pancreas-derived insulin is essential to the normal maintenance of body protein balance is well known. The importance of altered endocrine pancreas function in microgravity is not yet established. We proposed to examine the influence of a microgravity model system, the High Aspect Ratio Vessel (HARV) upon islets of Langerhans from Wistar Furth rats. Islets were cultured in the HARV for 48 hr in Medium-199 and contrasted to static control islets (PLATE). Nitrogenous compounds elaborated into the media (micromoles/ml) were analyzed at 0 and 48 hr of culture and compared to PLATE with a 2-way ANOVA (HARV vs Hour).
Seeing your way to health: the visual pedagogy of Bess Mensendieck's physical culture system.
Veder, Robin
This essay examines the images and looking practices central to Bess M. Mensendieck's (c.1866-1959) 'functional exercise' system, as documented in physical culture treatises published in Germany and the United States between 1906 and 1937. Believing that muscular realignment could not occur without seeing how the body worked, Mensendieck taught adult non-athletes to see skeletal alignment and muscular movement in their own and others' bodies. Three levels of looking practices are examined: didactic sequences; penetrating inspection and appreciation of physiological structures; and ideokinetic visual metaphors for guiding movement. With these techniques, Mensendieck's work bridged the body cultures of German Nacktkultur (nudism), American labour efficiency and the emerging physical education profession. This case study demonstrates how sport historians could expand their analyses to include practices of looking as well as questions of visual representation.
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Elaine da Silva
Full Text Available Introduction: Emphasizes the role of innovation in the Information and Knowledge Society as a favorable element of developing regional, national and global levels. Objective: The aim is reflect about the coming of innovation in the context of production systems and their respective role for the development of society. Methodology: The research sets up a qualitative approach to literature and exploratory nature. Results: As from selected literature review, presents conceptual aspects of innovation in the context of production systems and analyzes its contribution to the national innovation system and the role of organizational culture focused on innovation. Conclusions: Think about innovation from the systemic approach, based on the interaction of the different relevant agents to influence and contribute to the generation of innovation is fundamental in the current conjuncture, characterized by the agility of the transformations in social, economic, political and technological contexts.
Joint-Service Integration: An Organizational Culture Study of the United States Department of Defense Voluntary Education System
Benson, Martin K.
The purpose of the descriptive case study with a multiple case framework was to (a) describe the organizational cultures of education programs and leaders in the United States (U.S.) Department of Defense (DoD) voluntary education system on Oahu, Hawaii; (b) determine if an overlapping common organizational culture exists; and (c) assess the…
Comparing the Cultural Dimensions and Learners' Perceived Effectiveness of Online Learning Systems (OLS) among American and Malaysian Learners
Keng, Seng C.
With the rapid and exponential growth of Internet use worldwide, online learning has become one of the most widely used learning paradigms in the education environment. Yet despite the rapidly increasing cultural diversity of online learners, few studies have investigated the effectiveness of cross-cultural Online Learning Systems (OLS) using a…
Oil-free culture system for in vitro bovine embryo production
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Paulo B.D. Gonçalves
Full Text Available The use of oil to avoid water evaporation from cell culture has several disadvantages, amongst which there is the migration of compounds from media to oil and from oil to media. The aim of this study was to evaluate the osmolality of a culture system using four-well plates with water in the central hole as an alternative to in vitro bovine embryo production (IVP. In addition, the osmolality changes of the oocyte washing medium were assessed in 35mm dishes with or without 2 mL of silicon oil overlay. Osmolality of oocyte washing medium changed a great deal over time after 60 minutes on a 39°C heated plate (291 mOsm kg-1, which was not detected when the medium was overlaid with silicon oil (280 mOsm kg-1; P0.05. Blastocyst rates were higher when embryos were cultured in presence of water or oil (29.7 and 29.9% for water and 33% in oil conventional microdrop system, except in the group that oocytes were washed in hyperosmotic washing medium (15.1%; P<0.05. Groups cultured in absence of water in the central hole had lower blastocyst rates (P<0.05 independently of exposure (15.5% or not (16.2 and 16.8% to hyperosmotic washing medium. In conclusion, four-well plates with water in the central hole can be an alternative to replace oil overlay for bovine IVP, maintaining stable osmolality and embryo development rates.
Improving safety culture in hospitals: Facilitators and barriers to implementation of Systemic Falls Investigative Method (SFIM).
Zecevic, Aleksandra A; Li, Alvin Ho-Ting; Ngo, Charity; Halligan, Michelle; Kothari, Anita
The purpose of this study was to assess the facilitators and barriers to implementation of the Systemic Falls Investigative Method (SFIM) on selected hospital units. A cross-sectional explanatory mixed methods design was used to converge results from a standardized safety culture survey with themes that emerged from interviews and focus groups. Findings were organized by six elements of the Ottawa Model of Research Use framework. A geriatric rehabilitation unit of an acute care hospital and a neurological unit of a rehabilitation hospital were selected purposefully due to the high frequency of falls. Hospital staff who took part in: surveys (n = 39), interviews (n = 10) and focus groups (n = 12), and 38 people who were interviewed during falls investigations: fallers, family, unit staff and hospital management. Implementation of the SFIM to investigate fall occurrences. Percent of positive responses on the Modified Stanford Patient Safety Culture Survey Instrument converged with qualitative themes on facilitators and barriers for intervention implementation. Both hospital units had an overall poor safety culture which hindered intervention implementation. Facilitators were hospital accreditation, strong emphasis on patient safety, infrastructure and dedicated champions. Barriers included heavy workloads, lack of time, lack of resources and poor communication. Successful implementation of SFIM requires regulatory and organizational support, committed frontline staff and allocation of resources to identify active causes and latent contributing factors to falls. System-wide adjustments show promise for promotion of safety culture in hospitals where falls happen regularly. © The Author 2017. Published by Oxford University Press in association with the International Society for Quality in Health Care. All rights reserved. For permissions, please e-mail: email@example.com
Menaquinone-4 enhances osteogenic potential of human amniotic fluid mesenchymal stem cells cultured in 2D and 3D dynamic culture systems.
Mandatori, Domitilla; Penolazzi, Letizia; Pipino, Caterina; Di Tomo, Pamela; Di Silvestre, Sara; Di Pietro, Natalia; Trevisani, Sara; Angelozzi, Marco; Ucci, Mariangela; Piva, Roberta; Pandolfi, Assunta
Menaquinones, also known as Vitamin K2 family, regulate calcium homeostasis in a 'bone-vascular cross-talk' and recently received particular attention for their positive effect on bone formation. Given that the correlation between menaquinones and bone metabolism to date is still unclear, the objective of our study was to investigate the possible role of menaquinone-4 (MK-4), an isoform of the menaquinones family, in the modulation of osteogenesis. For this reason, we used a model of human amniotic fluid mesenchymal stem cells (hAFMSCs) cultured both in two-dimensional (2D) and three-dimensional (3D; RCCS™bioreactor) in vitro culture systems. Furthermore, to mimic the 'bone remodelling unit' in vitro, hAFMSCs were co-cultured in the 3D system with human monocyte cells (hMCs) as osteoclast precursors. The results showed that in a conventional 2D culture system, hAFMSCs were responsive to the MK-4, which significantly improved the osteogenic process through γ-glutamyl carboxylase-dependent pathway. The same results were obtained in the 3D dynamic system where MK-4 treatment supported the osteoblast-like formation promoting the extracellular bone matrix deposition and the expression of the osteogenic-related proteins (alkaline phosphatase, osteopontin, collagen type-1 and osteocalcin). Notably, when the hAFMSCs were co-cultured in a 3D dynamic system with the hMCs, the presence of MK-4 supported the cellular aggregate formation as well as the osteogenic function of hAFMSCs, but negatively affected the osteoclastogenic process. Taken together, our results demonstrate that MK-4 supported the aggregate formation of hAFMSCs and increased the osteogenic functions. Specifically, our data could help to optimize bone regenerative medicine combining cell-based approaches with MK-4 treatment. Copyright © 2017 John Wiley & Sons, Ltd.
Optimization of chemically defined cell culture media--replacing fetal bovine serum in mammalian in vitro methods
DEFF Research Database (Denmark)
van der Valk, J; Brunner, D; De Smet, K
with an undefined and variable composition. Defined media supplements are commercially available for some cell types. However, information on the formulation by the companies is often limited and such supplements can therefore not be regarded as completely defined. The development of defined media is difficult......, reproducible and reduce the use of experimental animals. Good cell culture practice (GCCP) is an attempt to develop a common standard for in vitro methods. The implementation of the use of chemically defined media is part of the GCCP. This will decrease the dependence on animal serum, a supplement...... and often takes place in isolation. A workshop was organised in 2009 in Copenhagen to discuss strategies to improve the development and use of serum-free defined media. In this report, the results from the meeting are discussed and the formulation of a basic serum-free medium is suggested. Furthermore...
Effect of different culture systems on the production of foot and mouth disease trivalent vaccine
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Amr Ismail Hassan
Full Text Available Aim: This study aims to determine the effect of the stationary rawx, roller, and the suspension cell culture systems on the total virus yield infectivity and antigenicity. Materials and Methods: Three serotypes of foot and mouth disease virus (FMDV (serotype A, O and SAT-2 were inoculated separately into baby hamster kidney-21 cell line in rawx, roller, and suspension cultivation systems using multiplicity of infection (1:100. Samples were taken from the total virus yield from each system at 15, 18, 21, and 24 h post-inoculation. Testing the total virus yield infectivity through virus titration and antigenicity through estimation of complement fixing titer and 146S content and evaluation of the potency of the vaccine prepared from the different cultivation systems were done. Results: The results showed that the FMDV titer of serotype A, O, and SAT-2 obtained from the roller cultivation system showed the highest level followed by suspension cultivation system then the rawx cultivation system. The FMDV titer showed its highest level at 21 h post-inoculation in all the cultivation systems and then decline at 24 h post-inoculation. The antigenicity reached its highest value content at 18 h post-inoculation either by complement fixation test or by quantifying the 146S intact virion. Montanide ISA 206 oil inactivated trivalent vaccines were prepared from the tested serotypes (A Iran O5. O Panasia and SAT-2/EGY/2012 harvested at 18 h post-inoculation from the 3 culture systems. The results of tracing the antibody response showed that the mean antibody response from the roller cultivation system start its protective antibody titer earlier at 2 weeks post-vaccination (WPV than the vaccine prepared from the other two cultivation system and the immune protection period lasts longer for 36 WPV for the roller cultivation system vaccine than the other two cultivation systems. Conclusion: The best cultivation system used for the production of FMD vaccine
Quantifying the Metrics That Characterize Safety Culture of Three Engineered Systems
International Nuclear Information System (INIS)
Tucker, Julie; Ernesti, Mary; Tokuhiro, Akira
With potential energy shortages and increasing electricity demand, the nuclear energy option is being reconsidered in the United States. Public opinion will have a considerable voice in policy decisions that will 'road-map' the future of nuclear energy in this country. This report is an extension of the last author's work on the 'safety culture' associated with three engineered systems (automobiles, commercial airplanes, and nuclear power plants) in Japan and the United States. Safety culture, in brief is defined as a specifically developed culture based on societal and individual interpretations of the balance of real, perceived, and imagined risks versus the benefits drawn from utilizing a given engineered systems. The method of analysis is a modified scale analysis, with two fundamental Eigen-metrics, time- (t) and number-scales (N) that describe both engineered systems and human factors. The scale analysis approach is appropriate because human perception of risk, perception of benefit and level of (technological) acceptance are inherently subjective, therefore 'fuzzy' and rarely quantifiable in exact magnitude. Perception of risk, expressed in terms of the psychometric factors 'dread risk' and 'unknown risk', contains both time- and number-scale elements. Various engineering system accidents with fatalities, reported by mass media are characterized by t and N, and are presented in this work using the scale analysis method. We contend that level of acceptance infers a perception of benefit at least two orders larger magnitude than perception of risk. The 'amplification' influence of mass media is also deduced as being 100- to 1000-fold the actual number of fatalities/serious injuries in a nuclear-related accident. (authors)
Hetero-cellular prototyping by synchronized multi-material bioprinting for rotary cell culture system.
Snyder, Jessica; Son, Ae Rin; Hamid, Qudus; Wu, Honglu; Sun, Wei
Bottom-up tissue engineering requires methodological progress of biofabrication to capture key design facets of anatomical arrangements across micro, meso and macro-scales. The diffusive mass transfer properties necessary to elicit stability and functionality require hetero-typic contact, cell-to-cell signaling and uniform nutrient diffusion. Bioprinting techniques successfully build mathematically defined porous architecture to diminish resistance to mass transfer. Current limitations of bioprinted cell assemblies include poor micro-scale formability of cell-laden soft gels and asymmetrical macro-scale diffusion through 3D volumes. The objective of this work is to engineer a synchronized multi-material bioprinter (SMMB) system which improves the resolution and expands the capability of existing bioprinting systems by packaging multiple cell types in heterotypic arrays prior to deposition. This unit cell approach to arranging multiple cell-laden solutions is integrated with a motion system to print heterogeneous filaments as tissue engineered scaffolds and nanoliter droplets. The set of SMMB process parameters control the geometric arrangement of the combined flow's internal features and constituent material's volume fractions. SMMB printed hepatocyte-endothelial laden 200 nl droplets are cultured in a rotary cell culture system (RCCS) to study the effect of microgravity on an in vitro model of the human hepatic lobule. RCCS conditioning for 48 h increased hepatocyte cytoplasm diameter 2 μm, increased metabolic rate, and decreased drug half-life. SMMB hetero-cellular models present a 10-fold increase in metabolic rate, compared to SMMB mono-culture models. Improved bioprinting resolution due to process control of cell-laden matrix packaging as well as nanoliter droplet printing capability identify SMMB as a viable technique to improve in vitro model efficacy.
Hetero-cellular prototyping by synchronized multi-material bioprinting for rotary cell culture system
International Nuclear Information System (INIS)
Snyder, Jessica; Son, Ae Rin; Hamid, Qudus; Sun, Wei; Wu, Honglu
Bottom-up tissue engineering requires methodological progress of biofabrication to capture key design facets of anatomical arrangements across micro, meso and macro-scales. The diffusive mass transfer properties necessary to elicit stability and functionality require hetero-typic contact, cell-to-cell signaling and uniform nutrient diffusion. Bioprinting techniques successfully build mathematically defined porous architecture to diminish resistance to mass transfer. Current limitations of bioprinted cell assemblies include poor micro-scale formability of cell-laden soft gels and asymmetrical macro-scale diffusion through 3D volumes. The objective of this work is to engineer a synchronized multi-material bioprinter (SMMB) system which improves the resolution and expands the capability of existing bioprinting systems by packaging multiple cell types in heterotypic arrays prior to deposition. This unit cell approach to arranging multiple cell-laden solutions is integrated with a motion system to print heterogeneous filaments as tissue engineered scaffolds and nanoliter droplets. The set of SMMB process parameters control the geometric arrangement of the combined flow's internal features and constituent material's volume fractions. SMMB printed hepatocyte-endothelial laden 200 nl droplets are cultured in a rotary cell culture system (RCCS) to study the effect of microgravity on an in vitro model of the human hepatic lobule. RCCS conditioning for 48 h increased hepatocyte cytoplasm diameter 2 μm, increased metabolic rate, and decreased drug half-life. SMMB hetero-cellular models present a 10-fold increase in metabolic rate, compared to SMMB mono-culture models. Improved bioprinting resolution due to process control of cell-laden matrix packaging as well as nanoliter droplet printing capability identify SMMB as a viable technique to improve in vitro model efficacy. (paper)
The realistic performance achievable with mycobacterial automated culture systems in high and low prevalence settings
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Klatser Paul R
Full Text Available Abstract Background Diagnostic tests are generally used in situations with similar pre-test probability of disease to where they were developed. When these tests are applied in situations with very different pre-test probabilities of disease, it is informative to model the likely implications of known characteristics of test performance in the new situation. This is the case for automated Mycobacterium tuberculosis (MTB liquid culture systems for tuberculosis case detection which were developed and are widely used in low burden settings but are only beginning to be applied on a large scale in high burden settings. Methods Here we model the performance of MTB liquid culture systems in high and low tuberculosis (TB prevalence settings using detailed published data concentrating on the likely frequency of cross-contamination events. Results Our model predicts that as the TB prevalence in the suspect population increases there is an exponential increase in the risk of MTB cross-contamination events expected in otherwise negative samples, even with equivalent technical performance of the laboratories. Quality control and strict cross-contamination measures become increasingly critical as the burden of MTB infection among TB suspects increases. Even under optimal conditions the realistically achievable specificity of these systems in high burden settings will likely be significantly below that obtained in low TB burden laboratories. Conclusions Liquid culture systems can play a valuable role in TB case detection in laboratories in high burden settings, but laboratory workers, policy makers and clinicians should be aware of the increased risks, independent of laboratory proficiency, of cross-contamination events in high burden settings.
Fast Prototyping of Sensorized Cell Culture Chips and Microfluidic Systems with Ultrashort Laser Pulses
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Sebastian M. Bonk
Full Text Available We developed a confined microfluidic cell culture system with a bottom plate made of a microscopic slide with planar platinum sensors for the measurement of acidification, oxygen consumption, and cell adhesion. The slides were commercial slides with indium tin oxide (ITO plating or were prepared from platinum sputtering (100 nm onto a 10-nm titanium adhesion layer. Direct processing of the sensor structures (approximately three minutes per chip by an ultrashort pulse laser facilitated the production of the prototypes. pH-sensitive areas were produced by the sputtering of 60-nm Si3N4 through a simple mask made from a circuit board material. The system body and polydimethylsiloxane (PDMS molding forms for the microfluidic structures were manufactured by micromilling using a printed circuit board (PCB milling machine for circuit boards. The microfluidic structure was finally imprinted in PDMS. Our approach avoided the use of photolithographic techniques and enabled fast and cost-efficient prototyping of the systems. Alternatively, the direct production of metallic, ceramic or polymeric molding tools was tested. The use of ultrashort pulse lasers improved the precision of the structures and avoided any contact of the final structures with toxic chemicals and possible adverse effects for the cell culture in lab-on-a-chip systems.
Reducing unnecessary culturing: a systems approach to evaluating urine culture ordering and collection practices among nurses in two acute care settings
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Robert Redwood
Full Text Available Abstract Background Inappropriate ordering and acquisition of urine cultures leads to unnecessary treatment of asymptomatic bacteriuria (ASB. Treatment of ASB contributes to antimicrobial resistance particularly among hospital-acquired organisms. Our objective was to investigate urine culture ordering and collection practices among nurses to identify key system-level and human factor barriers and facilitators that affect optimal ordering and collection practices. Methods We conducted two focus groups, one with ED nurses and the other with ICU nurses. Questions were developed using the Systems Engineering Initiative for Patient Safety (SEIPS framework. We used iterative categorization (directed content analysis followed by summative content analysis to code and analyze the data both deductively (using SEIPS domains and inductively (emerging themes. Results Factors affecting optimal urine ordering and collection included barriers at the person, process, and task levels. For ED nurses, barriers included patient factors, physician communication, reflex culture protocols, the electronic health record, urinary symptoms, and ED throughput. For ICU nurses, barriers included physician notification of urinalysis results, personal protective equipment, collection technique, patient body habitus, and Foley catheter issues. Conclusions We identified multiple potential process barriers to nurse adherence with evidence-based recommendations for ordering and collecting urine cultures in the ICU and ED. A systems approach to identifying barriers and facilitators can be useful to design interventions for improving urine ordering and collection practices.
Using Remote Sensing Technology on the Delimitation of the Conservation Area for the Jianan Irrigation System Cultural Landsccape
Directory of Open Access Journals (Sweden)
C. H. Wang
Full Text Available In recent years the cultural landscape has become an important issue for cultural heritages throughout the world. It represents the "combined works of nature and of man" designated in Article 1 of the World Heritage Convention. When a landscape has a cultural heritage value, important features should be marked and mapped through the delimitation of a conservation area, which may be essential for further conservation work. However, a cultural landscape's spatial area is usually wider than the ordinary architectural type of cultural heritage, since various elements and impact factors, forming the cultural landscape's character, lie within a wide geographic area. It is argued that the conservation of a cultural landscape may be influenced by the delimitation of the conservation area, the corresponding land management measures, the limits and encouragements. The Jianan Irrigation System, an historical cultural landscape in southern Taiwan, was registered as a living cultural heritage site in 2009. However, the system's conservation should not be limited to just only the reservoir or canals, but expanded to irrigated areas where farmland may be the most relevant. Through the analysis process, only approximately 42,000 hectares was defined as a conservation area, but closely related to agricultural plantations and irrigated by the system. This is only half of the 1977 irrigated area due to urban sprawl and continuous industrial expansion.
Altered TNF-Alpha, Glucose, Insulin and Amino Acids in Islets Langerhans Cultured in a Microgravity Model System
Tobin, Brian W.; Leeper-Woodford, Sandra K.; Hashemi, Brian B.; Smith, Scott M.; Sams, Clarence F.
The present studies were designed to determine effects of a microgravity model system upon lipopolysaccharide (LPS) stimulated tumor necrosis factor alpha (TNF-alpha) activity and indices of insulin and fuel homeostasis of pancreatic islets of Langerhans. Islets (1726+/-1 17,150 u IEU) from Wistar Furth rats were treated as: 1) HARV (High Aspect Ratio Vessel cell culture) , 2) HARV plus LPS, 3) static culture, 4) static culture plus LPS. TNF-alpha (L929 cytotoxicity assay) was significantly increased in LPS-induced HARV and static cultures, yet the increase was more pronounced in the static culture group (palpha production of pancreatic islets of Langerhans, favoring a lesser TNF activity in the HARV. These alterations in fuel homeostasis may be promulgated by gravity averaged cell culture methods or by three dimensional cell assembly.
Voluntary organ donation system adapted to Chinese cultural values and social reality.
Huang, Jiefu; Millis, J Michael; Mao, Yilei; Millis, M Andrew; Sang, Xinting; Zhong, Shouxian
Organ donation and transplant systems have unique characteristics based on the local culture and socioeconomic context. China's transplant and organ donation systems developed without regulatory oversight until 2006 when regulation and policy were developed and then implemented over the next several years. Most recently, the pilot project of establishing a voluntary citizen-based deceased donor program was established. The pilot program addressed the legal, financial, and cultural barriers to organ donation in China. The pilot program has evolved into a national program. Significantly, it established a uniquely Chinese donor classification system. The Chinese donor classification system recognizes donation after brain death (category I), donation after circulatory death (category II), and donation after brain death followed by circulatory death (category III). Through August 2014, the system has identified 2326 donors and provided 6416 organs that have been allocated though a transparent organ allocation system. The estimated number of donors in 2014 is 1147. As China's attitudes toward organ donation have matured and evolved and as China, as a nation, is taking its place on the world stage, it is recognizing that its past practice of using organs from executed prisoners is not sustainable. It is time to recognize that the efforts to regulate transplantation and provide voluntary citizen-based deceased organ donation have been successful and that China should use this system to provide organs for all transplants in every province and hospital in China. At the national organ transplant congress on October 30, 2014, the Chairman of the China's national organ donation and transplantation committee, Jeifu Huang required all hospitals to stop using organs from executed prisoners immediately and the civilian organ donation will be sole source for organ transplant in China starting January 2015. © 2015 American Association for the Study of Liver Diseases.
Comparison of the clinical and microbiological characteristics of Campylobacter and Helicobacter bacteremia: the importance of time to blood culture positivity using the BACTEC blood culture systems.
Yamamoto, Kei; Hayakawa, Kayoko; Nagashima, Maki; Shimada, Kayo; Kutsuna, Satoshi; Takeshita, Nozomi; Kato, Yasuyuki; Kanagawa, Shuzo; Yamada, Koji; Mezaki, Kazuhisa; Kirikae, Teruo; Ohmagari, Norio
Campylobacter spp. and Helicobacter spp. are rare but important causes of bacteremia in humans. Distinguishing these bacteria is complicated because of their similar phenotypic profiles. We conducted clinical and microbiological investigations of Campylobacter spp. or Helicobacter spp. bacteremia. Patients diagnosed with bacteremia from 2008 to 2014 were included. The clinical and microbiological characteristics of Campylobacter spp. and Helicobacter spp. bacteremia were compared. The BACTEC system was used in blood cultures. A receiver operating characteristic curve was plotted based on the time to blood culture positivity. Sixteen cases of Helicobacter spp. bacteremia (patient age: 61 ± 18 years) and 14 cases of Campylobacter spp. bacteremia (patient age: 49 ± 21 years) were identified. Median time to blood culture positivity was longer for the Helicobacter spp. cases than the Campylobacter spp. cases (91.4 h vs 55.3 h, p culture positivity > 75 h predicted Helicobacter spp. bacteremia with a sensitivity of 0.88 and a specificity of 0.93 (area under the receiver operating characteristic curve of 0.90). In conclusion, a time to blood culture positivity was useful in distinguishing Helicobacter spp. bacteremia from Campylobacter spp. bacteremia.
A 3D human neural cell culture system for modeling Alzheimer's disease
Kim, Young Hye; Choi, Se Hoon; D'Avanzo, Carla; Hebisch, Matthias; Sliwinski, Christopher; Bylykbashi, Enjana; Washicosky, Kevin J.; Klee, Justin B.; Brüstle, Oliver; Tanzi, Rudolph E.; Kim, Doo Yeon
Stem cell technologies have facilitated the development of human cellular disease models that can be used to study pathogenesis and test therapeutic candidates. These models hold promise for complex neurological diseases such as Alzheimer's disease (AD) because existing animal models have been unable to fully recapitulate all aspects of pathology. We recently reported the characterization of a novel three-dimensional (3D) culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of β-amyloid and accumulation of hyperphosphorylated tau. Here we provide instructions for the generation and analysis of 3D human neural cell cultures, including the production of genetically modified human neural progenitor cells (hNPCs) with familial AD mutations, the differentiation of the hNPCs in a 3D matrix, and the analysis of AD pathogenesis. The 3D culture generation takes 1–2 days. The aggregation of β-amyloid is observed after 6-weeks of differentiation followed by robust tau pathology after 10–14 weeks. PMID:26068894
New method for rapid Susceptibility Testing on blood culture with HB&L system: preliminary data
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Vincenzo Rondinelli
Full Text Available Blood culture, although represents the gold standard in detecting the ethiological agent of sepsis, is rather rarely required in relation to the real diagnostic importance. The result of this test depends in fact on many factors (sample volume, time of collection, accuracy, antibiotic therapy, contamination, number of drawings, drawing site, interpretation difficulties, etc. that are often considered by many clinicians so limited as to doubt about their actual value. The disadvantages are therefore represented by the lack of standardization but also by the low sensitivity and above all by the technical times too long for the clinical needs. Blood culture begins with the drawing of samples from the "septic" patient followed incubation of the bottles in automatic thermostated systems. In case of positive result (36 hours, the culture is Gram stained and streaked on solid media in order to obtain isolated colonies for the identification and the susceptibility testing (48 hours from positive result. The long time required for pathogen identification and susceptibility testing involves empirical broad spectrum antibiotic therapy that can promote the increase of bacterial resistance but also patient management costs. A clinically useful report should be available on short notice in order to guide the clinician to choose the most appropriate antibiotic. The microbiologist has therefore the hard work of reviewing the organization and the management of the procedures.We have therefore started to consider the possibility of treating the blood as an biological liquid in order to quickly determine the susceptibility of bacteria to antibiotics.
a Restoration Oriented Hbim System for Cultural Heritage Documentation: the Case Study of Parma Cathedral
Bruno, N.; Roncella, R.
The need to safeguard and preserve Cultural Heritage (CH) is increasing and especially in Italy, where the amount of historical buildings is considerable, having efficient and standardized processes of CH management and conservation becomes strategic. At the time being, there are no tools capable of fulfilling all the specific functions required by Cultural Heritage documentation and, due to the complexity of historical assets, there are no solution as flexible and customizable as CH specific needs require. Nevertheless, BIM methodology can represent the most effective solution, on condition that proper methodologies, tools and functions are made available. The paper describes an ongoing research on the implementation of a Historical BIM system for the Parma cathedral, aimed at the maintenance, conservation and restoration. Its main goal was to give a concrete answer to the lack of specific tools required by Cultural Heritage documentation: organized and coordinated storage and management of historical data, easy analysis and query, time management, 3D modelling of irregular shapes, flexibility, user-friendliness, etc. The paper will describe the project and the implemented methodology, focusing mainly on survey and modelling phases. In describing the methodology, critical issues about the creation of a HBIM will be highlighted, trying to outline a workflow applicable also in other similar contexts.
Columbia River System Operation Review : Final Environmental Impact Statement, Appendix D: Cultural Resources.
Energy Technology Data Exchange (ETDEWEB)
Columbia River System Operation Review (U.S.)
This study attempts to identify and analyze the impacts of the System Operating Strategy (SOS) alternatives on cultural resources. The impacts include effects on Native American traditional cultural values, properties and practices. They also include effects on archeological or historic properties meeting the criteria of the National Register of Historic Places. In addition to responding to the requirements of the National Environmental Policy Act (NEPA), this analysis addresses the requirements of the National Historic Preservation Act (NHPA), the Archeological Resources Protection Act (ARPA), the Native American Graves Protection and Repatriation Act (NAGPRA), the Native American Religious Freedom Act (NARFA), and other relevant legislation. To meet their legally mandated cultural resources requirements, the SOR agencies will develop agreements and Implementation Plans with the appropriate State Historic Preservation Officers (SHPOs), Tribes, and the Advisory Council on Historic Preservation (ACHP) detailing the measures necessary to best manage the resource. The planning and implementation activities will be staged over a number of years in consultation with affected Tribes.
Studies on luminous, Vibrio harveyi associated with shrimp culture system rearing Penaeus monodon.
Kannapiran, E; Ravindran, J; Chandrasekar, R; Kalaiarasi, A
Microbiological studies in a modified extensive shrimp culture system at Nambuthalai, southeast coast of India were carried out fora period of 120 days. Population dynamics and distribution profile of luminous bacteria and total heterotrophic bacteria in the water sediment and animal samples were monitored. Luminous bacteria associated with exoskeleton, gills and gut were isolated and quantified. The total heterotrophic bacterial counts ranged from 1.3 x 10(4) to 25.3 x 10(4) CFU ml(-1) in water and 1.5 x 10(6) to 26.2 x 10(6) CFU g(-1) in sediment. The V. harveyi population density varied between 0.6 x 10(4) and 8.8 x 10(4) LCFU ml(-1) in water and from 1.2 x 10(6) to 10.4 x 10(6) LCFU g(-1) sediment respectively. The gut of the animal was found to harbor high density of V. harveyi than gills and exoskeleton. The total heterotrophic bacteria and V. harveyi population density showed increasing trend during the culture period. The high V harveyi density observed in this study at the end of the culture period correlated with the outbreak of white spot disease.
A management system integrating radiation protection and safety supporting safety culture in the hospital
International Nuclear Information System (INIS)
Almen, A.; Lundh, C.
Quality assurance has been identified as an important part of radiation protection and safety for a considerable time period. A rational expansion and improvement of quality assurance is to integrate radiation protection and safety in a management system. The aim of this study was to explore factors influencing the implementing strategy when introducing a management system including radiation protection and safety in hospitals and to outline benefits of such a system. The main experience from developing a management system is that it is possible to create a vast number of common policies and routines for the whole hospital, resulting in a cost-efficient system. One of the key benefits is the involvement of management at all levels, including the hospital director. Furthermore, a transparent system will involve staff throughout the organisation as well. A management system supports a common view on what should be done, who should do it and how the activities are reviewed. An integrated management system for radiation protection and safety includes key elements supporting a safety culture. (authors)
3-Dimensional culture systems for anti-cancer compound profiling and high-throughput screening reveal increases in EGFR inhibitor-mediated cytotoxicity compared to monolayer culture systems.
Howes, Amy L; Richardson, Robyn D; Finlay, Darren; Vuori, Kristiina
3-dimensional (3D) culture models have the potential to bridge the gap between monolayer cell culture and in vivo studies. To benefit anti-cancer drug discovery from 3D models, new techniques are needed that enable their use in high-throughput (HT) screening amenable formats. We have established miniaturized 3D culture methods robust enough for automated HT screens. We have applied these methods to evaluate the sensitivity of normal and tumorigenic breast epithelial cell lines against a panel of oncology drugs when cultured as monolayers (2D) and spheroids (3D). We have identified two classes of compounds that exhibit preferential cytotoxicity against cancer cells over normal cells when cultured as 3D spheroids: microtubule-targeting agents and epidermal growth factor receptor (EGFR) inhibitors. Further improving upon our 3D model, superior differentiation of EC50 values in the proof-of-concept screens was obtained by co-culturing the breast cancer cells with normal human fibroblasts and endothelial cells. Further, the selective sensitivity of the cancer cells towards chemotherapeutics was observed in 3D co-culture conditions, rather than as 2D co-culture monolayers, highlighting the importance of 3D cultures. Finally, we examined the putative mechanisms that drive the differing potency displayed by EGFR inhibitors. In summary, our studies establish robust 3D culture models of human cells for HT assessment of tumor cell-selective agents. This methodology is anticipated to provide a useful tool for the study of biological differences within 2D and 3D culture conditions in HT format, and an important platform for novel anti-cancer drug discovery.
Beyond opening up the black box: Investigating the role of algorithmic systems in Wikipedian organizational culture
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R Stuart Geiger
Full Text Available Scholars and practitioners across domains are increasingly concerned with algorithmic transparency and opacity, interrogating the values and assumptions embedded in automated, black-boxed systems, particularly in user-generated content platforms. I report from an ethnography of infrastructure in Wikipedia to discuss an often understudied aspect of this topic: the local, contextual, learned expertise involved in participating in a highly automated social–technical environment. Today, the organizational culture of Wikipedia is deeply intertwined with various data-driven algorithmic systems, which Wikipedians rely on to help manage and govern the "anyone can edit" encyclopedia at a massive scale. These bots, scripts, tools, plugins, and dashboards make Wikipedia more efficient for those who know how to work with them, but like all organizational culture, newcomers must learn them if they want to fully participate. I illustrate how cultural and organizational expertise is enacted around algorithmic agents by discussing two autoethnographic vignettes, which relate my personal experience as a veteran in Wikipedia. I present thick descriptions of how governance and gatekeeping practices are articulated through and in alignment with these automated infrastructures. Over the past 15 years, Wikipedian veterans and administrators have made specific decisions to support administrative and editorial workflows with automation in particular ways and not others. I use these cases of Wikipedia's bot-supported bureaucracy to discuss several issues in the fields of critical algorithms studies; critical data studies; and fairness, accountability, and transparency in machine learning—most principally arguing that scholarship and practice must go beyond trying to "open up the black box" of such systems and also examine sociocultural processes like newcomer socialization.
Comparison of radiometric and conventional culture systems in detecting Haemophilus influenzae type b bacteremia in rats
International Nuclear Information System (INIS)
Mitchell, M.J.; Zwahlen, A.; Elliott, H.L.; Ford, N.K.; Charache, F.P.; Moxon, E.R.
To compare the efficiency of detecting Haemophilus influenzae type b bacteremia by the BACTEC radiometric system and a conventional Trypticase soy broth blood culture system, the authors developed an in vivo model of bacteremia in rats. After intravenous injection of 50 to 200 CFU into adult rats, there was a linear logarithmic increase in CFU per milliliter of rat blood during the first 10 h (r = 0.98), allowing accurate prediction of the level of bacteremia with time. Culture bottles were inoculated with 0.5 ml of blood obtained by cardiac puncture and processed as clinical samples in the microbiology laboratory with RS and conventional protocols. They found the following. (i) The first detection of bacteremia by RS was similar to that by TSB if a Gram stain of the TSB was done on day 1 and was superior if that smear was omitted (P less than 0.01). (ii) The detection times in both systems were comparable at different magnitudes of bacteremia (10(1) to 10(4) CFU/ml). (iii) Supplementation of inoculated bottles with 2 ml of sterile rat blood interfered with Gram stain detection in TSB but resulted in increased 14 CO 2 production in RS. (iv) No difference in detection time was found between RS and TSB for four different clinical isolates. These studies show that, in a biologically relevant model, the detection of positive blood cultures for H. influenzae type b by RS was comparable to or better than detection by TSB when blood was processed analogously to clinical specimens
Review of vascularised bone tissue-engineering strategies with a focus on co-culture systems.
Liu, Yuchun; Chan, Jerry K Y; Teoh, Swee-Hin
Poor angiogenesis within tissue-engineered grafts has been identified as a main challenge limiting the clinical introduction of bone tissue-engineering (BTE) approaches for the repair of large bone defects. Thick BTE grafts often exhibit poor cellular viability particularly at the core, leading to graft failure and lack of integration with host tissues. Various BTE approaches have been explored for improving vascularisation in tissue-engineered constructs and are briefly discussed in this review. Recent investigations relating to co-culture systems of endothelial and osteoblast-like cells have shown evidence of BTE efficacy in increasing vascularization in thick constructs. This review provides an overview of key concepts related to bone formation and then focuses on the current state of engineered vascularized co-culture systems using bone repair as a model. It will also address key questions regarding the generation of clinically relevant vascularized bone constructs as well as potential directions and considerations for research with the objective of pursuing engineered co-culture systems in other disciplines of vascularized regenerative medicine. The final objective is to generate serious and functional long-lasting vessels for sustainable angiogenesis that will enable enhanced cellular survival within thick voluminous bone grafts, thereby aiding in bone formation and remodelling in the long term. However, more evidence about the quality of blood vessels formed and its associated functional improvement in bone formation as well as a mechanistic understanding of their interactions are necessary for designing better therapeutic strategies for translation to clinical settings. Copyright © 2012 John Wiley & Sons, Ltd.
A New Acquisition and Imaging System for Environmental Measurements: An Experience on the Italian Cultural Heritage
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Fabio Leccese
Full Text Available A new acquisition system for remote control of wall paintings has been realized and tested in the field. The system measures temperature and atmospheric pressure in an archeological site where a fresco has been put under control. The measuring chain has been designed to be used in unfavorable environments where neither electric power nor telecommunication infrastructures are available. The environmental parameters obtained from the local monitoring are then transferred remotely allowing an easier management by experts in the field of conservation of cultural heritage. The local acquisition system uses an electronic card based on microcontrollers and sends the data to a central unit realized with a Raspberry-Pi. The latter manages a high quality camera to pick up pictures of the fresco. Finally, to realize the remote control at a site not reached by internet signals, a WiMAX connection based on different communication technologies such as WiMAX, Ethernet, GPRS and Satellite, has been set up.
Supplementation of serum free media with HT is not sufficient to restore growth properties of DHFR-/- cells in fed-batch processes - Implications for designing novel CHO-based expression platforms.
Florin, Lore; Lipske, Carolin; Becker, Eric; Kaufmann, Hitto
DHFR-deficient CHO cells are the most commonly used host cells in the biopharmaceutical industry and over the years, individual substrains have evolved, some have been engineered with improved properties and platform technologies have been designed around them. Unexpectedly, we have observed that different DHFR-deficient CHO cells show only poor growth in fed-batch cultures even in HT supplemented medium, whereas antibody producer cells derived from these hosts achieved least 2-3 fold higher peak cell densities. Using a set of different expression vectors, we were able to show that this impaired growth performance was not due to the selection procedure possibly favouring fast growing clones, but a direct consequence of DHFR deficiency. Re-introduction of the DHFR gene reproducibly restored the growth phenotype to the level of wild-type CHO cells or even beyond which seemed to be dose-dependent. The requirement for a functional DHFR gene to achieve optimal growth under production conditions has direct implications for cell line generation since it suggests that changing to a selection system other than DHFR would require another CHO host which - especially for transgenic CHO strains and tailor-suited process platforms - this could mean significant investments and potential changes in product quality. In these cases, DHFR engineering of the current CHO-DG44 or DuxB11-based host could be an attractive alternative. Copyright © 2011 Elsevier B.V. All rights reserved.
Enrichment of skin-derived neural precursor cells from dermal cell populations by altering culture conditions.
Bayati, Vahid; Gazor, Rohoullah; Nejatbakhsh, Reza; Negad Dehbashi, Fereshteh
As stem cells play a critical role in tissue repair, their manipulation for being applied in regenerative medicine is of great importance. Skin-derived precursors (SKPs) may be good candidates for use in cell-based therapy as the only neural stem cells which can be isolated from an accessible tissue, skin. Herein, we presented a simple protocol to enrich neural SKPs by monolayer adherent cultivation to prove the efficacy of this method. To enrich neural SKPs from dermal cell populations, we have found that a monolayer adherent cultivation helps to increase the numbers of neural precursor cells. Indeed, we have cultured dermal cells as monolayer under serum-supplemented (control) and serum-supplemented culture, followed by serum free cultivation (test) and compared. Finally, protein markers of SKPs were assessed and compared in both experimental groups and differentiation potential was evaluated in enriched culture. The cells of enriched culture concurrently expressed fibronectin, vimentin and nestin, an intermediate filament protein expressed in neural and skeletal muscle precursors as compared to control culture. In addition, they possessed a multipotential capacity to differentiate into neurogenic, glial, adipogenic, osteogenic and skeletal myogenic cell lineages. It was concluded that serum-free adherent culture reinforced by growth factors have been shown to be effective on proliferation of skin-derived neural precursor cells (skin-NPCs) and drive their selective and rapid expansion.
Recent advances of rearing cabinet instrumentation and control system for insect stock culture
Hermawan, Wawan; Kasmara, Hikmat; Melanie, Panatarani, Camellia; Joni, I. Made
Helicoverpa armigera (Hubner) is one of a serious pest of horticulture in Indonesia. Helicoverpa armigera Nuclear Polyhedrovirus (HaNPV) has attracted interest for many researchers as a pest control for larvae of this species. Currently, we investigating the agrochemical formulations of HaNPV by introducing nanotechnology. Thus it is required an acceptable efficiency of insect stock cultures equipped with advance instruments to resolve the difficulties on insect stock seasons dependency. In addition, it is important to improve the insect survival with the aid of artificial natural environment and gain high insect production. This paper reports the rearing cabinet used as preparation of stock culture includes air-conditioning system, lighting, i.e. day and night control, and the main principles on recent technical and procedural advances apparatus of the system. The rearing system was moveable, designed and build by allowing air-conditioned cabinet for rearing insects, air motion and distribution as well as temperature and humidity being precisely controlled. The air was heated, humidified, and dehumidified respectively using a heater and ultrasonic nebulizer as actuators. Temperature and humidity can be controlled at any desired levels from room temperature (20°C) to 40 ± 1°C and from 0 to 80% RH with an accuracy of ±3% R.H. It is concluded that the recent design has acceptable performance based on the defined requirement for insect rearing and storage.
Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems
Radmer, R.; Cox, J.; Lieberman, D.; Behrens, P.; Arnett, K.
Algal cultures can be very rapid and efficient means to generate biomass and regenerate the atmosphere for closed environmental life support systems. However, as in the case of most higher plants, a significant fraction of the biomass produced by most algae cannot be directly converted to a useful food product by standard food technology procedures. This waste biomass will serve as an energy drain on the overall system unless it can be efficiently recycled without a significant loss of its energy content. Experiments are reported in which cultures of the alga Scenedesmus obliquus were grown in the light and at the expense of an added carbon source, which either replaced or supplemented the actinic light. As part of these experiments, hydrolyzed waste biomass from these same algae were tested to determine whether the algae themselves could be made part of the biological recycling process. Results indicate that hydrolyzed algal (and plant) biomass can serve as carbon and energy sources for the growth of these algae, suggesting that the efficiency of the closed system could be significantly improved using this recycling process.
ADASIR system: a Cuban learning experience from accidents and promotion of radiation safety culture
International Nuclear Information System (INIS)
Ferro Fernandez, Ruben; Ilizastigui Perez, Fidel; Fuente Puch, Andres de la
The Cuban Regulatory Authority is carrying out a National Program for fostering and development of Safety Culture taking into account the wide recognition of the Human factors contribution to radiological events. The program includes the introduction of new regulatory practices and initiatives in order to increase the safety culture in Cuban facilities. The most recent of those initiatives is the System for Radiological Event Analysis, Dissemination and Learning, called ADASIR (Initials of the System's name in Spanish). The main purpose of this system is to provide a better understanding and knowledge of any radiological event reported both in the country and abroad reducing, this way, the possibility of a new occurrence of a similar event in our facilities. A team of regulatory and external experts make together a detailed review of all available information about the event and identify possible root causes, failed barriers and other important data of national interest. The results of such review are documented and sent to any Cuban facility or organization with potential for similar occurrence according to the equipment and practices they have. The document includes an specific checklist which could be used by the facilities to make a self-assessment and evaluate theirs strength and weakness in regards to a similar event. This paper presents the main characteristics and results of this new experience of Cuban Regulatory Authority. (author)
Principles for the development of Aboriginal health interventions: culturally appropriate methods through systemic empathy.
Kendall, Elizabeth; Barnett, Leda
To increase Aboriginal participation in mainstream health services, it is necessary to understand the factors that influence health service usage. This knowledge can contribute to the development of culturally appropriate health services that respect Aboriginal ways of being. We used a community-based participatory approach to examine the reasons for underutilization of health services by Aboriginal Australians. Based on three focus groups and 18 interviews with Aboriginal health professionals, leaders, and community members in rural, regional, and urban settings, we identified five factors that influenced usage, including (1) negative historical experiences, (2) cultural incompetence, (3) inappropriate communication, (4) a collective approach to health, and (5) a more holistic approach to health. Given that these factors have shaped negative Aboriginal responses to health interventions, they are likely to be principles by which more appropriate solutions are generated. Although intuitively sensible and well known, these principles remain poorly understood by non-Aboriginal health systems and even less well implemented. We have conceptualized these principles as the foundation of an empathic health system. Without empathy, health systems in Australia, and internationally, will continue to face the challenge of building effective services to improve the state of health for all minority populations.
Photoperiod effect on commercial fishes cultured in different types of experimental systems
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Aragón-Flores, E.A.
Full Text Available The influence on the abiotic factors in aquaculture has been widely studied in order to control growing variables. Studies have been made to assess the positive or negative influence of photoperiod in the different life stages of some fish species. The photoperiod can influence feed visualization and social behavior in fishes; these responses influence fish growth and survival. Similarly, photoperiod plays an important role in the release of hormones that stimulate sexual development and reproduction. The aim of this review was to research the influence of photoperiod in different commercial fish species cultured in different types of experimental systems.
Radon exposure system for mammalian cells in culture: Design, operation, and dosimetry
International Nuclear Information System (INIS)
Seed, T.M.; Kretz, N.D.; Schlenker, R.A.
A novel system for Rn gas exposure of mammalian cells in culture has been designed, constructed, and used to directly assess both the magnitude and the nature of chronic, low-dose Rn/Rn daughter toxicity of exposed vital lung cells isolated from normal pulmonary tissue, propagated and exposed in vitro. Direct correlations between atmospheric Rn concentrations, alpha-particle fluences, and macro- and microdoses of absorbed radiation doses by lung cells provide for a heretofore unavailable assessment of critical doses to vital cells
Documentation of Cultural Heritages Using a GIS Based Information and Management System; Case Study of Safranbolu
Seker, D. Z.; Alkan, M.; Kutoglu, S. S.; Akcin, H.
Documentation of the cultural heritage sites is extremely important for monitoring and preserves them from natural disasters and human made activities. Due to its very rich historical background from the first human settlements in Catalhoyuk and Alacahoyuk and civilizations such as Byzantine, Seljuk and Ottoman, there are lots of cultural heritage sites in Turkey. 3D modeling and recording of historical buildings using modern tools and techniques in several locations of Turkey have been conducted and still continuing. The nine cultural sites in Turkey are included in the protection list of UNESCO as cultural heritage and one of them is the township of Safranbolu, which is the one of the most outstanding example of the traditional Turkish Architecture and also unique itself in terms of conservation of the human settlement in their authentic environmental motif up till now. In this study outcomes and further studies of a research project related to study area which is supported by the Turkish National Research Center (TUBITAK) with the project number 106Y157, will be presented in details. The basic aim of the study is development a GIS based information and management system for the city of Safranbolu. All historical buildings which are registered are assigned with the database. 3D modeling some of the selected building among the buildings which are registered as historical monuments using different data comes from different sources similar to their original constructions were realized and then it will be distributed via internet by a web-based information system designed during the project. Also some of the buildings were evaluated using close range photogrammetric technique to obtain their façade reliefs, were also assigned with the database. Designed database consists of 3D models, locations, historical information, cadastral and land register data of the selected buildings together with the other data collected during the project related to buildings. Using this
Usability, Communicability and Cultural Tourism in Interactive Systems: Trends, Economic Effects and Social Impact
Cipolla Ficarra, Francisco V.; Nicol, Emma; Cipolla-Ficarra, Miguel
We have developed a set of techniques and an analysis methodology aimed at boosting the quality of interactive tourism systems. The details of it will be presented in full and with real examples which have yielded interesting results in the last few years, both from the social and economical point of view, but with a huge wealth of cultural and natural heritage. We will also present a first guidelines to foster tourism in those villages that are willing to promote themselves in the national and international market at a low cost.
Multicenter Clinical Evaluation of BacT/Alert Virtuo Blood Culture System.
Jacobs, Michael R; Mazzulli, Tony; Hazen, Kevin C; Good, Caryn E; Abdelhamed, Ayman M; Lo, Pauline; Shum, Bianche; Roman, Katharine P; Robinson, Danielle C
BacT/Alert Virtuo is an advanced, automated blood culture system incorporating improved automation and an enhanced detection algorithm to shorten time to detection. A multicenter study of the investigational Virtuo system (bioMérieux, Inc., Durham, NC) compared to BacT/Alert 3D (BTA3D) for detection of bacteremia/fungemia in four bottle types, SA and FA Plus (aerobic) and SN and FN Plus (anaerobic), was performed in a clinical setting with patient samples in a matched system design clinical trial. Blood was added to paired aerobic or anaerobic bottles, with the volume in each bottle in each pair required to be ≤10 ml and with the volumes required to be within 30% of each other. Of 5,709 bottle sets (52.5% aerobic pairs and 47.5% anaerobic pairs), 430 (7.5%) were positive for bacterial or fungal growth, with 342 (6.0%) clinically significant and 83 (1.5%) contaminated. A total of 3,539 sets (62.0%) were volume compliant, with 203 sets (5.7%) clinically significant. The positivity rates for volume-compliant bottle pairs determined by the two systems were comparable, with 68.7% of clinically significant isolates detected by both instruments, 15.7% by Virtuo only, and 15.7% by BTA3D only. Virtuo detected microbial growth nearly 2 h sooner overall than BTA3D (mean, 15.9 h versus 17.7 h). Shorter time to detection by Virtuo was related to organism group, with the time to detection being significantly shorter for enteric Gram-negative bacilli and enterococci (means, 3.6 h and 2.3 h shorter, respectively). This large clinical study demonstrated that the Virtuo blood culture system produced results comparable to those seen with the long-established BTA3D system, with significantly shorter time to detection. Copyright © 2017 Jacobs et al.
First births with a simplified culture system for clinical IVF and embryo transfer.
Van Blerkom, Jonathan; Ombelet, Willem; Klerkx, Elke; Janssen, Mia; Dhont, Nathalie; Nargund, Geeta; Campo, Rudi
This study reports the outcome results from a pilot clinical trial using a simplified laboratory method for human IVF. This system reproducibly generates de novo the atmospheric and culture conditions that support normal fertilization and preimplantation embryogenesis to the hatched blastocyst stage without the need for specialized medical-grade gases or equipment. Development from insemination to the hatched blastocyst stage occurs undisturbed in a completely closed system that enables timed performance assessments for embryo selection in situ that, in this study, involved single-embryo transfers on day 3. With the simplified culture system, 8/23 embryos implanted, one miscarried at 8weeks of gestation and seven healthy babies have been born. The methodology and results are discussed with regard to how this simplified system can be adopted worldwide to meet the growing need for accessible and affordable IVF. A common notion concerning the demographics of infertility is that it is largely a phenomenon associated with developed countries, where infertility treatments are commonplace. In fact, most infertile couples reside in developing/low-resource countries where infertility diagnosis and treatment is nonexistent, inaccessible or unaffordable by the vast majority of young men and women in need. The irony of this situation is that bilateral tubal occlusions, for which IVF was originally indicated and is the most effective treatment, is by far the most common cause of their infertility. We have addressed one aspect of this issue, the IVF laboratory, as part of a wider effort by the Walking Egg Project to design and establish small, dedicated centres in developing countries to provide assisted reproduction technologies that are affordable and accessible to a wider proportion of the population in need. The methods for conventional IVF designed to addresses tubal obstructions are relatively simple and free of complex instrumentation and the highly developed
Development of an in situ evaluation system for neural cells using extracellular matrix-modeled gel culture.
Nagai, Takayuki; Ikegami, Yasuhiro; Mizumachi, Hideyuki; Shirakigawa, Nana; Ijima, Hiroyuki
Two-dimensional monolayer culture is the most popular cell culture method. However, the cells may not respond as they do in vivo because the culture conditions are different from in vivo conditions. However, hydrogel-embedding culture, which cultures cells in a biocompatible culture substrate, can produce in vivo-like cell responses, but in situ evaluation of cells in a gel is difficult. In this study, we realized an in vivo-like environment in vitro to produce cell responses similar to those in vivo and established an in situ evaluation system for hydrogel-embedded cell responses. The extracellular matrix (ECM)-modeled gel consisted of collagen and heparin (Hep-col) to mimic an in vivo-like environment. The Hep-col gel could immobilize growth factors, which is important for ECM functions. Neural stem/progenitor cells cultured in the Hep-col gel grew and differentiated more actively than in collagen, indicating an in vivo-like environment in the Hep-col gel. Second, a thin-layered gel culture system was developed to realize in situ evaluation of the gel-embedded cells. Cells in a 200-μm-thick gel could be evaluated clearly by a phase-contrast microscope and immunofluorescence staining through reduced optical and diffusional effects. Finally, we found that the neural cells cultured in this system had synaptic connections and neuronal action potentials by immunofluorescence staining and Ca 2+ imaging. In conclusion, this culture method may be a valuable evaluation system for neurotoxicity testing. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.
Testing a theory of organizational culture, climate and youth outcomes in child welfare systems: a United States national study.
Williams, Nathaniel J; Glisson, Charles
Theories of organizational culture and climate (OCC) applied to child welfare systems hypothesize that strategic dimensions of organizational culture influence organizational climate and that OCC explains system variance in youth outcomes. This study provides the first structural test of the direct and indirect effects of culture and climate on youth outcomes in a national sample of child welfare systems and isolates specific culture and climate dimensions most associated with youth outcomes. The study applies multilevel path analysis (ML-PA) to a U.S. nationwide sample of 2,380 youth in 73 child welfare systems participating in the second National Survey of Child and Adolescent Well-being. Youths were selected in a national, two-stage, stratified random sample design. Youths' psychosocial functioning was assessed by caregivers' responses to the Child Behavior Checklist at intake and at 18-month follow-up. OCC was assessed by front-line caseworkers' (N=1,740) aggregated responses to the Organizational Social Context measure. Comparison of the a priori and subsequent trimmed models confirmed a reduced model that excluded rigid organizational culture and explained 70% of the system variance in youth outcomes. Controlling for youth- and system-level covariates, systems with more proficient and less resistant organizational cultures exhibited more functional, more engaged, and less stressful climates. Systems with more proficient cultures and more engaged, more functional, and more stressful climates exhibited superior youth outcomes. Findings suggest child welfare administrators can support service effectiveness with interventions that improve specific dimensions of culture and climate. Copyright © 2013 Elsevier Ltd. All rights reserved.
A 3D Sphere Culture System Containing Functional Polymers for Large-Scale Human Pluripotent Stem Cell Production
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Tomomi G. Otsuji
Full Text Available Utilizing human pluripotent stem cells (hPSCs in cell-based therapy and drug discovery requires large-scale cell production. However, scaling up conventional adherent cultures presents challenges of maintaining a uniform high quality at low cost. In this regard, suspension cultures are a viable alternative, because they are scalable and do not require adhesion surfaces. 3D culture systems such as bioreactors can be exploited for large-scale production. However, the limitations of current suspension culture methods include spontaneous fusion between cell aggregates and suboptimal passaging methods by dissociation and reaggregation. 3D culture systems that dynamically stir carrier beads or cell aggregates should be refined to reduce shearing forces that damage hPSCs. Here, we report a simple 3D sphere culture system that incorporates mechanical passaging and functional polymers. This setup resolves major problems associated with suspension culture methods and dynamic stirring systems and may be optimal for applications involving large-scale hPSC production.
International Safety Management – Safety Management Systems and the Challenges of Changing a Culture
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Gregory Hanchrow
Full Text Available Over the past generation, the ISM code has brought forth tremendous opportunities to investigate and enhance the human factor in shipping through the implementation of Safety Management Systems. One of the critical factors to this implementation has been mandatory compliance and a requirement for obtaining a Document of Compliance (DOC for vessels operating globally or at least internationally. A primary objective of these systems is to maintain them as "living" or "dynamic" systems that are always evolving. As the ISM code has evolved, there have been instances where large organizations have opted to maintain a voluntary DOC from their respective class society. This has been accomplished with a large human factor element as typically an organizational culture does not always accept change readily especially if there is not a legal requirement to do so. In other words, when considering maritime training is it possible that organizations may represent cultural challenges? The intent of this paper will be to research large maritime operations that have opted for a document of compliance voluntarily and compare them to similar organizations that have been mandated by international law to do the same. The result should be to gain insight into the human factors that must contribute to a culture change in the organization for the purposes of a legal requirement versus the human factors that contribute to a voluntary establishment of a safety management system. This analysis will include both the executive decision making that designs a system implementation and the operational sector that must execute its implementation. All success and failures of education and training can be determined by the outcome. Did the training achieve its goal? Or has the education prepared the students to embrace a new idea in conjunction with a company goal or a new regulatory scheme? In qualifying the goal of a successful ISM integration by examining both
FGF1 and IGF1-conditioned 3D culture system promoted the amplification and cancer stemness of lung cancer cells.
Liu, Pengpeng; Zhang, Rui; Yu, Wenwen; Ye, Yingnan; Cheng, Yanan; Han, Lei; Dong, Li; Chen, Yongzi; Wei, Xiyin; Yu, Jinpu
Lung cancer stem cells (LCSCs) are considered as the cellular origins of metastasis and relapse of lung cancer. However, routine two-dimensional culture system (2D-culture) hardly mimics the growth and functions of LCSCs in vivo and therefore significantly decreases the stemness activity of LCSCs. In this study, we constructed a special BME-based three-dimensional culture system (3D-culture) to amplify LCSCs in human lung adenocarcinoma cell line A549 cells and found 3D-culture promoted the enrichment and amplification of LCSCs in A549 cells displaying higher proliferation potential and invasion activity, but lower apoptosis. The expression and secretion levels of FGF1 and IGF1 were dramatically elevated in 3D-culture compared to 2D-culture. After growing in FGF1 and IGF1-conditioned 3D-culture, the proportion of LCSCs with specific stemness phenotypes in A549 cells significantly increased compared to that in conventional 3D suspension culture system. Further results indicated that FGF1 and IGF1 promoted the amplification and cancer stemness of LCSCs dependent on MAPK signaling pathway. Our data firstly established a growth factors-conditioned 3D-culture for LCSCs and demonstrated the effects of FGF1 and IGF1 in promoting the enrichment and amplification of LCSCs which might provide a feasible cell model in vitro for both mechanism study and translational research on lung cancer. Copyright © 2017 Elsevier Ltd. All rights reserved.
A system to preserve pedicab as cultural heritage in Solo city, Indonesia
Sari, Wahyu Apri W.; Handayani, Cahyaning D.; Putri, Yulina Rahayu L.; Dewa, Ratna T.
Pedicab, a traditional three-wheeled transportation operated manually, has become the cultural heritage for decades, or even before the Indonesia Independence Day. Today, this kind of traditional transportation ought to compete with other modern transportation modes, such as bus, motorcycle, and taxi. This research investigated the possibility to divert the function of pedicab from conventional usage to the tourism usage that can provide more additional value to compete with other means of transportations. Descriptive qualitative was used as the method to gather the information, in addition to literature studies on the previous researches and the fundamental theories. We also did some interviews to the both sides, namely the public transportation users and the pedicab drivers. However, a city will be recognized if it has many uniqueness and cultural senses. By this consideration, it is expected that the pedicab drivers will have a capacity to approach and introduce their services directly to the tourists, which is supported by additional support systems, such as information and training system. Hopefully, the existence of pedicab can still be preserved for many decades ahead.
Two sides of the same coin? The (techno)epistemic cultures of systems and synthetic biology.
Kastenhofer, Karen
Systems and synthetic biology both emerged around the turn of this century as labels for new research approaches. Although their disciplinary status as well as their relation to each other is rarely discussed in depth, now and again the idea is invoked that both approaches represent 'two sides of the same coin'. The following paper focuses on this general notion and compares it with empirical findings concerning the epistemic cultures prevalent in the two contexts. Drawing on interviews with researchers from both fields, on participatory observation in conferences and courses and on documentary analysis, this paper delineates differences and similarities, incompatibilities and blurred boundaries. By reconstructing systems and synthetic biology's epistemic cultures, this paper argues that they represent two 'communities of vision', encompassing heterogeneous practices. Understanding the relation of the respective visions of understanding nature and engineering life is seen as indispensible for the characterisation of (techno)science in more general terms. Depending on the conceptualisation of understanding and construction (or: science and engineering), related practices such as in silico modelling for enhancing understanding or enabling engineering can either be seen as incommensurable or 'two sides of one coin'. Copyright © 2013 Elsevier Ltd. All rights reserved.
Critical Incident Stress Management (CISM) in complex systems: cultural adaptation and safety impacts in healthcare.
Müller-Leonhardt, Alice; Mitchell, Shannon G; Vogt, Joachim; Schürmann, Tim
In complex systems, such as hospitals or air traffic control operations, critical incidents (CIs) are unavoidable. These incidents can not only become critical for victims but also for professionals working at the "sharp end" who may have to deal with critical incident stress (CIS) reactions that may be severe and impede emotional, physical, cognitive and social functioning. These CIS reactions may occur not only under exceptional conditions but also during every-day work and become an important safety issue. In contrast to air traffic management (ATM) operations in Europe, which have readily adopted critical incident stress management (CISM), most hospitals have not yet implemented comprehensive peer support programs. This survey was conducted in 2010 at the only European general hospital setting which implemented CISM program since 2004. The aim of the article is to describe possible contribution of CISM in hospital settings framed from the perspective of organizational safety and individual health for healthcare professionals. Findings affirm that daily work related incidents also can become critical for healthcare professionals. Program efficiency appears to be influenced by the professional culture, as well as organizational structure and policies. Overall, findings demonstrate that the adaptation of the CISM program in general hospitals takes time but, once established, it may serve as a mechanism for changing professional culture, thereby permitting the framing of even small incidents or near misses as an opportunity to provide valuable feedback to the system. Copyright © 2014 Elsevier Ltd. All rights reserved.
First report of Fusarium oxysporum species complex infection in zebrafish culturing system.
Kulatunga, D C M; Dananjaya, S H S; Park, B K; Kim, C-H; Lee, J; De Zoysa, M
Fusarium oxysporum species complex (FOSC) is a highly diverse fungus. Recently, F. oxysporum infection was identified from zebrafish (Danio rerio) culturing system in Korea. Initially, a rapid whitish smudge was appeared in the water with the fungal blooming on walls of fish tanks. Microscopic studies were conducted on fungal hyphae, colony pigmentation and chlamydospore formation and the presence of macro- and microspores confirmed that the isolated fungus as F. oxysporum. Furthermore, isolated F. oxysporum was confirmed by internal transcribed spacer sequencing which matched (100%) to nine F. oxysporum sequences available in GenBank. Experimental hypodermic injection of F. oxysporum into adult zebrafish showed the development of fungal mycelium and pathogenicity similar to signs observed. Histopathologic results revealed a presence of F. oxysporum hyphae in zebrafish muscle. Fusarium oxysporum growth was increased with sea salt in a concentration-dependent manner. Antifungal susceptibility results revealed that F. oxysporum is resistant to copper sulphate (up to 200 μg mL -1 ) and sensitive to nystatin (up to 40 μg mL -1 ). This is the first report of FOSC from zebrafish culture system, suggesting it appears as an emerging pathogen, thus posing a significant risk on zebrafish facilities in the world. © 2016 John Wiley & Sons Ltd.
Customization of ¹³C-MFA strategy according to cell culture system.
Quek, Lake-Ee; Nielsen, Lars K
(13)C-MFA is far from being a simple assay for quantifying metabolic activity. It requires considerable up-front experimental planning and familiarity with the cell culture system in question, as well as optimized analytics and adequate computation frameworks. The success of a (13)C-MFA experiment is ultimately rated by the ability to accurately quantify the flux of one or more reactions of interest. In this chapter, we describe the different (13)C-MFA strategies that have been developed for the various fermentation or cell culture systems, as well as the limitations of the respective strategies. The strategies are affected by many factors and the (13)C-MFA modeling and experimental strategy must be tailored to conditions. The prevailing philosophy in the computation process is that any metabolic processes that produce significant systematic bias in the labeling pattern of the metabolites being measured must be described in the model. It is equally important to plan a labeling strategy by analytical screening or by heuristics.
Management Systems and Safety Culture in the Nuclear Energy Sector (ISO 9001 & GS-R-3)
International Nuclear Information System (INIS)
Smetnik, A.; Murlis, D.
Nowadays, the enterprises of the Rosatom State Nuclear Energy Corporation that provides products and services to foreign customers should rely on the requirements to the management systems established by the IAEA Standard GS-R-3 "The management system for facilities and activities". This results from the fact that in order to enter foreign markets, Russian suppliers have to meet foreign requirements related to quality assurance, protection of the environment, nuclear and radiation safety, etc. For instance, the Finnish customer "Fennovoima" requires full compliance of the management systems of the Russian companies involved in the construction of the Hanhikivi-1 NPP with the GS-R-3 Standard. ISO 9001 quality management systems were widely implemented in the nuclear industry enterprises in Russia. The assessment of compliance of the quality management systems with the established requirements is carried out by the certification bodies. The same relates to the environmental management systems that are implemented at the majority of nuclear industry facilities in Russia. But due to their uniqueness and associated significant risks, the nuclear industry enterprises have to meet current safety requirements and principles established in the IAEA Safety Standards, such as safety culture and risk management.
Retinoid production using metabolically engineered Escherichia coli with a two-phase culture system.
Jang, Hui-Jeong; Yoon, Sang-Hwal; Ryu, Hee-Kyung; Kim, Jung-Hun; Wang, Chong-Long; Kim, Jae-Yean; Oh, Deok-Kun; Kim, Seon-Won
Retinoids are lipophilic isoprenoids composed of a cyclic group and a linear chain with a hydrophilic end group. These compounds include retinol, retinal, retinoic acid, retinyl esters, and various derivatives of these structures. Retinoids are used as cosmetic agents and effective pharmaceuticals for skin diseases. Retinal, an immediate precursor of retinoids, is derived by β-carotene 15,15'-mono(di)oxygenase (BCM(D)O) from β-carotene, which is synthesized from the isoprenoid building blocks isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Retinoids are chemically unstable and biologically degraded via retinoic acid. Although extensive studies have been performed on the microbial production of carotenoids, retinoid production using microbial metabolic engineering has not been reported. Here, we report retinoid production using engineered Escherichia coli that express exogenous BCM(D)O and the mevalonate (MVA) pathway for the building blocks synthesis in combination with a two-phase culture system using a dodecane overlay. Among the BCM(D)O tested in E. coli, the synthetic retinoid synthesis protein (SR), based on bacteriorhodopsin-related protein-like homolog (Blh) of the uncultured marine bacteria 66A03, showed the highest β-carotene cleavage activity with no residual intracellular β-carotene. By introducing the exogenous MVA pathway, 8.7 mg/L of retinal was produced, which is 4-fold higher production than that of augmenting the MEP pathway (dxs overexpression). There was a large gap between retinal production and β-carotene consumption using the exogenous MVA pathway; therefore, the retinal derivatives were analyzed. The derivatives, except for retinoic acid, that formed were identified, and the levels of retinal, retinol, and retinyl acetate were measured. Amounts as high as 95 mg/L retinoids were obtained from engineered E. coli DH5α harboring the synthetic SR gene and the exogenous MVA pathway in addition to dxs overexpression, which
Application of three-dimensional culture systems to study mammalian spermatogenesis, with an emphasis on the rhesus monkey (Macaca mulatta
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Mahmoud Huleihel
Full Text Available In vitro culture of spermatogonial stem cells (SSCs has generally been performed using two-dimensional (2D culture systems; however, such cultures have not led to the development of complete spermatogenesis. It seems that 2D systems do not replicate optimal conditions of the seminiferous tubules (including those generated by the SSC niche and necessary for spermatogenesis. Recently, one of our laboratories has been able to induce proliferation and differentiation of mouse testicular germ cells to meiotic and postmeiotic stages including generation of sperm in a 3D soft agar culture system (SACS and a 3D methylcellulose culture system (MCS. It was suggested that SACS and MCS form a special 3D microenvironment that mimics germ cell niche formation in the seminiferous tubules, and thus permits mouse spermatogenesis in vitro. In this review, we (1 provide a brief overview of the differences in spermatogenesis in rodents and primates, (2 summarize data related to attempts to generate sperm in vitro, (3 report for the first time formation of colonies/clusters of cells and differentiation of meiotic (expression of CREM-1 and postmeiotic (expression of acrosin germ cells from undifferentiated spermatogonia isolated from the testis of prepubertal rhesus monkeys and cultured in SACS and MCS, and (4 indicate research needed to optimize 3D systems for in vitroprimate spermatogenesis and for possible future application to man.
Cell fiber-based three-dimensional culture system for highly efficient expansion of human induced pluripotent stem cells.
Ikeda, Kazuhiro; Nagata, Shogo; Okitsu, Teru; Takeuchi, Shoji
Human pluripotent stem cells are a potentially powerful cellular resource for application in regenerative medicine. Because such applications require large numbers of human pluripotent stem cell-derived cells, a scalable culture system of human pluripotent stem cell needs to be developed. Several suspension culture systems for human pluripotent stem cell expansion exist; however, it is difficult to control the thickness of cell aggregations in these systems, leading to increased cell death likely caused by limited diffusion of gases and nutrients into the aggregations. Here, we describe a scalable culture system using the cell fiber technology for the expansion of human induced pluripotent stem (iPS) cells. The cells were encapsulated and cultured within the core region of core-shell hydrogel microfibers, resulting in the formation of rod-shaped or fiber-shaped cell aggregations with sustained thickness and high viability. By encapsulating the cells with type I collagen, we demonstrated a long-term culture of the cells by serial passaging at a high expansion rate (14-fold in four days) while retaining its pluripotency. Therefore, our culture system could be used for large-scale expansion of human pluripotent stem cells for use in regenerative medicine.
Publishable Manuscript on Cross-cultural Comparison of Systems of Provision in the Food Sector
DEFF Research Database (Denmark)
Reisch, Lucia A.; Gwozdz, Wencke; Rito, Ana
The goal of this report is to analyse and compare cross-culturally systems of provision of food, e.g., retail structure, eating out (school lunches, restaurants, catering) in Europe (Task 7.3.4). The basic assumption is that a highly concentrated retail structure as well as children's immediate...... food environments are overwhelmingly "obesogenic" and hence contribute to the obesity pandemic. The report starts with a general overview of the respective systems of food provision, namely the retail structures and the market environment in Europe and the I.Family countries in particular....... It then looks into families' eating out and discusses the causes and consequences of out-of-home eating in schools, restaurants and other places. Wrapping up, the report then reviews policies for healthier out-of-home eating in general and addresses the school food environment in particular. We conclude...
Successful long-term maintenance of Mansonella perstans in an in vitro culture system
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Abdel Jelil Njouendou
Full Text Available Abstract Background Approximately 114 million people are infected with Mansonella perstans in large proportions of Africa. In contrast to other filariae that infect humans, M. perstans-infected individuals show no distinct pathology or specific clinical picture, indicating a well-tuned adaptation to the host. In addition, since M. perstans adult worms reside in serous cavities which are difficult to access, research has been hindered and there is a paucity of knowledge about the biology of M. perstans, especially the development of the different life stages as well as M. perstans-driven immune responses. Thus in this study, an in vitro culture system was developed which allows an in-depth analysis of M. perstans. Results Culicoides species were caught in Ediki (Kumba, Southwest Region within Cameroon following a blood meal on a microfilaremic donor that had 1500 microfilariae/ml of peripheral blood and kept in captivity for 12 days at 23 °C. In a pilot experiment, 15 infective larvae were obtained from the midges and co-cultured with a confluent monolayer of monkey kidney epithelial cells (LLC-MK2 in DMEM medium supplemented with 10% FBS for up to 77 days. The resulting survival rates of 33% revealed that the cell-conditioned medium was suitable for long-term maintenance of M. perstans worms. To confirm these preliminary observations, 249 infective larvae were cultured for 50 days and their development was monitored daily and microscopically graded for motility. In total, 170 (68.3% filariae survived and 124 (49.8% larvae moulted between days 21–30 to become L5 stage larvae which were motile and showed continuous vigorous movement. Conclusion We have established an in vitro culture system for the generation and long-term maintenance of viable M. perstans worms. This technique will be an important tool to study parasite biology and development, the role in host immunity, and might be helpful to discover novel treatment strategies against
Incorporating Hofstede' National Culture in Human Factor Analysis and Classification System (HFACS: Cases of Indonesian Aviation Safety
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Pratama Gradiyan Budi
Full Text Available National culture plays an important role in the application of ergonomics and safety. This research examined role of national culture in accident analysis of Indonesian aviation using framework of Human Factors Analysis and Classification System (HFACS. 53 Indonesian aviation accidents during year of 2001-2012 were analyzed using the HFACS framework by authors and were validated to 14 air-transport experts in Indonesia. National culture is viewed with Hofstede' lens of national culture. Result shows that high collectivistic, low uncertainty avoidance, high power distance, and masculinity dimension which are characteristics of Indonesian culture, play an important role in Indonesian aviation accident and should be incorporated within HFACS. Result is discussed in relation with HFACS and Indonesian aviation accident analysis.
Introduction to polyphasic dispersed systems theory application to open systems of microorganisms' culture
Thierie, Jacques
This book introduces a new paradigm in system description and modelling. The author shows the theoretical and practical successes of his approach, which involves replacing a traditional uniform description with a polyphasic description. This change of perspective reveals new fluxes that are cryptic in the classical description. Several case studies are given in this book, which is of interest of those working with biotechnology and green chemistry.
Mouse embryonic stem cell culture for generation of three-dimensional retinal and cortical tissues.
Eiraku, Mototsugu; Sasai, Yoshiki
Generation of compound tissues with complex structures is a major challenge in cell biology. In this article, we describe a protocol for mouse embryonic stem cell (ESC) culture for in vitro generation of three-dimensional retinal tissue, comparing it with the culture protocol for cortical tissue generation. Dissociated ESCs are reaggregated in a 96-well plate with reduced cell-plate adhesion and cultured as floating aggregates. Retinal epithelium is efficiently generated when ESC aggregates are cultured in serum-free medium containing extracellular matrix proteins, spontaneously forming hemispherical vesicles and then progressively transforming into a shape reminiscent of the embryonic optic cup in 9-10 d. In long-term culture, the ESC-derived optic cup generates a fully stratified retinal tissue consisting of all major neural retinal components. In contrast, the cortical differentiation culture can be started without exogenous extracellular matrix proteins, and it generates stratified cortical epithelia consisting of four distinct layers in 13 d.
Large-scale infection of the ascidian Ciona intestinalis by the gregarine Lankesteria ascidiae in an inland culture system.
Mita, Kaoru; Kawai, Narudo; Rueckert, Sonja; Sasakura, Yasunori
An important way to keep transgenic and mutant lines of the ascidian Ciona intestinalis, a model system for e.g. genetic functions, in laboratories is via culturing systems. Here we report a disease of C. intestinalis observed in an inland culturing system. The disease, called 'long feces syndrome,' is expressed in affected animals by the following characteristic symptoms of the digestive system: (1) excretion of long and thin feces, (2) pale color of the stomach, and (3) congestion of the digestive tube by digested material. Severely diseased animals usually die within a week after the first symptoms occur, implying a high risk of this disease for ascidian culturing systems. The digestive tubes of the diseased animals are occupied by the gregarine apicomplexan parasite Lankesteria ascidiae, suggesting that large-scale infection by this parasite is the cause of long feces syndrome.
Changing the system by changing the workforce: employing consumers to increase access, cultural diversity, and engagement.
Wenz-Gross, Melodie; Irsfeld, Toni DuBrino; Twomey, Tammy; Perez, Ana; Thompson, Judith; Wally, Martha; Colleton, Barbara; Kroell, Christine; McKeown, Steven K; Metz, Peter
Services to families have traditionally been delivered in a medical model. This presents challenges including workforce shortages, lack of cultural diversity, lack of training in strength-based work, and difficulty in successfully engaging and retaining families in the therapy process. The system of care (SOC) effort has worked to establish formal roles for caregivers in SOC to improve services. This paper provides an example of one community's efforts to change the SOC by expanding the roles available to caregivers in creating systems change. It describes the model developed by Communities of Care (CoC), a SOC in Central Massachusetts, and its evolution over a 10 year period. First person accounts by system partners, caregivers hired into professional roles as well as a family receiving services, demonstrate how hiring caregivers at all levels can change systems and change lives, not only for those being served but for the caregiver/professionals doing the work. It also demonstrates, however, that change at the system level is incremental, takes time, and can be fleeting unless an ongoing effort is made to support and sustain those changes.
Navigating oceans and cultures: Polynesian and European navigation systems in the late eighteenth century
Walker, M.
Significant differences in the rotation of the celestial dome between the tropical and temperate zones did not stop the peoples of either the tropical Pacific or temperate Europe from using geocentric astronomy to guide exploration of the oceans. Although the differences in the night sky contributed to differences between the Pacific Island and European systems for navigation at sea, the two navigation systems exhibit substantial similarities. Both systems define positions on the surface of the Earth using two coordinates that vary at right angles to each other and use stars, and to a lesser extent the sun, to determine directions. This essay explores similarities and differences in the use of geocentric astronomy for navigation at sea by the peoples of Polynesia and Europe in the late eighteenth century. Captain Cook's orders to discover the unknown southern continent after observing the transit of Venus combined with differences in language and culture to obscure the deeper similarities between the navigation systems used by Cook and the Polynesians. Although it was a further 200 years before anthropologists studied Pacific navigation, collaborations in voyaging with communities in Oceania demonstrated the effectiveness of Pacific navigation systems, revived interest in traditional voyaging in island communities around the Pacific, and potentially open the way for further collaborations in other areas.
Establishment of a Novel Lingual Organoid Culture System: Generation of Organoids Having Mature Keratinized Epithelium from Adult Epithelial Stem Cells
Hisha, Hiroko; Tanaka, Toshihiro; Kanno, Shohei; Tokuyama, Yoko; Komai, Yoshihiro; Ohe, Shuichi; Yanai, Hirotsugu; Omachi, Taichi; Ueno, Hiroo
Despite the strong need for the establishment of a lingual epithelial cell culture system, a simple and convenient culture method has not yet been established. Here, we report the establishment of a novel lingual epithelium organoid culture system using a three-dimensional matrix and growth factors. Histological analyses showed that the generated organoids had both a stratified squamous epithelial cell layer and a stratum corneum. Very recently, we showed via a multicolor lineage tracing method that Bmi1-positive stem cells exist at the base of the epithelial basal layer in the interpapillary pit. Using our new culture system, we found that organoids could be generated by single Bmi1-positive stem cells and that in the established organoids, multiple Bmi1-positive stem cells were generated at the outermost layer. Moreover, we observed that organoids harvested at an early point in culture could be engrafted and maturate in the tongue of recipient mice and that the organoids generated from carcinogen-treated mice had an abnormal morphology. Thus, this culture system presents valuable settings for studying not only the regulatory mechanisms of lingual epithelium but also lingual regeneration and carcinogenesis.
Enhanced removal of petroleum hydrocarbons using a bioelectrochemical remediation system with pre-cultured anodes
International Nuclear Information System (INIS)
Venkidusamy, Krishnaveni; Megharaj, Mallavarapu; Marzorati, Massimo; Lockington, Robin; Naidu, Ravi
Bioelectrochemical remediation (BER) systems such as microbial fuel cells (MFCs) have recently emerged as a green technology for the effective remediation of petroleum hydrocarbon contaminants (PH) coupled with simultaneous energy recovery. Recent research has shown that biofilms previously enriched for substrate degrading bacteria resulted in excellent performance in terms of substrate removal and electricity generation but the effects on hydrocarbon contaminant degradation were not examined. Here we investigate the differences between enriched biofilm anodes and freshly inoculated new anodes in diesel fed single chamber mediatorless microbial fuel cells (DMFC) using various techniques for the enhancement of PH contaminant remediation with concomitant electricity generation. An anodophilic microbial consortium previously selected for over a year through continuous culturing with a diesel concentration of about 800 mg l"−"1 and which now showed complete removal of this concentration of diesel within 30 days was compared to that of a freshly inoculated new anode MFC (showing 83.4% removal of diesel) with a simultaneous power generation of 90.81 mW/m"2 and 15.04 mW/m"2 respectively. The behaviour of pre-cultured anodes at a higher concentration of PH (8000 mg l"−"1) was also investigated. Scanning electron microscopy observation revealed a thick biofilm covering the pre-cultured anodic electrode but not the anode from the freshly inoculated MFC. High resolution imaging showed the presence of thin 60 nm diametre pilus-like projections emanating from the cells. Anodic microbial community profiling confirmed that the selection for diesel degrading exoelectrogenic bacteria had occurred. Identification of a biodegradative gene (alkB) provided strong evidence of the catabolic pathway used for diesel degradation in the DMFCs.