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Characterization of an extracellular serine protease of Leishmania (Leishmania) amazonensis.
Silva-Lopez, R E; Coelho, M G Pinto; De Simone, S G
A serine protease was purified 942-fold from culture supernatant of L. amazonensis promastigotes using (NH4)2SO4 precipitation followed by affinity chromatography on aprotinin-agarose and continuous elution electrophoresis by Prep Cell, yielding a total recovery of 61%. The molecular mass of the active enzyme estimated by SDS-PAGE under conditions of reduction was 56 kDa and 115 kDa under conditions of non-reduction, suggesting that the protease is a dimeric protein. Additionally, it was found to be a non-glycosylated enzyme, with a pI of 5.0. The optimal pH and temperature of the enzyme were 7.5 and 28 degrees C respectively, using alpha-N-rho-tosyl-L-arginine-methyl ester (L-TAME) as substrate. Assays of thermal stability indicated that 61% of the enzyme activity was preserved after 1 h of pre-treatment at 42 degrees C. Haemoglobin, bovine serum albumin (BSA), ovalbumin, fibrinogen, collagen, gelatin and peptide substrates containing arginine in an ester bond and amide substrates containing hydrophobic residues at the P1 site were hydrolysed by this extracellular protease. The insulin beta-chain was also hydrolysed by the enzyme and many peptidic bonds were susceptible to the protease action, and 4 of them (L11-V12, E3-A14, L15-Y16 and Y16-L17) were identified. Inhibition studies suggested that the enzyme belongs to the serine protease class inhibited by calcium and manganese and activated by zinc. These findings show that this enzyme of L. amazonensis is a novel serine protease, which differs from all known flagellate proteases characterized. PMID:16038400
The Comparative Genomics and Phylogenomics of Leishmania amazonensis Parasite
Tschoeke, Diogo A; Nunes, Gisele L; Jardim, Rodrigo; Lima, Joana; Dumaresq, Aline SR; Gomes, Monete R; de Mattos Pereira, Leandro; Loureiro, Daniel R; Stoco, Patricia H; de Matos Guedes, Herbert Leonel; de Miranda, Antonio Basilio; Ruiz, Jeronimo; Pitaluga, André; Silva, Floriano P; Probst, Christian M; Dickens, Nicholas J; Mottram, Jeremy C; Grisard, Edmundo C; Dávila, Alberto MR
Leishmaniasis is an infectious disease caused by Leishmania species. Leishmania amazonensis is a New World Leishmania species belonging to the Mexicana complex, which is able to cause all types of leishmaniasis infections. The L. amazonensis reference strain MHOM/BR/1973/M2269 was sequenced identifying 8,802 codifying sequences (CDS), most of them of hypothetical function. Comparative analysis using six Leishmania species showed a core set of 7,016 orthologs. L. amazonensis and Leishmania mexicana share the largest number of distinct orthologs, while Leishmania braziliensis presented the largest number of inparalogs. Additionally, phylogenomic analysis confirmed the taxonomic position for L. amazonensis within the "Mexicana complex", reinforcing understanding of the split of New and Old World Leishmania. Potential non-homologous isofunctional enzymes (NISE) were identified between L. amazonensis and Homo sapiens that could provide new drug targets for development. PMID:25336895
Leishmania amazonensis: PKC-like protein kinase modulates the (Na++K+)ATPase activity.
Almeida-Amaral, Elmo Eduardo de; Caruso-Neves, Celso; Lara, Lucienne Silva; Pinheiro, Carla Mônica; Meyer-Fernandes, José Roberto
The present study aimed to identify the presence of protein kinase C-like (PKC-like) in Leishmania amazonensis and to elucidate its possible role in the modulation of the (Na(+)+K(+))ATPase activity. Immunoblotting experiments using antibody against a consensus sequence (Ac 543-549) of rabbit protein kinase C (PKC) revealed the presence of a protein kinase of 80 kDa in L. amazonensis. Measurements of protein kinase activity showed the presence of both (Ca(2+)-dependent) and (Ca(2+)-independent) protein kinase activity in plasma membrane and cytosol. Phorbol ester (PMA) activation of the Ca(2+)-dependent protein kinase stimulated the (Na(+)+K(+))ATPase activity, while activation of the Ca(2+)-independent protein kinase was inhibitory. Both effects of protein kinase on the (Na(+)+K(+))ATPase of the plasma membrane were lower than that observed in intact cells. PMA induced the translocation of protein kinase from cytosol to plasma membrane, indicating that the maximal effect of protein kinase on the (Na(+)+K(+))ATPase activity depends on the synergistic action of protein kinases from both plasma membrane and cytosol. This is the first demonstration of a protein kinase activated by PMA in L. amazonensis and the first evidence for a possible role in the regulation of the (Na(+)+K(+))ATPase activity in this trypanosomatid. Modulation of the (Na(+)+K(+))ATPase by protein kinase in a trypanosomatid opens up new possibilities to understand the regulation of ion homeostasis in this parasite. PMID:17475255
Characterization of an antigen from Leishmania amazonensis amastigotes able to elicit protective responses in a murine model.
Beyrodt, C G; Pinto, A R; Freymüller, E; Barbiéri, C L
Lymphoproliferative responses to an antigen from Leishmania amazonensis amastigotes with an apparent molecular mass of 30 kDa, termed p30, were evaluated with BALB/c mice. The p30 antigen was purified after separation of parasite extracts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroelution. Lymphoproliferative responses to p30 were obtained by subcutaneous immunization of animals with L. amazonensis amastigote extracts, and maximal stimulation indices were observed at an antigen concentration of 5 microg/ml. Induction of lymphoproliferation by p30 is stage specific, and no differences in the responses to this antigen between mice susceptible and resistant to L. amazonensis were detected. The predominant T cells characterized in the lymphocyte cultures were CD4+. Lymphokine analysis of the supernatants from these cultures indicated that Th1 is the subset involved in the lymphoproliferative responses to the antigen. BALB/c mice immunized with p30 and challenged with L. amazonensis amastigotes showed a very low level of infection, indicating a protective role for p30 and a correlation between Th1 and protection. Further biochemical characterization studies showed that this antigen presents cysteine proteinase activity. PMID:9169731
Immunoproteomic and bioinformatic approaches to identify secreted Leishmania amazonensis, L. braziliensis, and L. infantum proteins with specific reactivity using canine serum.
Lima, B S S; Fialho, L C; Pires, S F; Tafuri, W L; Andrade, H M
Leishmania spp have a wide range of hosts, and each host can harbor several Leishmania species. Dogs, for example, are frequently infected by Leishmania infantum, where they constitute its main reservoir, but they also serve as hosts for L. braziliensis and L. amazonensis. Serological tests for antibody detection are valuable tools for diagnosis of L. infantum infection due to the high levels of antibodies induced, unlike what is observed in L. amazonensis and L. braziliensis infections. Likewise, serology-based antigen-detection can be useful as an approach to diagnose any Leishmania species infection using different corporal fluid samples. Immunogenic and secreted proteins constitute powerful targets for diagnostic methods in antigen detection. As such, we performed immunoproteomic (2-DE, western blot and mass spectrometry) and bioinformatic screening to search for reactive and secreted proteins from L. amazonensis, L. braziliensis, and L. infantum. Twenty-eight non-redundant proteins were identified, among which, six were reactive only in L. amazonensis extracts, 10 in L. braziliensis extracts, and seven in L. infantum extracts. After bioinformatic analysis, seven proteins were predicted to be secreted, two of which were reactive only in L. amazonensis extracts (52kDa PDI and the glucose-regulated protein 78), one in L. braziliensis extracts (pyruvate dehydrogenase E1 beta subunit) and three in L. infantum extracts (two conserved hypothetical proteins and elongation factor 1-beta). We propose that proteins can be suitable targets for diagnostic methods based on antigen detection. PMID:27198787
Leishmania amazonensis: characterization of an ecto-pyrophosphatase activity.
Freitas-Mesquita, Anita Leocadio; Fonseca-de-Souza, André Luiz; Meyer-Fernandes, José Roberto
Several ecto-enzymatic activities have been described in the plasma membrane of the protozoan Leishmania amazonensis, which is the major etiological agent of diffuse cutaneous leishmaniasis in South America. These enzymes, including ecto-phosphatases, contribute to the survival of the parasite by participating in phosphate metabolism. This work identifies and characterizes the extracellular hydrolysis of inorganic pyrophosphate related to an ecto-pyrophosphatase activity of the promastigote form of L. amazonensis. This ecto-pyrophosphatase activity is insensitive to MnCl2 but is strongly stimulated by MgCl2. This stimulation was not observed during the hydrolysis of p-nitrophenyl phosphate (p-NPP) or β-glycerophosphate, two substrates for different ecto-phosphatases present in the L. amazonensis plasma membrane. Furthermore, extracellular PPi hydrolysis is more efficient at alkaline pHs, while p-NPP hydrolysis occurs mainly at acidic pHs. These results led us to conclude that extracellular PPi is hydrolyzed not by non-specific ecto-phosphatases but rather by a genuine ecto-pyrophosphatase. In the presence of 5mM MgCl2, the ecto-pyrophosphatase activity from L. amazonensis is sensitive to micromolar concentrations of NaF and millimolar concentrations of CaCl2. Moreover, this activity is significantly higher during the first days of L. amazonensis culture, which suggests a possible role for this enzyme in parasite growth. PMID:24316462
Chronic infection by Leishmania amazonensis mediated through MAPK ERK mechanisms
Martinez, Pedro A.; Petersen, Christine A.
Leishmania amazonensis is an intracellular protozoan parasite responsible for chronic cutaneous leishmaniasis (CL). CL is a neglected tropical disease responsible for infecting millions of people worldwide. L. amazonensis promotes alteration of various signaling pathways that are essential for host cell survival. Specifically, through parasite-mediated phosphorylation of extracellular signal regulated kinase (ERK), L. amazonensis inhibits cell-mediated parasite killing and promotes its own survival by co-opting multiple host cell functions. In this review we highlight Leishmania-host cell signaling alterations focusing on those specific to 1) motor proteins, 2) prevention of NADPH subunit phosphorylation impairing reactive oxygen species production (ROS), and 3) localized endosomal signaling to up-regulate ERK phosphorylation. This review will focus upon mechanisms and possible explanations as to how Leishmania spp. evades the various layers of defense employed by the host immune response. PMID:24838145
LaRbp38: A Leishmania amazonensis protein that binds nuclear and kinetoplast DNAs
Lira, C.B.B.; Siqueira Neto, J.L.; Giardini, M.A.; Winck, F.V.; Ramos, C.H.I.; Cano, M.I.N. . E-mail: email@example.com
Leishmania amazonensis causes a wide spectrum of leishmaniasis. There are no vaccines or adequate treatment for leishmaniasis, therefore there is considerable interest in the identification of new targets for anti-leishmania drugs. The central role of telomere-binding proteins in cell maintenance makes these proteins potential targets for new drugs. In this work, we used a combination of purification chromatographies to screen L. amazonensis proteins for molecules capable of binding double-stranded telomeric DNA. This approach resulted in the purification of a 38 kDa polypeptide that was identified by mass spectrometry as Rbp38, a trypanosomatid protein previously shown to stabilize mitochondrial RNA and to associate with nuclear and kinetoplast DNAs. Western blotting and supershift assays confirmed the identity of the protein as LaRbp38. Competition and chromatin immunoprecipitation assays confirmed that LaRbp38 interacted with kinetoplast and nuclear DNAs in vivo and suggested that LaRbp38 may have dual cellular localization and more than one function.
HSP70 of Leishmania amazonensis alters resistance to different stresses and mitochondrial bioenergetics
Codonho, Bárbara Santoni; Costa, Solange dos Santos; Peloso, Eduardo de Figueiredo; Joazeiro, Paulo Pinto; Gadelha, Fernanda Ramos; Giorgio, Selma
The 70 kDa heat shock protein (HSP70) is a molecular chaperone that assists the parasite Leishmania in returning to homeostasis after being subjected to different types of stress during its life cycle. In the present study, we evaluated the effects of HSP70 transfection of L. amazonensis promastigotes (pTEX-HSP70) in terms of morphology, resistance, infectivity and mitochondrial bioenergetics. The pTEX-HSP70 promastigotes showed no ultrastructural morphological changes compared to control parasites. Interestingly, the pTEX-HSP70 promastigotes are resistant to heat shock, H2O2-induced oxidative stress and hyperbaric environments. Regarding the bioenergetics parameters, the pTEX-HSP70 parasites had higher respiratory rates and released less H2O2 than the control parasites. Nevertheless, the infectivity capacity of the parasites did not change, as verified by the infection of murine peritoneal macrophages and human macrophages, as well as the infection of BALB/c mice. Together, these results indicate that the overexpression of HSP70 protects L. amazonensis from stress, but does not interfere with its infective capacity. PMID:27304024
HSP70 of Leishmania amazonensis alters resistance to different stresses and mitochondrial bioenergetics.
Codonho, Bárbara Santoni; Costa, Solange Dos Santos; Peloso, Eduardo de Figueiredo; Joazeiro, Paulo Pinto; Gadelha, Fernanda Ramos; Giorgio, Selma
The 70 kDa heat shock protein (HSP70) is a molecular chaperone that assists the parasite Leishmania in returning to homeostasis after being subjected to different types of stress during its life cycle. In the present study, we evaluated the effects of HSP70 transfection of L. amazonensis promastigotes (pTEX-HSP70) in terms of morphology, resistance, infectivity and mitochondrial bioenergetics. The pTEX-HSP70 promastigotes showed no ultrastructural morphological changes compared to control parasites. Interestingly, the pTEX-HSP70 promastigotes are resistant to heat shock, H2O2-induced oxidative stress and hyperbaric environments. Regarding the bioenergetics parameters, the pTEX-HSP70 parasites had higher respiratory rates and released less H2O2 than the control parasites. Nevertheless, the infectivity capacity of the parasites did not change, as verified by the infection of murine peritoneal macrophages and human macrophages, as well as the infection of BALB/c mice. Together, these results indicate that the overexpression of HSP70 protects L. amazonensis from stress, but does not interfere with its infective capacity. PMID:27304024
Activity of Cuban Plants Extracts against Leishmania amazonensis
García, Marley; Monzote, Lianet; Scull, Ramón; Herrera, Pedro
Natural products have long been providing important drug leads for infectious diseases. Leishmaniasis is a major health problem worldwide that affects millions of people especially in the developing nations. There is no immunoprophylaxis (vaccination) available for Leishmania infections, and conventional treatments are unsatisfactory; therefore, antileishmanial drugs are urgently needed. In this work, 48 alcoholic extracts from 46 Cuban plants were evaluated by an in vitro bioassay against Leishmania amazonensis. Furthermore, their toxicity was assayed against murine macrophage. The three most potent extracts against the amastigote stage of Leishmania amazonensis were from Hura crepitans, Bambusa vulgaris, and Simarouba glauca. PMID:22530133
Activity of Cuban Plants Extracts against Leishmania amazonensis.
García, Marley; Monzote, Lianet; Scull, Ramón; Herrera, Pedro
Natural products have long been providing important drug leads for infectious diseases. Leishmaniasis is a major health problem worldwide that affects millions of people especially in the developing nations. There is no immunoprophylaxis (vaccination) available for Leishmania infections, and conventional treatments are unsatisfactory; therefore, antileishmanial drugs are urgently needed. In this work, 48 alcoholic extracts from 46 Cuban plants were evaluated by an in vitro bioassay against Leishmania amazonensis. Furthermore, their toxicity was assayed against murine macrophage. The three most potent extracts against the amastigote stage of Leishmania amazonensis were from Hura crepitans, Bambusa vulgaris, and Simarouba glauca. PMID:22530133
Verbascoside Inhibits Promastigote Growth and Arginase Activity of Leishmania amazonensis.
Maquiaveli, Claudia C; Lucon-Júnior, João F; Brogi, Simone; Campiani, Giuseppe; Gemma, Sandra; Vieira, Paulo C; Silva, Edson R
Verbascoside (1) is a phenylethanoid glycoside that has antileishmanial activity against Leishmania infantum and Leishmania donovani. In this study, we verified the activity of 1 on Leishmania amazonensis and arginase inhibition. Compound 1 showed an EC50 of 19 μM against L. amazonensis promastigotes and is a competitive arginase inhibitor (Ki = 0.7 μM). Docking studies were performed to assess the interaction of 1 with arginase at the molecular level. Arginase is an enzyme of the polyamine biosynthesis pathway that is important to parasite infectivity, and the results of our study suggest that 1 could be useful to develop new approaches for treating leishmaniasis. PMID:27096224
Leishmania amazonensis Engages CD36 to Drive Parasitophorous Vacuole Maturation
Okuda, Kendi; Tong, Mei; Dempsey, Brian; Moore, Kathryn J.; Gazzinelli, Ricardo T.; Silverman, Neal
Leishmania amastigotes manipulate the activity of macrophages to favor their own success. However, very little is known about the role of innate recognition and signaling triggered by amastigotes in this host-parasite interaction. In this work we developed a new infection model in adult Drosophila to take advantage of its superior genetic resources to identify novel host factors limiting Leishmania amazonensis infection. The model is based on the capacity of macrophage-like cells, plasmatocytes, to phagocytose and control the proliferation of parasites injected into adult flies. Using this model, we screened a collection of RNAi-expressing flies for anti-Leishmania defense factors. Notably, we found three CD36-like scavenger receptors that were important for defending against Leishmania infection. Mechanistic studies in mouse macrophages showed that CD36 accumulates specifically at sites where the parasite contacts the parasitophorous vacuole membrane. Furthermore, CD36-deficient macrophages were defective in the formation of the large parasitophorous vacuole typical of L. amazonensis infection, a phenotype caused by inefficient fusion with late endosomes and/or lysosomes. These data identify an unprecedented role for CD36 in the biogenesis of the parasitophorous vacuole and further highlight the utility of Drosophila as a model system for dissecting innate immune responses to infection. PMID:27280707
Trypanosoma cruzi Differentiates and Multiplies within Chimeric Parasitophorous Vacuoles in Macrophages Coinfected with Leishmania amazonensis.
Pessoa, Carina Carraro; Ferreira, Éden Ramalho; Bayer-Santos, Ethel; Rabinovitch, Michel; Mortara, Renato Arruda; Real, Fernando
The trypanosomatids Leishmania amazonensis and Trypanosoma cruzi are excellent models for the study of the cell biology of intracellular protozoan infections. After their uptake by mammalian cells, the parasitic protozoan flagellates L. amazonensis and T. cruzi lodge within acidified parasitophorous vacuoles (PVs). However, whereas L. amazonensis develops in spacious, phagolysosome-like PVs that may enclose numerous parasites, T. cruzi is transiently hosted within smaller vacuoles from which it soon escapes to the host cell cytosol. To investigate if parasite-specific vacuoles are required for the survival and differentiation of T. cruzi, we constructed chimeric vacuoles by infection of L. amazonensis amastigote-infected macrophages with T. cruzi epimastigotes (EPIs) or metacyclic trypomastigotes (MTs). These chimeric vacuoles, easily observed by microscopy, allowed the entry and fate of T. cruzi in L. amazonensis PVs to be dynamically recorded by multidimensional imaging of coinfected cells. We found that although T. cruzi EPIs remained motile and conserved their morphology in chimeric vacuoles, T. cruzi MTs differentiated into amastigote-like forms capable of multiplying. These results demonstrate that the large adaptive vacuoles of L. amazonensis are permissive to T. cruzi survival and differentiation and that noninfective EPIs are spared from destruction within the chimeric PVs. We conclude that T. cruzi differentiation can take place in Leishmania-containing vacuoles, suggesting this occurs prior to their escape into the host cell cytosol. PMID:26975994
Immune responses associated with susceptibility of C57BL/10 mice to Leishmania amazonensis.
Afonso, L C; Scott, P
Leishmaniae are protozoans which, depending upon both the host and parasite species, can cause either a healing or nonhealing infection. While C57BL/10 mice are able to heal following infection with Leishmania major, they fail to heal following infection with Leishmania amazonensis. In order to address the role of Th1 and Th2 cell responses in the outcome of these infections in C57BL/10 mice, gamma interferon (IFN-gamma) and interleukin-4 (IL-4) production was assessed. While cells from L. major-infected C57BL/10 mice produced high levels of IFN-gamma, cells from L. amazonensis-infected animals produced little or no IFN-gamma. On the other hand, IL-4 was produced only by cells from L. amazonensis-infected C57BL/10 mice, but this production was restricted to the first few weeks of infection. Later in infection, when lesions were evident, no IL-4 was detected. Treatment of BALB/c mice with a monoclonal antibody (11B11) directed against IL-4 induced a dramatic reduction in L. amazonensis lesions. This reduction was associated with a decrease in IL-4 levels and an increase in IFN-gamma production. However, only a slight reduction in lesion sizes and parasite numbers was observed when anti-IL-4-treated C57BL/10 mice were infected with L. amazonensis. These results suggest that IL-4 may have an important role in mediating susceptibility to L. amazonensis in BALB/c mice, as previously demonstrated for L. major. More importantly, however, the data suggest that susceptibility to L. amazonensis in C57BL/10 mice is due to the absence of a Th1 cell response, rather than to the presence of a Th2 cell response. PMID:8514400
A putative role for inosine 5' monophosphate dehydrogenase (IMPDH) in Leishmania amazonensis programmed cell death.
Pitaluga, A N; Moreira, M E C; Traub-Csekö, Y M
Leishmania amazonensis undergoes apoptosis-like programmed cell death (PCD) under heat shock conditions. We identified a potential role for inosine 5' monophosphate dehydrogenase (IMPDH) in L. amazonensis PCD. Trypanosomatids do not have a "de novo" purine synthesis pathway, relying on the salvage pathway for survival. IMPDH, a key enzyme in the purine nucleotide pathway, is related to cell growth and apoptosis. Since guanine nucleotide depletion triggers cell cycle arrest and apoptosis in several organisms we analyzed the correlation between IMPDH and apoptosis-like death in L. amazonensis. The L. amazonensis IMPDH inhibition effect on PCD was evaluated through gene expression analysis, mitochondrial depolarization and detection of Annexin-V labeled parasites. We demonstrated a down-regulation of impdh expression under heat shock treatment, which mimics the natural mammalian host infection. Also, IMPDH inhibitors ribavirin and mycophenolic acid (MPA) prevented cell growth and generated an apoptosis-like phenotype in sub-populations of L. amazonensis promastigotes. Our results are in accordance with previous results showing that a subpopulation of parasites undergoes apoptosis-like cell death in the nutrient poor environment of the vector gut. Here, we suggest the involvement of purine metabolism in previously observed apoptosis-like cell death during Leishmania infection. PMID:25499513
The genome sequence of Leishmania (Leishmania) amazonensis: functional annotation and extended analysis of gene models.
Real, Fernando; Vidal, Ramon Oliveira; Carazzolle, Marcelo Falsarella; Mondego, Jorge Maurício Costa; Costa, Gustavo Gilson Lacerda; Herai, Roberto Hirochi; Würtele, Martin; de Carvalho, Lucas Miguel; Carmona e Ferreira, Renata; Mortara, Renato Arruda; Barbiéri, Clara Lucia; Mieczkowski, Piotr; da Silveira, José Franco; Briones, Marcelo Ribeiro da Silva; Pereira, Gonçalo Amarante Guimarães; Bahia, Diana
We present the sequencing and annotation of the Leishmania (Leishmania) amazonensis genome, an etiological agent of human cutaneous leishmaniasis in the Amazon region of Brazil. L. (L.) amazonensis shares features with Leishmania (L.) mexicana but also exhibits unique characteristics regarding geographical distribution and clinical manifestations of cutaneous lesions (e.g. borderline disseminated cutaneous leishmaniasis). Predicted genes were scored for orthologous gene families and conserved domains in comparison with other human pathogenic Leishmania spp. Carboxypeptidase, aminotransferase, and 3'-nucleotidase genes and ATPase, thioredoxin, and chaperone-related domains were represented more abundantly in L. (L.) amazonensis and L. (L.) mexicana species. Phylogenetic analysis revealed that these two species share groups of amastin surface proteins unique to the genus that could be related to specific features of disease outcomes and host cell interactions. Additionally, we describe a hypothetical hybrid interactome of potentially secreted L. (L.) amazonensis proteins and host proteins under the assumption that parasite factors mimic their mammalian counterparts. The model predicts an interaction between an L. (L.) amazonensis heat-shock protein and mammalian Toll-like receptor 9, which is implicated in important immune responses such as cytokine and nitric oxide production. The analysis presented here represents valuable information for future studies of leishmaniasis pathogenicity and treatment. PMID:23857904
Inhibitory effects promoted by 5'-nucleotides on the ecto-3'-nucleotidase activity of Leishmania amazonensis.
Freitas-Mesquita, Anita Leocadio; Gomes, Marta T; Vieira, Danielle P; Paes-Vieira, Lisvane; Nascimento, Michelle T C; Lopes, Angela H C S; Meyer-Fernandes, José Roberto
The protozoan parasite Leishmania amazonensis is the etiological agent of cutaneous leishmaniasis. During its life cycle, the flagellated metacyclic promastigote forms are transmitted to vertebrate hosts by sandfly bites, and they develop into amastigotes inside macrophages, where they multiply. L. amazonensis possesses a bifunctional enzyme, called 3'-nucleotidase/nuclease (3'NT/NU), which is able to hydrolyze extracellular 3'-monophosphorylated nucleosides and nucleic acids. 3'NT/NU plays an important role in the generation of extracellular adenosine and has been described as a key enzyme in the acquisition of purines by trypanosomatids. Furthermore, it has been observed that 3'NT/NU also plays a valuable role in the establishment of parasitic infection. In this context, this study aimed to investigate the modulation of the 3'-nucleotidase (3'NT) activity of L. amazonensis by several nucleotides. It was observed that 3'NT activity is inhibited by micromolar concentrations of guanosine and guanine nucleotides. The inhibition promoted by 5'-GMP on the 3'NT activity of L. amazonensis is reversible and uncompetitive because the addition of the inhibitor decreased the kinetic parameters Km and Vmax. Finally, we found that the addition of 5'-GMP is able to reverse the stimulation promoted by 3'-AMP in a macrophage-parasite interaction assay. The determination of compounds that can inhibit the 3'NT activity of Leishmania is very important because this enzyme does not occur in mammals, making it a potential therapeutic target. PMID:27531705
Leishmania amazonensis: Biological and biochemical characterization of ecto-nucleoside triphosphate diphosphohydrolase activities.
Pinheiro, Carla M; Martins-Duarte, Erica S; Ferraro, Rodrigo B; Fonseca de Souza, André Luíz; Gomes, Marta T; Lopes, Angela H C S; Vannier-Santos, Marcos A; Santos, André L S; Meyer-Fernandes, José R
The presence of Leishmania amazonensis ecto-nucleoside triphosphate triphosphohydrolase activities was demonstrated using antibodies against different NTPDase members by Western blotting, flow cytometry, and immunoelectron microscopy analysis. Living promastigote cells sequentially hydrolyzed the ATP molecule generating ADP, AMP, and adenosine, indicating that this surface enzyme may play a role in the salvage of purines from the extracellular medium. The L. amazonensis ecto-NTPDase activities were insensitive to Triton X-100, but they were enhanced by divalent cations, such as Mg(2+). In addition, the ecto-NTPDase activities decreased with time for 96 h when promastigotes were grown in vitro. On the other hand, these activities increased considerably when measured in living amastigote forms. Furthermore, the treatment with adenosine, a mediator of several relevant biological phenomena, induced a decrease in the reactivity with anti-CD39 antibody, raised against mammalian E-NTPDase, probably because of down regulation in the L. amazonensis ecto-NTPDase expression. Also, adenosine and anti-NTPDase antibodies induced a significant diminishing in the interaction between promastigotes of L. amazonensis and mouse peritoneal macrophages. PMID:16603157
E-NTPDase (ecto-nucleoside triphosphate diphosphohydrolase) of Leishmania amazonensis inhibits macrophage activation.
Gomes, Rodrigo Saar; de Carvalho, Luana Cristina Faria; de Souza Vasconcellos, Raphael; Fietto, Juliana Lopes Rangel; Afonso, Luís Carlos Crocco
Leishmania amazonensis, the causal agent of diffuse cutaneous leishmaniasis, is known for its ability to modulate the host immune response. Because a relationship between ectonucleotidase activity and the ability of Leishmania to generate injury in C57BL/6 mice has been demonstrated, in this study we evaluated the involvement of ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) activity of L. amazonensis in the process of infection of J774-macrophages. Our results show that high-activity parasites show increased survival rate in LPS/IFN-γ-activated cells, by inhibiting the host-cell NO production. Conversely, inhibition of E-NTPDase activity reduces the parasite survival rates, an effect associated with increased macrophage NO production. E-NTPDase activity generates substrate for the production of extracellular adenosine, which binds to A2B receptors and reduces IL-12 and TNF-α produced by activated macrophages, thus inhibiting NO production. These results indicate that E-NTPDase activity is important for survival of L. amazonensis within macrophages, showing the role of the enzyme in modulating macrophage response and lower NO production, which ultimately favors infection. Our results point to a new mechanism of L. amazonensis infection that may pave the way for the development of new treatments for this neglected disease. PMID:25554487
Leishmania amazonensis: Increase in ecto-ATPase activity and parasite burden of vinblastine-resistant protozoa.
Giarola, Naira Lígia Lima; Silveira, Thaís Souza; Inacio, Job Domingos Filho; Vieira, Lisvane Paes; Almeida-Amaral, Elmo Eduardo; Meyer-Fernandes, José Roberto
Leishmania amazonensis is a protozoan parasite that induces mucocutaneous and diffuse cutaneous lesions upon infection. An important component in treatment failure is the emergence of drug-resistant parasites. It is necessary to clarify the mechanism of resistance that occurs in these parasites to develop effective drugs for leishmaniasis treatment. Promastigote forms of L. amazonensis were selected by gradually increasing concentrations of vinblastine and were maintained under continuous drug pressure (resistant cells). Vinblastine-resistant L. amazonensis proliferated similarly to control parasites. However, resistant cells showed changes in the cell shape, irregular flagella and a decrease in rhodamine 123 accumulation, which are factors associated with the development of resistance, suggesting the MDR phenotype. The Mg-dependent-ecto-ATPase, an enzyme located on cell surface of Leishmania parasites, is involved in the acquisition of purine and participates in the adhesion and infectivity process. We compared control and resistant L. amazonensis ecto-enzymatic activities. The control and resistant Leishmania ecto-ATPase activities were 16.0 ± 1.5 nmol Pi × h(-1) × 10(-7) cells and 40.0 ± 4.4 nmol Pi × h(-1) × 10(-7)cells, respectively. Interestingly, the activity of other ecto-enzymes present on the L. amazonensis cell surface, the ecto-5' and 3'-nucleotidases and ecto-phosphatase, did not increase. The level of ecto-ATPase modulation is related to the degree of resistance of the cell. Cells resistant to 10 μM and 60 μM of vinblastine have ecto-ATPase activities of 22.7 ± 0.4 nmol Pi × h(-1) × 10(-7) cells and 33.8 ± 0.8 nmol Pi × h(-1) × 10(-7)cells, respectively. In vivo experiments showed that both lesion size and parasite burden in mice infected with resistant parasites are greater than those of L. amazonensis control cells. Furthermore, our data established a relationship between the increase in ecto-ATPase activity and greater infectivity and
A comparison of two distinct murine macrophage gene expression profiles in response to Leishmania amazonensis infection
Background The experimental murine model of leishmaniasis has been widely used to characterize the immune response against Leishmania. CBA mice develop severe lesions, while C57BL/6 present small chronic lesions under L. amazonensis infection. Employing a transcriptomic approach combined with biological network analysis, the gene expression profiles of C57BL/6 and CBA macrophages, before and after L. amazonensis infection in vitro, were compared. These strains were selected due to their different degrees of susceptibility to this parasite. Results The genes expressed by C57BL/6 and CBA macrophages, before and after infection, differ greatly, both with respect to absolute number as well as cell function. Uninfected C57BL/6 macrophages express genes involved in the deactivation pathway of macrophages at lower levels, while genes related to the activation of the host immune inflammatory response, including apoptosis and phagocytosis, have elevated expression levels. Several genes that participate in the apoptosis process were also observed to be up-regulated in C57BL/6 macrophages infected with L. amazonensis, which is very likely related to the capacity of these cells to control parasite infection. By contrast, genes involved in lipid metabolism were found to be up-regulated in CBA macrophages in response to infection, which supports the notion that L. amazonensis probably modulates parasitophorous vacuoles in order to survive and multiply in host cells. Conclusion The transcriptomic profiles of C57BL/6 macrophages, before and after infection, were shown to be involved in the macrophage pathway of activation, which may aid in the control of L. amazonensis infection, in contrast to the profiles of CBA cells. PMID:22321871
Reprogramming Neutral Lipid Metabolism in Mouse Dendritic Leucocytes Hosting Live Leishmania amazonensis Amastigotes
Lecoeur, Hervé; Giraud, Emilie; Prévost, Marie-Christine; Milon, Geneviève; Lang, Thierry
Background After loading with live Leishmania (L) amazonensis amastigotes, mouse myeloid dendritic leucocytes/DLs are known to undergo reprogramming of their immune functions. In the study reported here, we investigated whether the presence of live L. amazonensis amastigotes in mouse bone marrow-derived DLs is able to trigger re-programming of DL lipid, and particularly neutral lipid metabolism. Methodology/Principal Findings Affymetrix-based transcriptional profiles were determined in C57BL/6 and DBA/2 mouse bone marrow-derived DLs that had been sorted from cultures exposed or not to live L. amazonensis amastigotes. This showed that live amastigote-hosting DLs exhibited a coordinated increase in: (i) long-chain fatty acids (LCFA) and cholesterol uptake/transport, (ii) LCFA and cholesterol (re)-esterification to triacyl-sn-glycerol (TAG) and cholesteryl esters (CE), respectively. As these neutral lipids are known to make up the lipid body (LB) core, oleic acid was added to DL cultures and LB accumulation was compared in live amastigote-hosting versus amastigote-free DLs by epi-fluorescence and transmission electron microscopy. This showed that LBs were both significantly larger and more numerous in live amastigote-hosting mouse dendritic leucocytes. Moreover, many of the larger LB showed intimate contact with the membrane of the parasitophorous vacuoles hosting the live L. amazonensis amastigotes. Conclusions/Significance As leucocyte LBs are known to be more than simple neutral lipid repositories, we set about addressing two related questions. Could LBs provide lipids to live amastigotes hosted within the DL parasitophorous vacuole and also deliver? Could LBs impact either directly or indirectly on the persistence of L. amazonensis amastigotes in rodent skin? PMID:23785538
Eupomatenoid-5 Isolated from Leaves of Piper regnellii Induces Apoptosis in Leishmania amazonensis.
Garcia, Francielle Pelegrin; Lazarin-Bidóia, Danielle; Ueda-Nakamura, Tânia; Silva, Sueli de Oliveira; Nakamura, Celso Vataru
Leishmania spp. are protozoa responsible for leishmaniasis, a neglected disease that kills up to 50,000 people every year. Current therapies mainly rely on antimonial drugs that are inadequate because of their poor efficacy and safety and increased drug resistance. An urgent need exists to find new and more affordable drugs. Our previous study demonstrated the antileishmanial activity of eupomatenoid-5, a neolignan obtained from leaves of Piper regnellii var. pallescens. The aim of the present study was to clarify the mode of action of eupomatenoid-5 against L. amazonensis. We used biochemical and morphological techniques and demonstrated that eupomatenoid-5 induced cell death in L. amazonensis promastigotes, sharing some phenotypic features observed in metazoan apoptosis, including increased reactive oxygen species production, hypopolarization of mitochondrial potential, phosphatidylserine exposure, decreased cell volume, and G0/G1 phase cell cycle arrest. PMID:23573160
Bioactivity of flavonoids isolated from Lychnophora markgravii against Leishmania amazonensis amastigotes.
Salvador, Marcos José; Sartori, Fabiana Terezinha; Sacilotto, Ana Claudia B C; Pral, Elizabeth M F; Alfieri, Silvia Celina; Vichnewski, Walter
The bioactivity of the flavonoids pinostrobin (1), pinocembrin (2), tectochrysin (3), galangin 3-methyl ether (4), and tiliroside (5) isolated from Lychnophora markgravii aerial parts was investigated in vitro against amastigote stages of Leishmania amazonensis. The compounds were isolated by several chromatographic techniques and their chemical structures were established by ESI-MS and NMR spectroscopic data. The flavonoids 1 and 3 were the most active compounds; they markedly reduced the viability of Leishmania amastigotes. PMID:19791501
CrATP interferes in the promastigote-macrophage interaction in Leishmania amazonensis infection.
Ennes-Vidal, V; Castro, R O S; Britto, C; Barrabin, H; D'Avila-Levy, C M; Moreira, O C
Recent have shown the relationship between Ecto-Nucleoside-Triphosphate-Diphosphohydrolases (Ecto-NTPDases or ecto-nucleotidases) and virulence and infectivity in trypanosomatids. In this work, the inhibition of the ecto-ATPase activities and promastigote growth of Leishmania amazonensis by CrATP was characterized. Furthermore, this compound was used to investigate the role of ecto-nucleotidase in the interaction of L. amazonensis with resident peritoneal macrophages obtained from BALB/c mice. CrATP partially inhibits the ecto-ATPase activity, presenting Ki values of 575·7±199·1 and 383·5±79·0 μm, in the presence or absence of 5 mm MgCl2, respectively. The apparent Kms for ATP (2·9±0·5 mm to Mg2+-dependent ecto-ATPase and 0·4±0·2 mm to Mg2+-independent ecto-ATPase activities) are not significantly altered by CrATP, suggesting a reversible non-competitive inhibition of both enzymes. When CrATP was added to the cultivation medium at 500 μm, it drastically inhibited the cellular growth. The interaction of promastigote forms of L. amazonensis with BALB/c peritoneal macrophages is strongly affected by CrATP. When the parasites were treated with 500 μm CrATP before interacting with macrophages, the adhesion and endocytic indices were strongly reduced to 53·0±14·8% and 39·8±1·1%, respectively. These results indicate that ecto-nucleotidase plays an important role in the infection process caused by Leishmania amazonensis. PMID:21679488
PvD1 defensin, a plant antimicrobial peptide with inhibitory activity against Leishmania amazonensis.
do Nascimento, Viviane V; Mello, Érica de O; Carvalho, Laís P; de Melo, Edésio J T; Carvalho, André de O; Fernandes, Katia V S; Gomes, Valdirene M
Plant defensins are small cysteine-rich peptides and exhibit antimicrobial activity against a variety of both plant and human pathogens. Despite the broad inhibitory activity that plant defensins exhibit against different micro-organisms, little is known about their activity against protozoa. In a previous study, we isolated a plant defensin named PvD1 from Phaseolus vulgaris (cv. Pérola) seeds, which was seen to be deleterious against different yeast cells and filamentous fungi. It exerted its effects by causing an increase in the endogenous production of ROS (reactive oxygen species) and NO (nitric oxide), plasma membrane permeabilization and the inhibition of medium acidification. In the present study, we investigated whether PvD1 could act against the protozoan Leishmania amazonensis. Our results show that, besides inhibiting the proliferation of L. amazonensis promastigotes, the PvD1 defensin was able to cause cytoplasmic fragmentation, formation of multiple cytoplasmic vacuoles and membrane permeabilization in the cells of this organism. Furthermore, we show, for the first time, that PvD1 defensin was located within the L. amazonensis cells, suggesting the existence of a possible intracellular target. PMID:26285803
Protein kinase A of Leishmania amazonensis as a potential target for methoxy-amidine.
Genestra, M; Echevarria, A; Cysne-Finkelstein, L; Leon, L L
Cyclic 3',5'-adenosine monophosphate (cAMP) is one of the most important signaling molecules for cell growth and differentiation in several systems including protozoal parasites such as Trypanosoma cruzi and Leishmania species. The most important event during Leishmania developmental cycle is the differentiation of procyclic into metacyclic promastigotes, which is associated with the appearance of pathogenicity. As previously demonstrated Leishmania amazonensis metacyclogenesis is associated with an increase of a protein kinase A activity, and therefore further studies on the activity of this phosphorylating enzyme as a target for chemotherapy were performed. Among several amidine derivatives tested by the authors against trypanosomatids (T. cruzi, T. evansi and L. amazonensis) the most effective compounds was defined as that with a methoxy group as substituent. In this work the inhibitory effect of this derivative on the phosphorylating activity of cAMP-dependent protein kinase (PKA) of promastigotes (containing high amounts of metacyclic forms) and axenic amastigotes of L. amazonensis is demonstrated. Soluble fractions (SF) and enriched membrane fractions (MF) were submitted to anion exchange chromatography in a DEAE-cellulose column and the collected fractions used to evaluate the phosphorylating activity associated with cAMP, in the presence/absence of methoxy-amidine and pentamidine (CAS 100-33-4), the latter being used as reference drug. PMID:11765595
C5 induces different cell death pathways in promastigotes of Leishmania amazonensis.
Mendes, Edevi Arbonelli; Desoti, Vânia Cristina; Silva, Sueli de Oliveira; Ueda-Nakamura, Tânia; Dias Filho, Benedito Prado; Yamada-Ogatta, Sueli Fumie; Sarragiotto, Maria Helena; Nakamura, Celso Vataru
Leishmaniasis is a neglected infection that is caused by Leishmania protozoa, affecting millions of people worldwide, mainly in tropical and subtropical regions. This disease has different clinical forms: cutaneous, mucocutaneous, and visceral. The drugs that are currently available for the treatment of this infection have limitations, such as high toxicity, long-term treatment, and leads to drug-resistant strains. Numerous studies, in various experimental models, have sought to develop more effective and less toxic chemotherapeutic agents against leishmaniasis. In the present study, we evaluated the mechanism of cell death that is induced by n-benzyl 1-(4-methoxy)phenyl-9H-β-carboline-3-carboxamide (C5) against Leishmania amazonensis. C5 increased reactive oxygen species production, depolarization of the mitochondrial membrane, DNA fragmentation, decrease of cell volume, lipoperoxidation, the accumulation of lipid bodies, and acidic vesicular organelles (AVOs) and caused the intense formation of autophagic compartments in L. amazonensis promastigotes. The results indicate that C5 causes L. amazonensis death through different pathways. PMID:27317947
Efficacy of miltefosine treatment in Leishmania amazonensis-infected BALB/c mice.
Godinho, Joseane Lima Prado; Simas-Rodrigues, Cíntia; Silva, Rosane; Ürmenyi, Turán Peter; de Souza, Wanderley; Rodrigues, Juliany Cola Fernandes
Leishmaniasis is one of the most serious worldwide diseases caused by protozoan parasites of the Leishmania genus, affecting millions of people around the world. All currently available treatments present severe toxic side effects, require long-term compliance, cause serious side effects and are uncomfortable for patients. Leishmania amazonensis, a species endemic to Brazil, causes severe localised or diffuse skin lesions in humans. Owing to the unsatisfactory nature of the currently available chemotherapies, new approaches have been assessed for improved therapeutic intervention strategies against leishmaniasis. Miltefosine is an alkylphospholipid analogue that exhibits potent activity against the different clinical manifestations of leishmaniasis. Thus, the aim of this study was to investigate the long-term efficacy of miltefosine in BALB/c mice infected with L. amazonensis owing to the lack of a profound study demonstrating its dose-dependent and long-term effects. It was observed that animals treated with 20-50 mg/kg/day of miltefosine exhibited a significant dose-dependent reduction in lesion size; furthermore, in mice receiving higher doses, lesions disappeared after the end of treatment. To confirm a possible parasitological cure, mice up to 250 days after the end of treatment were analysed. No lesions or presence of parasite DNA were found in mice treated with 30, 40 and 50 mg/kg/day of miltefosine. In summary, these results show that miltefosine may be used to treat cutaneous leishmaniasis caused by L. amazonensis, alone or as combination therapy. PMID:22226653
Leishmania amazonensis: characterization of an ouabain-insensitive Na+-ATPase activity.
de Almeida-Amaral, Elmo Eduardo; Caruso-Neves, Celso; Pires, Vanessa Maria Pereira; Meyer-Fernandes, José Roberto
We characterized ouabain-insensitive Na+-ATPase activity present in the plasma membrane of Leishmania amazonensis and investigated its possible role in the growth of the parasite. An increase in Na+ concentration in the presence of 1mM ouabain, increased the ATPase activity with a V(max) of 154.1+/-13.5nmol Pi x h(-1) x mg(-1) and a K0.5 of 28.9+/-7.7mM. Furosemide and sodium orthovanadate inhibited the Na+-stimulated ATPase activity with an IC(50) of 270microM and 0.10microM, respectively. Furosemide inhibited the growth of L. amazonensis after 48h incubation, with maximal effect after 96h. The IC50 for furosemide was 840. On the other hand, ouabain (1mM) did not change the growth of the parasite. Taken together, these results show that L. amazonensis expresses a P-type, ouabain-insensitive Na+-ATPase that could be involved with the growth of the parasite. PMID:17825292
Leishmania amazonensis Arginase Compartmentalization in the Glycosome Is Important for Parasite Infectivity
da Silva, Maria Fernanda Laranjeira; Zampieri, Ricardo Andrade; Muxel, Sandra M.; Beverley, Stephen M.; Floeter-Winter, Lucile M.
In Leishmania, de novo polyamine synthesis is initiated by the cleavage of L-arginine to urea and L-ornithine by the action of arginase (ARG, E.C. 126.96.36.199). Previous studies in L. major and L. mexicana showed that ARG is essential for in vitro growth in the absence of polyamines and needed for full infectivity in animal infections. The ARG protein is normally found within the parasite glycosome, and here we examined whether this localization is required for survival and infectivity. First, the localization of L. amazonensis ARG in the glycosome was confirmed in both the promastigote and amastigote stages. As in other species, arg− L. amazonensis required putrescine for growth and presented an attenuated infectivity. Restoration of a wild type ARG to the arg− mutant restored ARG expression, growth and infectivity. In contrast, restoration of a cytosol-targeted ARG lacking the glycosomal SKL targeting sequence (argΔSKL) restored growth but failed to restore infectivity. Further study showed that the ARGΔSKL protein was found in the cytosol as expected, but at very low levels. Our results indicate that the proper compartmentalization of L. amazonensis arginase in the glycosome is important for enzyme activity and optimal infectivity. Our conjecture is that parasite arginase participates in a complex equilibrium that defines the fate of L-arginine and that its proper subcellular location may be essential for this physiological orchestration. PMID:22479507
PvD1 defensin, a plant antimicrobial peptide with inhibitory activity against Leishmania amazonensis
do Nascimento, Viviane V.; Mello, Érica de O.; Carvalho, Laís P.; de Melo, Edésio J.T.; Carvalho, André de O.; Fernandes, Katia V.S.; Gomes, Valdirene M.
Plant defensins are small cysteine-rich peptides and exhibit antimicrobial activity against a variety of both plant and human pathogens. Despite the broad inhibitory activity that plant defensins exhibit against different micro-organisms, little is known about their activity against protozoa. In a previous study, we isolated a plant defensin named PvD1 from Phaseolus vulgaris (cv. Pérola) seeds, which was seen to be deleterious against different yeast cells and filamentous fungi. It exerted its effects by causing an increase in the endogenous production of ROS (reactive oxygen species) and NO (nitric oxide), plasma membrane permeabilization and the inhibition of medium acidification. In the present study, we investigated whether PvD1 could act against the protozoan Leishmania amazonensis. Our results show that, besides inhibiting the proliferation of L. amazonensis promastigotes, the PvD1 defensin was able to cause cytoplasmic fragmentation, formation of multiple cytoplasmic vacuoles and membrane permeabilization in the cells of this organism. Furthermore, we show, for the first time, that PvD1 defensin was located within the L. amazonensis cells, suggesting the existence of a possible intracellular target. PMID:26285803
Altered Dendritic Cell Phenotype in Response to Leishmania amazonensis Amastigote Infection Is Mediated by MAP Kinase, ERK
Boggiatto, Paola Mercedes; Jie, Fei; Ghosh, Mousumi; Gibson-Corley, Katherine Nicole; Ramer-Tait, Amanda Ellen; Jones, Douglas Elliot; Petersen, Christine Anne
Initiation of productive immune responses against Leishmania depends on the successful transition of dendritic cells (DC) from an immature to a mature phenotype. This process is characterized by high CD40 surface expression as well as interleukin-12 production, which are frequently seen in response to L. major infection. In vivo footpad infection of C3HeB/FeJ mice for 7 days with L. amazonensis promoted an immature CD11c+ DC phenotype characterized by both significantly low CD40 surface expression and significantly decreased interleukin-12p40 production compared with L. major infection of these same mice. In vitro infection of bone marrow-derived dendritic cells with L. amazonensis amastigotes resulted in rapid and significant phosphorylation of the mitogen activated protein kinase, extracellular signal-regulated kinase 1/2, observed within minutes of exposure to the parasite. Infection with L. amazonensis promastigotes led to increased 1/2 phosphorylation after 4 hours of infection compared with L. major infection, which correlated with promastigote transformation into amastigotes. Treatment of bone marrow-derived dendritic cells with a mitogen activated protein kinase kinase-specific inhibitor, PD98059, led to regained surface CD40 expression and interleukin-12p40 production following L. amazonensis amastigote infection compared with non-treated, infected DC. Treatment of L. amazonensis-infected mice with the highly-specific mitogen activated protein kinase kinase inhibitor, CI-1040, enhanced surface CD40 expression on CD11c+ DC obtained from the draining lymph node. L. amazonensis amastigotes, through activation of extracellular signal-regulated kinase 1/2, inhibit the ability of DC to undergo proper maturation both in vitro and in vivo. PMID:19349356
Identification and characterization of new Leishmania promastigote surface antigens, LaPSA-38S and LiPSA-50S, as major immunodominant excreted/secreted components of L. amazonensis and L. infantum.
Bras-Gonçalves, Rachel; Petitdidier, Elodie; Pagniez, Julie; Veyrier, Renaud; Cibrelus, Prisca; Cavaleyra, Mireille; Maquaire, Sarah; Moreaux, Jérôme; Lemesre, Jean-Loup
We have previously demonstrated that sera from dogs vaccinated with excreted/secreted antigens (ESA) of Leishmania infantum promastigotes (LiESAp) mainly recognized an immunodominant antigen of 54 kDa. An anti-LiESAp-specific IgG2 humoral response was observed and associated to Th1-type response in vaccinated dogs. This response was highly correlated with a long-lasting and strong LiESAp-vaccine protection toward L. infantum experimental infection. In addition, it was also shown that dogs from the vaccinated group developed a selective IgG2 response against an immunodominant antigen of 45 kDa of Leishmania amazonensis ESA promastigotes (LaESAp). In order to identify and characterize these immunodominant antigens, a mouse monoclonal antibody (mAb F5) was produced by immunization against LaESAp. It was found to recognize the major antigenic targets of both LaESAp and LiESAp. Analysis with mAb F5 of L. amazonensis amastigote and promastigote cDNA expression libraries enabled the identification of clones encoding proteins with significant structural homology to the promastigote surface antigens named PSA-2/gp-46. Among them, one clone presented a full-length cDNA and encoded a novel L. amazonensis protein of 38.6 kDa calculated molecular mass (LaPSA-38S) sharing an amino acid sequence consistent with that of the PSA polymorphic family and a N-terminal signal peptide, characteristic of a secreted protein. We then screened a L. infantum promastigote DNA cosmid library using a cDNA probe derived from the LaPSA-38S gene and identified a full-length clone of a novel excreted/secreted protein of L. infantum with a calculated molecular mass of 49.2 kDa and named LiPSA-50S. The fact that a significant immunological reactivity was observed against PSA, suggests that these newly identified proteins could have an important immunoregulatory influence on the immune response. This hypothesis is supported by the fact that (i) these proteins were naturally excreted/secreted by viable
The Calpain Inhibitor MDL28170 Induces the Expression of Apoptotic Markers in Leishmania amazonensis Promastigotes
Marinho, Fernanda A.; Gonçalves, Keyla C. S.; Oliveira, Simone S. C.; Gonçalves, Diego S.; Matteoli, Filipe P.; Seabra, Sergio H.; Oliveira, Ana Carolina S.; Bellio, Maria; Oliveira, Selma S.; Souto-Padrón, Thaïs; d'Avila-Levy, Claudia M.; Santos, André L. S.; Branquinha, Marta H.
Background Human cutaneous leishmaniasis is caused by distinct species, including Leishmania amazonensis. Treatment of cutaneous leishmaniasis is far from satisfactory due to increases in drug resistance and relapses, and toxicity of compounds to the host. As a consequence for this situation, the development of new leishmanicidal drugs and the search of new targets in the parasite biology are important goals. Methodology/Principal Findings In this study, we investigated the mechanism of death pathway induced by the calpain inhibitor MDL28170 on Leishmania amazonensis promastigote forms. The combined use of different techniques was applied to contemplate this goal. MDL28170 treatment with IC50 (15 µM) and two times the IC50 doses induced loss of parasite viability, as verified by resazurin assay, as well as depolarization of the mitochondrial membrane, which was quantified by JC-1 staining. Scanning and transmission electron microscopic images revealed drastic alterations on the parasite morphology, some of them resembling apoptotic-like death, including cell shrinking, surface membrane blebs and altered chromatin condensation pattern. The lipid rearrangement of the plasma membrane was detected by Annexin-V labeling. The inhibitor also induced a significant increase in the proportion of cells in the sub-G0/G1 phase, as quantified by propidium iodide staining, as well as genomic DNA fragmentation, detected by TUNEL assay. In cells treated with MDL28170 at two times the IC50 dose, it was also possible to observe an oligonucleossomal DNA fragmentation by agarose gel electrophoresis. Conclusions/Significance The data presented in the current study suggest that MDL28170 induces apoptotic marker expression in promastigotes of L. amazonensis. Altogether, the results described in the present work not only provide a rationale for further exploration of the mechanism of action of calpain inhibitors against trypanosomatids, but may also widen the investigation of the
Reactive Oxygen Species Production and Mitochondrial Dysfunction Contribute to Quercetin Induced Death in Leishmania amazonensis
Fonseca-Silva, Fernanda; Inacio, Job D. F.; Canto-Cavalheiro, Marilene M.; Almeida-Amaral, Elmo Eduardo
Background Leishmaniasis, a parasitic disease caused by protozoa of the genus Leishmania, affects more than 12 million people worldwide. Quercetin has generated considerable interest as a pharmaceutical compound with a wide range of therapeutic activities. One such activity is exhibited against the bloodstream parasite Trypanosoma brucei and amastigotes of Leishmania donovani. However, the mechanism of protozoan action of quercetin has not been studied. Methodology/Principal Findings In the present study, we report here the mechanism for the antileishmanial activity of quercetin against Leishmania amazonensis promastigotes. Quercetin inhibited L. amazonensis promastigote growth in a dose- and time- dependent manner beginning at 48 hours of treatment and with maximum growth inhibition observed at 96 hours. The IC50 for quercetin at 48 hours was 31.4 µM. Quercetin increased ROS generation in a dose-dependent manner after 48 hours of treatment. The antioxidant GSH and NAC each significantly reduced quercetin-induced cell death. In addition, quercetin caused mitochondrial dysfunction due to collapse of mitochondrial membrane potential. Conclusions/Significance The effects of several drugs that interfere directly with mitochondrial physiology in parasites such as Leishmania have been described. The unique mitochondrial features of Leishmania make this organelle an ideal drug target while minimizing toxicity. Quercetin has been described as a pro-oxidant, generating ROS which are responsible for cell death in some cancer cells. Mitochondrial membrane potential loss can be brought about by ROS added directly in vitro or induced by chemical agents. Taken together, our results demonstrate that quercetin eventually exerts its antileishmanial effect on L. amazonensis promastigotes due to the generation of ROS and disrupted parasite mitochondrial function. PMID:21346801
Functional characterization of LIT1, the Leishmania amazonensis ferrous iron transporter
Jacques, Ismaele; Andrews, Norma W.; Huynh, Chau
Leishmania amazonensis LIT1 was identified based on homology with IRT1, a ferrous iron transporter from Arabidopsis thaliana. Δlit1 Leishmania amazonensis are defective in intracellular replication and lesion formation in vivo, a virulence phenotype attributed to defective intracellular iron acquisition. Here we functionally characterize LIT1, directly demonstrating that it functions as a ferrous iron membrane transporter from the ZIP family. Conserved residues in the predicted transmembrane domains II, IV, V and VII of LIT1 are essential for iron transport in yeast, including histidines that were proposed to function as metal ligands in ZIP transporters. LIT1 also contains two regions within the predicted intracellular loop that are not found in Arabidopsis IRT1. Deletion of region I inhibited LIT1 expression on the surface of Leishmania promastigotes. Deletion of region II did not interfere with LIT1 trafficking to the surface, but abolished its iron transport capacity when expressed in yeast. Mutagenesis revealed two motifs within region II, HGHQH and TPPRDM, that are independently required for iron transport by LIT1. D263 was identified as a key residue required for iron transport within the TPPRDM motif, while P260 and P261 were dispensable. Deletion of proline-rich regions within region I and between regions I and II did not affect iron transport in yeast, but in Leishmania amazonensis were not able to rescue the intracellular growth of Δlit1 parasites, or their ability to form lesions in mice. These results are consistent with a potential role of the unique intracellular loop of LIT1 in intracellular regulation by Leishmania-specific factors. PMID:20025906
Evaluation of Macroalgae Sulfated Polysaccharides on the Leishmania (L.) amazonensis Promastigote
Pires, Camila Lehnhardt; Rodrigues, Selma Dzimidas; Bristot, Daniel; Gaeta, Henrique Hessel; de Oliveira Toyama, Daniela; Farias, Wladimir Ronald Lobo; Toyama, Marcos Hikari
The sulfated polysaccharides from Solieria filiformis (Sf), Botryocladia occidentalis (Bo), Caulerpa racemosa (Cr) and Gracilaria caudata (Gc) were extracted and extensively purified. These compounds were then subjected to in vitro assays to evaluate the inhibition of these polysaccharides on the growth of Leishmania (L.) amazonensis promastigotes. Under the same assay conditions, only three of the four sulfated polysaccharides were active against L. amazonensis, and the polysaccharide purified from Cr was the most potent (EC50 value: 34.5 μg/mL). The polysaccharides derived from Bo and Sf demonstrated moderate anti-leishmanial activity (EC50 values of 63.7 μg/mL and 137.4 μg/mL). In addition, we also performed in vitro cytotoxic assays toward peritoneal macrophages and J774 macrophages. For the in vitro cytotoxicity assay employing J774 cells, all of the sulfated polysaccharides decreased cell survival, with CC50 values of 27.3 μg/mL, 49.3 μg/mL, 73.2 μg/mL, and 99.8 μg/mL for Bo, Cr, Gc, and Sf, respectively. However, none of the sulfated polysaccharides reduced the cell growth rate of the peritoneal macrophages. These results suggest that macroalgae contain compounds with various chemical properties that can control specific pathogens. According to our results, the assayed sulfated polysaccharides were able to modulate the growth rate and cell survival of Leishmania (L.) amazonensis promastigotes in in vitro assays, and these effects involved the interaction of the sulfated polysaccharides on the cell membrane of the parasites. PMID:23519148
A Trypanosomatid Iron Transporter that Regulates Mitochondrial Function Is Required for Leishmania amazonensis Virulence.
Mittra, Bidyottam; Laranjeira-Silva, Maria Fernanda; Perrone Bezerra de Menezes, Juliana; Jensen, Jennifer; Michailowsky, Vladimir; Andrews, Norma W
Iron, an essential co-factor of respiratory chain proteins, is critical for mitochondrial function and maintenance of its redox balance. We previously reported a role for iron uptake in differentiation of Leishmania amazonensis into virulent amastigotes, by a mechanism that involves reactive oxygen species (ROS) production and is independent of the classical pH and temperature cues. Iron import into mitochondria was proposed to be essential for this process, but evidence supporting this hypothesis was lacking because the Leishmania mitochondrial iron transporter was unknown. Here we describe MIT1, a homolog of the mitochondrial iron importer genes mrs3 (yeast) and mitoferrin-1 (human) that is highly conserved among trypanosomatids. MIT1 expression was essential for the survival of Trypanosoma brucei procyclic but not bloodstream forms, which lack functional respiratory complexes. L. amazonensis LMIT1 null mutants could not be generated, suggesting that this mitochondrial iron importer is essential for promastigote viability. Promastigotes lacking one LMIT1 allele (LMIT1/Δlmit1) showed growth defects and were more susceptible to ROS toxicity, consistent with the role of iron as the essential co-factor of trypanosomatid mitochondrial superoxide dismutases. LMIT1/Δlmit1 metacyclic promastigotes were unable to replicate as intracellular amastigotes after infecting macrophages or cause cutaneous lesions in mice. When induced to differentiate axenically into amastigotes, LMIT1/Δlmit1 showed strong defects in iron content and function of mitochondria, were unable to upregulate the ROS-regulatory enzyme FeSOD, and showed mitochondrial changes suggestive of redox imbalance. Our results demonstrate the importance of mitochondrial iron uptake in trypanosomatid parasites, and highlight the role of LMIT1 in the iron-regulated process that orchestrates differentiation of L. amazonensis into infective amastigotes. PMID:26741360
A Trypanosomatid Iron Transporter that Regulates Mitochondrial Function Is Required for Leishmania amazonensis Virulence
Mittra, Bidyottam; Laranjeira-Silva, Maria Fernanda; Perrone Bezerra de Menezes, Juliana; Jensen, Jennifer; Michailowsky, Vladimir; Andrews, Norma W.
Iron, an essential co-factor of respiratory chain proteins, is critical for mitochondrial function and maintenance of its redox balance. We previously reported a role for iron uptake in differentiation of Leishmania amazonensis into virulent amastigotes, by a mechanism that involves reactive oxygen species (ROS) production and is independent of the classical pH and temperature cues. Iron import into mitochondria was proposed to be essential for this process, but evidence supporting this hypothesis was lacking because the Leishmania mitochondrial iron transporter was unknown. Here we describe MIT1, a homolog of the mitochondrial iron importer genes mrs3 (yeast) and mitoferrin-1 (human) that is highly conserved among trypanosomatids. MIT1 expression was essential for the survival of Trypanosoma brucei procyclic but not bloodstream forms, which lack functional respiratory complexes. L. amazonensis LMIT1 null mutants could not be generated, suggesting that this mitochondrial iron importer is essential for promastigote viability. Promastigotes lacking one LMIT1 allele (LMIT1/Δlmit1) showed growth defects and were more susceptible to ROS toxicity, consistent with the role of iron as the essential co-factor of trypanosomatid mitochondrial superoxide dismutases. LMIT1/Δlmit1 metacyclic promastigotes were unable to replicate as intracellular amastigotes after infecting macrophages or cause cutaneous lesions in mice. When induced to differentiate axenically into amastigotes, LMIT1/Δlmit1 showed strong defects in iron content and function of mitochondria, were unable to upregulate the ROS-regulatory enzyme FeSOD, and showed mitochondrial changes suggestive of redox imbalance. Our results demonstrate the importance of mitochondrial iron uptake in trypanosomatid parasites, and highlight the role of LMIT1 in the iron-regulated process that orchestrates differentiation of L. amazonensis into infective amastigotes. PMID:26741360
Specific immunization of mice against Leishmania mexicana amazonensis using solubilized promastigotes
NASA Technical Reports Server (NTRS)
Barral-Netto, M.; Sadigursky, M.; Reed, S. G.; Sonnenfeld, G.
In this work, it was demonstrated that mice (BALB/c strain) highly susceptible to Leishmania mexicana amazonensis can be protected against infection by this parasite by being preimmunized with whole solubilized (in a buffer that contained EDTA, NP-40, and SDS) promastigotes; the use of adjuvant or intact inactivated parasite cells is shown to be not necessary. The best immunization schedule consisted of three intravenous injections of 5 x 10 to the 7th parasite equivalents, administered one to eight weeks before infection. Immunized mice exhibited a marked inhibition of primary lesion development, reduced numbers of parasites in the spleen, and reduced death rate.
Complete set of mitochondrial pan-edited mRNAs in Leishmania mexicana amazonensis LV78
Maslov, Dmitri A.
Editing of mRNA transcribed from the mitochondrial cryptogenes ND8 (G1), ND9 (G2), G3, G4, ND3 (G5), RPS12 (G6) was investigated in Leishmania mexicana amazonensis, strain LV78, by amplification of the cDNA, cloning and sequencing. For each of these genes, extensively and partially edited transcripts were found to be relatively abundant compared to the respective pre-edited molecules. Moreover, the editing patterns observed in a majority of transcripts of each gene were consistent among themselves which allowed for inferring consensus editing sequences. The open reading frames contained in the consensus sequences were predicted to encode polypeptides that were highly similar to their counterparts in other species of Trypanosomatidae. Several kinetoplast DNA minicircles from this species available in the public domain were found to contain genes for guide RNAs which mediate editing of some of the mRNAs. The results indicate that the investigated strain of L. m. amazonensis has preserved its full editing capacity in spite of the long-term maintenance in culture. This property differs drastically from the other Leishmania species which lost some or all of the G1–G5 mRNA editing ability in culture. PMID:20546801
Effect of Elatol, Isolated from Red Seaweed Laurencia dendroidea, on Leishmania amazonensis
dos Santos, Adriana Oliveira; Veiga-Santos, Phercyles; Ueda-Nakamura, Tânia; Filho, Benedito Prado Dias; Sudatti, Daniela Bueno; Bianco, Éverson Miguel; Pereira, Renato Crespo; Nakamura, Celso Vataru
In the present study, we investigated the antileishmanial activity of sesquiterpene elatol, the major constituent of the Brazilian red seaweed Laurencia dendroidea (Hudson) J.V. Lamouroux, against L. amazonensis. Elatol after 72 h of treatment, showed an IC50 of 4.0 μM and 0.45 μM for promastigote and intracellular amastigote forms of L. amazonensis, respectively. By scanning and transmission electron microscopy, parasites treated with elatol revealed notable changes compared with control cells, including: pronounced swelling of the mitochondrion; appearance of concentric membrane structures inside the organelle; destabilization of the plasma membrane; and formation of membrane structures, apparently an extension of the endoplasmic reticulum, which is suggestive of an autophagic process. A cytotoxicity assay showed that the action of the isolated compound is more specific for protozoa, and it is not toxic to macrophages. Our studies indicated that elatol is a potent antiproliferative agent against promastigote and intracellular amastigote forms, and may have important advantages for the development of new anti-leishamanial chemotherapies. PMID:21139841
Resveratrol Is Active against Leishmania amazonensis: In Vitro Effect of Its Association with Amphotericin B
Ferreira, Christian; Soares, Deivid Costa; do Nascimento, Michelle Tanny Cunha; Pinto-da-Silva, Lucia Helena; Sarzedas, Carolina Galvão; Tinoco, Luzineide Wanderley
Resveratrol is a polyphenol found in black grapes and red wine and has many biological activities. In this study, we evaluated the effect of resveratrol alone and in association with amphotericin B (AMB) against Leishmania amazonensis. Our results demonstrate that resveratrol possesses both antipromastigote and antiamastigote effects, with 50% inhibitory concentrations (IC50s) of 27 and 42 μM, respectively. The association of resveratrol with AMB showed synergy for L. amazonensis amastigotes, as demonstrated by the mean sums of fractional inhibitory index concentration (mean ΣFIC) of 0.483, although for promastigotes, this association was indifferent. Treatment with resveratrol increased the percentage of promastigotes in the sub-G0/G1 phase of the cell cycle, reduced the mitochondrial potential, and showed an elevated choline peak and CH2-to-CH3 ratio in the nuclear magnetic resonance (NMR) spectroscopy analysis; all these features indicate parasite death. Resveratrol also decreased the activity of the enzyme arginase in uninfected and infected macrophages with and without stimulation with interleukin-4 (IL-4), also implicating arginase inhibition in parasite death. The anti-Leishmania effect of resveratrol and its potential synergistic association with AMB indicate that these compounds should be subjected to further studies of drug association therapy in vivo. PMID:25114129
Cell death in amastigote forms of Leishmania amazonensis induced by parthenolide
Background Leishmania amazonensis infection results in diverse clinical manifestations: cutaneous, mucocutaneous or visceral leishmaniasis. The arsenal of drugs available for treating Leishmania infections is limited. Therefore, new, effective, and less toxic leishmaniasis treatments are still needed. We verified cell death in amastigote forms of Leishmania amazonensis induced by the sesquiterpene lactone parthenolide. Results The tested compound was able to concentration-dependently affect axenic and intracellular amastigotes, with IC50 values of 1.3 μM and 2.9 μM, respectively after 72 h incubation. No genotoxic effects were observed in a micronucleus test in mice. Parthenolide induced morphological and ultrastructural changes in axenic amastigotes, including a loss of membrane integrity, swelling of the mitochondrion, cytoplasmic vacuoles, and intense exocytic activity in the region of the flagellar pocket. These results led us to investigate the occurrence of autophagic vacuoles with monodansylcadaverine and the integrity of the plasma membrane and mitochondrial membrane potential using flow cytometry. In all of the tests, parthenolide had positive results. Conclusions Our results indicate that the antileishmanial action of parthenolide is associated with autophagic vacuole appearance, a reduction of fluidity, a loss of membrane integrity, and mitochondrial dysfunction. Considering the limited repertoire of existing antileishmanial compounds, the products derived from medicinal plants has been one the greatest advances to help develop new chemotherapeutic approaches. PMID:24913205
Individual and combined antiparasitic effect of six plant metabolites against Leishmania amazonensis and Trypanosoma cruzi.
Sandjo, Louis P; de Moraes, Milene H; Kuete, Victor; Kamdoum, Blaise C; Ngadjui, Bonaventure T; Steindel, Mario
Six plant metabolites including isobavachalcone (1), 4-hydroxylonchocarpine (2), and (E)-1-(2,2-dimethyl-2H-chromen-6-yl)-3-(4-hydroxyphenyl)prop-2-en-1-one (3), 6,8-(di-3-methyl-but-2-enyl)eriodictyol (4), damnacanthal (5), and buesgenine (6) were evaluated for their leishmanicidal and trypanocidal activities against intracellular amastigotes of Leishmania amazonensis and Trypanosoma cruzi. Compounds 2-4 and 6 displayed antileishmanial activity while 3 and 5 showed trypanocidal effect. The leishmanicidal activity of 6 was expressed with the lowest IC50 (5.70μg/mL) whereas the most trypanocidal metabolite (5) showed its activity with IC50 at 11.14μg/mL. In addition, antiprotozoal effect of mixtures of 1-6 prepared at different ratios (3:1, 1:1, and 1:3) was also investigated. Interestingly, 1 and 2 initially inactive against T. cruzi, displayed trypanocidal activities when mixed together. This activity increased when 3 (13.63μg/mL) was combined with 1 in ratios 1:1 (10.01μg/mL) and 3:1 (7.78μg/mL). Moreover, the leishmanicidal effect of 4 against L. amazonensis increased in the mixture 6/4 (1:3). PMID:26906638
Benzaldehyde Thiosemicarbazone Derived from Limonene Complexed with Copper Induced Mitochondrial Dysfunction in Leishmania amazonensis
Britta, Elizandra Aparecida; Barbosa Silva, Ana Paula; Ueda-Nakamura, Tânia; Dias-Filho, Benedito Prado; Silva, Cleuza Conceição; Sernaglia, Rosana Lázara; Nakamura, Celso Vataru
Background Leishmaniasis is a major health problem that affects more than 12 million people. Treatment presents several problems, including high toxicity and many adverse effects, leading to the discontinuation of treatment and emergence of resistant strains. Methodology/Principal Findings We evaluated the in vitro antileishmanial activity of benzaldehyde thiosemicarbazone derived from limonene complexed with copper, termed BenzCo, against Leishmania amazonensis. BenzCo inhibited the growth of the promastigote and axenic amastigote forms, with IC50 concentrations of 3.8 and 9.5 µM, respectively, with 72 h of incubation. Intracellular amastigotes were inhibited by the compound, with an IC50 of 10.7 µM. BenzCo altered the shape, size, and ultrastructure of the parasites. Mitochondrial membrane depolarization was observed in protozoa treated with BenzCo but caused no alterations in the plasma membrane. Additionally, BenzCo induced lipoperoxidation and the production of mitochondrial superoxide anion radicals in promastigotes and axenic amastigotes of Leishmania amazonensis. Conclusion/Significance Our studies indicated that the antileishmania activity of BenzCo might be associated with mitochondrial dysfunction and oxidative damage, leading to parasite death. PMID:22870222
Understanding the Mechanisms Controlling Leishmania amazonensis Infection In Vitro: The Role of LTB4 Derived From Human Neutrophils
Tavares, Natalia Machado; Araújo-Santos, Théo; Afonso, Lilian; Nogueira, Paula Monalisa; Lopes, Ulisses Gazos; Soares, Rodrigo Pedro; Bozza, Patrícia Torres; Bandeira-Melo, Christianne; Borges, Valeria Matos; Brodskyn, Cláudia
Neutrophils are rapidly recruited to the site of Leishmania infection and play an active role in capturing and killing parasites. They are the main source of leukotriene B4 (LTB4), a potent proinflammatory lipid mediator. However, the role of LTB4 in neutrophil infection by Leishmania amazonensis is not clear. In this study, we show that L. amazonensis or its lipophosphoglycan can induce neutrophil activation, degranulation, and LTB4 production. Using pharmacological inhibitors of leukotriene synthesis, our findings reveal an LTB4-driven autocrine/paracrine regulatory effect. In particular, neutrophil-derived LTB4 controls L. amazonensis killing, degranulation, and reactive oxygen species production. In addition, L. amazonensis infection induces an early increase in Toll-like receptor 2 expression, which facilitates parasite internalization. Nuclear factor kappa B (NFkB) pathway activation represents a required upstream event for L. amazonensis–induced LTB4 synthesis. These leishmanicidal mechanisms mediated by neutrophil-derived LTB4 act through activation of its receptor, B leukotriene receptor 1 (BLT1). PMID:24634497
IL-27 enhances Leishmania amazonensis infection via ds-RNA dependent kinase (PKR) and IL-10 signaling.
Barreto-de-Souza, Victor; Ferreira, Pedro L C; Vivarini, Aislan de Carvalho; Calegari-Silva, Teresa; Soares, Deivid Costa; Regis, Eduardo G; Pereira, Renata M S; Silva, Aristóbolo M; Saraiva, Elvira M; Lopes, Ulisses G; Bou-Habib, Dumith Chequer
The protozoan parasite Leishmania infects and replicates in macrophages, causing a spectrum of diseases in the human host, varying from cutaneous to visceral clinical forms. It is known that cytokines modulate the immunological response against Leishmania and are relevant for infection resolution. Here, we report that Interleukin (IL)-27 increases Leishmania amazonensis replication in human and murine macrophages and that the blockage of the IL-10 receptor on the surface of infected cells abolished the IL-27-mediated enhancement of Leishmania growth. IL-27 induced the activation/phosphorylation of protein kinase R (PKR) in macrophages, and PKR blockage or PKR gene deletion abrogated the enhancement of the parasite growth driven by IL-27, as well as the L. amazonensis-induced macrophage production of IL-27. We also observed that L. amazonensis-induced expression of IL-27 depends on type I interferon signaling and the engagement of Toll-like receptor 2. Treatment of Leishmania-infected mice with IL-27 increased lesion size and parasite loads in the footpad and lymph nodes of infected animals, indicating that this cytokine exerts a local and a systemic effect on parasite growth and propagation. In conclusion, we show that IL-27 is a L. amazonensis-enhancing factor and that the PKR/IFN1 axis and IL-10 are critical mediators of this IL-27 induced effect. PMID:25466588
In vitro evaluation of (-)α-bisabolol as a promising agent against Leishmania amazonensis.
Rottini, Mariana Margatto; Amaral, Ana Claudia Fernandes; Ferreira, Jose Luiz Pinto; Silva, Jefferson Rocha de Andrade; Taniwaki, Noemi Nosomi; Souza, Celeste da Silva Freitas de; d'Escoffier, Luiz Ney; Almeida-Souza, Fernando; Hardoim, Daiana de Jesus; Gonçalves da Costa, Sylvio Celso; Calabrese, Kátia da Silva
Current treatments for leishmaniasis present some difficulties due to their toxicity, the use of the intravenous route for administration and therapy duration, which may lead to treatment discontinuation. The aim of this study is to investigate new treatment alternatives to improve patients well being. Therefore, we evaluated the inhibitory effect of (-)α-bisabolol, a sesquiterpene alcohol found in various essential oils of different plant species, against the promastigotes and intracellular amastigotes forms of Leishmania amazonensis, as well as the cytotoxic, morphological and ultrastructural alterations of treated cells. Promastigotes forms of L. amazonensis were incubated with (-)α-bisabolol to determine the antileishmanial activity of this compound. The cytotoxicity effect was evaluated by testing against J774.G8 cells. After these tests, the infected and uninfected cells with L. amazonensis were used to determine if the (-)α-bisabolol was able to kill intracellular parasites and to cause some morphological changes in the cells. The (-)α-bisabolol compound showed significant antileishmanial activity against promastigotes with a 50% effective concentration of 8.07 µg/ml (24 h) and 4.26 µg/ml (48 h). Against intracellular amastigotes the IC50 (inhibitory concentration) of (-)α-bisabolol (24 h) was 4.15 µg/ml. The (-)α-bisabolol also showed a cytotoxic effect against the macrophage strain J774.G8. The value of 50% cytotoxic concentration was 14.82 µg/ml showing that (-)α-bisabolol is less toxic to macrophages than to the parasite. Ultrastructural studies of treated promastigotes and amastigotes showed several alterations, such as loss of cytoplasmic organelles, including the nucleus, and the presence of lipid inclusions. This study showed that (-)α-bisabolol has promising antileishmanial properties, as it can act against the promastigote forms and is able to penetrate the cell, and is also active against the amastigote forms. About 69
Modulation of Na+/K+ ATPase Activity by Hydrogen Peroxide Generated through Heme in L. amazonensis.
Rocco-Machado, Nathália; Cosentino-Gomes, Daniela; Meyer-Fernandes, José Roberto
Leishmania amazonensis is a protozoan parasite that occurs in many areas of Brazil and causes skin lesions. Using this parasite, our group showed the activation of Na+/K+ ATPase through a signaling cascade that involves the presence of heme and protein kinase C (PKC) activity. Heme is an important biomolecule that has pro-oxidant activity and signaling capacity. Reactive oxygen species (ROS) can act as second messengers, which are required in various signaling cascades. Our goal in this work is to investigate the role of hydrogen peroxide (H2O2) generated in the presence of heme in the Na+/K+ ATPase activity of L. amazonensis. Our results show that increasing concentrations of heme stimulates the production of H2O2 in a dose-dependent manner until a concentration of 2.5 μM heme. To confirm that the effect of heme on the Na+/K+ ATPase is through the generation of H2O2, we measured enzyme activity using increasing concentrations of H2O2 and, as expected, the activity increased in a dose-dependent manner until a concentration of 0.1 μM H2O2. To investigate the role of PKC in this signaling pathway, we observed the production of H2O2 in the presence of its activator phorbol 12-myristate 13-acetate (PMA) and its inhibitor calphostin C. Both showed no effect on the generation of H2O2. Furthermore, we found that PKC activity is increased in the presence of H2O2, and that in the presence of calphostin C, H2O2 is unable to activate the Na+/K+ ATPase. 100 μM of Mito-TEMPO was capable of abolishing the stimulatory effect of heme on Na+/K+ ATPase activity, indicating that mitochondria might be the source of the hydrogen peroxide production induced by heme. The modulation of L. amazonensis Na+/K+ ATPase by H2O2 opens new possibilities for understanding the signaling pathways of this parasite. PMID:26070143
Detection of Leishmania (Leishmania) amazonensis and Leishmania (Leishmania) infantum chagasi in Brazilian bats.
Savani, Elisa San Martin Mouriz; de Almeida, Marilene Fernandes; de Oliveira Camargo, Maria Cecília Gibrail; D'Auria, Sandra Regina Nicoletti; Silva, Miriam Martos Sodré; de Oliveira, Maria Lúcia; Sacramento, Débora
Although bats are one of the most abundant mammals in the new world and are present in virtually all ecosystems, including urban and peri-urban environments, few studies have investigated the role of these animals in the epidemiological chain of leishmaniosis. Here, we report a study of 683 bats captured in São Paulo county (southeastern from Brazil), which were screened by serology, parasitologic methods and polymerase chain reaction (PCR) for trypanosomatids. The indirect immunofluorescent antibody test demonstrated that 0.9% of bats react positively for leishmaniosis and PCR detected the presence of DNA of Leishmania (Leishmania) amazonensis in 18 bats and Leishmania (Leishmania) infantum chagasi in 3 specimens. These results indicate that further studies are necessary to evaluate the role of bats in maintenance of the Leishmania life cycle, especially in areas where these diseases are endemic. PMID:19939568
Leishmania amazonensis: Anionic currents expressed in oocytes upon microinjection of mRNA from the parasite.
Lagos M, Luisa F; Moran, Oscar; Camacho, Marcela
Transport mechanisms involved in pH homeostasis are relevant for the survival of Leishmania parasites. The presence of chloride conductive pathways in Leishmania has been anticipated since anion channel inhibitors limit the proton extrusion mediated by the H+ATPase, which is the major regulator of intracellular pH in amastigotes. In this study, we used Xenopus laevis oocytes as a heterologous expression system in which to study the expression of ion channels upon microinjection of polyA mRNA from Leishmania amazonensis. After injection of polyA mRNA into the oocytes, we measured three different types of currents. We discuss the possible origin of each, and propose that Type 3 currents could be the result of the heterologous expression of proteins from Leishmania since they show different pharmacological and biophysical properties as compared to endogenous oocyte currents. PMID:17328895
Evaluation of Antileishmanial Activity of Albaha Medicinal Plants against Leishmania amazonensis
Al-Sokari, Saeed S.; Ali, Nasser A. Awadh; Monzote, Lianet; Al-Fatimi, Mohamed A.
Sixteen methanolic extracts obtained from thirteen plant species, selected either from ethnobotanical or chemotaxonomical data, were screened for their antileishmanial activity against Leishmania amazonensis. The cytotoxic activity against normal peritoneal macrophages from normal BALB/c mice was also determined. Eight extracts had IC50 values ranging from <12.5 to 37.8 µg/mL against promastigotes. Achillea biebersteinii flower, Euphorbia helioscopia, and Solanum incanum leaf extracts showed antileishmanial activities with IC50 between <12.5–26.9 µg/mL and acceptable selectivity indices of 8–5. The other leishmanicidal plant extracts, with IC50 ranging from 18.0 to 29.5 µg/mL, exhibited low selectivity indices. PMID:26357662
In Vitro Activity of the Antifungal Azoles Itraconazole and Posaconazole against Leishmania amazonensis
de Macedo-Silva, Sara Teixeira; Urbina, Julio A.; de Souza, Wanderley; Rodrigues, Juliany Cola Fernandes
Leishmaniasis, caused by protozoan parasites of the Leishmania genus, is one of the most prevalent neglected tropical diseases. It is endemic in 98 countries, causing considerable morbidity and mortality. Pentavalent antimonials are the first line of treatment for leishmaniasis except in India. In resistant cases, miltefosine, amphotericin B and pentamidine are used. These treatments are unsatisfactory due to toxicity, limited efficacy, high cost and difficult administration. Thus, there is an urgent need to develop drugs that are efficacious, safe, and more accessible to patients. Trypanosomatids, including Leishmania spp. and Trypanosoma cruzi, have an essential requirement for ergosterol and other 24-alkyl sterols, which are absent in mammalian cells. Inhibition of ergosterol biosynthesis is increasingly recognized as a promising target for the development of new chemotherapeutic agents. The aim of this work was to investigate the antiproliferative, physiological and ultrastructural effects against Leishmania amazonensis of itraconazole (ITZ) and posaconazole (POSA), two azole antifungal agents that inhibit sterol C14α-demethylase (CYP51). Antiproliferative studies demonstrated potent activity of POSA and ITZ: for promastigotes, the IC50 values were 2.74 µM and 0.44 µM for POSA and ITZ, respectively, and for intracellular amastigotes, the corresponding values were 1.63 µM and 0.08 µM, for both stages after 72 h of treatment. Physiological studies revealed that both inhibitors induced a collapse of the mitochondrial membrane potential (ΔΨm), which was consistent with ultrastructural alterations in the mitochondrion. Intense mitochondrial swelling, disorganization and rupture of mitochondrial membranes were observed by transmission electron microscopy. In addition, accumulation of lipid bodies, appearance of autophagosome-like structures and alterations in the kinetoplast were also observed. In conclusion, our results indicate that ITZ and POSA are potent
Potent In Vitro Antiproliferative Synergism of Combinations of Ergosterol Biosynthesis Inhibitors against Leishmania amazonensis
de Macedo-Silva, S. T.; Visbal, G.; Urbina, J. A.; de Souza, W.
Leishmaniases comprise a spectrum of diseases caused by protozoan parasites of the Leishmania genus. Treatments available have limited safety and efficacy, high costs, and difficult administration. Thus, there is an urgent need for safer and more-effective therapies. Most trypanosomatids have an essential requirement for ergosterol and other 24-alkyl sterols, which are absent in mammalian cells. In previous studies, we showed that Leishmania amazonensis is highly susceptible to aryl-quinuclidines, such as E5700, which inhibit squalene synthase, and to the azoles itraconazole (ITZ) and posaconazole (POSA), which inhibit C-14α-demethylase. Herein, we investigated the antiproliferative, ultrastructural, and biochemical effects of combinations of E5700 with ITZ and POSA against L. amazonensis. Potent synergistic antiproliferative effects were observed against promastigotes, with fractional inhibitory concentration (FIC) ratios of 0.0525 and 0.0162 for combinations of E5700 plus ITZ and of E5700 plus POSA, respectively. Against intracellular amastigotes, FIC values were 0.175 and 0.1125 for combinations of E5700 plus ITZ and E5700 plus POSA, respectively. Marked alterations of the ultrastructure of promastigotes treated with the combinations were observed, in particular mitochondrial swelling, which was consistent with a reduction of the mitochondrial transmembrane potential, and an increase in the production of reactive oxygen species. We also observed the presence of vacuoles similar to autophagosomes in close association with mitochondria and an increase in the number of lipid bodies. Both growth arrest and ultrastructural/biochemical alterations were strictly associated with the depletion of the 14-desmethyl endogenous sterol pool. These results suggest the possibility of a novel combination therapy for the treatment of leishmaniasis. PMID:26239973
Potent In Vitro Antiproliferative Synergism of Combinations of Ergosterol Biosynthesis Inhibitors against Leishmania amazonensis.
de Macedo-Silva, S T; Visbal, G; Urbina, J A; de Souza, W; Rodrigues, J C F
Leishmaniases comprise a spectrum of diseases caused by protozoan parasites of the Leishmania genus. Treatments available have limited safety and efficacy, high costs, and difficult administration. Thus, there is an urgent need for safer and more-effective therapies. Most trypanosomatids have an essential requirement for ergosterol and other 24-alkyl sterols, which are absent in mammalian cells. In previous studies, we showed that Leishmania amazonensis is highly susceptible to aryl-quinuclidines, such as E5700, which inhibit squalene synthase, and to the azoles itraconazole (ITZ) and posaconazole (POSA), which inhibit C-14α-demethylase. Herein, we investigated the antiproliferative, ultrastructural, and biochemical effects of combinations of E5700 with ITZ and POSA against L. amazonensis. Potent synergistic antiproliferative effects were observed against promastigotes, with fractional inhibitory concentration (FIC) ratios of 0.0525 and 0.0162 for combinations of E5700 plus ITZ and of E5700 plus POSA, respectively. Against intracellular amastigotes, FIC values were 0.175 and 0.1125 for combinations of E5700 plus ITZ and E5700 plus POSA, respectively. Marked alterations of the ultrastructure of promastigotes treated with the combinations were observed, in particular mitochondrial swelling, which was consistent with a reduction of the mitochondrial transmembrane potential, and an increase in the production of reactive oxygen species. We also observed the presence of vacuoles similar to autophagosomes in close association with mitochondria and an increase in the number of lipid bodies. Both growth arrest and ultrastructural/biochemical alterations were strictly associated with the depletion of the 14-desmethyl endogenous sterol pool. These results suggest the possibility of a novel combination therapy for the treatment of leishmaniasis. PMID:26239973
Parasitophorous vacuoles of Leishmania amazonensis-infected macrophages maintain an acidic pH.
Antoine, J C; Prina, E; Jouanne, C; Bongrand, P
Leishmania amastigotes are intracellular protozoan parasites of mononuclear phagocytes which reside within parasitophorous vacuoles of phagolysosomal origin. The pH of these compartments was studied with the aim of elucidating strategies used by these microorganisms to evade the microbicidal mechanisms of their host cells. For this purpose, rat bone marrow-derived macrophages were infected with L. amazonensis amastigotes. Intracellular acidic compartments were localized by using the weak base 3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine as a probe. This indicator, which can be detected by light microscopy by using immunocytochemical methods, mainly accumulated in perinuclear lysosomes of uninfected cells, whereas in infected cells, it was essentially localized in parasitophorous vacuoles, which thus appeared acidified. Phagolysosomal pH was estimated quantitatively in living cells loaded with the pH-sensitive endocytic tracer fluoresceinated dextran. After a 15- to 20-h exposure, the tracer was mainly detected in perinuclear lysosomes and parasitophorous vacuoles of uninfected and infected macrophages, respectively. Fluorescence intensities were determined from digitized video images of single cells after processing and automatic subtraction of background. We found statistically different mean pH values of 5.17 to 5.48 for lysosomes and 4.74 to 5.26 for parasitophorous vacuoles. As for lysosomes of monensin-treated cells, the pH gradient of parasitophorous vacuoles collapsed after monensin was added. This very likely indicates that these vacuoles maintain an acidic internal pH by an active process. These results show that L. amazonensis amastigotes are acidophilic and opportunistic organisms and suggest that these intracellular parasites have evolved means for survival under these harsh conditions and have acquired plasma membrane components compatible with the environment. Images PMID:1689700
Antisense phosphorothioate oligonucleotides: selective killing of the intracellular parasite Leishmania amazonensis.
Ramazeilles, C; Mishra, R K; Moreau, S; Pascolo, E; Toulmé, J J
We targeted the mini-exon sequence, present at the 5' end of every mRNA of the protozoan parasite Leishmania amazonensis, by phosphorothioate oligonucleotides. A complementary 16-mer (16PS) was able to kill amastigotes--the intracellular stage of the parasite--in murine macrophages in culture. After 24 hr of incubation with 10 microM 16PS, about 30% infected macrophages were cured. The oligomer 16PS acted through antisense hybridization in a sequence-dependent way; no effect on parasites was observed with noncomplementary phosphorothioate oligonucleotides. The antisense oligonucleotide 16PS was a selective killer of the protozoans without any detrimental effect to the host macrophage. Using 16PS linked to a palmitate chain, which enabled it to complex with low density lipoproteins, improved the leishmanicidal efficiency on intracellular amastigotes, probably due to increased endocytosis. Phosphorothioate oligonucleotides complementary to the intron part of the mini-exon pre-RNA were also effective, suggesting that antisense oligomers could prevent trans-splicing in these parasites. Images PMID:8058724
Modulation of Leishmania (L.) amazonensis Growth in Cultured Mouse Macrophages by Prostaglandins and Platelet Activating Factor
Lonardoni, M. V. C.; Barbieri, C. L.; Russo, M.
The role of endogenously synthesized PAF and prostaglandins on the infection of mouse macrophages by Letsbmanta (L.) amazonensis was investigated, as well as the possible correlation between the effects of these inflammatory mediators with nitric oxide production. It was found that pretreatment of macrophages with 10−5 M of the PAF antagonists, BN-52021 or WEB-2086, increased macrophage infection by 17 and 59%, respectively. The cyclooxygenase inhibitor, indomethacin (10 μg/ml), induced a significant inhibition which was reversed by addition of PGE (10-3 M) to the culture medium. These results suggested that the infection of macrophages by leisbmanla is inhibited by PAF and enhanced by prostaglandins and that these mediators are produced by macrophages during this infection. This was confirmed by addition of these mediators to the culture medium before infection; PAF (10−6, 10−9 and 10−12M) reduced significantly the infection whereas PGE2 (10−5 M) induced a marked enhancement. This effect of exogenous PAF on macrophage infection was reversed by the two PAF antagonists used in this study as well as by the inhibitor of nitric oxide synthesis, L-arginine methyl ester (100 mM). Taken together the data suggest that endogenous production of PAF and PGE2 exert opposing effects on Lesbmana–macrophage interaction and that nitric oxide may be involved in the augmented destruction of parasites induced by PAF. PMID:18472932
Protein kinase A activity is associated with metacyclogenesis in Leishmania amazonensis.
Genestra, Marcelo; Cysne-Finkelstein, Léa; Leon, Leonor
Because of the importance of cell signalling processes in proliferation and differentiation, the adenylate cyclase pathway was studied, specifically the protein kinase A (PKA) in Leishmania amazonensis. The PKAs of soluble (SF) and enriched membrane fractions (MF) from infective/non-infective promastigotes and axenic amastigotes were assayed. In order to purify the PKA molecule, fractions were chromatographed on DEAE-cellulose columns and the phosphorylative activity was evaluated using [gamma(32)P]-ATP as the phosphate source. These experiments were performed in the presence of cyclic adenosine monophosphate (cAMP) and an inhibitor of PKA. Our data demonstrated that the PKA activity was significantly higher (about two times) in SF from promastigotes with a high concentration of metacyclic forms, when compared with the non-infective promastigotes, suggesting an association of this activity and the metacyclogenesis process. A discrete phosphorylative activity in axenic amastigotes was observed. As the adenylate cyclase/cAMP pathway would be involved in the parasite-host interiorization, the PKA activity may constitute a good intracellular target for studies of leishmanicidal drugs. PMID:15338471
Determination of femto Newton forces and fluid viscosity using optical tweezers: application to Leishmania amazonensis
NASA Astrophysics Data System (ADS)
Fontes, Adriana; Giorgio, Selma; de Castro, Archimedes B., Jr.; Neto, Vivaldo M.; Pozzo, Liliana d. Y.; Marques, Gustavo P.; Barbosa, Luiz C.; Cesar, Carlos L.
The objective of this research is to use the displacements of a polystyrene microsphere trapped by an optical tweezers (OT) as a force transducer in mechanical measurements in life sciences. To do this we compared the theoretical optical and hydrodynamic models with experimental data under a broad variation of parameters such as fluid viscosity, refractive index, drag velocity and wall proximities. The laser power was measured after the objective with an integration sphere because normal power meters do not provide an accurate measurement for beam with high numerical apertures. With this careful laser power determination the plot of the optical force (calculated by the particle displacement) versus hydrodynamic force (calculated by the drag velocity) under very different conditions shows an almost 45 degrees straight line. This means that hydrodynamic models can be used to calibrate optical forces and vice-versa. With this calibration we observed the forces of polystyrene bead attached to the protozoa Leishmania amazonensis, responsible for a serious tropical disease. The force range is from 200 femto Newtons to 4 pico Newtons and these experiments shows that OT can be used for infection mechanism and chemotaxis studies in parasites. The other application was to use the optical force to measure viscosities of few microliters sample. Our result shows 5% accuracy measurements.
Extracellular matrix alterations in experimental Leishmania amazonensis infection in susceptible and resistant mice
Leishmania is inoculated, by the bite of an infected sandfly, into the skin of the host, where the promastigotes are phagocyted by dermal macrophages. The dermal region comprises cells and abundant extracellular matrix. Studies show that matrix metalloproteinases play an important role in host defense responses against pathogens in mammals and that their activities lead to the production of antimicrobial peptides. The aim of this study is to evaluate the changes in the distribution of fibronectin and laminin as well as in the elastic system fibres during the course of infection caused by Leishmania amazonensis in mice with distinct genetic backgrounds of susceptibility to this parasite. The results showed that BALB/c presented an enhancement of fibronectin during the course of infection when compared to their control group while the infected or non-infected C3H.He showed a decrease of this protein at end of the experiment. Laminin, on the other hand, remained unaltered in both strains. Also in both BALB/c and C3H.He mice the elastic and elaunin fibres remained unchanged while the oxytalan fibres decreased along the experiment. Ninety days after the infection C3H.He mice had recovered their capacity to produce oxytalan fibres. PMID:22316002
The in vitro biological activity of the Brazilian brown seaweed Dictyota mertensii against Leishmania amazonensis.
dos Santos Aliança, Amanda Silva; dos Anjos, Keicyanne Fernanda Lessa; de Vasconcelos Reis, Thiago Nogueira; Higino, Taciana Mirely Maciel; Brelaz-de-Castro, Maria Carolina Accioly; Bianco, Éverson Miguel; de Figueiredo, Regina Celia Bressan Queiroz
Seaweeds present a wide variety of interesting bioactive molecules. In the present work we evaluated the biological activity of the dichloromethane/methanol (2:1) extract (DME) from the brown seaweed Dictyota mertensii against Leishmania amazonensis and its cytotoxic potential on mammalian cells. The extract showed significant inhibitory effect on the growth of promastigote forms (IC50=71.60 μg/mL) and low toxicity against mammalian cells (CC50=233.10 μg/mL). The DME was also efficient in inhibiting the infection in macrophages, with CC50 of 81.4 μg/mL and significantly decreased the survival of amastigote forms within these cells. The selectivity index showed that DME was more toxic to both promastigote (SI=3.25) and amastigote (SI=2.86) forms than to macrophages. Increased NO production was observed in treated macrophages suggesting that besides acting directly on the parasites, the DME also shows an immunomodulatory effect on macrophages. Drastic ultrastructural alterations consistent with loss of viability and cell death were observed in treated parasites. Confocal microscopy and cytometry analyzes showed no significant impairment of plasma membrane integrity, whereas an intense depolarization of mitochondrial membrane could be observed by using propidium iodide and rhodamine 123 staining, respectively. The low toxicity to mammalian cells and the effective activity against promastigotes and amastigotes, point to the use of DME as a promising agent for the treatment of cutaneous leishmaniasis. PMID:25207712
Distinct Macrophage Fates after in vitro Infection with Different Species of Leishmania: Induction of Apoptosis by Leishmania (Leishmania) amazonensis, but Not by Leishmania (Viannia) guyanensis
DaMata, Jarina Pena; Mendes, Bárbara Pinheiro; Maciel-Lima, Kátia; Menezes, Cristiane Alves Silva; Dutra, Walderez Ornelas; Sousa, Lirlândia Pires; Horta, Maria Fátima
Leishmania is an intracellular parasite in vertebrate hosts, including man. During infection, amastigotes replicate inside macrophages and are transmitted to healthy cells, leading to amplification of the infection. Although transfer of amastigotes from infected to healthy cells is a crucial step that may shape the outcome of the infection, it is not fully understood. Here we compare L. amazonensis and L. guyanensis infection in C57BL/6 and BALB/c mice and investigate the fate of macrophages when infected with these species of Leishmania in vitro. As previously shown, infection of mice results in distinct outcomes: L. amazonensis causes a chronic infection in both strains of mice (although milder in C57BL/6), whereas L. guyanensis does not cause them disease. In vitro, infection is persistent in L. amazonensis-infected macrophages whereas L. guyanensis growth is controlled by host cells from both strains of mice. We demonstrate that, in vitro, L. amazonensis induces apoptosis of both C57BL/6 and BALB/c macrophages, characterized by PS exposure, DNA cleavage into nucleosomal size fragments, and consequent hypodiploidy. None of these signs were seen in macrophages infected with L. guyanensis, which seem to die through necrosis, as indicated by increased PI-, but not Annexin V-, positive cells. L. amazonensis-induced macrophage apoptosis was associated to activation of caspases-3, -8 and -9 in both strains of mice. Considering these two species of Leishmania and strains of mice, macrophage apoptosis, induced at the initial moments of infection, correlates with chronic infection, regardless of its severity. We present evidence suggestive that macrophages phagocytize L. amazonensis-infected cells, which has not been verified so far. The ingestion of apoptotic infected macrophages by healthy macrophages could be a way of amastigote spreading, leading to the establishment of infection. PMID:26513474
Leishmania amazonensis promastigotes in 3D Collagen I culture: an in vitro physiological environment for the study of extracellular matrix and host cell interactions
Rodrigues, Juliany C.F.; Viana, Nathan B.; Pontes, Bruno; Pereira, Camila F.A.; Silva-Filho, Fernando C.
Leishmania amazonensis is the causative agent of American cutaneous leishmaniasis, an important neglected tropical disease. Once Leishmania amazonensis is inoculated into the human host, promastigotes are exposed to the extracellular matrix (ECM) of the dermis. However, little is known about the interaction between the ECM and Leishmania promastigotes. In this study we established L. amazonensis promastigote culture in a three-dimensional (3D) environment mainly composed of Collagen I (COL I). This 3D culture recreates in vitro some aspects of the human host infection site, enabling the study of the interaction mechanisms of L. amazonensis with the host ECM. Promastigotes exhibited "freeze and run" migration in the 3D COL I matrix, which is completely different from the conventional in vitro swimming mode of migration. Moreover, L. amazonensis promastigotes were able to invade, migrate inside, and remodel the 3D COL I matrix. Promastigote trans-matrix invasion and the freeze and run migration mode were also observed when macrophages were present in the matrix. At least two classes of proteases, metallo- and cysteine proteases, are involved in the 3D COL I matrix degradation caused by Leishmania. Treatment with a mixture of protease inhibitors significantly reduced promastigote invasion and migration through this matrix. Together our results demonstrate that L. amazonensis promastigotes release proteases and actively remodel their 3D environment, facilitating their migration. This raises the possibility that promastigotes actively interact with their 3D environment during the search for their cellular "home"—macrophages. Supporting this hypothesis, promastigotes migrated faster than macrophages in a novel 3D co-culture model. PMID:24765565
Increased tau phosphorylation and receptor for advanced glycation endproducts (RAGE) in the brain of mice infected with Leishmania amazonensis.
Gasparotto, Juciano; Senger, Mario Roberto; Kunzler, Alice; Degrossoli, Adriana; de Simone, Salvatore Giovanni; Bortolin, Rafael Calixto; Somensi, Nauana; Girardi, Carolina Saibro; de Souza, Celeste da Silva Freitas; Calabrese, Kátia da Silva; Dal-Pizzol, Felipe; Moreira, José Claudio Fonseca; Silva-Jr, Floriano Paes; Gelain, Daniel Pens
Leishmaniasis is a parasitosis caused by several species of the genus Leishmania, an obligate intramacrophagic parasite. Although neurologic symptoms have been observed in human cases of leishmaniasis, the manifestation of neurodegenerative processes is poorly studied. The aim of the present work was to investigate if peripheral infection of BALB/c mice with Leishmania amazonensis affects tau phosphorylation and RAGE protein content in the brain, which represent biochemical markers of neurodegenerative processes observed in diseases with a pro-inflammatory component, including Alzheimer's disease and Down syndrome. Four months after a single right hind footpad subcutaneous injection of L. amazonensis, the brain cortex of BALB/c mice was isolated. Western blot analysis indicated an increase in tau phosphorylation (Ser(396)) and RAGE immunocontent in infected animals. Brain tissue TNF-α, IL-1β, and IL-6 levels were not different from control animals; however, increased protein carbonylation, decreased IFN-γ levels and impairment in antioxidant defenses were detected. Systemic antioxidant treatment (NAC 20mg/kg, i.p.) inhibited tau phosphorylation and recovered IFN-γ levels. These data, altogether, indicate an association between impaired redox state, tau phosphorylation and RAGE up-regulation in the brain cortex of animals infected with L. amazonensis. In this context, it is possible that neurologic symptoms associated to chronic leishmaniasis are associated to disruptions in the homeostasis of CNS proteins, such as tau and RAGE, as consequence of oxidative stress. This is the first demonstration of alterations in biochemical parameters of neurodegeneration in an experimental model of Leishmania infection. PMID:25014011
Combined effect of the essential oil from Chenopodium ambrosioides and antileishmanial drugs on promastigotes of Leishmania amazonensis.
Monzote, Lianet; Montalvo, Ana Margarita; Scull, Ramón; Miranda, Migdalia; Abreu, Juan
To date, there are no vaccines against Leishmania, and chemotherapy remains the mainstay for the control of leishmaniasis. The drugs of choice used for leishmaniasis therapy are significantly toxic, expensive and with a growing frequency of refractory infections. Because of these limitations, a combination therapy is the better hope. This work demonstrates that the essential oil from Chenopodium ambrosioides shows a synergic activity after incubation in conjunction with pentamidine against promastigotes of Leishmania amazonensis. However, an indifferent effect has been found for combinations of meglumine antimoniate or amphotericin B and the essential oil. PMID:17823757
Functional complementation of Leishmania (Leishmania) amazonensis AP endonuclease gene (lamap) in Escherichia coli mutant strains challenged with DNA damage agents
Verissimo-Villela, Erika; Kitahara-Oliveira, Milene Yoko; dos Reis, Ana Beatriz de Bragança; Albano, Rodolpho Mattos; Da-Cruz, Alda Maria; Bello, Alexandre Ribeiro
During its life cycle Leishmania spp. face several stress conditions that can cause DNA damages. Base Excision Repair plays an important role in DNA maintenance and it is one of the most conserved mechanisms in all living organisms. DNA repair in trypanosomatids has been reported only for Old World Leishmania species. Here the AP endonuclease from Leishmania (L.) amazonensis was cloned, expressed in Escherichia coli mutants defective on the DNA repair machinery, that were submitted to different stress conditions, showing ability to survive in comparison to the triple null mutant parental strain BW535. Phylogenetic and multiple sequence analyses also confirmed that LAMAP belongs to the AP endonuclease class of proteins. PMID:27223868
Shewanella amazonensis sp. nov., a novel metal-reducing facultative anaerobe from Amazonian shelf muds
NASA Technical Reports Server (NTRS)
Venkateswaran, K.; Dollhopf, M. E.; Aller, R.; Stackebrandt, E.; Nealson, K. H.
A new bacterial species belonging to the genus Shewanella is described on the basis of phenotypic characterization and sequence analysis of its 16S rRNA-encoding and gyrase B (gyrB) genes. This organism, isolated from shallow-water marine sediments derived from the Amazon River delta, is a Gram-negative, motile, polarly flagellated, facultatively anaerobic, rod-shaped eubacterium and has a G&C content of 51.7 mol%. Strain SB2BT is exceptionally active in the anaerobic reduction of iron, manganese and sulfur compounds. SB2BT grows optimally at 35 degrees C, with 1-3% NaCl and over a pH range of 7-8. Analysis of the 16S rDNA sequence revealed a clear affiliation between strain SB2BT and members of the gamma subclass of the class Proteobacteria. High similarity values were found with certain members of the genus Shewanella, especially with Shewanella putrefaciens, and this was supported by cellular fatty acid profiles and phenotypic characterization. DNA-DNA hybridization between strain SB2BT and its phylogenetically closest relatives revealed low similarity values (24.6-42.7%) which indicated species status for strain SB2BT. That SB2BT represents a distinct bacterial species within the genus Shewanella is also supported by gyrB sequence analysis. Considering the source of the isolate, the name Shewanella amazonensis sp. nov. is proposed and strain SB2BT (= ATCC 700329T) is designated as the type strain.
In Vitro and In Vivo Activities of 2,3-Diarylsubstituted Quinoxaline Derivatives against Leishmania amazonensis.
Kaplum, Vanessa; Cogo, Juliana; Sangi, Diego Pereira; Ueda-Nakamura, Tânia; Corrêa, Arlene Gonçalves; Nakamura, Celso Vataru
Leishmaniasis is endemic in 98 countries and territories worldwide. The therapies available for leishmaniasis have serious side effects, thus prompting the search for new therapies. The present study investigated the antileishmanial activities of 2,3-diarylsubstituted quinoxaline derivatives against Leishmania amazonensis The antiproliferative activities of 6,7-dichloro-2,3-diphenylquinoxaline (LSPN329) and 2,3-di-(4-methoxyphenyl)-quinoxaline (LSPN331) against promastigotes and intracellular amastigotes were assessed, and the cytotoxicities of LSPN329 and LSPN331 were determined. Morphological and ultrastructural alterations were examined by electron microscopy, and biochemical alterations, reflected by the mitochondrial membrane potential (ΔΨm), mitochondrial superoxide anion (O2·(-)) concentration, the intracellular ATP concentration, cell volume, the level of phosphatidylserine exposure on the cell membrane, cell membrane integrity, and lipid inclusions, were evaluated. In vivo antileishmanial activity was evaluated in a murine cutaneous leishmaniasis model. Compounds LSPN329 and LSPN331 showed significant selectivity for promastigotes and intracellular amastigotes and low cytotoxicity. In promastigotes, ultrastructural alterations were observed, including an increase in lipid inclusions, concentric membranes, and intense mitochondrial swelling, which were associated with hyperpolarization of ΔΨm, an increase in the O2·(-) concentration, decreased intracellular ATP levels, and a decrease in cell volume. Phosphatidylserine exposure and DNA fragmentation were not observed. The cellular membrane remained intact after treatment. Thus, the multifactorial response that was responsible for the cellular collapse of promastigotes was based on intense mitochondrial alterations. BALB/c mice treated with LSPN329 or LSPN331 showed a significant decrease in lesion thickness in the infected footpad. Therefore, the antileishmanial activity and mitochondrial mechanism of
Determination of fluid viscosity and femto Newton forces of Leishmania amazonensis using optical tweezers
NASA Astrophysics Data System (ADS)
Fontes, Adriana; Giorgio, Selma; de Castro, Archimedes, Jr.; Neto, Vivaldo M.; de Y. Pozzo, Liliana; de Thomaz, Andre A.; Barbosa, Luiz C.; Cesar, Carlos L.
The displacements of a polystyrene microsphere trapped by an optical tweezers (OT) can be used as a force transducer for mechanical measurements in life sciences such as the measurement of forces of living microorganisms or the viscosity of local fluids. The technique we used allowed us to measure forces on the 200 femto Newtons to 4 pico Newtons range of the protozoa Leishmania amazonensis, responsible for a serious tropical disease. These observations can be used to understand the infection mechanism and chemotaxis of these parasites. The same technique was used to measure viscosities of few microliters sample with agreement with known samples better than 5%. To calibrate the force as a function of the microsphere displacement we first dragged the microsphere in a fluid at known velocity for a broad range of different optical and hydrodynamical parameters. The hydrodynamical model took into account the presence of two walls and the force depends on drag velocity, fluid viscosity and walls proximities, while the optical model in the geometric optics regime depends on the particle and fluid refractive indexes and laser power. To measure the high numerical (NA) aperture laser beam power after the objective we used an integration sphere to avoid the systematic errors of usual power meters for high NA beams. After this careful laser power measurement we obtained an almost 45 degrees straight line for the plot of the optical force (calculated by the particle horizontal displacement) versus hydrodynamic force (calculated by the drag velocity) under variation of all the parameters described below. This means that hydrodynamic models can be used to calibrate optical forces, as we have done for the parasite force measurement, or vice-versa, as we did for the viscosity measurements.
Leishmania amazonensis: effects of heat shock on ecto-ATPase activity.
Peres-Sampaio, Carlos Eduardo; de Almeida-Amaral, Elmo Eduardo; Giarola, Naira Ligia Lima; Meyer-Fernandes, José Roberto
In this work we demonstrated that promastigotes of Leishmania amazonensis exhibit an Mg-dependent ecto-ATPase activity, which is stimulated by heat shock. The Mg-dependent ATPase activity of cells grown at 22 and 28 degrees C was 41.0+/-5.2 nmol Pi/h x 10(7)cells and 184.2+/-21.0 nmol Pi/h x 10(7)cells, respectively. When both promastigotes were pre-incubated at 37 degrees C for 2h, the ATPase activity of cells grown at 22 degrees C was increased to 136.4+/-10.6 nmol Pi/h x 10(7) whereas that the ATPase activity of cells grown at 28 degrees C was not modified by the heat shock (189.8+/-10.3 nmol Pi/h x 10(7)cells). It was observed that Km of the enzyme from cells grown at 22 degrees C (Km=980.2+/-88.6 microM) was the same to the enzyme from cells grown at 28 degrees C (Km=901.4+/-91.9 microM). In addition, DIDS (4,4'-diisothiocyanatostilbene 2,2'-disulfonic acid) and suramin, two inhibitors of ecto-ATPases, also inhibited similarly the ATPase activities from promastigotes grown at 22 and 28 degrees C. We also observed that cells grown at 22 degrees C exhibit the same ecto-phosphatase and ecto 3'- and 5'-nucleotidase activities than cells grown at 28 degrees C. Interestingly, cycloheximide, an inhibitor of protein synthesis, suppressed the heat-shock effect on ecto-ATPase activity of cells grown at 22 degrees C were exposed at 37 degrees C for 2h. A comparison between the stimulation of the Mg-dependent ecto-ATPase activity of virulent and avirulent promastigotes by the heat shock showed that avirulent promastigotes had a higher stimulation than virulent promastigotes after heat stress. PMID:18295760
Photodynamic therapy for American cutaneous leishmaniasis: the efficacy of methylene blue in hamsters experimentally infected with Leishmania (Leishmania) amazonensis.
Peloi, Lílian Somenci; Biondo, Carlos Eduardo Guerino; Kimura, Elza; Politi, Mario José; Lonardoni, Maria Valdrinez Campana; Aristides, Sandra Mara Alessi; Dorea, Regina Coeli Cunha; Hioka, Noboru; Silveira, Thaís Gomes Verzignassi
The aim of this study was to investigate the effectiveness of Photodynamic Therapy (PDT) using Methylene Blue (MB) as the photosensitizing compound and a Light-Emitting Diode (LED) in American cutaneous leishmaniasis (ACL). Hamsters were experimentally infected with Leishmania (Leishmania) amazonensis. After the development of the lesions in the footpad, the animals were treated with MB three times a week for 3 months. Ten minutes after each application of MB, the lesions were irradiated with LED for 1 h. The lesions were evaluated weekly by the measurement of the hamster footpad thickness. At the end of the treatment the parasitic load was quantified in the regional lymph node of the hamsters. The treatment promoted a decrease in the thickness of infected footpad (P=0.0001) and reduction in the parasitic load in the regional lymph node (P=0.0007) of the animals from group treated with MB+LED. PDT using MB+LED in ACL caused by L. amazonensis shows a strong photodynamic effect. This therapy is very promising, once it is an inexpensive system and the own patient can apply it in their wound and in their house without the need of technical assistance. PMID:21575635
Leishmania amazonensis DNA in wild females of Lutzomyia cruzi (Diptera: Psychodidae) in the state of Mato Grosso do Sul, Brazil
de Oliveira, Everton Falcão; Casaril, Aline Etelvina; Mateus, Nathália Lopes Fontoura; Murat, Paula Guerra; Fernandes, Wagner Souza; Oshiro, Elisa Teruya; de Oliveira, Alessandra Gutierrez; Galati, Eunice Aparecida Bianchi
Studies on natural infection by Leishmania spp of sandflies collected in endemic and nonendemic areas can provide important information on the distribution and intensity of the transmission of these parasites. This study sought to investigate the natural infection by Leishmaniain wild female sandflies. The specimens were caught in the city of Corumbá, state of Mato Grosso do Sul (Brazil) between October 2012-March 2014, and dissected to investigate flagellates and/or submitted to molecular analysis to detect Leishmania DNA. A total of 1,164 females (77.56% of which were Lutzomyia cruzi) representing 11 species were investigated using molecular analysis; 126 specimens of Lu. cruziwere dissected and also submitted to molecular analysis. The infection rate based on the presence of Leishmania DNA considering all the sandfly species analysed was 0.69%; only Leishmania (Leishmania) amazonensis was identified in Lu. cruzi by the molecular analysis. The dissections were negative for flagellates. This is the first record of the presence of L. (L.) amazonensis DNA in Lu. cruzi, and the first record of this parasite in this area. These findings point to the need for further investigation into the possible role of this sandfly as vector of this parasite. PMID:26602870
Neutrophils have a protective role during early stages of Leishmania amazonensis infection in BALB/c mice.
Sousa, L M A; Carneiro, M B H; Resende, M E; Martins, L S; Dos Santos, L M; Vaz, L G; Mello, P S; Mosser, D M; Oliveira, M A P; Vieira, L Q
Neutrophils are involved in the early stages of immune responses to pathogens. Here, we investigated the role of neutrophils during the establishment of Leishmania amazonensis infection in BALB/c and C57BL/6 mice. First, we showed an accumulation of neutrophils between 6 and 24 h post-infection, followed by a reduction in neutrophil numbers after 72 h. Next, we depleted neutrophils prior to infection using RB6-8C5 or 1A8 mAb. Neutrophil depletion led to faster lesion development, increased parasite numbers and higher arginase activity during the first week of infection in BALB/c mice, but not in C57BL/6 mice. Increased susceptibility was accompanied by augmented levels of anti-L. amazonensis IgG and increased production of IL-10 and IL-17. Because IL-10 is a mediator of susceptibility to Leishmania infection, we blocked IL-10 signalling in neutrophil-depleted mice using anti-IL-10R. Interestingly, inhibition of IL-10 signalling abrogated the increase in parasite loads observed in neutrophil-depleted mice, suggesting that parasite proliferation is at least partially mediated by IL-10. Additionally, we tested the effect of IL-17 in inflammatory macrophages and observed that IL-17 increased arginase activity and favoured parasite growth. Taken together, our data indicate that neutrophils control parasite numbers and limit lesion development during the first week of infection in BALB/c mice. PMID:24102495
In vivo and in vitro phagocytosis of Leishmania (Leishmania) amazonensis promastigotes by B-1 cells.
Geraldo, M M; Costa, C R; Barbosa, F M C; Vivanco, B C; Gonzaga, W F K M; Novaes E Brito, R R; Popi, A F; Lopes, J D; Xander, P
Leishmaniasis is caused by Leishmania parasites that infect several cell types. The promastigote stage of Leishmania is internalized by phagocytic cells and transformed into the obligate intracellular amastigote form. B-1 cells are a subpopulation of B cells that are able to differentiate in vitro and in vivo into mononuclear phagocyte-like cells with phagocytic properties. B-1 cells use several receptors for phagocytosis, such as the mannose receptor and third complement receptor. Leishmania binds to the same receptors on macrophages. In this study, we demonstrated that phagocytes derived from B-1 cells (B-1 CDP) were able to internalize promastigotes of L. (L.) amazonensis in vitro. The internalized promastigotes differentiated into amastigotes. Our results showed that the phagocytic index was higher in B-1 CDP compared to peritoneal macrophages and bone marrow-derived macrophages. The in vivo phagocytic ability of B-1 cells was also demonstrated. Parasites were detected inside purified B-1 cells after intraperitoneal infection with L. (L.) amazonensis promastigotes. Intraperitoneal stimulation with the parasites led to an increase in both IL-10 and TNF-α. These results highlight the importance of studying B-1 CDP cells as phagocytic cells that can participate and contribute to immunity to parasites. PMID:27084328
Distinct courses of infection with Leishmania (L.) amazonensis are observed in BALB/c, BALB/c nude and C57BL/6 mice.
Velasquez, Leonardo G; Galuppo, Mariana K; DE Rezende, Eloiza; Brandão, Wesley N; Peron, Jean Pierre; Uliana, Silvia R B; Duarte, Maria Irma; Stolf, Beatriz S
Leishmania (L.) amazonensis [L. (L.) amazonensis] is widely distributed in Brazil and its symptomatic infections usually lead to few localized lesions and sometimes to diffuse cutaneous form, with nodules throughout the body, anergy to parasite antigens and poor therapeutic response. The variability of these manifestations draws attention to the need for studies on the pathophysiology of infection by this species. In this study, we analysed the course and immunological aspects of L. (L.) amazonensis infection in BALB/c and C57BL/6 strains, both susceptible, but displaying different clinical courses, and athymic BALB/c nude, to illustrate the role of T cell dependent responses. We analysed footpad thickness and parasite burden by in vivo imaging. Furthermore, we evaluated the cellular profile and cytokine production in lymph nodes and the inflammatory infiltrates of lesions. Nude mice showed delayed lesion development and less inflammatory cells in lesions, but higher parasite burden than BALB/c and C57BL/6. BALB/c and C57BL/6 mice had similar parasite burdens, lesion sizes and infiltrates until 6 weeks after infection, and after that C57BL/6 mice controlled the infection. Small differences in parasite numbers were observed in C57BL/6 macrophages in vitro, indicating that in vivo milieu accounts for most differences in infection. We believe our results shed light on the role of host immune system in the course of L. (L.) amazonensis infection by comparing three mouse strains that differ in parasitaemia and inflammatory cells. PMID:26892342
The Effect of Ursolic Acid on Leishmania (Leishmania) amazonensis Is Related to Programed Cell Death and Presents Therapeutic Potential in Experimental Cutaneous Leishmaniasis
Yamamoto, Eduardo S.; Campos, Bruno L. S.; Jesus, Jéssica A.; Laurenti, Márcia D.; Ribeiro, Susan P.; Kallás, Esper G.; Rafael-Fernandes, Mariana; Santos-Gomes, Gabriela; Silva, Marcelo S.; Sessa, Deborah P.; Lago, João H. G.; Levy, Débora; Passero, Luiz F. D.
Among neglected tropical diseases, leishmaniasis is one of the most important ones, affecting more than 12 million people worldwide. The available treatments are not well tolerated, and present diverse side effects, justifying the search for new therapeutic compounds. In the present study, the activity of ursolic acid (UA) and oleanolic acid (OA) were assayed in experimental cutaneous leishmaniasis (in vitro and in vivo). Promastigote forms of L. amazonensis were incubated with OA and UA for 24h, and effective concentration 50% (EC50) was estimated. Ultraestructural alterations in Leishmania amazonensis promastigotes after UA treatment were evaluated by transmission electron microscopy, and the possible mode of action was assayed through Annexin V and propidium iodide staining, caspase 3/7 activity, DNA fragmentation and transmembrane mitochondrial potential. The UA potential was evaluated in intracellular amastigotes, and its therapeutic potential was evaluated in L. amazonensis infected BALB/c mice. UA eliminated L. amazonensis promastigotes with an EC50 of 6.4 μg/mL, comparable with miltefosine, while OA presented only a marginal effect on promastigote forms at 100 μg/mL. The possible mechanism by which promastigotes were eliminated by UA was programmed cell death, independent of caspase 3/7, but it was highly dependent on mitochondria activity. UA was not toxic for peritoneal macrophages from BALB/c mice, and it was able to eliminate intracellular amastigotes, associated with nitric oxide (NO) production. OA did not eliminate amastigotes nor trigger NO. L. amazonensis infected BALB/c mice submitted to UA treatment presented lesser lesion size and parasitism compared to control. This study showed, for the first time, that UA eliminate promastigote forms through a mechanism associated with programed cell death, and importantly, was effective in vivo. Therefore, UA can be considered an interesting candidate for future tests as a prototype drug for the treatment
Leishmania donovani Nucleoside Hydrolase Terminal Domains in Cross-Protective Immunotherapy Against Leishmania amazonensis Murine Infection
Nico, Dirlei; Gomes, Daniele Crespo; Palatnik-de-Sousa, Iam; Morrot, Alexandre; Palatnik, Marcos; Palatnik-de-Sousa, Clarisa Beatriz
Nucleoside hydrolases of the Leishmania genus are vital enzymes for the replication of the DNA and conserved phylogenetic markers of the parasites. Leishmania donovani nucleoside hydrolase (NH36) induced a main CD4+ T cell driven protective response against L. chagasi infection in mice which is directed against its C-terminal domain. In this study, we used the three recombinant domains of NH36: N-terminal domain (F1, amino acids 1–103), central domain (F2 aminoacids 104–198), and C-terminal domain (F3 amino acids 199–314) in combination with saponin and assayed their immunotherapeutic effect on Balb/c mice previously infected with L. amazonensis. We identified that the F1 and F3 peptides determined strong cross-immunotherapeutic effects, reducing the size of footpad lesions to 48 and 64%, and the parasite load in footpads to 82.6 and 81%, respectively. The F3 peptide induced the strongest anti-NH36 antibody response and intradermal response (IDR) against L. amazonenis and a high secretion of IFN-γ and TNF-α with reduced levels of IL-10. The F1 vaccine, induced similar increases of IgG2b antibodies and IFN-γ and TNF-α levels, but no IDR and no reduction of IL-10. The multiparameter flow cytometry analysis was used to assess the immune response after immunotherapy and disclosed that the degree of the immunotherapeutic effect is predicted by the frequencies of the CD4+ and CD8+ T cells producing IL-2 or TNF-α or both. Total frequencies and frequencies of double-cytokine CD4 T cell producers were enhanced by F1 and F3 vaccines. Collectively, our multifunctional analysis disclosed that immunotherapeutic protection improved as the CD4 responses progressed from 1+ to 2+, in the case of the F1 and F3 vaccines, and as the CD8 responses changed qualitatively from 1+ to 3+, mainly in the case of the F1 vaccine, providing new correlates of immunotherapeutic protection against cutaneous leishmaniasis in mice based on T-helper TH1 and CD8+ mediated immune responses
Identification of Differentially Expressed Proteins from Leishmania amazonensis Associated with the Loss of Virulence of the Parasites
Magalhães, Rubens D. M.; Duarte, Mariana C.; Mattos, Eliciane C.; Martins, Vivian T.; Lage, Paula S.; Chávez-Fumagalli, Miguel A.; Lage, Daniela P.; Menezes-Souza, Daniel; Régis, Wiliam C. B.; Manso Alves, Maria J.; Soto, Manuel; Tavares, Carlos A. P.; Nagen, Ronaldo A. P.; Coelho, Eduardo A. F.
Background The present study analyzed whether or not the in vitro cultivation for long periods of time of pre-isolated Leishmania amazonensis from lesions of chronically infected BALB/c mice was able to interfere in the parasites' infectivity using in vivo and in vitro experiments. In addition, the proteins that presented a significant decrease or increase in their protein expression content were identified applying a proteomic approach. Methodology/Principal Findings Parasites were cultured in vitro for 150 days. Aliquots were collected on the day 0 of culture (R0), as well as after ten (R10; 50 days of culture), twenty (R20; 100 days of culture), and thirty (R30; 150 days of culture) passages, and were used to analyze the parasites' in vitro and in vivo infectivity, as well as to perform the proteomic approach. Approximately 837, 967, 935, and 872 spots were found in 2-DE gels prepared from R0, R10, R20, and R30 samples, respectively. A total of 37 spots presented a significant decrease in their intensity of expression, whereas a significant increase in protein content during cultivation could be observed for 19 proteins (both cases >2.0 folds). Some of these identified proteins can be described, such as diagnosis and/or vaccine candidates, while others are involved in the infectivity of Leishmania. It is interesting to note that six proteins, considered hypothetical in Leishmania, showed a significant decrease in their expression and were also identified. Conclusions/Significance The present study contributes to the understanding that the cultivation of parasites over long periods of time may well be related to the possible loss of infectivity of L. amazonensis. The identified proteins that presented a significant decrease in their expression during cultivation, including the hypothetical, may also be related to this loss of parasites' infectivity, and applied in future studies, including vaccine candidates and/or immunotherapeutic targets against leishmaniasis
Surface Zn-proteinase as a molecule for defense of Leishmania mexicana amazonensis promastigotes against cytolysis inside macrophage phagolysosomes.
Seay, M B; Heard, P L; Chaudhuri, G
The role of the surface membrane Zn-proteinase in protecting the cellular integrity of the macrophage parasite Leishmania mexicana amazonensis from intraphagolysosomal cytolysis was studied. These cells lose their infectivity to host macrophages after prolonged cultivation in axenic growth medium. The virulent and attenuated variants of the parasite cells were cloned. Failure of these attenuated parasite cells to survive inside macrophage phagolysosomes is associated with 20- to 50-fold reduction in the expression of surface gp63 protein. In situ inhibition of gp63 proteinase activity inside Leishmania-infected macrophage phagolysosomes with targeted delivery of an inhibitor of gp63 proteinase activity, 1,10-phenanthroline, selectively eliminated intracellular Leishmania amastigotes, further suggesting the importance of this proteinase in phagolysosomal survival of the parasite. An upstream sequence (US) of the gp63 gene was cloned in front of the bacterial chloramphenicol acetyltransferase (CAT) gene in plasmid pCATbasic. Transfection of L. mexicana amazonensis cells with this recombinant plasmid showed that expression of the CAT gene from this US is 15- to 20-fold higher in virulent clones than in avirulent clones of the parasite. Band shift analysis with the cloned US also showed that binding of protein(s) was 15- to 20-fold higher in virulent cell extract than in avirulent cell extract. Coating of attenuated cells or liposomes with proteolytically active gp63 protects them from degradation inside macrophage phagolysosomes. These results suggest a novel mechanism of survival of this phagolysosomal parasite with the help of its surface Zn-proteinase. PMID:8945556
Phage-fused epitopes from Leishmania infantum used as immunogenic vaccines confer partial protection against Leishmania amazonensis infection.
Costa, Lourena Emanuele; Chávez-Fumagalli, Miguel Angel; Martins, Vivian Tamietti; Duarte, Mariana Costa; Lage, Daniela Pagliara; Lima, Mayara I S; Pereira, Nathália Cristina De Jesus; Soto, Manuel; Tavares, Carlos Alberto Pereira; Goulart, Luiz Ricardo; Coelho, Eduardo Antonio Ferraz
Two mimotopes of Leishmania infantum identified by phage display were evaluated as vaccine candidates in BALB/c mice against Leishmania amazonensis infection. The epitope-based immunogens, namely B10 and C01, presented as phage-fused peptides; were used without association of a Th1 adjuvant, and they were administered isolated or in combination into animals. Both clones showed a specific production of interferon-gamma (IFN-γ), interleukin-12 (IL-12) and granulocyte/macrophage colony-stimulating factor (GM-CSF) after in vitro spleen cells stimulation, and they were able to induce a partial protection against infection. Significant reductions of parasite load in the infected footpads, liver, spleen, bone marrow and paws' draining lymph nodes were observed in the immunized mice, in comparison with the control groups (saline, saponin, wild-type and non-relevant clones). Protection was associated with an IL-12-dependent production of IFN-γ, mediated mainly by CD8(+) T cells, against parasite proteins. Protected mice also presented low levels of IL-4 and IL-10, as well as increased levels of parasite-specific IgG2a antibodies. The association of both clones resulted in an improved protection in relation to their individual use. More importantly, the absence of adjuvant did not diminish the cross-protective efficacy against Leishmania spp. infection. This study describes for the first time two epitope-based immunogens selected by phage display technology against L. infantum infected dogs sera, which induced a partial protection in BALB/c mice infected with L. amazonensis. PMID:26099574
Co-infection by Leishmania amazonensis and L. infantum/L. chagasi in a case of diffuse cutaneous leishmaniasis in Bolivia.
Martinez, E; Mollinedo, S; Torrez, M; Muñoz, M; Bañuls, A L; Le Pont, F
We present the first report of a co-infection by Leishmania amazonensis and L. infantum/L. chagasi isolated in 1993 from a patient with diffuse cutaneous leishmaniasis (DCL), living in the sub-Andean region of Bolivia. This is the third reported case of DCL in Bolivia, but the first one with isoenzymatic identification of the aetiological agents involved and the first one giving evidence for a mixed infection by 2 Leishmania parasites in the same lesion. PMID:12474481
Pre-exposure with low-dose UVA suppresses lesion development and enhances Th1 response in BALB/c mice infected with Leishmania (Leishmania) amazonensis.
Khaskhely, N M; Maruno, M; Takamiyagi, A; Uezato, H; Kasem, K M; Hosokawa, A; Kariya, K; Hashiguchi, Y; Landires, E A; Nonaka, S
This study was conducted to determine whether exposing mice to ultraviolet (UV) radiation would alter the pathogenesis of infection with Leishmania (Leishmania) amazonensis (L. amazonensis) which causes progressive cutaneous disease in susceptible mouse strains. BALB/c mice were irradiated with 10 and 30 J/cm(2) UVA on shaved skin of the back from Dermaray (M-DMR-100) for 4 consecutive days before infection with Leishmania promastigotes. The course of disease was recorded by measuring the size of lesions at various times after infection. Mice groups irradiated with UVA 10 and 30 J/cm(2) showed significantly suppressed lesion development compared with the non-irradiated mice. Light and electron microscopy revealed a few parasites at the site of inoculation in UVA-irradiated subjects. Sandwich enzyme-linked-immunosorbent-assay (ELISA) examination of sera showed dose dependently upregulated interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12, and downregulated interleukin (IL)-4 and interleukin (IL)-10 levels in UVA-irradiated as compared with the non-irradiated mice. Positive signals for IFN-gamma mRNA in irradiated mice were obtained by RT-PCR, while non-irradiated mice showed negative results. None of the examined samples showed signal for IL-4 mRNA. The present study disclosed that exposure of mice to different low-doses of UVA irradiation prior to infection may interfere with immunity to L. amazonensis in the murine model. This indicates that the cell-mediated response switch from Th2 to Th1 pattern suppressed the cutaneous lesions of L. amazonensis. PMID:11390207
Cross-Protective Immunity to Leishmania amazonensis is Mediated by CD4+ and CD8+ Epitopes of Leishmania donovani Nucleoside Hydrolase Terminal Domains.
Nico, Dirlei; Gomes, Daniele Crespo; Alves-Silva, Marcus Vinícius; Freitas, Elisangela Oliveira; Morrot, Alexandre; Bahia, Diana; Palatnik, Marcos; Rodrigues, Mauricio M; Palatnik-de-Sousa, Clarisa B
The nucleoside hydrolase (NH) of Leishmania donovani (NH36) is a phylogenetic marker of high homology among Leishmania parasites. In mice and dog vaccination, NH36 induces a CD4+ T cell-driven protective response against Leishmania chagasi infection directed against its C-terminal domain (F3). The C-terminal and N-terminal domain vaccines also decreased the footpad lesion caused by Leishmania amazonensis. We studied the basis of the crossed immune response using recombinant generated peptides covering the whole NH36 sequence and saponin for mice prophylaxis against L. amazonensis. The F1 (amino acids 1-103) and F3 peptide (amino acids 199-314) vaccines enhanced the IgG and IgG2a anti-NH36 antibodies to similar levels. The F3 vaccine induced the strongest DTH response, the highest proportions of NH36-specific CD4+ and CD8+ T cells after challenge and the highest expression of IFN-γ and TNF-α. The F1 vaccine, on the other hand, induced a weaker but significant DTH response and a mild enhancement of IFN-γ and TNF-α levels. The in vivo depletion with anti-CD4 or CD8 monoclonal antibodies disclosed that cross-protection against L. amazonensis infection was mediated by a CD4+ T cell response directed against the C-terminal domain (75% of reduction of the size of footpad lesion) followed by a CD8+ T cell response against the N-terminal domain of NH36 (57% of reduction of footpad lesions). Both vaccines were capable of inducing long-term cross-immunity. The amino acid sequence of NH36 showed 93% identity to the sequence of the NH A34480 of L. amazonensis, which also showed the presence of completely conserved predicted epitopes for CD4+ and CD8+ T cells in F1 domain, and of CD4+ epitopes differing by a single amino acid, in F1 and F3 domains. The identification of the C-terminal and N-terminal domains as the targets of the immune response to NH36 in the model of L. amazonensis infection represents a basis for the rationale development of a bivalent vaccine against
Lipophosphoglycans from Leishmania amazonensis Strains Display Immunomodulatory Properties via TLR4 and Do Not Affect Sand Fly Infection
Nogueira, Paula M.; Assis, Rafael R.; Torrecilhas, Ana C.; Saraiva, Elvira M.; Pessoa, Natália L.; Campos, Marco A.; Marialva, Eric F.; Ríos-Velasquez, Cláudia M.; Pessoa, Felipe A.; Secundino, Nágila F.; Rugani, Jerônimo N.; Nieves, Elsa; Turco, Salvatore J.; Melo, Maria N.
The immunomodulatory properties of lipophosphoglycans (LPG) from New World species of Leishmania have been assessed in Leishmania infantum and Leishmania braziliensis, the causative agents of visceral and cutaneous leishmaniasis, respectively. This glycoconjugate is highly polymorphic among species with variation in sugars that branch off the conserved Gal(β1,4)Man(α1)-PO4 backbone of repeat units. Here, the immunomodulatory activity of LPGs from Leishmania amazonensis, the causative agent of diffuse cutaneous leishmaniasis, was evaluated in two strains from Brazil. One strain (PH8) was originally isolated from the sand fly and the other (Josefa) was isolated from a human case. The ability of purified LPGs from both strains was investigated during in vitro interaction with peritoneal murine macrophages and CHO cells and in vivo infection with Lutzomyia migonei. In peritoneal murine macrophages, the LPGs from both strains activated TLR4. Both LPGs equally activate MAPKs and the NF-κB inhibitor p-IκBα, but were not able to translocate NF-κB. In vivo experiments with sand flies showed that both stains were able to sustain infection in L. migonei. A preliminary biochemical analysis indicates intraspecies variation in the LPG sugar moieties. However, they did not result in different activation profiles of the innate immune system. Also those polymorphisms did not affect infectivity to the sand fly. PMID:27508930
Novel 3,4-methylenedioxyde-6-X-benzaldehyde-thiosemicarbazones: Synthesis and antileishmanial effects against Leishmania amazonensis.
de Melos, Jorge Luiz R; Torres-Santos, Eduardo Caio; Faiões, Viviane dos S; Del Cistia, Catarina de Nigris; Sant'Anna, Carlos Maurício R; Rodrigues-Santos, Cláudio Eduardo; Echevarria, Aurea
A series of eleven 3,4-methylenedioxyde-6-X-benzaldehyde-thiosemicarbazones (16-27) was synthesised as part of a study to search for potential new drugs with a leishmanicidal effect. The thiosemicarbazones, ten of which are new compounds, were prepared in good yields (85-98%) by the reaction of 3,4-methylenedioxyde-6-benzaldehydes (6-X-piperonal), previously synthesised for this work by several methodologies, and thiosemicarbazide in ethanol with a few drops of H2SO4. These compounds were evaluated against Leishmania amazonensis promastigotes, and derivatives where X = I (22) and X = CN (23) moieties showed impressive results, having IC₅₀ = 20.74 μM and 16.40 μM, respectively. The intracellular amastigotes assays showed IC₅₀ = 22.00 μM (22) and 17.00 μM (23), and selectivity index >5.7 and >7.4, respectively, with a lower toxicity compared to pentamidine (positive control, SI = 4.5). The results obtained from the preliminary QSAR study indicated the hydrophobicity (log P) as a fundamental parameter for the 2D-QSAR linear model. A molecular docking study demonstrated that both compounds interact with flavin mononucleotide (FMN), important binding site of NO synthase. PMID:26375353
Lipophosphoglycans from Leishmania amazonensis Strains Display Immunomodulatory Properties via TLR4 and Do Not Affect Sand Fly Infection.
Nogueira, Paula M; Assis, Rafael R; Torrecilhas, Ana C; Saraiva, Elvira M; Pessoa, Natália L; Campos, Marco A; Marialva, Eric F; Ríos-Velasquez, Cláudia M; Pessoa, Felipe A; Secundino, Nágila F; Rugani, Jerônimo N; Nieves, Elsa; Turco, Salvatore J; Melo, Maria N; Soares, Rodrigo P
The immunomodulatory properties of lipophosphoglycans (LPG) from New World species of Leishmania have been assessed in Leishmania infantum and Leishmania braziliensis, the causative agents of visceral and cutaneous leishmaniasis, respectively. This glycoconjugate is highly polymorphic among species with variation in sugars that branch off the conserved Gal(β1,4)Man(α1)-PO4 backbone of repeat units. Here, the immunomodulatory activity of LPGs from Leishmania amazonensis, the causative agent of diffuse cutaneous leishmaniasis, was evaluated in two strains from Brazil. One strain (PH8) was originally isolated from the sand fly and the other (Josefa) was isolated from a human case. The ability of purified LPGs from both strains was investigated during in vitro interaction with peritoneal murine macrophages and CHO cells and in vivo infection with Lutzomyia migonei. In peritoneal murine macrophages, the LPGs from both strains activated TLR4. Both LPGs equally activate MAPKs and the NF-κB inhibitor p-IκBα, but were not able to translocate NF-κB. In vivo experiments with sand flies showed that both stains were able to sustain infection in L. migonei. A preliminary biochemical analysis indicates intraspecies variation in the LPG sugar moieties. However, they did not result in different activation profiles of the innate immune system. Also those polymorphisms did not affect infectivity to the sand fly. PMID:27508930
Antibody subclass profile against Leishmania braziliensis and Leishmania amazonensis in the diagnosis and follow-up of mucosal leishmaniasis.
Junqueira Pedras, Mariana; Orsini, Marcela; Castro, Marcelo; Passos, Valéria M A; Rabello, Ana
Sensitivities and specificities of IgG subclasses-ELISA and IgG-indirect fluorescent antibody test (IFAT) against Leishmania braziliensis (Lb) and L. amazonensis (La) antigens were determined in 17 patients with mucosal (ML) and 19 with muco-cutaneous (MCL) leishmaniasis. Using IFAT-IgG both antigens gave high sensitivities and were statistically similar, being 89.5% with La and 100% with Lb. Using ELISA, the highest sensitivity was achieved with total IgG for ML (94.7% with both antigens) and MCL (100% with both antigens). Cross-reactivity, observed with Chagas disease and malaria sera reduced the specificity of the IgG-based assays, being 50 to 70% with IFAT and 40 to 70% with ELISA. An increase in specificity was obtained with IgG1-ELISA (90% with Lb and 100% with La). Serum levels of anti-Lb-IgG and IgG3 dropped 90 days after treatment. IgG subclasses antibody detection constitute an valuable alternative to increase the efficiency of sorological diagnostics of ML/MCL. PMID:14596966
Time-course analysis of the Shewanella amazonensis SB2B proteome in response to sodium chloride shock
Parnell, John J.; Callister, Stephen J.; Rompato, Giovanni; Nicora, Carrie D.; Pasa-Tolic, Ljiljana; Williamson, Ashley; Pfrender, Michael E.
Organisms in the genus Shewanella have become models for response to environmental stress. One of the most important environmental stresses is change in osmolarity. In this study, we experimentally determine the response mechanisms of Shewanella amazonensis SB2B during osmotic stress. Osmotic stress in SB2B was induced through exposure to NaCl, and the time-course proteomics response was measured using liquid chromatography mass spectrometry. Protein trends were qualitatively compared to gene expression trends and to phenotypic characterization. Osmotic stress affects motility, and has also been associated with a change in the membrane fatty acid composition (due to induction of branched chain amino acid degradation pathways); however, we show this is not the case for SB2B. Although proteins and genes involved with branched chain amino acid degradation are induced, fatty acid degradation pathways are not induced and no change in the fatty acid profile occurs in SB2B as a result of osmotic shock. The most extensive response of SB2B over the time course of acclimation to high salt involves an orchestrated sequence of events comprising increased expression of signal transduction associated with motility and restricted cell division and DNA replication. After SB2B has switched to increased branched chain amino acid degradation, motility, and cellular replication proteins return to pre-perturbed levels.
Aureobasidium-Derived Soluble Branched (1,3-1,6) β-Glucan (Sophy β-glucan) Enhances Natural Killer Activity in Leishmania amazonensis-Infected Mice
Yatawara, Lalani; Wickramasinghe, Susiji; Nagataki, Mitsuru; Takamoto, Misa; Nomura, Haruka; Ikeue, Yasunori; Watanabe, Yoshiya
The β-glucans derived from yeast cell walls have been reported for having many immunomodulatory activities in vivo and in vitro. In this study, Aureobasidium-derived soluble branched (1,3-1,6) β-glucan (Sophy β-glucan) was checked for natural killer (NK) activity and for the production of IFN-γ and IL-4 in Leishmania amazonensis infection. The main experiment was performed with a group of female C57BL/6 and BALB/c mice, orally supplemented with 5% of Sophy β-glucan and infected with promastogotes of L. amazonensis (1 × 107) into the footpad. Increase in the footpad thickness with time was observed in BALB/c mice in spite of the oral Sophy β-glucan supplement, but it was less in C57BL/6 mice. The difference in overall mean footpad thickness between 'infection only' versus 'infection + glucan' groups was statistically significant (P < 0.001). High NK activity in C57BL/6 than BALB/c mice was observed in 'glucan only' group compared to the control group and also in 'infection + glucan' group compared to 'infection only' group. The difference in the NK activity among these groups was significant (P < 0.05). The IFN-γ level increased at weeks 7 and 8 post-infection in C57BL/6 mice and was significantly high in 'infection + glucan' group compared to the 'infection only' group (P < 0.05). IL-4 levels did not increase up to detectable levels throughout the study. The results led a conclusion that Sophy β-glucan enhances NK activity and cellular immunity in L. amazonensis-infected mice. PMID:19967081
In Vitro Activities of ER-119884 and E5700, Two Potent Squalene Synthase Inhibitors, against Leishmania amazonensis: Antiproliferative, Biochemical, and Ultrastructural Effects▿
Fernandes Rodrigues, Juliany Cola; Concepcion, Juan Luis; Rodrigues, Carlos; Caldera, Aura; Urbina, Julio A.; de Souza, Wanderley
ER-119884 and E5700, novel arylquinuclidine derivatives developed as cholesterol-lowering agents, were potent in vitro growth inhibitors of both proliferative stages of Leishmania amazonensis, the main causative agent of cutaneous leishmaniasis in South America, with the 50% inhibitory concentrations (IC50s) being in the low-nanomolar to subnanomolar range. The compounds were very potent noncompetitive inhibitors of native L. amazonensis squalene synthase (SQS), with inhibition constants also being in the nanomolar to subnanomolar range. Growth inhibition was strictly associated with the depletion of the parasite's main endogenous sterols and the concomitant accumulation of exogenous cholesterol. Using electron microscopy, we identified the intracellular structures affected by the compounds. A large number of lipid inclusions displaying different shapes and electron densities were observed after treatment with both SQS inhibitors, and these inclusions were associated with an intense disorganization of the membrane that surrounds the cell body and flagellum, as well as the endoplasmic reticulum and the Golgi complex. Cells treated with ER-119884 but not those treated with E5700 had an altered cytoskeleton organization due to an abnormal distribution of tubulin, and many were arrested at cytokinesis. A prominent contractile vacuole and a phenotype typical of programmed cell death were frequently found in drug-treated cells. The selectivity of the drugs was demonstrated with the JC-1 mitochondrial fluorescent label and by trypan blue exclusion tests with macrophages, which showed that the IC50s against the host cells were 4 to 5 orders of magnitude greater that those against the intracellular parasites. Taken together, our results show that ER-119884 and E5700 are unusually potent and selective inhibitors of the growth of Leishmania amazonensis, probably because of their inhibitory effects on de novo sterol biosynthesis at the level of SQS, but some of our
Morinda citrifolia Linn. fruit (Noni) juice induces an increase in NO production and death of Leishmania amazonensis amastigotes in peritoneal macrophages from BALB/c.
Almeida-Souza, Fernando; de Souza, Celeste da Silva Freitas; Taniwaki, Noemi Nosomi; Silva, João José Mendes; de Oliveira, Renata Mondêgo; Abreu-Silva, Ana Lúcia; Calabrese, Kátia da Silva
Leishmaniasis is a complex disease that is considered a serious public health problem. Due to the absence of an effective vaccine and debilitating chemotherapy better therapies are urgently needed. This situation has stimulated the search for alternative treatments such as the use of herbal medicines. Several studies conducted with Morinda citrifolia Linn. have shown various biological activities such as antitumor, immunomodulation and antileishmanial activity, however its mechanisms of action are still unknown. This study aimed to analyze the activity of M. citrifolia fruit juice against Leishmania amazonensis and its action on peritoneal macrophages from BALB/c infected with L. amazonensis. Activity against the promastigote forms showed IC50 at 275.3 μg/mL. Transmission electron microscopy was used to evaluate the ultrastructural alterations in the promastigotes treated with the juice and the results showed cytoplasmic vacuolization, lipid inclusion and increased activity of exocytosis. The juice treatment presented an IC50 at 208.4 μg/mL against intracellular amastigotes and led to an increased nitrite production in infected and non-infected macrophages. When macrophages were pre-treated with iNOS inhibitors, aminoguanidine or 1400W, the intracellular amastigotes increased, demonstrating the important role of NO production in M. citrifolia fruit activity. In conclusion, our results reveal that treatment with M. citrifolia fruit juice can increase NO production in peritoneal macrophages and this ability has an important role in the killing of L. amazonensis intracellular amastigotes. PMID:27328771
Morinda citrifolia Linn. Reduces Parasite Load and Modulates Cytokines and Extracellular Matrix Proteins in C57BL/6 Mice Infected with Leishmania (Leishmania) amazonensis.
Almeida-Souza, Fernando; Cardoso, Flávia de Oliveira; Souza, Bruno Vinicius da Conceição; do Valle, Tânia Zaverucha; de Sá, Joicy Cortez; Oliveira, Iara Dos Santos da Silva; de Souza, Celeste da Silva Freitas; Moragas Tellis, Carla Junqueira; Chagas, Maria do Socorro Dos Santos; Behrens, Maria Dutra; Abreu-Silva, Ana Lúcia; Calabrese, Kátia da Silva
The absence of an effective vaccine and the debilitating chemotherapy for Leishmaniasis demonstrate the need for developing alternative treatments. Several studies conducted with Morinda citrifolia have shown various biological activities, including antileishmanial activity, however its mechanisms of action are unknown. This study aimed to analyze the in vivo activity of M. citrifolia fruit juice (Noni) against Leishmania (Leishmania) amazonensis in C57BL/6 mice. M. citrifolia fruit juice from the Brazilian Amazon has shown the same constitution of other juices produced around the world and liquid chromatography-mass spectrometry analysis identified five compounds: deacetylasperulosidic acid, asperulosidic acid, rutin, nonioside B and nonioside C. Daily intragastric treatment with Noni was carried out after 55 days of L. (L.) amazonensis infection in C57BL/6 mice. Parasitic loads, cytokine and extracellular protein matrix expressions of the lesion site were analyzed by qPCR. Histopathology of the lesion site, lymph nodes and liver were performed to evaluate the inflammatory processes. Cytokines and biochemical parameters of toxicity from sera were also evaluated. The Noni treatment at 500 mg.kg-1.day-1 for 60 days decreased the lesion size and parasitic load in the footpad infected with L. (L.) amazonensis. The site of infection also showed decreased inflammatory infiltrates and decreased cytokine expressions for IL-12, TNF-α, TGF-β and IL-10. On the other hand, Noni treatment enhanced the extracellular matrix protein expressions of collagen IV, fibronectin and laminin in the infected footpad as well collagen I and II, fibronectin and laminin in the mock-infected footpads. No toxicity was observed at the end of treatment. These data show the efficacy of Noni treatment. PMID:27579922
Recombinant Forms of Leishmania amazonensis Excreted/Secreted Promastigote Surface Antigen (PSA) Induce Protective Immune Responses in Dogs.
Petitdidier, Elodie; Pagniez, Julie; Papierok, Gérard; Vincendeau, Philippe; Lemesre, Jean-Loup; Bras-Gonçalves, Rachel
Preventive vaccination is a highly promising strategy for interrupting leishmaniasis transmission that can, additionally, contribute to elimination. A vaccine formulation based on naturally excreted secreted (ES) antigens was prepared from L. infantum promastigote culture supernatant. This vaccine achieved successful results in Phase III trials and was licensed and marketed as CaniLeish. We recently showed that newly identified ES promastigote surface antigen (PSA), from both viable promastigotes and axenically-grown amastigotes, represented the major constituent and the highly immunogenic antigen of L. infantum and L. amazonensis ES products. We report here that three immunizations with either the recombinant ES LaPSA-38S (rPSA) or its carboxy terminal part LaPSA-12S (Cter-rPSA), combined with QA-21 as adjuvant, confer high levels of protection in naive L. infantum-infected Beagle dogs, as checked by bone marrow parasite absence in respectively 78.8% and 80% of vaccinated dogs at 6 months post-challenge. The parasite burden in infected vaccinated dogs was significantly reduced compared to placebo group, as measured by q-PCR. Moreover, our results reveal humoral and cellular immune response clear-cut differences between vaccinated and control dogs. An early increase in specific IgG2 antibodies was observed in rPSA/QA-21- and Cter-rPSA/QA-21-immunized dogs only. They were found functionally active in vitro and were highly correlated with vaccine protection. In vaccinated protected dogs, IFN-γ and NO productions, as well as anti-leishmanial macrophage activity, were increased. These data strongly suggest that ES PSA or its carboxy-terminal part, in recombinant forms, induce protection in a canine model of zoonotic visceral leishmaniasis by inducing a Th1-dominant immune response and an appropriate specific antibody response. These data suggest that they could be considered as important active components in vaccine candidates. PMID:27223609
Recombinant Forms of Leishmania amazonensis Excreted/Secreted Promastigote Surface Antigen (PSA) Induce Protective Immune Responses in Dogs
Petitdidier, Elodie; Pagniez, Julie; Papierok, Gérard; Vincendeau, Philippe; Lemesre, Jean-Loup; Bras-Gonçalves, Rachel
Preventive vaccination is a highly promising strategy for interrupting leishmaniasis transmission that can, additionally, contribute to elimination. A vaccine formulation based on naturally excreted secreted (ES) antigens was prepared from L. infantum promastigote culture supernatant. This vaccine achieved successful results in Phase III trials and was licensed and marketed as CaniLeish. We recently showed that newly identified ES promastigote surface antigen (PSA), from both viable promastigotes and axenically-grown amastigotes, represented the major constituent and the highly immunogenic antigen of L. infantum and L. amazonensis ES products. We report here that three immunizations with either the recombinant ES LaPSA-38S (rPSA) or its carboxy terminal part LaPSA-12S (Cter-rPSA), combined with QA-21 as adjuvant, confer high levels of protection in naive L. infantum-infected Beagle dogs, as checked by bone marrow parasite absence in respectively 78.8% and 80% of vaccinated dogs at 6 months post-challenge. The parasite burden in infected vaccinated dogs was significantly reduced compared to placebo group, as measured by q-PCR. Moreover, our results reveal humoral and cellular immune response clear-cut differences between vaccinated and control dogs. An early increase in specific IgG2 antibodies was observed in rPSA/QA-21- and Cter-rPSA/QA-21-immunized dogs only. They were found functionally active in vitro and were highly correlated with vaccine protection. In vaccinated protected dogs, IFN-γ and NO productions, as well as anti-leishmanial macrophage activity, were increased. These data strongly suggest that ES PSA or its carboxy-terminal part, in recombinant forms, induce protection in a canine model of zoonotic visceral leishmaniasis by inducing a Th1-dominant immune response and an appropriate specific antibody response. These data suggest that they could be considered as important active components in vaccine candidates. PMID:27223609
Antileishmanial activity and mechanism of action from a purified fraction of Zingiber officinalis Roscoe against Leishmania amazonensis.
Duarte, Mariana C; Tavares, Grasiele S V; Valadares, Diogo G; Lage, Daniela P; Ribeiro, Tatiana G; Lage, Letícia M R; Rodrigues, Marcella R; Faraco, André A G; Soto, Manuel; da Silva, Eduardo S; Chávez Fumagalli, Miguel A; Tavares, Carlos A P; Leite, João Paulo V; Oliveira, Jamil S; Castilho, Rachel O; Coelho, Eduardo A F
In recent years, considerable attention has been given to identify new antileishmanial products derived from medicinal plants, although, to date, no new effective compound has been recently applied to treat leishmaniasis. In the present study, the antileishmanial activity of a water extract from Zingiber officinalis Roscoe (ginger) was investigated and a purified fraction, named F10, was identified as responsible by this biological activity. The chemical characterization performed for this fraction showed that it is mainly composed by flavonoids and saponins. The water extract and the F10 fraction presented IC50 values of 125.5 and 49.8 μg/mL, respectively. Their selectivity indexes (SI) were calculated and values were seven and 40 times higher, respectively, in relation to the value found for amphotericin B, which was used as a control. Additional studies were performed to evaluate the toxicity of these compounds in human red blood cells, besides of the production of nitrite, as an indicator of nitric oxide (NO), in treated and infected macrophages. The results showed that both F10 fraction and water extract were not toxic to human cells, and they were able to stimulate the nitrite production, with values of 13.6 and 5.4 μM, respectively, suggesting that their biological activity could be due to macrophages activation via NO production. In conclusion, the present study shows that a purified fraction from ginger could be evaluated in future works as a therapeutic alternative, on its own or in association with other drugs, to treat disease caused by L. amazonensis. PMID:27013260
Leishmania amazonensis chemotaxis under glucose gradient studied by the strength and directionality of forces measured with optical tweezers
NASA Astrophysics Data System (ADS)
de Ysasa Pozzo, Liliana; Fontes, Adriana; de Thomaz, André A.; Barbosa, Luiz Carlos; Ayres, Diana Copi; Giorgio, Selma; Cesar, Carlos Lenz
Chemotaxis is the mechanism microorganisms use to sense the environment surrounding them and to direct their movement towards attractive, or away from the repellent, chemicals. The biochemical sensing is almost the only way for communication between unicellular organisms. Prokaryote and Eukaryote chemotaxis has been mechanically studied mainly by observing the directionality and timing of the microorganisms movements subjected to a chemical gradient, but not through the directionality and strength of the forces it generates. To observe the vector force of microorganisms under a chemical gradient we developed a system composed of two large chambers connected by a tiny duct capable to keep the chemical gradient constant for more than ten hours. We also used the displacements of a microsphere trapped in an Optical Tweezers as the force transducer to measure the direction and the strength of the propulsion forces of flagellum of the microorganism under several gradient conditions. A 9μm diameter microsphere particle was trapped with a Nd:YAG laser and its movement was measured through the light scattered focused on a quadrant detector. We observed the behavior of the protozoa Leishmania amazonensis (eukaryote) under several glucose gradients. This protozoa senses the gradient around it by swimming in circles for three to five times following by tumbling, and not by the typical straight swimming/tumbling of bacteria. Our results also suggest that force direction and strength are also used to control its movement, not only the timing of swimming/tumbling, because we observed a higher force strength clearly directed towards the glucose gradient.
Localization of major histocompatibility complex class II molecules in phagolysosomes of murine macrophages infected with Leishmania amazonensis.
Antoine, J C; Jouanne, C; Lang, T; Prina, E; de Chastellier, C; Frehel, C
Leishmania-infected macrophages are potential antigen-presenting cells for CD4+ T lymphocytes, which recognize parasite antigens bound to major histocompatibility complex class II molecules (Ia). However, the intracellular sites where Ia and antigens may interact are far from clear, since parasites grow within the modified lysosomal compartment of the host cell, whereas Ia molecules seem to be targeted to endosomes. To address this question, the expression and fate of Ia molecules were studied by immunocytochemistry in Leishmania amazonensis-infected murine macrophages stimulated with gamma interferon. In uninfected macrophages, Ia molecules were localized on the plasma membrane and in perinuclear vesicles, but they underwent a dramatic redistribution after infection, since most of the intracellular staining was then associated with the periphery of the parasitophorous vacuoles (p.v.) and quite often polarized towards amastigote-binding sites. The Ii invariant chain, which is transiently associated with Ia during their intracellular transport, although well expressed in infected macrophages, apparently did not reach the p.v. Similar findings were observed with macrophages from mice either resistant or highly susceptible to Leishmania infection. In order to determine the origin of p.v.-associated Ia, the fate of plasma membrane, endosomal, and lysosomal markers, detected with specific antibodies, was determined after infection. At 48 h after infection, p.v. was found to exhibit a membrane composition typical of mature lysosomes. Overall, these data suggest that (i) Ia located in p.v. originate from secondary lysosomes involved in the biogenesis of this compartment or circulate in several endocytic organelles, including lysosomes and (ii) p.v. could play a role in antigen processing and presentation. Alternatively, the presence of high amounts of Ia in p.v. could be due to a Leishmania-induced mechanism by means of which this organism may evade the immune response
Leishmania amazonensis Amastigotes Highly Express a Tryparedoxin Peroxidase Isoform That Increases Parasite Resistance to Macrophage Antimicrobial Defenses and Fosters Parasite Virulence
Henard, Calvin A.; Carlsen, Eric D.; Hay, Christie; Kima, Peter E.; Soong, Lynn
Professional phagocytes generate a myriad of antimicrobial molecules to kill invading microorganisms, of which nitrogen oxides are integral in controlling the obligate intracellular pathogen Leishmania. Although reactive nitrogen species produced by the inducible nitric oxide synthase (iNOS) can promote the clearance of intracellular parasites, some Leishmania species/stages are relatively resistant to iNOS-mediated antimicrobial activity. The underlying mechanism for this resistance remains largely uncharacterized. Here, we show that the amastigote form of L. amazonensis is hyper-resistant to the antimicrobial actions of cytokine-activated murine and human macrophages as compared to its promastigote counterpart. Amastigotes exhibit a marked ability to directly counter the cytotoxicity of peroxynitrite (ONOO−), a leishmanicidal oxidant that is generated during infection through the combined enzymatic activities of NADPH oxidase and iNOS. The enhanced antinitrosative defense of amastigotes correlates with the increased expression of a tryparedoxin peroxidase (TXNPx) isoform that is also upregulated in response to iNOS enzymatic activity within infected macrophages. Accordingly, ectopic over-expression of the TXNPx isoform by L. amazonensis promastigotes significantly enhances parasite resistance against ONOO− cytotoxicity. Moreover, TXNPx-overexpressing parasites exhibit greater intra-macrophage survival, and increased parasite growth and lesion development in a murine model of leishmaniasis. Our investigations indicate that TXNPx isoforms contribute to Leishmania's ability to adapt to and antagonize the hostile microenvironment of cytokine-activated macrophages, and provide a mechanistic explanation for persistent infection in experimental and human leishmaniasis. PMID:25033301
Contrasting human cytokine responses to promastigote whole-cell extract and the Leishmania analogue receptor for activated C kinase antigen of L. amazonensis in natural infection versus immunization
Azeredo-Coutinho, R B G; Matos, D C S; Armôa, G G R; Maia, R M; Schubach, A; Mayrink, W; Mendonça, S C F
It is known that the same antigen can induce different immune responses, depending upon the way that it is presented to the immune system. The objective of this study was to compare cytokine responses of peripheral blood mononuclear cells (PBMC) from cutaneous leishmaniasis patients and subjects immunized with a first-generation candidate vaccine composed of killed Leishmania amazonensis promastigotes to a whole-cell promastigote antigen extract (La) and to the recombinant protein LACK (Leishmania analogue receptor for activated C kinase), both from L. amazonensis. Thirty-two patients, 35 vaccinees and 13 healthy subjects without exposure to Leishmania, were studied. Cytokine production was assessed by enzyme-linked immunosorbent assay and enzyme-linked immunospot assay. The interferon (IFN)-γ levels stimulated by La were significantly higher and the levels of interleukin (IL)-10 significantly lower than those stimulated by LACK in the patient group, while LACK induced a significantly higher IFN-γ production and a significantly lower IL-10 production compared with those induced by La in the vaccinated group. LACK also induced a significantly higher frequency of IFN-γ-producing cells than did La in the vaccinated group. The contrast in the cytokine responses stimulated by LACK and La in PBMC cultures from vaccinated subjects versus patients indicates that the human immune response to crude and defined Leishmania antigens as a consequence of immunization differs from that induced by natural infection. PMID:18627399
Ecological Niche Modelling Predicts Southward Expansion of Lutzomyia (Nyssomyia) flaviscutellata (Diptera: Psychodidae: Phlebotominae), Vector of Leishmania (Leishmania) amazonensis in South America, under Climate Change.
Carvalho, Bruno M; Rangel, Elizabeth F; Ready, Paul D; Vale, Mariana M
Vector borne diseases are susceptible to climate change because distributions and densities of many vectors are climate driven. The Amazon region is endemic for cutaneous leishmaniasis and is predicted to be severely impacted by climate change. Recent records suggest that the distributions of Lutzomyia (Nyssomyia) flaviscutellata and the parasite it transmits, Leishmania (Leishmania) amazonensis, are expanding southward, possibly due to climate change, and sometimes associated with new human infection cases. We define the vector's climatic niche and explore future projections under climate change scenarios. Vector occurrence records were compiled from the literature, museum collections and Brazilian Health Departments. Six bioclimatic variables were used as predictors in six ecological niche model algorithms (BIOCLIM, DOMAIN, MaxEnt, GARP, logistic regression and Random Forest). Projections for 2050 used 17 general circulation models in two greenhouse gas representative concentration pathways: "stabilization" and "high increase". Ensemble models and consensus maps were produced by overlapping binary predictions. Final model outputs showed good performance and significance. The use of species absence data substantially improved model performance. Currently, L. flaviscutellata is widely distributed in the Amazon region, with records in the Atlantic Forest and savannah regions of Central Brazil. Future projections indicate expansion of the climatically suitable area for the vector in both scenarios, towards higher latitudes and elevations. L. flaviscutellata is likely to find increasingly suitable conditions for its expansion into areas where human population size and density are much larger than they are in its current locations. If environmental conditions change as predicted, the range of the vector is likely to expand to southeastern and central-southern Brazil, eastern Paraguay and further into the Amazonian areas of Bolivia, Peru, Ecuador, Colombia and Venezuela
Ecological Niche Modelling Predicts Southward Expansion of Lutzomyia (Nyssomyia) flaviscutellata (Diptera: Psychodidae: Phlebotominae), Vector of Leishmania (Leishmania) amazonensis in South America, under Climate Change
Carvalho, Bruno M.; Ready, Paul D.
Vector borne diseases are susceptible to climate change because distributions and densities of many vectors are climate driven. The Amazon region is endemic for cutaneous leishmaniasis and is predicted to be severely impacted by climate change. Recent records suggest that the distributions of Lutzomyia (Nyssomyia) flaviscutellata and the parasite it transmits, Leishmania (Leishmania) amazonensis, are expanding southward, possibly due to climate change, and sometimes associated with new human infection cases. We define the vector's climatic niche and explore future projections under climate change scenarios. Vector occurrence records were compiled from the literature, museum collections and Brazilian Health Departments. Six bioclimatic variables were used as predictors in six ecological niche model algorithms (BIOCLIM, DOMAIN, MaxEnt, GARP, logistic regression and Random Forest). Projections for 2050 used 17 general circulation models in two greenhouse gas representative concentration pathways: "stabilization" and "high increase". Ensemble models and consensus maps were produced by overlapping binary predictions. Final model outputs showed good performance and significance. The use of species absence data substantially improved model performance. Currently, L. flaviscutellata is widely distributed in the Amazon region, with records in the Atlantic Forest and savannah regions of Central Brazil. Future projections indicate expansion of the climatically suitable area for the vector in both scenarios, towards higher latitudes and elevations. L. flaviscutellata is likely to find increasingly suitable conditions for its expansion into areas where human population size and density are much larger than they are in its current locations. If environmental conditions change as predicted, the range of the vector is likely to expand to southeastern and central-southern Brazil, eastern Paraguay and further into the Amazonian areas of Bolivia, Peru, Ecuador, Colombia and
The human parasite Leishmania amazonensis downregulates iNOS expression via NF-κB p50/p50 homodimer: role of the PI3K/Akt pathway
Calegari-Silva, Teresa C.; Vivarini, Áislan C.; Miqueline, Marina; Dos Santos, Guilherme R. R. M.; Teixeira, Karina Luiza; Saliba, Alessandra Mattos; Nunes de Carvalho, Simone; de Carvalho, Laís; Lopes, Ulisses G.
Leishmania amazonensis activates the NF-κB transcriptional repressor homodimer (p50/p50) and promotes nitric oxide synthase (iNOS) downregulation. We investigated the role of PI3K/Akt in p50/p50 NF-κB activation and the effect on iNOS expression in L. amazonensis infection. The increased occupancy of p50/p50 on the iNOS promoter of infected macrophages was observed and we demonstrated that both p50/p50 NF-κB induction and iNOS downregulation in infected macrophages depended on PI3K/Akt activation. Importantly, the intracellular growth of the parasite was also impaired during PI3K/Akt signalling inhibition and in macrophages knocked-down for Akt 1 expression. It was also observed that the increased nuclear levels of p50/p50 in L. amazonensis-infected macrophages were associated with reduced phosphorylation of 907 Ser p105, the precursor of p50. Corroborating these data, we demonstrated the increased levels of phospho-9 Ser GSK3β in infected macrophages, which is associated with GSK3β inhibition and, consequently, its inability to phosphorylate p105. Remarkably, we found that the levels of pPTEN 370 Ser, a negative regulator of PI3K, increased due to L. amazonensis infection. Our data support the notion that PI3K/Akt activity is sustained during the parasite infection, leading to NF-κB 105 phosphorylation and further processing to originate p50/p50 homodimers and the consequent downregulation of iNOS expression. PMID:26400473
The human parasite Leishmania amazonensis downregulates iNOS expression via NF-κB p50/p50 homodimer: role of the PI3K/Akt pathway.
Calegari-Silva, Teresa C; Vivarini, Áislan C; Miqueline, Marina; Dos Santos, Guilherme R R M; Teixeira, Karina Luiza; Saliba, Alessandra Mattos; Nunes de Carvalho, Simone; de Carvalho, Laís; Lopes, Ulisses G
Leishmania amazonensis activates the NF-κB transcriptional repressor homodimer (p50/p50) and promotes nitric oxide synthase (iNOS) downregulation. We investigated the role of PI3K/Akt in p50/p50 NF-κB activation and the effect on iNOS expression in L. amazonensis infection. The increased occupancy of p50/p50 on the iNOS promoter of infected macrophages was observed and we demonstrated that both p50/p50 NF-κB induction and iNOS downregulation in infected macrophages depended on PI3K/Akt activation. Importantly, the intracellular growth of the parasite was also impaired during PI3K/Akt signalling inhibition and in macrophages knocked-down for Akt 1 expression. It was also observed that the increased nuclear levels of p50/p50 in L. amazonensis-infected macrophages were associated with reduced phosphorylation of 907 Ser p105, the precursor of p50. Corroborating these data, we demonstrated the increased levels of phospho-9 Ser GSK3β in infected macrophages, which is associated with GSK3β inhibition and, consequently, its inability to phosphorylate p105. Remarkably, we found that the levels of pPTEN 370 Ser, a negative regulator of PI3K, increased due to L. amazonensis infection. Our data support the notion that PI3K/Akt activity is sustained during the parasite infection, leading to NF-κB 105 phosphorylation and further processing to originate p50/p50 homodimers and the consequent downregulation of iNOS expression. PMID:26400473
DFT/PCM, QTAIM, 1H NMR conformational studies and QSAR modeling of thirty-two anti-Leishmania amazonensis Morita-Baylis-Hillman Adducts
NASA Astrophysics Data System (ADS)
Filho, Edilson B. A.; Moraes, Ingrid A.; Weber, Karen C.; Rocha, Gerd B.; Vasconcellos, Mário L. A. A.
Morita-Baylis-Hillman Adducts (MBHA) has been recently synthesized and bio-evaluated by our research group against Leishmania amazonensis, parasite that causes cutaneous and mucocutaneous leishmaniasis. We present here a theoretical conformational study of thirty-two leismanicidal MBHA by B3LYP/6-31+g(d) calculations with Polarized Continuum Model (PCM) to simulate water influence. Intramolecular Hydrogen Bonds (IHBs) indicated to control the most conformational preferences of MBHA. Quantum Theory Atoms in Molecules (QTAIM) calculations were able to characterize these interactions at Bond Critical Point level. Compounds presenting an unusual seven member IHB between NO2 group and hydroxyl moiety, supported by experimental spectroscopic data, showed a considerable improvement of biological activity (lower IC50 values). These results are in accordance to redox NO2 mechanism of action. Based on structural observations, some molecular descriptors were calculated and submitted to Quantitative Structure-Activity Relationship (QSAR) studies through the PLS Regression Method. These studies provided a model with good validation parameters values (R2 = 0.71, Q2 = 0.61 and Qext2 = 0.92).
Leishmania amazonensis: heme stimulates (Na(+)+K(+))ATPase activity via phosphatidylinositol-specific phospholipase C/protein kinase C-like (PI-PLC/PKC) signaling pathways.
Almeida-Amaral, Elmo Eduardo; Cardoso, Viviane Carrozino; Francioli, Fernanda Gomes; Meyer-Fernandes, José Roberto
In the present paper we studied the involvement of the phosphatidylinositol-specific PLC (PI-PLC)/protein kinase C (PKC) pathway in (Na(+)+K(+))ATPase stimulation by heme in Leishmania amazonensis promastigotes. Heme stimulated the PKC-like activity with a concentration of 50nM. Interestingly, the maximal stimulation of the PKC-like activity promoted by phorbol ester was of the same magnitude promoted by heme. However, the stimulatory effect of heme is completely abolished by ET-18-OCH(3) and U73122, specific inhibitors of PI-PLC. (Na(+)+K(+))ATPase activity is increased in the presence of increased concentrations of heme, being maximally affected at 50nM. This effect was completely reversed by 10nM calphostin C, an inhibitor of PKC. Thus, the effect of 50nM heme on (Na(+)+K(+))ATPase activity is completely abolished by ET-18-OCH(3) and U73122. Taken together, these results demonstrate that the heme receptor mediates the stimulatory effect of heme on the (Na(+)+K(+))ATPase activity through a PI-PLC/PKC signaling pathway. PMID:20045694
Abortive infection of Lutzomyia longipalpis insect vectors by aflagellated LdARL-3A-Q70L overexpressing Leishmania amazonensis parasites.
Cuvillier, Armelle; Miranda, Jose Carlos; Ambit, Audrey; Barral, Aldina; Merlin, Gilles
Leishmania donovani ADP-ribosylation factor-like protein 3A (LdARL-3A) is a small G protein isolated from the protozoan parasite L. donovani with no defined physiological function. Previously [Cuvillier, A., Redon, F., Antoine, J.-C., Chardin, P., DeVos, T., and Merlin, G. (2000) J Cell Sci 113: 2065-2074] we have shown that overexpression in L. amazonensis promastigotes of the mutated protein LdARL-3A-Q70L, which remains constitutively associated with GTP, leads to the disappearance of the flagellum but does not impair cell viability or growth. Here we report that parasites overexpressing LdARL-3A-Q70L can invade in vitro cultivated macrophages to the same extent as control cells, demonstrating that the flagellum is not necessary for attachment to or engulfment into macrophages. These infections are productive because amastigotes differentiate and multiply. However, aflagellated LdARL-3A-Q70L-overexpressing Leishmania promastigotes could not survive in experimentally infected Lutzomyia longipalpis insect vectors, in contrast to untransfected or native LdARL-3A-overexpressing cells. Overexpression of the native and mutated proteins did not modify in vitro procyclic to metacyclic lipophosphoglycan maturation or differentiation from procyclic to metacyclic promastigotes, nevertheless there is a block in transmission of Leishmania. Better understanding of LdARL-3A pathways, notably those regarding flagellum biogenesis, may lead to the future development of Leishmania-specific drugs, which may stop parasite transmission in nature without affecting other species. PMID:12969377
Activity, toxicity and analysis of resistance of essential oil from Chenopodium ambrosioides after intraperitoneal, oral and intralesional administration in BALB/c mice infected with Leishmania amazonensis: a preliminary study.
Monzote, Lianet; Montalvo, Ana M; Scull, Ramón; Miranda, Migdalia; Abreu, Juan
The World Health Organization has classified the leishmaniasis as a major tropical disease. Current therapy is toxic, expensive and cause several adverse effects. The majority of people in endemic areas of leishmaniasis depend of natural and traditional medicine. This study was developed to examine the activity of the essential oil from Chenopodium ambrosioides in BALB/c mice infected with Leishmania amazonensis. The infected animals received two cycle of treatment by different routes (intraperitoneal, oral or intralesional route). The intraperitoneal administration of the essential oil at dose of 30 mg/Kg prevented lesion development and decrease the parasite burden. Oral administration retarded the infection in the experimental model compared with untreated mice, although it was less effective that the intraperitoneal route. The administration by intralesional route did not show activity. Intraperitoneal and oral treatment at 30 mg/Kg with the essential oil had better antileishmanial effect that treatment with the reference drug, amphotericin B at 1 mg/Kg. Preliminarily, we examined the toxicity and the resistance after treatment. Signs of toxicity were evident only in the animals treated by intraperitoneal route. No resistance was detected in L. amazonensis isolates obtained from treated mice. These data clearly demonstrated that this natural product could be an alternative for the development of a new drug against cutaneous leishmaniasis based in the ethnomedical information. PMID:17254746
Regulation of Leishmania (L.) amazonensis Protein Expression by Host T Cell Dependent Responses: Differential Expression of Oligopeptidase B, Tryparedoxin Peroxidase and HSP70 Isoforms in Amastigotes Isolated from BALB/c and BALB/c Nude Mice
Teixeira, Priscila Camillo; Velasquez, Leonardo Garcia; Lepique, Ana Paula; de Rezende, Eloiza; Bonatto, José Matheus Camargo; Barcinski, Marcello Andre; Cunha-Neto, Edecio; Stolf, Beatriz Simonsen
Leishmaniasis is an important disease that affects 12 million people in 88 countries, with 2 million new cases every year. Leishmania amazonensis is an important agent in Brazil, leading to clinical forms varying from localized (LCL) to diffuse cutaneous leishmaniasis (DCL). One interesting issue rarely analyzed is how host immune response affects Leishmania phenotype and virulence. Aiming to study the effect of host immune system on Leishmania proteins we compared proteomes of amastigotes isolated from BALB/c and BALB/c nude mice. The athymic nude mice may resemble patients with diffuse cutaneous leishmaniasis, considered T-cell hyposensitive or anergic to Leishmania´s antigens. This work is the first to compare modifications in amastigotes' proteomes driven by host immune response. Among the 44 differentially expressed spots, there were proteins related to oxidative/nitrosative stress and proteases. Some correspond to known Leishmania virulence factors such as OPB and tryparedoxin peroxidase. Specific isoforms of these two proteins were increased in parasites from nude mice, suggesting that T cells probably restrain their posttranslational modifications in BALB/c mice. On the other hand, an isoform of HSP70 was increased in amastigotes from BALB/c mice. We believe our study may allow identification of potential virulence factors and ways of regulating their expression. PMID:25692783
Homologues of the 24-kDa flagellar Ca(2+)-binding protein gene of Trypanosoma cruzi are present in other members of the Trypanosomatidae family.
Maldonado, R A; Linss, J; Thomaz, N; Olson, C L; Engman, D M; Goldenberg, S
A full-length cDNA encoding the 24-kDa flagellar Ca(2+)-binding protein (FCaBP) of the Dm28c clone of Trypanosoma cruzi was cloned and characterized. Comparison of the deduced amino acid sequence with those of the FCaBPs of other T. cruzi strains revealed greater than 97% sequence conservation. FCaBP-like genes are found in Trypanosoma conorhini, Trypanosoma freitasi, Trypanosoma lewisi, Herpetomonas megaseliae, Leptomonas seymouri, and Phytomonas serpens, but not in Crithidia deanei, Leishmania amazonensis, or Endotrypanum schaudinni: Among various T. cruzi strains, FCaBP genes are located on chromosomes of different size, although all strains possess multiple FCaBP genes organized as tandemly arranged gene families. Northern and Western blot analyses revealed that FCaBP mRNAs are produced in all organisms possessing FCaBP-hybridizing sequences, indicating that expression of FCaBP or an FCaBP-like protein is common to a number of trypanosomatid species. PMID:9225770
Bernardes Filho, Fred; Avelleira, João Carlos Regazzi
Brazilian physician and researcher Henrique da Rocha Lima was born in 1879 in the city of Rio de Janeiro, where he studied medicine and obtained the degree of M.D. in 1901. He specialized in Clinical Medicine in Germany and was the ambassador in European countries of the scientific medicine that emerged from the Oswaldo Cruz Institute in the early twentieth century. Rocha Lima has discovered the causative agent of typhus and had a major contribution to the studies of yellow fever, Chagas disease, Carrión's disease and histoplasmosis. His genius, his research and his discoveries projected his name, and, with it, the image of Brazil in the international scientific scene. PMID:26131867
Bernardes Filho, Fred; Avelleira, João Carlos Regazzi
Brazilian physician and researcher Henrique da Rocha Lima was born in 1879 in the city of Rio de Janeiro, where he studied medicine and obtained the degree of M.D. in 1901. He specialized in Clinical Medicine in Germany and was the ambassador in European countries of the scientific medicine that emerged from the Oswaldo Cruz Institute in the early twentieth century. Rocha Lima has discovered the causative agent of typhus and had a major contribution to the studies of yellow fever, Chagas disease, Carrión's disease and histoplasmosis. His genius, his research and his discoveries projected his name, and, with it, the image of Brazil in the international scientific scene. PMID:26131867
Leonardo da Vinci and the Downburst.
NASA Astrophysics Data System (ADS)
Gedzelman, Stanley David
Evidence from the drawings, experiments, and writings of Leonardo da Vinci are presented to demonstrate that da Vinci recognized and, possibly, discovered the downburst and understood its associated airflow. Other early references to vortex flows resembling downbursts are mentioned.
The major histocompatibility complex genes impact pain response in DA and DA.1U rats.
Guo, Yuan; Yao, Fan-Rong; Cao, Dong-Yuan; Li, Li; Wang, Hui-Sheng; Xie, Wen; Zhao, Yan
Our recent studies have shown that the difference in basal pain sensitivity to mechanical and thermal stimulation between Dark-Agouti (DA) rats and a novel congenic DA.1U rats is major histocompatibility complex (MHC) genes dependent. In the present study, we further used DA and DA.1U rats to investigate the role of MHC genes in formalin-induced pain model by behavioral, electrophysiological and immunohistochemical methods. Behavioral results showed biphasic nociceptive behaviors increased significantly following the intraplantar injection of formalin in the hindpaw of DA and DA.1U rats. The main nociceptive behaviors were lifting and licking, especially in DA rats (P<0.001 and P<0.01). The composite pain scores (CPS) in DA rats were significantly higher than those in DA.1U rats in both phases of the formalin test (P<0.01). Electrophysiological results also showed the biphasic increase in discharge rates of C and Aδ fibers of L5 dorsal root in the two strains, and the net change of the discharge rate of DA rats was significantly higher than that of DA.1U rats (P<0.05). The mechanical thresholds decreased after formalin injection in both strains (P<0.01), and the net change in the mechanical threshold in DA was greater than that in DA.1U rats (P<0.05). The expression of RT1-B, representation of MHC class II molecule, in laminae I-II of L4/5 spinal cord in DA rats was significantly higher than that in DA.1U rats in the respective experimental group (P<0.05). These results suggested that both DA and DA.1U rats exhibited nociceptive responses in formalin-induced pain model and DA rats were more sensitive to noxious chemical stimulus than DA.1U rats, indicating that MHC genes might contribute to the difference in pain sensitivity. PMID:25861730
Fluorine substitution enhanced photovoltaic performance of a D-A(1)-D-A(2) copolymer.
Dang, Dongfeng; Chen, Weichao; Yang, Renqiang; Zhu, Weiguo; Mammo, Wendimagegn; Wang, Ergang
A new alternating donor-acceptor (D-A1-D-A2) copolymer containing two electron-deficient moieties, isoindigo and quinoxaline, was synthesized. The photovoltaic performance of this polymer could be improved by incorporating fluorine atoms into the quinoxaline units, resulting in an efficiency of 6.32%. This result highlights the attractive promise of D-A1-D-A2 copolymers for high-performance bulk heterojunction solar cells. PMID:24000353
Autocorrelation descriptor improvements for QSAR: 2DA_Sign and 3DA_Sign.
Sliwoski, Gregory; Mendenhall, Jeffrey; Meiler, Jens
Quantitative structure-activity relationship (QSAR) is a branch of computer aided drug discovery that relates chemical structures to biological activity. Two well established and related QSAR descriptors are two- and three-dimensional autocorrelation (2DA and 3DA). These descriptors encode the relative position of atoms or atom properties by calculating the separation between atom pairs in terms of number of bonds (2DA) or Euclidean distance (3DA). The sums of all values computed for a given small molecule are collected in a histogram. Atom properties can be added with a coefficient that is the product of atom properties for each pair. This procedure can lead to information loss when signed atom properties are considered such as partial charge. For example, the product of two positive charges is indistinguishable from the product of two equivalent negative charges. In this paper, we present variations of 2DA and 3DA called 2DA_Sign and 3DA_Sign that avoid information loss by splitting unique sign pairs into individual histograms. We evaluate these variations with models trained on nine datasets spanning a range of drug target classes. Both 2DA_Sign and 3DA_Sign significantly increase model performance across all datasets when compared with traditional 2DA and 3DA. Lastly, we find that limiting 3DA_Sign to maximum atom pair distances of 6 Å instead of 12 Å further increases model performance, suggesting that conformational flexibility may hinder performance with longer 3DA descriptors. Consistent with this finding, limiting the number of bonds in 2DA_Sign from 11 to 5 fails to improve performance. PMID:26721261
Virtual observatory publishing with DaCHS
NASA Astrophysics Data System (ADS)
Demleitner, M.; Neves, M. C.; Rothmaier, F.; Wambsganss, J.
The Data Center Helper Suite DaCHS is an integrated publication package for building VO and Web services, supporting the entire workflow from ingestion to data mapping to service definition. It implements all major data discovery, data access, and registry protocols defined by the VO. DaCHS in this sense works as glue between data produced by the data providers and the standard protocols and formats defined by the VO. This paper discusses central elements of the design of the package and gives two case studies of how VO protocols are implemented using DaCHS' concepts.
Origin of the DA and non-DA white dwarf stars
NASA Technical Reports Server (NTRS)
Shipman, Harry L.
Various proposals for the bifurcation of the white dwarf cooling sequence are reviewed. 'Primordial' theories, in which the basic bifurcation of the white dwarf sequence is rooted in events predating the white dwarf stage of stellar evolution, are discussed, along with the competing 'mixing' theories in which processes occurring during the white dwarf stage are responsible for the existence of DA or non-DA stars. A new proposal is suggested, representing a two-channel scenario. In the DA channel, some process reduces the hydrogen layer mass to the value of less than 10 to the -7th. The non-DA channel is similar to that in the primordial scenario. These considerations suggest that some mechanism operates in both channels to reduce the thickness of the outermost layer of the white dwarf. It is also noted that accretion from the interstellar medium has little to do with whether a particular white dwarf becomes a DA or a non-DA star.
Measurement of D->A Momentum Aperture and Test of D->A Field Qualities
Halling, Mike
The data presented here were taken during two shifts dedicated to D->A studies. The goal during both of these study periods was a test of the field quality of the D->A channel devices, but for the first study period on 10/21/91 the TBT system was not operational so we simply measured the transfer efficiency as a function of momentum. The conclusion from these measurements is as follows: (1) The momentum aperture of the D->A channel is smaller than expected. (2) The restriction is in physical space is somewhere near A:IKIK. (3) The field quality of the injection channel devices is adequate.
The local effect of octreotide on mechanical pain sensitivity is more sensitive in DA rats than DA.1U rats.
Yao, Fan-Rong; Wang, Hui-Sheng; Guo, Yuan; Zhao, Yan
A recent study by the authors indicated that major histocompatibility complex (MHC) genes are associated with the differences in basal pain sensitivity and in formalin model between Dark-Agouti (DA) and novel congenic DA.1U rats, which have the same genetic background as DA rats except for the u alleles of MHC. The objective of the present study is to investigate whether there is a difference in the pristane-induced arthritis (PIA) model and local analgesic effect of octreotide (OCT) between DA and DA.1U rats. The hindpaw mechanical withdrawal threshold (MWT) and heat withdrawal latency (HWL) were observed. The C unit firings of the tibial nerve evoked by non-noxious and noxious toe movements were recorded by electrophysiological methods in normal and PIA models in DA and DA.1U rats before and after local OCT administration. The expression of somatostatin receptor 2A (SSTR2A) was observed by immunohistochemistry. The results demonstrate that DA rats have a higher mechanical sensitivity than DA.1U rats after PIA. Local OCT administration significantly elevated MWT in DA rats under normal and PIA sate, but not in DA.1U rats. The electrophysiological experiments showed OCT significantly attenuated the firings of C units evoked by non-noxious and noxious stimulation in DA rats more than those in DA.1U rats both in normal and PIA states. In addition, the expression of SSTR2A in the dorsal horn of the spinal cord was significantly higher in DA than in DA.1U rats. All of the findings suggest a higher local analgesic effect of OCT in DA rats than DA.1U rats, which might be associated with the MHC genes. PMID:26606866
Common colds on Tristan da Cunha
Shibli, M.; Gooch, S.; Lewis, H. E.; Tyrrell, D. A. J.
Eight epidemics of respiratory disease have been observed among islanders of Tristan da Cunha. They seem to be initiated by the arrival of ships and transmission seemed to occur as a result of close human contact but could not always be traced. Islanders suffered from less colds than those in less isolated communities. PMID:5282927
The PAN-DA data acquisition system
Petravick, D.; Berg, D.; Berman, E.; Bernett, M.; Constanta-Fanourakis, P.; Dorries, T.; Haire, M.; Kaczar, K; MacKinnon, B.; Moore, C.; Nicinski, T.; Oleynik, G.; Pordes, R.; Sergey, G.; Votava, M.; White, V.
The Online and Data Acquisition software groups at Fermi National Accelerator Laboratory have extended the VAXONLINE data acquisition package to include a VME based data path. The resulting environment, PAN-DA, provides a high throughput for logging, filtering, formatting and selecting events. 10 refs., 1 fig.
Hidden sketches by Leonardo da Vinci revealed
NASA Astrophysics Data System (ADS)
Dumé, Belle
Three drawings on the back of Leonardo da Vinci's The Virgin and Child with St Anne (circa 1508) have been discovered by researchers led by Michel Menu from the Centre de Recherche et de Restauration des Musées de France (C2RMF) and the Louvre Museum in Paris.
How to Think Like Leonardo da Vinci
ERIC Educational Resources Information Center
Caouette, Ralph
To be effective and relevant in twenty-first-century learning, art needs to be more inclusive. In this article, the author discusses how teachers can find a good example in Leonardo da Vinci for building an art program. His art, design, and curiosity are the perfect foundation for any art program, at any level. (Contains 3 resources and 3 online…
40 CFR 60.51Da - Reporting requirements.
Code of Federal Regulations, 2013 CFR
... Generating Units § 60.51Da Reporting requirements. (a) For SO2, NOX, PM, and NOX plus CO emissions, the... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Reporting requirements. 60.51Da Section 60.51Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS...
40 CFR 60.47Da - Commercial demonstration permit.
Code of Federal Regulations, 2012 CFR
... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Commercial demonstration permit. 60.47Da Section 60.47Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS... Steam Generating Units § 60.47Da Commercial demonstration permit. (a) An owner or operator of...
40 CFR 60.51Da - Reporting requirements.
Code of Federal Regulations, 2014 CFR
... Generating Units § 60.51Da Reporting requirements. (a) For SO2, NOX, PM, and NOX plus CO emissions, the... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Reporting requirements. 60.51Da Section 60.51Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS...
40 CFR 60.47Da - Commercial demonstration permit.
Code of Federal Regulations, 2014 CFR
... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Commercial demonstration permit. 60.47Da Section 60.47Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS... Steam Generating Units § 60.47Da Commercial demonstration permit. (a) An owner or operator of...
40 CFR 60.51Da - Reporting requirements.
Code of Federal Regulations, 2012 CFR
... Generating Units § 60.51Da Reporting requirements. (a) For SO2, NOX, PM, and NOX plus CO emissions, the... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Reporting requirements. 60.51Da Section 60.51Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS...
40 CFR 60.47Da - Commercial demonstration permit.
Code of Federal Regulations, 2013 CFR
... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Commercial demonstration permit. 60.47Da Section 60.47Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS... Steam Generating Units § 60.47Da Commercial demonstration permit. (a) An owner or operator of...
Code of Federal Regulations, 2011 CFR
... 32 National Defense 3 2011-07-01 2009-07-01 true DA Form 4. 516.25 Section 516.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY AID OF CIVIL AUTHORITIES AND PUBLIC RELATIONS LITIGATION Reporting Legal Proceedings to HQDA § 516.25 DA Form 4. (a) General. The DA Form 4 (See figure...
Code of Federal Regulations, 2010 CFR
... 32 National Defense 3 2010-07-01 2010-07-01 true DA Form 4. 516.25 Section 516.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY AID OF CIVIL AUTHORITIES AND PUBLIC RELATIONS LITIGATION Reporting Legal Proceedings to HQDA § 516.25 DA Form 4. (a) General. The DA Form 4 (See figure...
A Day in the Life at DaVita Academy
ERIC Educational Resources Information Center
Weinstein, Margery
When a company name means "giving life," the bar for learning and development programs is held high. In this article, the author describes what it takes to graduate from DaVita Academy, the soft skills training program dialysis services company DaVita offers all its employees. DaVita's chief executive officer, Kent Thiry, states that the Academy…
Burattini, Emilio; Cinque, Gianfelice; Dabagov, Sultan; Grilli, Antonio; Marcelli, Augusto; Pace, Elisabetta; Piccinini, Massimo; Raco, Agostino; Monti, Francesca
The new Synchrotron Radiation facility at Frascati exploits the intense photon emission from DA{phi}NE, the 0,51 GeV storage ring circulating over 1 A of electrons. Among the three beamlines commissioned, the Synchrotron INfrared Beamline At Da{phi}ne (SINBAD) is fully operational by a brilliant SR beam spanning the entire IR. Recently, the soft X-ray beamline has been characterized and, once implemented the double-crystal monocromator, X-ray Absorption Spectroscopy is applied on material standards in the distinguishing energy region below 4 keV. An UltraViolet line, presently dedicated to photobiology dosimetry, has also given first results on cell irradiation in the UVB band.
The Real Code of Leonardo da Vinci
Ose, Leiv
Leonardo da Vinci was born in Italy. Among the researchers and scientists, he is favourably known for his remarkable efforts in scientific work. His investigations of atherosclerosis judiciously combine three separate fields of research. In 1506, he finished his masterpiece, painting of Mona Lisa. A careful clinical examination of the famous painting reveals a yellow irregular leather-like spot at the inner end of the left upper eyelid and a soft bumpy well-defined swelling of the dorsum of the right hand beneath the index finger about 3 cm long. This is probably the first case of familial hypercholesterolemia (FH). The FH code of Leonardo da Vinci was given immense consideration by scientists like Carl Muller, who described the xanthomas tuberosum and angina pectoris. On the contrary, Akira Endo searched for microbial metabolites that would inhibit HMG-CoA reductase, the rate-limiting enzyme in the synthesis of cholesterol and finally, Michael Brown and Joseph Goldstein published a remarkable series of elegant and insightful papers in the 70s and 80s. They established that the cellular uptake of low-density lipoprotein (LDL) essentially requires the LDL receptor. In conclusion: this was the real Code of Leonardo da Vinci. PMID:19924278
40 CFR 60.52Da - Recordkeeping requirements.
Code of Federal Regulations, 2010 CFR
... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Recordkeeping requirements. 60.52Da Section 60.52Da Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Electric Utility Steam Generating Units for...
Asteroid 2012 DA14 To Whiz Past Earth Safely
The small near-Earth asteroid 2012 DA14 will pass very close to Earth on Feb. 15, 2013. Asteroid 2012 DA14 will be closest to Earth at about 11:24 a.m. PST (2:24 p.m. EST and 1924 UTC), on Feb. 15,...
The Case: Bunche-Da Vinci Learning Partnership Academy
ERIC Educational Resources Information Center
Eisenberg, Nicole; Winters, Lynn; Alkin, Marvin C.
The Bunche-Da Vinci case described in this article presents a situation at Bunche Elementary School that four theorists were asked to address in their evaluation designs (see EJ791771, EJ719772, EJ791773, and EJ792694). The Bunche-Da Vinci Learning Partnership Academy, an elementary school located between an urban port city and a historically…
Da Costa's syndrome or neurocirculatory asthenia.
Paul, O
The syndrome variously called Da Costa's syndrome, effort syndrome, neurocirculatory asthenia, etc has been studied for more than 100 years by many distinguished physicians. Originally identified in men in wartime, it has been widely recognised as a common chronic condition in both sexes in civilian life. Although the symptoms may seem to appear after infections and various physical and psychological stresses, neurocirculatory asthenia is most often encountered as a familial disorder that is unrelated to these factors, although they may aggravate an existing tendency. Respiratory complaints (including breathlessness, with and without effort, and smothering sensations) are almost universal, and palpitation, chest discomfort, dizziness and faintness, and fatigue are common. The physical examination is normal. The aetiology is obscure but patients usually have a normal life span. Reassurance and measures to improve physical fitness are helpful. PMID:3314950
75 FR 52292 - Airworthiness Directives; Diamond Aircraft Industries GmbH Models DA 40 and DA 40F Airplanes
Federal Register 2010, 2011, 2012, 2013, 2014
... ``significant rule'' under the DOT Regulatory Policies and Procedures (44 FR 11034, February 26, 1979); and 3... Federal Aviation Administration 14 CFR Part 39 RIN 2120-AA64 Airworthiness Directives; Diamond Aircraft... new airworthiness directive (AD) for all Diamond Aircraft Industries GmbH Models DA 40 and DA...
75 FR 75868 - Airworthiness Directives; Diamond Aircraft Industries GmbH Models DA 40 and DA 40F Airplanes
Federal Register 2010, 2011, 2012, 2013, 2014
... Register on August 25, 2010 (75 FR 52292). That NPRM proposed to require a retrofit of the rear passenger... 12866, (2) Is not a ``significant rule'' under DOT Regulatory Policies and Procedures (44 FR 11034... Aircraft Industries GmbH Models DA 40 and DA 40F Airplanes AGENCY: Federal Aviation Administration,...
Metal Lines in DA White Dwarfs
NASA Astrophysics Data System (ADS)
Zuckerman, B.; Koester, D.; Reid, I. N.; Hünsch, M.
We report Keck telescope HIRES echelle observations of DA white dwarfs in a continuation of an extensive search for metals. These spectra are supplemented with new JHK magnitudes that are used to determine improved atmospheric parameters. Of the DA white dwarfs not in binary or common proper motion systems, about 25% show Ca II lines. For these, Ca abundances are determined from comparison with theoretical equivalent widths from model atmosphere calculations; in a few cases we also obtain Mg, Fe, Si, and Al abundances. If Ca is not observed, we generally determine very stringent upper limits. We compare the data to predictions of previously published models involving the accretion/diffusion of interstellar matter and of comets. The derived abundances are not obviously compatible with the predictions of either model, which up to now could only be tested with traces of metals in helium-rich white dwarfs. By modifying certain assumptions in the published interstellar accretion model we are able to match the distribution of the elements in the white dwarf atmospheres, but, even so, tests of other expectations from this scenario are less successful. Because comet accretion appears unlikely to be the primary cause of the DAZ phenomenon, the data suggest that no more than about 20% of F-type main-sequence stars are accompanied by Oort-like comet clouds. This represents the first observational estimate of this fraction. A plausible alternative to the accretion of cometary or interstellar matter is disruption and accretion of asteroidal material, a model first suggested in 1990 to explain excess near-infrared emission from the DAZ G29-38. An asteroidal debris model to account for the general DAZ phenomenon does not presently disagree with the HIRES data, but neither is there any compelling evidence in support of such a model. The HIRES data indicate that in close red dwarf/white dwarf binaries not known to be cataclysmic variables there is, nonetheless, significant mass
Der Telemanipulator daVinci als mechanisches Trackingsystem
NASA Astrophysics Data System (ADS)
Käst, Johannes; Neuhaus, Jochen; Nickel, Felix; Kenngott, Hannes; Engel, Markus; Short, Elaine; Reiter, Michael; Meinzer, Hans-Peter; Maier-Hein, Lena
Der Telemanipulator daVinci (Intuitive Surgical, Sunnyvale, Kalifornien) ist ein M aster-Slave System für roboterassistierte minimalinvasive Chirurgie. Da er über integrierte Gelenksensoren verfügt, kann er unter Verwendung der daVinci-API als mechanisches Trackingsystem verwendet werden. In dieser Arbeit evaluieren wir die Präzision und Genauigkeit eines daVinci mit Hilfe eines Genauigkeitsphantoms mit bekannten Maßen. Der ermittelte Positionierungsfehler liegt in der Größenordnung von 6 mm und ist somit für einen Großteil der medizinischen Fragestellungen zu hoch. Zur Reduktion des Fehlers schlagen wir daher eine Kalibrierung der Gelenksensoren vor.
40 CFR 60.49Da - Emission monitoring.
Code of Federal Regulations, 2013 CFR
... affected unit that qualifies as a gas-fired or oil-fired unit, as defined in 40 CFR 72.2, may use, as an... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Emission monitoring. 60.49Da Section 60... Generating Units § 60.49Da Emission monitoring. (a) An owner or operator of an affected facility subject...
40 CFR 60.49Da - Emission monitoring.
Code of Federal Regulations, 2010 CFR
... affected unit that qualifies as a gas-fired or oil-fired unit, as defined in 40 CFR 72.2, may use, as an... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Emission monitoring. 60.49Da Section 60... Generating Units for Which Construction is Commenced After September 18, 1978 § 60.49Da Emission...
40 CFR 60.49Da - Emission monitoring.
Code of Federal Regulations, 2014 CFR
... affected unit that qualifies as a gas-fired or oil-fired unit, as defined in 40 CFR 72.2, may use, as an... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Emission monitoring. 60.49Da Section 60... Generating Units § 60.49Da Emission monitoring. (a) An owner or operator of an affected facility subject...
Tree Branching: Leonardo da Vinci's Rule versus Biomechanical Models
Minamino, Ryoko; Tateno, Masaki
This study examined Leonardo da Vinci's rule (i.e., the sum of the cross-sectional area of all tree branches above a branching point at any height is equal to the cross-sectional area of the trunk or the branch immediately below the branching point) using simulations based on two biomechanical models: the uniform stress and elastic similarity models. Model calculations of the daughter/mother ratio (i.e., the ratio of the total cross-sectional area of the daughter branches to the cross-sectional area of the mother branch at the branching point) showed that both biomechanical models agreed with da Vinci's rule when the branching angles of daughter branches and the weights of lateral daughter branches were small; however, the models deviated from da Vinci's rule as the weights and/or the branching angles of lateral daughter branches increased. The calculated values of the two models were largely similar but differed in some ways. Field measurements of Fagus crenata and Abies homolepis also fit this trend, wherein models deviated from da Vinci's rule with increasing relative weights of lateral daughter branches. However, this deviation was small for a branching pattern in nature, where empirical measurements were taken under realistic measurement conditions; thus, da Vinci's rule did not critically contradict the biomechanical models in the case of real branching patterns, though the model calculations described the contradiction between da Vinci's rule and the biomechanical models. The field data for Fagus crenata fit the uniform stress model best, indicating that stress uniformity is the key constraint of branch morphology in Fagus crenata rather than elastic similarity or da Vinci's rule. On the other hand, mechanical constraints are not necessarily significant in the morphology of Abies homolepis branches, depending on the number of daughter branches. Rather, these branches were often in agreement with da Vinci's rule. PMID:24714065
Migration of ATLAS PanDA to CERN
NASA Astrophysics Data System (ADS)
Stewart, Graeme Andrew; Klimentov, Alexei; Koblitz, Birger; Lamanna, Massimo; Maeno, Tadashi; Nevski, Pavel; Nowak, Marcin; Emanuel De Castro Faria Salgado, Pedro; Wenaus, Torre
The ATLAS Production and Distributed Analysis System (PanDA) is a key component of the ATLAS distributed computing infrastructure. All ATLAS production jobs, and a substantial amount of user and group analysis jobs, pass through the PanDA system, which manages their execution on the grid. PanDA also plays a key role in production task definition and the data set replication request system. PanDA has recently been migrated from Brookhaven National Laboratory (BNL) to the European Organization for Nuclear Research (CERN), a process we describe here. We discuss how the new infrastructure for PanDA, which relies heavily on services provided by CERN IT, was introduced in order to make the service as reliable as possible and to allow it to be scaled to ATLAS's increasing need for distributed computing. The migration involved changing the backend database for PanDA from MySQL to Oracle, which impacted upon the database schemas. The process by which the client code was optimised for the new database backend is discussed. We describe the procedure by which the new database infrastructure was tested and commissioned for production use. Operations during the migration had to be planned carefully to minimise disruption to ongoing ATLAS offline computing. All parts of the migration were fully tested before commissioning the new infrastructure and the gradual migration of computing resources to the new system allowed any problems of scaling to be addressed.
PKC phosphorylates residues in the N-terminal of the DA transporter to regulate amphetamine-induced DA efflux.
Wang, Qiang; Bubula, Nancy; Brown, Jason; Wang, Yunliang; Kondev, Veronika; Vezina, Paul
The DA transporter (DAT), a phosphoprotein, controls extracellular dopamine (DA) levels in the central nervous system through transport or reverse transport (efflux). Multiple lines of evidence support the claim that PKC significantly contributes to amphetamine-induced DA efflux. Other signaling pathways, involving CaMKII and ERK, have also been shown to regulate DAT mediated efflux. Here we assessed the contribution of putative PKC residues (S4, S7, S13) in the N-terminal of the DAT to amphetamine-induced DA efflux by transfecting DATs containing different serine to alanine (S-A) point mutations into DA pre-loaded HEK-293 cells and incubating these cells in amphetamine (2μM). The effects of a S-A mutation at the non-PKC residue S12 and a threonine to alanine (T-A) mutation at the ERK T53 residue were also assessed for comparison. WT-DATs were used as controls. In an initial experiment, we confirmed that inhibiting PKC with Go6976 (130nM) significantly reduced amphetamine-induced DA efflux. In subsequent experiments, cells transfected with the S4A, S12A, S13A, T53A and S4,7,13A mutants showed a reduction in amphetamine-induced DA efflux similar to that observed with Go6976. Interestingly, cells transfected with the S7A mutant, identified by some as a PKC-PKA residue, showed unperturbed WT-DAT levels of amphetamine-induced DA efflux. These results indicate that phosphorylation by PKC of select residues in the DAT N-terminal can regulate amphetamine-induced efflux. PKC can act either independently or in concert with other kinases such as ERK to produce this effect. PMID:27113203
Destruction of Leishmania mexicana amazonensis amastigotes within macrophages by lysosomotropic amino acid esters
Leishmania amastigotes parasitize almost exclusively the mononuclear phagocytes of mammals. The organisms survive and multiply within acidified vacuoles (parasitophorous vacuoles; p.v.) akin to phagolysosomes. Certain amino acid esters are known to accumulate in and disrupt lysosomes. We postulated that, since Leishmania possess lysosome-like organelles, they may be susceptible to the potentially high ester concentrations attained in the p.v. We report here that L- amino acid esters can rapidly destroy intracellular Leishmania at concentrations that do not appear to damage the host cells. L-leu-OMe, which cured greater than or equal to 90% of infected macrophages at 0.8 mM concentrations, was used in most of the experiments. L-leu-OMe was only active after infection, implying inefficient transfer from secondary lysosomes to the p.v. Parasite destruction had several features in common with lysosomal and leukocyte damage induced by the esters, i.e., inactivity of D-amino acid esters, a marked pH dependence and increased killing after ester pulses at lower temperatures. Killing depended on the amino acid and on the ester substitution. The most active of the methyl esters assayed was that of leucine, followed by those of tryptophan, glutamic acid, methionine, phenylalanine, and tyrosine. Methyl esters of seven other amino acids were inactive when tested at up to 10 mM concentrations. Among leucine esters studied, benzyl ester was sixfold more active than the methyl homolog. The dipeptide L-leu-leu-OMe produced 90% cure at 0.08 mM concentrations. Leishmanicidal activity could be related to penetration of the parasites by the esters or to toxic ester hydrolysis products released in the p.v. The first hypothesis is supported by the pH-dependent destruction of isolated amastigotes by the esters. Furthermore, relatively high concentrations of L-leucine, methanol, or benzyl alcohol were not demonstrably toxic to the amastigotes. We postulate that ester concentrations sufficient to damage the intracellular amastigotes may be obtained within the p.v. after exposure of infected macrophages to the esters. Esters preferentially hydrolyzed by parasite enzymes may be expected to be leishmanicidal, but less damaging to the host. PMID:3950542
Chemotaxis study using optical tweezers to observe the strength and directionality of forces of Leishmania amazonensis
NASA Astrophysics Data System (ADS)
Pozzo, Liliana d. Y.; Fontes, Adriana; de Thomaz, André A.; Barbosa, Luiz C.; Ayres, Diana C.; Giorgio, Selma; Cesar, Carlos L.
The displacements of a dielectric microspheres trapped by an optical tweezers (OT) can be used as a force transducer for mechanical measurements in life sciences. This system can measure forces on the 50 femto Newtons to 200 pico Newtons range, of the same order of magnitude of a typical forces induced by flagellar motion. The process in which living microorganisms search for food and run away from poison chemicals is known is chemotaxy. Optical tweezers can be used to obtain a better understanding of chemotaxy by observing the force response of the microorganism when placed in a gradient of attractors and or repelling chemicals. This report shows such observations for the protozoa Leishmania amazomenzis, responsible for the leishmaniasis, a serious tropical disease. We used a quadrant detector to monitor the movement of the protozoa for different chemicals gradient. This way we have been able to observe both the force strength and its directionality. The characterization of the chemotaxis of these parasites can help to understand the infection mechanics and improve the diagnosis and the treatments employed for this disease.
Leishmania amazonensis promastigotes induce and are killed by neutrophil extracellular traps
Guimarães-Costa, Anderson B.; Nascimento, Michelle T. C.; Froment, Giselle S.; Soares, Rodrigo P. P.; Morgado, Fernanda N.; Conceição-Silva, Fátima; Saraiva, Elvira M.
Neutrophils are short-lived leukocytes that die by apoptosis, necrosis, and NETosis. Upon death by NETosis, neutrophils release fibrous traps of DNA, histones, and granule proteins named neutrophil extracellular traps (NETs), which can kill bacteria and fungi. Inoculation of the protozoan Leishmania into the mammalian skin causes local inflammation with neutrophil recruitment. Here, we investigated the release of NETs by human neutrophils upon their interaction with Leishmania parasites and NETs' ability to kill this protozoan. The NET constituents DNA, elastase, and histones were detected in traps associated to promastigotes by immunofluorescence. Electron microscopy revealed that Leishmania was ensnared by NETs released by neutrophils. Moreover, Leishmania and its surface lipophosphoglycan induced NET release by neutrophils in a parasite number- and dose-dependent manner. Disruption of NETs by DNase treatment during Leishmania–neutrophil interaction increased parasite survival, evidencing NETs' leishmanicidal effect. Leishmania killing was also elicited by NET-rich supernatants from phorbol 12-myristate 13-acetate-activated neutrophils. Immunoneutralization of histone during Leishmania–neutrophil interaction partially reverted Leishmania killing, and purified histone killed the parasites. Meshes composed of DNA and elastase were evidenced in biopsies of human cutaneous leishmaniasis. NET is an innate response that might contribute to diminish parasite burden in the Leishmania inoculation site. PMID:19346483
A cosmologia no ensino da geografia
NASA Astrophysics Data System (ADS)
Santos, S. C.; Chiaradia, A. P. M.
O principal objetivo deste trabalho é auxiliar o professor de Geografia em sala de aula no ensino de tópicos relacionados com a Cosmologia. A idéia deste trabalho surgiu quando foi constatado que o professor de Geografia tem dificuldades de ensinar este tópico. Esta constatação foi feita por uma das autoras ao lecionar este tópico no ensino fundamental e em discussões com outros professores de Geografia. Da mesma maneira que ocorria desde os tempos mais antigos, os alunos têm muito interesse em conhecer os fenômenos que ocorrem no Cosmo, porém os livros didáticos de Geografia utilizados em sala de aula não são ricos em informações sobre este assunto. Assim, o professor de Geografia tem poucas informações para discutir este assunto em sala de aula e não dá a devida importância para este tópico. Então, foi desenvolvido um material de apoio para professores de Geografia sobre a origem do Universo, sua evolução e seu possível futuro evolutivo segundo as mais recentes teorias, com base em perguntas feitas pelos alunos de ensino fundamental e as informações trazidas nos livros didáticos Não cabe a este material inovar e tão pouco trazer uma metodologia de ensino de Cosmologia. Neste material o professor de Geografia pode encontrará um banco de informações, que constitui no estabelecimento de conceitos, teorias e hipóteses, sobre a Cosmologia, em linguagem simples e de fácil entendimento. Para desenvolvê-lo, foram feitas pesquisas não exaustivas em livros e revistas científicas, compilação e discussão em forma cronológica das teorias aceitas sobre modelos cosmológicos. Portanto, este material será apresentado neste trabalho.
Overview of ATLAS PanDA Workload Management
Maeno T.; De K.; Wenaus T.; Nilsson P.; Stewart G. A.; Walker R.; Stradling A.; Caballero J.; Potekhin M.; Smith D.
The Production and Distributed Analysis System (PanDA) plays a key role in the ATLAS distributed computing infrastructure. All ATLAS Monte-Carlo simulation and data reprocessing jobs pass through the PanDA system. We will describe how PanDA manages job execution on the grid using dynamic resource estimation and data replication together with intelligent brokerage in order to meet the scaling and automation requirements of ATLAS distributed computing. PanDA is also the primary ATLAS system for processing user and group analysis jobs, bringing further requirements for quick, flexible adaptation to the rapidly evolving analysis use cases of the early datataking phase, in addition to the high reliability, robustness and usability needed to provide efficient and transparent utilization of the grid for analysis users. We will describe how PanDA meets ATLAS requirements, the evolution of the system in light of operational experience, how the system has performed during the first LHC data-taking phase and plans for the future.
The ATLAS PanDA Monitoring System and its Evolution
NASA Astrophysics Data System (ADS)
Klimentov, A.; Nevski, P.; Potekhin, M.; Wenaus, T.
The PanDA (Production and Distributed Analysis) Workload Management System is used for ATLAS distributed production and analysis worldwide. The needs of ATLAS global computing imposed challenging requirements on the design of PanDA in areas such as scalability, robustness, automation, diagnostics, and usability for both production shifters and analysis users. Through a system-wide job database, the PanDA monitor provides a comprehensive and coherent view of the system and job execution, from high level summaries to detailed drill-down job diagnostics. It is (like the rest of PanDA) an Apache-based Python application backed by Oracle. The presentation layer is HTML code generated on the fly in the Python application which is also responsible for managing database queries. However, this approach is lacking in user interface flexibility, simplicity of communication with external systems, and ease of maintenance. A decision was therefore made to migrate the PanDA monitor server to Django Web Application Framework and apply JSON/AJAX technology in the browser front end. This allows us to greatly reduce the amount of application code, separate data preparation from presentation, leverage open source for tools such as authentication and authorization mechanisms, and provide a richer and more dynamic user experience. We describe our approach, design and initial experience with the migration process.
Overview of ATLAS PanDA Workload Management
NASA Astrophysics Data System (ADS)
Maeno, T.; De, K.; Wenaus, T.; Nilsson, P.; Stewart, G. A.; Walker, R.; Stradling, A.; Caballero, J.; Potekhin, M.; Smith, D.; ATLAS Collaboration
The Production and Distributed Analysis System (PanDA) plays a key role in the ATLAS distributed computing infrastructure. All ATLAS Monte-Carlo simulation and data reprocessing jobs pass through the PanDA system. We will describe how PanDA manages job execution on the grid using dynamic resource estimation and data replication together with intelligent brokerage in order to meet the scaling and automation requirements of ATLAS distributed computing. PanDA is also the primary ATLAS system for processing user and group analysis jobs, bringing further requirements for quick, flexible adaptation to the rapidly evolving analysis use cases of the early datataking phase, in addition to the high reliability, robustness and usability needed to provide efficient and transparent utilization of the grid for analysis users. We will describe how PanDA meets ATLAS requirements, the evolution of the system in light of operational experience, how the system has performed during the first LHC data-taking phase and plans for the future.
Geomagnetic observations on tristan da cunha, south atlantic ocean
Matzka, J.; Olsen, N.; Maule, C.F.; Pedersen, L.W.; Berarducci, A.M.; Macmillan, S.
Few geomagnetic ground observations exist of the Earth's strongest core field anomaly, the South Atlantic Anomaly (SAA). The geomagnetic repeat station on the island Tristan da Cunha, located half-way between South Africa and South America at 37?? 05' S, 12?? 18' W, is therefore of crucial importance. We have conducted several sets of repeat station measurements during magnetically quiet conditions (Kp 2o or less) in 2004. The procedures are described and the results are compared to those from earlier campaigns and to the predictions of various global field models. Features of the local crustal bias field and the solar quiet daily variation are discussed. We also evaluate the benefit of continuous magnetic field recordings from Tristan da Cunha, and argue that such a data set is a very valuable addition to geomagnetic satellite data. Recently, funds were set up to establish and operate a magnetometer station on Tristan da Cunha during the Swarm magnetic satellite mission (2011-2014).
Recent Improvements in the ATLAS PanDA Pilot
NASA Astrophysics Data System (ADS)
Nilsson, P.; Caballero Bejar, J.; Compostella, G.; Contreras, C.; De, K.; Dos Santos, T.; Maeno, T.; Potekhin, M.; Wenaus, T.
The Production and Distributed Analysis system (PanDA) in the ATLAS experiment uses pilots to execute submitted jobs on the worker nodes. The pilots are designed to deal with different runtime conditions and failure scenarios, and support many storage systems. This talk will give a brief overview of the PanDA pilot system and will present major features and recent improvements including CernVM File System integration, the job retry mechanism, advanced job monitoring including JEM technology, and validation of new pilot code using the HammerCloud stress-testing system. PanDA is used for all ATLAS distributed production and is the primary system for distributed analysis. It is currently used at over 130 sites worldwide. We analyze the performance of the pilot system in processing LHC data on the OSG, EGI and Nordugrid infrastructures used by ATLAS, and describe plans for its further evolution.
Situação da Mulher na Astronomia Brasileira
NASA Astrophysics Data System (ADS)
Silva, Adriana V. R.
O conteúdo desse texto surgiu de uma apresentação de mesmo título que fiz na XXXI Reunião Anual da Sociedade Astronômica Brasileira (SAB) em 2005. Esse tema foi inspirado originalmente pela minha participação no "2nd UIPAP International Conference on Women in Physics" realizado entre 23 e 25 de maio de 2005 no Rio de Janeiro. Essa é uma conferência internacional que acontece de três em três anos, sendo que a primeira ocorreu em 2002 na cidade de Paris, França. Participei dessa conferência como membro da delegação da Sociedade Brasileira de Física e um dos trabalhos que apresentei versava sobre a situação das mulheres na Astronomia brasileira, cujos resultados principais discorro a seguir. A situação das astrônomas, baseada nos dados dos sócios da SAB coletados no final de 2004, é comparada com a das físicas brasileiras e também com as nossas colegas americanas. Os dados identificam ainda uma maior evasão da carreira por parte das mulheres do que os homens. Alguns dos possíveis motivos da evasão são discutidos, como o desejo de constituir família e/ou isolamento. Resultados um tanto preocupantes com relação à distribuição de bolsas de produtividade do CNPq também são apresentados. As principais discussões e estratégias recomendadas nesse congresso são mencionadas de forma resumida ao final.
The future of PanDA in ATLAS distributed computing
NASA Astrophysics Data System (ADS)
De, K.; Klimentov, A.; Maeno, T.; Nilsson, P.; Oleynik, D.; Panitkin, S.; Petrosyan, A.; Schovancova, J.; Vaniachine, A.; Wenaus, T.
Experiments at the Large Hadron Collider (LHC) face unprecedented computing challenges. Heterogeneous resources are distributed worldwide at hundreds of sites, thousands of physicists analyse the data remotely, the volume of processed data is beyond the exabyte scale, while data processing requires more than a few billion hours of computing usage per year. The PanDA (Production and Distributed Analysis) system was developed to meet the scale and complexity of LHC distributed computing for the ATLAS experiment. In the process, the old batch job paradigm of locally managed computing in HEP was discarded in favour of a far more automated, flexible and scalable model. The success of PanDA in ATLAS is leading to widespread adoption and testing by other experiments. PanDA is the first exascale workload management system in HEP, already operating at more than a million computing jobs per day, and processing over an exabyte of data in 2013. There are many new challenges that PanDA will face in the near future, in addition to new challenges of scale, heterogeneity and increasing user base. PanDA will need to handle rapidly changing computing infrastructure, will require factorization of code for easier deployment, will need to incorporate additional information sources including network metrics in decision making, be able to control network circuits, handle dynamically sized workload processing, provide improved visualization, and face many other challenges. In this talk we will focus on the new features, planned or recently implemented, that are relevant to the next decade of distributed computing workload management using PanDA.
The role of transparency in da Vinci stereopsis.
Zannoli, Marina; Mamassian, Pascal
The majority of natural scenes contains zones that are visible to one eye only. Past studies have shown that these monocular regions can be seen at a precise depth even though there are no binocular disparities that uniquely constrain their locations in depth. In the so-called da Vinci stereopsis configuration, the monocular region is a vertical line placed next to a binocular rectangular occluder. The opacity of the occluder has been mentioned to be a necessary condition to obtain da Vinci stereopsis. However, this opacity constraint has never been empirically tested. In the present study, we tested whether da Vinci stereopsis and perceptual transparency can interact using a classical da Vinci configuration in which the opacity of the occluder varied. We used two different monocular objects: a line and a disk. We found no effect of the opacity of the occluder on the perceived depth of the monocular object. A careful analysis of the distribution of perceived depth revealed that the monocular object was perceived at a depth that increased with the distance between the object and the occluder. The analysis of the skewness of the distributions was not consistent with a double fusion explanation, favoring an implication of occlusion geometry in da Vinci stereopsis. A simple model that includes the geometry of the scene could account for the results. In summary, the mechanism responsible to locate monocular regions in depth is not sensitive to the material properties of objects, suggesting that da Vinci stereopsis is solved at relatively early stages of disparity processing. PMID:21906614
Publishing Planetary Data to the VO using DaCHS
NASA Astrophysics Data System (ADS)
Demleitner, M.
Publishing to the Virtual Observatory has many advantages, but between standards compliance, registration, data organisation, and server operation it is also requires a nontrivial effort if starting from scratch. The DaCHS software from the German Astrophysical Virtual Observatory offers a packaged solution letting publishers concentrate on working with their own data and metadata as much as possible. For the planetary community, publishing is further facilitated by built-in support for the EPN-TAP data model. This contribution will show a typical publishing workflow together with a brief overview of how DaCHS meshes in with the VO and Web-based data usage.
Asteroid 1986 DA - Radar evidence for a metallic composition
NASA Technical Reports Server (NTRS)
Ostro, S. J.; Rosema, K. D.; Campbell, D. B.; Chandler, J. F.; Hine, A. A.; Hudson, R. S.
Echoes from the near-earth object 1986 DA show it to be significantly more reflective than other radar-detected asteroids. This result supports the hypothesis that 1986 DA is a piece of NiFe metal derived from the interior of a much larger object that melted, differentiated, cooled, and subsequently was disrupted in a catastrophic collision. This 2-kilometer asteroid, which appears smooth at centimeter to meter scales but extremely irregular at 10- to 100-meter scales, might be (or have been a part of) the parent body of some iron meteorites.
Identification of domains on the extrinsic 23 kDa protein possibly involved in electrostatic interaction with the extrinsic 33 kDa protein in spinach photosystem II.
Tohri, Akihiko; Dohmae, Naoshi; Suzuki, Takehiro; Ohta, Hisataka; Inoue, Yasunori; Enami, Isao
To elucidate the domains on the extrinsic 23 kDa protein involved in electrostatic interaction with the extrinsic 33 kDa protein in spinach photosystem II, we modified amino or carboxyl groups of the 23 kDa protein to uncharged methyl ester groups with N-succinimidyl propionate or glycine methyl ester in the presence of a water-soluble carbodiimide, respectively. The N-succinimidyl propionate-modified 23 kDa protein did not bind to the 33 kDa protein associated with PSII membranes, whereas the glycine methyl ester-modified 23 kDa protein completely bound. This indicates that positive charges on the 23 kDa protein are important for electrostatic interaction with the 33 kDa protein associated with the PSII membranes. Mapping of the N-succinimidyl propionate-modified sites of the 23 kDa protein was performed using Staphylococcus V8 protease digestion of the modified protein followed by determination of the mass of the resultant peptide fragments with MALDI-TOF MS. The results showed that six domains (Lys11-Lys14, Lys27-Lys38, Lys40, Lys90-Lys96, Lys143-Lys152, Lys166-Lys174) were modified with N-succinimidyl propionate. In these domains, Lys11, Lys13, Lys33, Lys38, Lys143, Lys166, Lys170 and Lys174 were wholly conserved in the 23 kDa protein from 12 species of higher plants. These positively charged lysyl residues on the 23 kDa protein may be involved in electrostatic interactions with the negatively charged carboxyl groups on the 33 kDa protein, the latter has been suggested to be important for the 23 kDa binding [Bricker, T.M. & Frankel, L.K. (2003) Biochemistry42, 2056-2061]. PMID:15009208
EPA promulgated minimum quality assurance (QA) requirements for Continuous Emission Monitoring Systems (CEMS) in 40 CFR Part 60 Appendix F. Appendix F requires Da source owners to develop site-specific QA plans and report the results of EPA specified QA activities each calendar q...
Asteroid 1986 DA: Radar evidence for a metallic composition
NASA Technical Reports Server (NTRS)
Ostro, Steven J.; Campbell, D. B.; Chandler, J. F.; Hine, A. A.; Hudson, R. S.; Rosema, K. D.; Shapiro, Irwin I.
Radar observations of the near-Earth asteroid 1986 DA were carried out at the Arecibo Observatory in April 1986, two months after its discovery. Radar results are consistent with the hypothesis that 1986 HA is a piece of NiFe metal derived from the interior of a much larger object that melted, differentiated, cooled and subsequently was disrupted in a catastrophic collision. This 2-km asteroid might be (or have been part of) the parent body of some iron meteorites. Or 1986 DA might share the parentage and/or part of the dynamical history of some meteorites without ever having contributed any of its own ejecta to our meteorite sample. Analysis of the samples returned from 1986 DA might ultimately involve economic considerations. Meteoritic metal is mostly iron with about 8 percent nickel, but also contains substantial concentrations of precious and strategic metals, including approx. 1 ppm of gold and approx. 10 ppm of platinum group elements. If these abundances apply to 1986 DA, it contains some 10(exp 16) g of iron, 10 (exp 15) g of nickel, 10(exp 11) g of platinum group metals, and 10(exp 10) g of gold.
40 CFR 60.45Da - Standard for mercury (Hg).
Code of Federal Regulations, 2010 CFR
... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Standard for mercury (Hg). 60.45Da... for mercury (Hg). (a) For each coal-fired electric utility steam generating unit other than an IGCC... gases that contain mercury (Hg) emissions in excess of each Hg emissions limit in paragraphs...
Training and Health. Leonardo da Vinci Series: Good Practices.
ERIC Educational Resources Information Center
Commission of the European Communities, Brussels (Belgium). Directorate-General for Education and Culture.
This document profiles programs in the fields of health and medicine that are offered through the European Commission's Leonardo da Vinci program. The following programs are profiled: (1) CYTOTRAIN (a transnational vocational training program in cervical cancer screening); (2) Apollo (a program of open and distance learning for paramedical…
The DaVinci Project: Multimedia in Art and Chemistry.
ERIC Educational Resources Information Center
Simonson, Michael; Schlosser, Charles
Provides an overview of the DaVinci Project, a collaboration of students, teachers, and researchers in chemistry and art to develop multimedia materials for grades 3-12 visualizing basic concepts in chemistry and visual art. Topics addressed include standards in art and science; the conceptual framework for the project; and project goals,…
The Potential da Vinci in All of Us
ERIC Educational Resources Information Center
Petto, Sarah; Petto, Andrew
The study of the human form is fundamental to both science and art curricula. For vertebrates, perhaps no feature is more important than the skeleton to determine observable form and function. As Leonard da Vinci's famous Proportions of the Human Figure (Virtruvian Man) illustrates, the size, shape, and proportions of the human body are defined by…
40 CFR 60.47Da - Commercial demonstration permit.
Code of Federal Regulations, 2010 CFR
... potential combustion concentration (80 percent reduction) for each 24-hour period of steam generator... owner or operator of a fluidized bed combustion electric utility steam generator (atmospheric or... Steam Generating Units for Which Construction is Commenced After September 18, 1978 § 60.47Da...
40 CFR 60.47Da - Commercial demonstration permit.
Code of Federal Regulations, 2011 CFR
... potential combustion concentration (80 percent reduction) for each 24-hour period of steam generator... owner or operator of a fluidized bed combustion electric utility steam generator (atmospheric or... Steam Generating Units for Which Construction is Commenced After September 18, 1978 § 60.47Da...
Women and Technical Professions. Leonardo da Vinci Series: Good Practices.
ERIC Educational Resources Information Center
Commission of the European Communities, Brussels (Belgium). Directorate-General for Education and Culture.
This document profiles programs for women in technical professions that are offered through the European Commission's Leonardo da Vinci program. The following programs are profiled: (1) Artemis and Diana (vocational guidance programs to help direct girls toward technology-related careers); (2) CEEWIT (an Internet-based information and…
Studying and Working Abroad. Leonardo da Vinci Series: Good Practices.
ERIC Educational Resources Information Center
Commission of the European Communities, Brussels (Belgium). Directorate-General for Education and Culture.
This document profiles recent successful examples of students studying and working abroad as part of the European Commission's Leonardo da Vinci program, which is designed to give students across the European Union the opportunity to experience vocational training in a foreign country. The following examples are presented: (1) 3 Finnish students…
Modelagem do vento e da fotosfera de AG Carinae
NASA Astrophysics Data System (ADS)
Groh, J. H.; Damineli, A.
A trajetória evolutiva das estrelas de alta massa depende fortemente de suas taxas de perda de massa. Apesar do rápido progresso no estudo destas estrelas, a taxa de perda de massa e outros parâmetros físicos básicos, como a temperatura superficial e a velocidade terminal do vento ainda não estão bem determinados. Isto ocorre devido à presença de ventos irregulares, rápidos e fortes ao redor destas estrelas, tornando a interpretação dos seus espectros uma tarefa difícil. Assim, a modelagem do vento e da fotosfera dessas estrelas está sendo cada vez mais usada para obter tais parâmetros a partir dos espectros. O aumento da taxa de perda de massa durante a fase LBV (Variáveis Luminosas Azuis), comparado com outros tipos de estrelas, tem sido atribuído a instabilidades do tipo S Doradus. Dispomos de uma base de dados espectroscópicos cobrindo 22 anos de observações de AG Carinae, incluindo um ciclo S Doradus completo, com espectros CCD em alta resolução na faixa óptica e infravermelha. Utilizamos o programa desenvolvido por Schmutz (1997) para uma análise preliminar desse ciclo, obtendo a taxa de perda de massa a partir da linha do Ha. Não existe uma correlação clara da taxa de perda de massa com mudanças da temperatura efetiva, do raio da estrela e do fluxo na banda V. A estrela atingiu seu mínimo fotométrico (raio mínimo) em 1990 e o máximo fotométrico (raio máximo) em 1995, enquanto que o fluxo máximo da linha do Ha ocorreu em 1996. Além disso a taxa de perda de massa não segue esse ciclo, contrariamente às idéias correntes. Para fazer um modelo mais realista estamos usando o programa CMFGEN (Hillier & Miller), que trata a fotosfera e o vento estelar de forma consistente, considerando a radiação fora do equilíbrio termodinâmico (NLTE) e com blanketting total de linhas. Simulamos o espectro de AG Carinae em duas épocas extremas do ciclo S Dor para testar os resultados obtidos com o modelo mais simplificado.
Roles and Delegation of Authority (R/DA) System
The processes of defining managerial roles and providing for delegation of authority are essential to any enterprise. At most large organizations, these processes are defined in policy manuals and through sets of standard operating procedures for many, if not all, business and administrative functions. Many of these staff-initiated, administrative functions require the routing of documents for approval to one or more levels of management. These employee-oriented, back office types of workflows tend to require more flexibility in determining to whom these documents should go to, while, at the same time, providing the responsible parties with the flexibility to delegate their approval authority or allow others to review their work. Although this practice is commonplace in manual, paper-based processes that exist in many organizations, it is difficult to provide the same flexibility in the more structured, electronic-based, workflow systems. The purpose of this report is to present a framework or architecture for creating a R/DA system and provide some insights associated with its design and utilization. To improve understanding and clarify subsequent discussion, the goals and requirements for the major R/DA system components, namely, the database and interface modules, are initially presented along with the identification of important concepts and the definition of critical terms. Next high-level functions relating the types of inputs to the outputs of the R/DA interface module are introduced and discussed. Then the relationships between the major R/DA modules and the primary components associated with its creation and maintenance are presented and analyzed. Finally, some conclusions are drawn relative to the advantages associated with developing a R/DA system for use in implementing an enterprise-wide, work-facilitating information system.
49 CFR 178.58 - Specification 4DA welded steel cylinders for aircraft use.
Code of Federal Regulations, 2012 CFR
... 49 Transportation 3 2012-10-01 2012-10-01 false Specification 4DA welded steel cylinders for...) SPECIFICATIONS FOR PACKAGINGS Specifications for Cylinders § 178.58 Specification 4DA welded steel cylinders for aircraft use. (a) Type, size, and service pressure. A DOT 4DA is a welded steel sphere (two...
49 CFR 178.58 - Specification 4DA welded steel cylinders for aircraft use.
Code of Federal Regulations, 2014 CFR
... 49 Transportation 3 2014-10-01 2014-10-01 false Specification 4DA welded steel cylinders for...) SPECIFICATIONS FOR PACKAGINGS Specifications for Cylinders § 178.58 Specification 4DA welded steel cylinders for aircraft use. (a) Type, size, and service pressure. A DOT 4DA is a welded steel sphere (two...
In vivo breakdown products of the 32 kDa thylakoid herbicide binding protein. [Spirodela oligorrhiza
Greenberg, B.M.; Mattoo, A.K.; Gaba, V.; Edelman, M.
The 32 kDa herbicide binding protein of PSII is degraded rapidly in the light. This phenomenon was investigated in Spirodela oligorrhiza. When fronds were radiolabeled in the presence of cycloheximide only the chloroplast-encoded proteins were labeled. Under these conditions, a breakdown product of 23.5 kDa was observed. This polypeptide was further degraded with kinetics similar to that of the 32 kDa protein. The 23.5 kDa polypeptide cross-reacted with an antibody specific to the 32 kDa protein. By protease digestion it was determined that the 23.5 kDa polypeptide is the intact N-terminal piece of the 32 kDa protein. Thus, this product corresponds to the membrane anchor of the 32 kDa protein. Using the same antibody, breakdown products of 16 kDa, 14 kDa and 12 kDa were observed. These products have also been observed in Zea mays and Solanum nigrum. Using DCMU during pulse-chase experiments at different light intensities, evidence was obtained that the 23.5 kDa breakdown product is generated in vivo.
JCoDA: a tool for detecting evolutionary selection
Background The incorporation of annotated sequence information from multiple related species in commonly used databases (Ensembl, Flybase, Saccharomyces Genome Database, Wormbase, etc.) has increased dramatically over the last few years. This influx of information has provided a considerable amount of raw material for evaluation of evolutionary relationships. To aid in the process, we have developed JCoDA (Java Codon Delimited Alignment) as a simple-to-use visualization tool for the detection of site specific and regional positive/negative evolutionary selection amongst homologous coding sequences. Results JCoDA accepts user-inputted unaligned or pre-aligned coding sequences, performs a codon-delimited alignment using ClustalW, and determines the dN/dS calculations using PAML (Phylogenetic Analysis Using Maximum Likelihood, yn00 and codeml) in order to identify regions and sites under evolutionary selection. The JCoDA package includes a graphical interface for Phylip (Phylogeny Inference Package) to generate phylogenetic trees, manages formatting of all required file types, and streamlines passage of information between underlying programs. The raw data are output to user configurable graphs with sliding window options for straightforward visualization of pairwise or gene family comparisons. Additionally, codon-delimited alignments are output in a variety of common formats and all dN/dS calculations can be output in comma-separated value (CSV) format for downstream analysis. To illustrate the types of analyses that are facilitated by JCoDA, we have taken advantage of the well studied sex determination pathway in nematodes as well as the extensive sequence information available to identify genes under positive selection, examples of regional positive selection, and differences in selection based on the role of genes in the sex determination pathway. Conclusions JCoDA is a configurable, open source, user-friendly visualization tool for performing evolutionary
The Da Vinci code dynamically de-coded.
Cohen, Mariam
The novel The Da Vinci Code, by Dan Brown has been on best-seller lists for over two years. An examination of Brown's previous novels reveals a well-designed plot line shared by all four novels that not only makes them good "thrillers" but also creates a mythological structure to the novels that draws on common unconscious fantasies in the same way that fairy tales do. One aspect of this mythological structure is the use of evil conspiracies (and benign ones as well) for the protagonist to overcome. In addition, The Da Vinci Code presents a religious theme involving legends about Mary Magdalene. This theme touches on the role of a feminine aspect to divinity in allowing for an erotic connection with the divine. PMID:16448349
[Lobectomy for lung cancer using the Da Vinci surgical system].
Nakamura, Hiroshige
Robot-assisted surgery using the da Vinci surgical system has attracted attention because of excellent operability without shaking by joint forceps under the clear vision of a three-dimensional high-definition camera in lung cancer surgery. Although this form of advanced medical care is not yet approved for insurance coverage, it is at the stage of clinical research and expected to be useful in hilar exposure, lymph node dissection, and suturing of the lung parenchyma or bronchus. Lung cancer surgery with the da Vinci system has the advantage of combining thoracotomy and minimally invasive surgery in video-assisted thoracic surgery. However, safety management, education, and significant cost are problems to be resolved. Several important issues such as sharing knowledge and technology of robotic surgery, education, training, development of new instruments, and acquisition of advanced medical insurance are discussed for the future development of robotic surgical systems. PMID:24946522
DaVinci's Mona Lisa entering the next dimension.
Carbon, Claus-Christian; Hesslinger, Vera M
For several of Leonardo da Vinci's paintings, such as The Virgin and Child with St Anne or the Mona Lisa, there exist copies produced by his own studio. In case of the Mona Lisa, a quite exceptional, rediscovered studio copy was presented to the public in 2012 by the Prado Museum in Madrid. Not only does it mirror its famous counterpart superficially; it also features the very same corrections to the lower layers, which indicates that da Vinci and the 'copyist' must have elaborated their panels simultaneously. On the basis of subjective (thirty-two participants estimated painter-model constellations) as well as objective data (analysis of trajectories between landmarks of both paintings), we revealed that both versions differ slightly in perspective. We reconstructed the original studio setting and found evidence that the disparity between both paintings mimics human binocular disparity. This points to the possibility that the two Giocondas together might represent the first stereoscopic image in world history. PMID:24303752
Da Rio, Nicola; Robberto, Massimo
We present the Tool for Astrophysical Data Analysis (TA-DA), a new software aimed to greatly simplify and improve the analysis of stellar photometric data in comparison with theoretical models, and allow the derivation of stellar parameters from multi-band photometry. Its flexibility allows one to address a number of such problems: from the interpolation of stellar models, or sets of stellar physical parameters in general, to the computation of synthetic photometry in arbitrary filters or units; from the analysis of observed color-magnitude diagrams to a Bayesian derivation of stellar parameters (and extinction) based on multi-band data. TA-DA is available as a pre-compiled Interactive Data Language widget-based application; its graphical user interface makes it considerably user-friendly. In this paper, we describe the software and its functionalities.
The da Vinci robot in right adrenalectomy: considerations on technique.
D'Annibale, Annibale; Fiscon, Valentino; Trevisan, Paolo; Pozzobon, Maurizia; Gianfreda, Valeria; Sovernigo, Gianna; Morpurgo, Emilio; Orsini, Camillo; Del Monte, Daniele
The da Vinci Robotic System (Intuitive Surgical, Mountain View, CA) became available at the General Surgery Department of Camposampiero Hospital in May 2001. From May 2001 to October 2002, 139 robotic operations were performed, one of which was a right adrenalectomy for a right adrenal mass. The progressive growth of the mass was the indication for surgical excision. Surgical adrenalectomy was successfully completed with da Vinci Robotic System using 5 ports (3 for the robotic system, 2 as service trocars). The wrist-like movements of the instrument's tip easily enabled the detachment of the right hepatic lobe from the gland and vessel isolation, while the 3-dimensional vision facilitated dissection of the veins from the vena cava. PMID:15259586
PanDaTox: a tool for accelerated metabolic engineering
Amitai, Gil; Sorek, Rotem
Metabolic engineering is often facilitated by cloning of genes encoding enzymes from various heterologous organisms into E. coli. Such engineering efforts are frequently hampered by foreign genes that are toxic to the E. coli host. We have developed PanDaTox (www.weizmann.ac.il/pandatox), a web-based resource that provides experimental toxicity information for more than 1.5 million genes from hundreds of different microbial genomes. The toxicity predictions, which were extensively experimentally verified, are based on serial cloning of genes into E. coli as part of the Sanger whole genome shotgun sequencing process. PanDaTox can accelerate metabolic engineering projects by allowing researchers to exclude toxic genes from the engineering plan and verify the clonability of selected genes before the actual metabolic engineering experiments are conducted.
40 CFR 60.49Da - Emission monitoring.
Code of Federal Regulations, 2011 CFR
... defined in 40 CFR 72.2, may use, as an alternative to the requirements specified in either paragraph (l... § 60.58b(i)(3) of subpart Eb of this part. (ii) Each 1-hour CO emissions average is calculated using... 40 Protection of Environment 6 2011-07-01 2011-07-01 false Emission monitoring. 60.49Da Section...
40 CFR 60.49Da - Emission monitoring.
Code of Federal Regulations, 2012 CFR
... affected unit that qualifies as a gas-fired or oil-fired unit, as defined in 40 CFR 72.2, may use, as an... § 60.58b(i)(3) of subpart Eb of this part. (ii) Each 1-hour CO emissions average is calculated using... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Emission monitoring. 60.49Da Section...
Surgical treatment of parietal defects with "da Vinci" surgical robot.
Vasilescu, D; Paun, S
The robotic surgery has come through the development of telemedicine and minimally invasive surgery concepts, being developed in the military medicine by NASA during the years 1970-1980. The purpose of this paper is to briefly present our experience in the new field of the robotic surgery, by analyzing the results obtained over a lot of 20 patients operated with the "da Vinci" robot within the last 5 years in the Clinical Emergency Hospital Bucharest for various abdominal defects. PMID:22802899
Improving Security in the ATLAS PanDA System
NASA Astrophysics Data System (ADS)
Caballero, J.; Maeno, T.; Nilsson, P.; Stewart, G.; Potekhin, M.; Wenaus, T.
The security challenges faced by users of the grid are considerably different to those faced in previous environments. The adoption of pilot jobs systems by LHC experiments has mitigated many of the problems associated with the inhomogeneities found on the grid and has greatly improved job reliability; however, pilot jobs systems themselves must then address many security issues, including the execution of multiple users' code under a common 'grid' identity. In this paper we describe the improvements and evolution of the security model in the ATLAS PanDA (Production and Distributed Analysis) system. We describe the security in the PanDA server which is in place to ensure that only authorized members of the VO are allowed to submit work into the system and that jobs are properly audited and monitored. We discuss the security in place between the pilot code itself and the PanDA server, ensuring that only properly authenticated workload is delivered to the pilot for execution. When the code to be executed is from a 'normal' ATLAS user, as opposed to the production system or other privileged actor, then the pilot may use an EGEE developed identity switching tool called gLExec. This changes the grid proxy available to the job and also switches the UNIX user identity to protect the privileges of the pilot code proxy. We describe the problems in using this system and how they are overcome. Finally, we discuss security drills which have been run using PanDA and show how these improved our operational security procedures.
Identification of an abundant 56 kDa protein implicated in food allergy as granule-bound starch synthase
Technology Transfer Automated Retrieval System (TEKTRAN)
Rice, the staple food of South and East Asian counties, is considered to be hypoallergenic. However, several clinical studies have documented rice-induced allergy in sensitive patients. Rice proteins with molecular weights of 14-16 kDa, 26 kDa, 33 kDa and 56 kDa have been identified as allergens. Re...
Amelioration of Auditory Response by DA9801 in Diabetic Mouse
Hong, Bin Na; Her, You Ri; Castañeda, Rodrigo; Moon, Hyo Won
Diabetes mellitus (DM) is a metabolic disease that involves disorders such as diabetic retinopathy, diabetic neuropathy, and diabetic hearing loss. Recently, neurotrophin has become a treatment target that has shown to be an attractive alternative in recovering auditory function altered by DM. The aim of this study was to evaluate the effect of DA9801, a mixture of Dioscorea nipponica and Dioscorea japonica extracts, in the auditory function damage produced in a STZ-induced diabetic model and to provide evidence of the mechanisms involved in enhancing these protective effects. We found a potential application of DA9801 on hearing impairment in the STZ-induced diabetic model, demonstrated by reducing the deterioration produced by DM in ABR threshold in response to clicks and normalizing wave I–IV latencies and Pa latencies in AMLR. We also show evidence that these effects might be elicited by inducing NGF related through Nr3c1 and Akt. Therefore, this result suggests that the neuroprotective effects of DA9801 on the auditory damage produced by DM may be affected by NGF increase resulting from Nr3c1 via Akt transformation. PMID:25878713
Amelioration of Auditory Response by DA9801 in Diabetic Mouse.
Lee, Yeong Ro; Hong, Bin Na; Her, You Ri; Castañeda, Rodrigo; Moon, Hyo Won; Kang, Tong Ho
Diabetes mellitus (DM) is a metabolic disease that involves disorders such as diabetic retinopathy, diabetic neuropathy, and diabetic hearing loss. Recently, neurotrophin has become a treatment target that has shown to be an attractive alternative in recovering auditory function altered by DM. The aim of this study was to evaluate the effect of DA9801, a mixture of Dioscorea nipponica and Dioscorea japonica extracts, in the auditory function damage produced in a STZ-induced diabetic model and to provide evidence of the mechanisms involved in enhancing these protective effects. We found a potential application of DA9801 on hearing impairment in the STZ-induced diabetic model, demonstrated by reducing the deterioration produced by DM in ABR threshold in response to clicks and normalizing wave I-IV latencies and Pa latencies in AMLR. We also show evidence that these effects might be elicited by inducing NGF related through Nr3c1 and Akt. Therefore, this result suggests that the neuroprotective effects of DA9801 on the auditory damage produced by DM may be affected by NGF increase resulting from Nr3c1 via Akt transformation. PMID:25878713
OpenDA-NEMO framework for ocean data assimilation
NASA Astrophysics Data System (ADS)
van Velzen, Nils; Altaf, Muhammad Umer; Verlaan, Martin
Data assimilation methods provide a means to handle the modeling errors and uncertainties in sophisticated ocean models. In this study, we have created an OpenDA-NEMO framework unlocking the data assimilation tools available in OpenDA for use with NEMO models. This includes data assimilation methods, automatic parallelization, and a recently implemented automatic localization algorithm that removes spurious correlations in the model based on uncertainties in the computed Kalman gain matrix. We have set up a twin experiment where we assimilate sea surface height (SSH) satellite measurements. From the experiments, we can conclude that the OpenDA-NEMO framework performs as expected and that the automatic localization significantly improves the performance of the data assimilation algorithm by successfully removing spurious correlations. Based on these results, it looks promising to extend the framework with new kinds of observations and work on improving the computational speed of the automatic localization technique such that it becomes feasible to include large number of observations.
The ATLAS PanDA Pilot in Operation
NASA Astrophysics Data System (ADS)
Nilsson, P.; Caballero, J.; De, K.; Maeno, T.; Stradling, A.; Wenaus, T.; ATLAS Collaboration
The Production and Distributed Analysis system (PanDA) [1-2] was designed to meet ATLAS [3] requirements for a data-driven workload management system capable of operating at LHC data processing scale. Submitted jobs are executed on worker nodes by pilot jobs sent to the grid sites by pilot factories. This paper provides an overview of the PanDA pilot [4] system and presents major features added in light of recent operational experience, including multi-job processing, advanced job recovery for jobs with output storage failures, gLExec [5-6] based identity switching from the generic pilot to the actual user, and other security measures. The PanDA system serves all ATLAS distributed processing and is the primary system for distributed analysis; it is currently used at over 100 sites worldwide. We analyze the performance of the pilot system in processing real LHC data on the OSG [7], EGI [8] and Nordugrid [9-10] infrastructures used by ATLAS, and describe plans for its evolution.
Common commercial cosmetic products induce arthritis in the DA rat.
Sverdrup, B; Klareskog, L; Kleinau, S
Many different agents, including mineral oil and silicone, have the capacity to act as immunological adjuvants, i.e., they can contribute to the activation of the immune system. Some adjuvants, including mineral oil, are known to induce arthritis in certain strains of rats after intradermal injection or percutaneous application. The aim of this study was to determine if common commercial cosmetic products containing mineral oil could induce arthritis in the highly susceptible DA (Dark Agouti) rat. Intradermal injection of five out of eight assayed cosmetic products without further additives resulted in arthritis with synovitis. One of the products induced a very aggressive arthritis, which had declined after 5-9 weeks. When this product was also assayed for arthritogenicity upon percutaneous administration, it induced a mild and transient arthritis in 5 out of 10 DA rats, whereas control animals showed no clinical signs of joint involvement. No arthritic reaction was seen in rats after peroral feeding with the most arthritogenic product or by intravaginal application of Freund's adjuvants. Silicone gel implants in DA rats did not cause arthritis. We conclude that mineral oils included in common commercially available products retain their adjuvant properties and are arthritogenic in the presently investigated arthritis-prone rat strain. There is yet no evidence that mineral oils present in cosmetics may contribute to arthritis in humans, but we suggest that this question should be subject to further investigation. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:9417771
OpenDA-NEMO framework for ocean data assimilation
NASA Astrophysics Data System (ADS)
van Velzen, Nils; Altaf, Muhammad Umer; Verlaan, Martin
Data assimilation methods provide a means to handle the modeling errors and uncertainties in sophisticated ocean models. In this study, we have created an OpenDA-NEMO framework unlocking the data assimilation tools available in OpenDA for use with NEMO models. This includes data assimilation methods, automatic parallelization, and a recently implemented automatic localization algorithm that removes spurious correlations in the model based on uncertainties in the computed Kalman gain matrix. We have set up a twin experiment where we assimilate sea surface height (SSH) satellite measurements. From the experiments, we can conclude that the OpenDA-NEMO framework performs as expected and that the automatic localization significantly improves the performance of the data assimilation algorithm by successfully removing spurious correlations. Based on these results, it looks promising to extend the framework with new kinds of observations and work on improving the computational speed of the automatic localization technique such that it becomes feasible to include large number of observations.
Basic principles of pharmaceutical science in Ayurvĕda.
Subhose, Varanasi; Srinivas, Pitta; Narayana, Ala
Pharmaceutical is one of the allied branches of science, which is closely associated with Medical science. Today pharmaceutical chemistry and pharmacognosy are playing important role in treatment for a disease and its prevention. Herbal medicines are being used by about 80% of the world population mostly in the developing countries in the primary health care. There has been an upsurge in demand for the Phyto-pharmaceutical products of Ayurvĕda in western nations, because of the fact that the synthetic drugs are considered to be unsafe. Due to this many national and multinational pharmaceutical companies are now concentrating on manufacturing of Ayurvĕdic Phyto-pharmaceutical products. Ayurvĕda is the Indian traditional system of medicine, which also deals about pharmaceutical science. The Ayurvĕdic knowledge of the pharmaceutical science is scattered in Ayurvĕdic classical texts. Săranghadhara Samhita, which is written by Săranghadhara, explain systematically about the information of the Ayurvĕdic pharmaceutical science and also updated it. Industrialized manufacturing of Ayurvĕdic dosage forms has brought in new challenges like deviation from basic concepts of medicine preparation. Săranghadhara Samrhită the devout text on pharmaceutics in Ayurvĕda comes handy to solve such problems, as the methods described are very lucid and easy to follow. PMID:17333665
[The Arabic influence in the "Colóquios dos simples e drogas da India" of Garcia da Orta].
Ricordel, Joëlle
The "Colóquios dos simples e drogas he cousas medicinais de Índia" (Conversations on the simples, drugs and medicinal substances of India) (1563) of Garcia da Orta is a botanical and pharmacognosy book. The author is a Portuguese physician who studied in the Spanish universities and practiced medicine mainly in India. He studies in short chapters presented in the form of dialogues about sixty simples. Sources to which he refers are indicative of a "classical" training, but also the mark of a curious and open mind to different cultures. The Arabic sources are numerous and mainly concern the identification of substances by abundant synonyms of their names in foreign languages and different medicinal uses that may have been done by the ancient physicians. However, Da Orta is critical with respect to these sources, seeking contradictions and differences of opinion among authors. He confronts them with the oral information collected thanks to a wide network of contacts. PMID:26529894
Seeking 'energy' vs. pain relief in spas in Brazil (Caldas da Imperatriz) and Portugal (Termas da Sulfurea).
Quintela, Maria Manuel
This paper is a comparative ethnography of the therapeutic practices at two different spa locations: Caldas da Imperatriz, SC, Brazil, and Termas da Sulfurea in Cabeco de Vide, Portugal. The comparison reveals the existence of contrasting 'explanatory models' held by the spa-goers as well as by the official medical systems. In the Portuguese context this model is highly medicalized; in the Brazilian case, spa treatments are viewed as 'alternative' or 'complementary' therapy and are also related to religious philosophies. Each model corresponds to a different idiom expressing certain experiences and world views, one focusing on 'pains' (dores) and the other on 'energy' (energia), the former leading to the rationale of 'curing', the latter to the notion of 'energizing'. In this paper the author intends to analyze and contrast the categories found in these models, which originate from different conceptions of health, illness and healing for Brazilian and Portuguese spa-goers. PMID:21563001
The 29-kDa proteins phosphorylated ion thrombin-activated human platelets are forms of the estrogen receptor-related 27-kDa heat shock protein
Mendelsohn, M.E.; Yan Zhu; O'Neill, S. )
Thrombin plays a critical role in platelet activation, hemostasis, and thrombosis. Cellular activation by thrombin leads to the phosphorylation of multiple proteins, most of which are unidentified. The authors have characterized several 29-kDa proteins that are rapidly phosphorylated following exposure of intact human platelets to thrombin. A murine monoclonal antibody raised to an unidentified estrogen receptor-related 29-kDa protein selectively recognized these proteins as well as a more basic, unphosphorylated 27-kDa protein. Cellular activation by thrombin led to a marked shift in the proportion of protein from the 27-kDa unphosphorylated form to the 29-kDa phosphoprotein species. Using this antibody, they isolated and sequenced a human cDNA clone encoding a protein that was identical to the mammalian 27-kDa heat shock protein (HSP27), a protein of uncertain function that is known to be phosphorylated to several forms and to be transcriptionally induced by estrogen. The 29-kDa proteins were confirmed to be phosphorylated forms of HSP27 by immunoprecipitation studies. Thus, the estrogen receptor-related protein is HSP27, and the three major 20-kDa proteins phosphorylated in thrombin-activated platelets are forms of HSP27. These data suggest a role for HSP27 in the signal transduction events of platelet activation.
Identification of sequence similarity between 60 kDa and 70 kDa molecular chaperones: evidence for a common evolutionary background?
Flores, A I; Cuezva, J M
Recent findings support the premise that chaperonins (60 kDa stress-proteins) and alpha-subunits of F-type ATPases (alpha-ATPase) are evolutionary related protein families. Two-dimensional gel patterns of synthesized proteins in unstressed and heat-shocked embryonic Drosophila melanogaster SL2 cells revealed that antibodies raised against the alpha-subunit of the F1-ATPase complex from rat liver recognize an inducible p71 member of the 70 kDa stress-responsive protein family. Molecular recognition of this stress-responsive 70 kDa protein by antibodies raised against the F1-ATPase alpha-subunit suggests the possibility of partial sequence similarity within these ATP-binding protein families. A multiple sequence alignment between alpha-ATPases and 60 kDa and 70 kDa molecular chaperones is presented. Statistical evaluation of sequence similarity reveals a significant degree of sequence conservation within the three protein families. The finding suggests a common evolutionary origin for the ATPases and molecular chaperone protein families of 60 kDa and 70 kDa, despite the lack of obvious structural resemblance between them. PMID:9065788
Stereoscopic augmented reality for da Vincii™ robotic biliary surgery
Volonté, Francesco; Buchs, Nicolas C.; Pugin, François; Spaltenstein, Joël; Jung, Minoa; Ratib, Osman; Morel, Philippe
INTRODUCTION New laparoscopic techniques put distance between the surgeon and his patient. PRESENTATION OF CASE 3D volume rendered images directly displayed in the da Vinci surgeon's console fill this gap by allowing the surgeon to fully immerse in its intervention. DISCUSSION During the robotic operation the surgeon has a greater control on the procedure because he can stay more focused not being obliged to turn is sight out of his operative field. Moreover, thanks to depth perception of the rendered images he had a precise view of important anatomical structures. CONCLUSION We describe our preliminary experience in the quest of computer-assisted robotic surgery. PMID:23466685
Ultratight crystal packing of a 10 kDa protein
Trillo-Muyo, Sergio; Chruszcz, Maksymilian; Minor, Wladek; Kuisiene, Nomeda
The crystal structure of the C-terminal domain of a putative U32 peptidase from G. thermoleovorans is reported; it is one of the most tightly packed protein structures reported to date. While small organic molecules generally crystallize forming tightly packed lattices with little solvent content, proteins form air-sensitive high-solvent-content crystals. Here, the crystallization and full structure analysis of a novel recombinant 10 kDa protein corresponding to the C-terminal domain of a putative U32 peptidase are reported. The orthorhombic crystal contained only 24.5% solvent and is therefore among the most tightly packed protein lattices ever reported.
A quasi-passive CMOS pipeline D/A converter
NASA Technical Reports Server (NTRS)
Wang, Fong-Jim; Temes, Gabor C.; Law, Simon
A novel pipeline digital-to-analog converter configuration, based on switched-capacitor techniques, is described. An n-bit D/A conversion can be implemented by cascading n + 1 unit cells. The device count of the circuit increases linearly, not exponentially, with the conversion accuracy. The new configuration can be pipelined. Hence, the conversion rate can be increased without requiring a higher clock rate. An experimental 10-bit DAC prototype has been fabricated using a 3-micron CMOS process. The results show that high-speed, high-accuracy, and low-power operation can be achieved without special process or postprocess trimming.
Leonardo da Vinci and the origin of semen
Noble, Denis; DiFrancesco, Dario; Zancani, Diego
It is well known that Leonardo da Vinci made several drawings of the human male anatomy. The early drawings (before 1500) were incorrect in identifying the origin of semen, where he followed accepted teaching of his time. It is widely thought that he did not correct this mistake, a view that is reflected in several biographies. In fact, he made a later drawing (after 1500) in which the description of the anatomy is remarkably accurate and must have been based on careful dissection. In addition to highlighting this fact, acknowledged previously in only one other source, this article reviews the background to Leonardo's knowledge of the relevant anatomy. PMID:27494016
Visual tracking of da Vinci instruments for laparoscopic surgery
NASA Astrophysics Data System (ADS)
Speidel, S.; Kuhn, E.; Bodenstedt, S.; Röhl, S.; Kenngott, H.; Müller-Stich, B.; Dillmann, R.
Intraoperative tracking of laparoscopic instruments is a prerequisite to realize further assistance functions. Since endoscopic images are always available, this sensor input can be used to localize the instruments without special devices or robot kinematics. In this paper, we present an image-based markerless 3D tracking of different da Vinci instruments in near real-time without an explicit model. The method is based on different visual cues to segment the instrument tip, calculates a tip point and uses a multiple object particle filter for tracking. The accuracy and robustness is evaluated with in vivo data.
Da Vinci's codex and the anatomy of healthcare.
Stephens-Borg, Keith
We usually display a laid-back approach to medical jargon throughout our theatre work. The word 'perioperative' is built from the Greek word 'peri' (around) and the Latin 'operari' (to work). Latin and Greek became the prefixed language of choice for Leonardo da Vinci, and his research was pivotal in determining the way in which surgical procedures are documented. Ancient manuscripts aided the unfolding of the secrets of anatomy, and Leonardo revealed that art was the key in expressive detailed explanation. PMID:23248927
Evolution of the ATLAS PanDA workload management system for exascale computational science
NASA Astrophysics Data System (ADS)
Maeno, T.; De, K.; Klimentov, A.; Nilsson, P.; Oleynik, D.; Panitkin, S.; Petrosyan, A.; Schovancova, J.; Vaniachine, A.; Wenaus, T.; Yu, D.; Atlas Collaboration
An important foundation underlying the impressive success of data processing and analysis in the ATLAS experiment [1] at the LHC [2] is the Production and Distributed Analysis (PanDA) workload management system [3]. PanDA was designed specifically for ATLAS and proved to be highly successful in meeting all the distributed computing needs of the experiment. However, the core design of PanDA is not experiment specific. The PanDA workload management system is capable of meeting the needs of other data intensive scientific applications. Alpha-Magnetic Spectrometer [4], an astro-particle experiment on the International Space Station, and the Compact Muon Solenoid [5], an LHC experiment, have successfully evaluated PanDA and are pursuing its adoption. In this paper, a description of the new program of work to develop a generic version of PanDA will be given, as well as the progress in extending PanDA's capabilities to support supercomputers and clouds and to leverage intelligent networking. PanDA has demonstrated at a very large scale the value of automated dynamic brokering of diverse workloads across distributed computing resources. The next generation of PanDA will allow other data-intensive sciences and a wider exascale community employing a variety of computing platforms to benefit from ATLAS' experience and proven tools.
Effects of DA-Phen, a dopamine-aminoacidic conjugate, on alcohol intake and forced abstinence.
Sutera, Flavia Maria; De Caro, Viviana; Cannizzaro, Carla; Giannola, Libero Italo; Lavanco, Gianluca; Plescia, Fulvio
The mesolimbic dopamine (DA) system plays a key role in drug reinforcement and is involved in the development of alcohol addiction. Manipulation of the DAergic system represents a promising strategy to control drug-seeking behavior. Previous studies on 2-amino-N-[2-(3,4-dihydroxy-phenyl)-ethyl]-3-phenyl-propionamide (DA-Phen) showed in vivo effects as a DA-ergic modulator. This study was aimed at investigate DA-Phen effects on operant behavior for alcohol seeking behavior, during reinstatement following subsequent periods of alcohol deprivation. For this purpose, male Wistar rats were tested in an operant paradigm of self-administration; behavioral reactivity and anxiety like-behavior during acute abstinence were evaluated. A characterization of DA-Phen CNS targeting by its quantification in the brain was also carried out. Our findings showed that DA-Phen administration was able to reduce relapse in alcohol drinking by 50% and reversed the alterations in behavioral reactivity and emotionality observed during acute abstinence. In conclusion, DA-Phen can reduce reinstatement of alcohol drinking in an operant-drinking paradigm following deprivation periods and reverse abstinence-induced behavioral phenotype. DA-Phen activity seems to be mediated by the modulation of the DAergic transmission. However further studies are needed to characterize DA-Phen pharmacodynamic and pharmacokinetic properties, and its potential therapeutic profile in alcohol addiction. PMID:27155501
[Textual Research on Chiding Disease and Matixiang Herb in Da he wai ke (External Medicine of Da He)].
Pu, Xiao-Lan; Zhang, Yi; Yang, Wen-Yu; Wu, Ya-Mei
Recorded in Da he wai ke (The External Medicine of Da He) in the Ming dynasty without a corresponding disease title in contemporary scholarly concepts of Traditional Chinese Medicine (TCM), Chiding disease was described as an acute symptomatic disease with inevitable serious conditions of convulsion-like or mania-like behavior, and as a refractory ailment. It was also considered as an alias of Hongsiding (namely the acute lymphangitis) in few ancient books. Matixiang, a Chinese herb for treatment of Chiding, was regarded as a nonsuch drug by Da he wai ke, and was considered as a take-then-cure drug by Chuang yang jing yan quan shu (Compendium for Experience in Sores Diseases). The present studies about Chiding and Matixiang show that: (1) the name of Chiding was first seen in Zhong zang jing (Master Hua's Classic of the Viscera) ; (2) Chiding was rarely mentioned in medical literature of the Song, Yuan and Qing Dynasty, but mostly seen in the Ming Dynasty; (3) Xinding and Huoyanding were found as synonyms of Chiding ; (4) the distinction between Chiding and Hongsiding (flaming sore) was revealed by comparison analysis of their etiology, affected part(s), symptoms, prognosis and syndromic pictures; (5) Chiding on the tongue is the same as "Tongue Ding" or "Sublingual Ding" in contemporary TCM and as "Sublingual Space Infection" in Western Medicine. Moreover, Chiding on the hands, feet or other skin parts could be classified as "Hands/Feet Ding" in contemporary TCM and as "Furuncle" (i.e., Acute Suppurative Folliculitis and Perifolliculitis or cellulites of the tongue) in Western Medicine; (6) Matixiang is probably deduced as Asarum forbesii Maxim., A. ichangense C. Y. Cheng et C. S. Yang, family Aristolochiaceae, or Valeriana jatamansi Jones., family Valerianaceae. PMID:23363849
Chandra Confirmation of a Pulsar Wind Nebula in DA 495
NASA Technical Reports Server (NTRS)
Arzoumanian, Z.; Safi-Harb, S.; Landecker, T.L.; Kothes, R.; Camilo, F.
As part of a multiwavelength study of the unusual radio supernova remnant DA 495, we present observations made with the Chandra X-ray Observatory. Imaging and spectroscopic analysis confirms the previously detected X-ray source at the heart of the annular radio nebula, establishing the radiative properties of two key emission components: a soft unresolved source with a blackbody temperature of 1 MK consistent with a neutron star, surrounded by a nontherma1 nebula 40" in diameter exhibiting a power-law spectrum with photon index Gamma = 1.63, typical of a pulsar wind nebula. Morphologically, the nebula appears to be slightly extended along a direction, in projection on the sky, previously demonstrated to be of significance in radio and ASCA observations; we argue that this represents the orientation of the pulsar spin axis. At smaller scales, a narrow X-ray feature is seen extending out 5" from the point source, but energetic arguments suggest that it is not the resolved termination shock of the pulsar wind against the ambient medium. Finally, we argue based on synchrotron lifetimes in the nebular magnetic field that DA 495 represents the first example of a pulsar wind nebula in which electromagnetic flux makes up a significant part, together with particle flux, of the neutron star's wind.
The DA+dMe Binary WD 0334-6400
NASA Astrophysics Data System (ADS)
Barrett, P. E.; Schlegel, E. M.; Dupuis, J.; Heathcote, B.
Multiwavelength observations of the star WD 0334-6400 (or Ret 1 in A Catalog and Altas of Cataclysmic Variables: Living Edition) show it to be a DA+dMe binary. Our analysis of the far-ultraviolet continuum and spectral lines reveal that Teff = 36500 K; log g = 7.7; and the photosphere contains C, N, O, Si, P, and S. C/H = 2 x 10-6, which is the highest abundance seen in a DA white dwarf at this temperature. The dMe star is seen as a flux excess in near-infrared photometry and appears to show occasional flaring of about one magnitude as seen in the Harvard plates. There is no evidence of periodic variability in the spectroscopic or photometric data. This paper is based on observations made with the NASA-CNES-CSA FUSE. This research is partially supported by NASA FUSE grants NAG5-12413 and NNG04GL18G.
The 37kDa/67kDa Laminin Receptor acts as a receptor for Aβ42 internalization
Da Costa Dias, Bianca; Jovanovic, Katarina; Gonsalves, Danielle; Moodley, Kiashanee; Reusch, Uwe; Knackmuss, Stefan; Weinberg, Marc S.; Little, Melvyn; Weiss, Stefan F. T.
Neuronal loss is a major neuropathological hallmark of Alzheimer's disease (AD). The associations between soluble Aβ oligomers and cellular components cause this neurotoxicity. The 37 kDa/67 kDa laminin receptor (LRP/LR) has recently been implicated in Aβ pathogenesis. In this study the mechanism underlying the pathological role of LRP/LR was elucidated. Försters Resonance Energy Transfer (FRET) revealed that LRP/LR and Aβ form a biologically relevant interaction. The ability of LRP/LR to form stable associations with endogenously shed Aβ was confirmed by pull down assays and Aβ-ELISAs. Antibody blockade of this association significantly lowered Aβ42 induced apoptosis. Furthermore, antibody blockade and shRNA mediated downregulation of LRP/LR significantly hampered Aβ42 internalization. These results suggest that LRP/LR is a receptor for Aβ42 internalization, mediating its endocytosis and contributing to the cytotoxicity of the neuropeptide by facilitating intra-cellular Aβ42 accumulation. These findings recommend anti-LRP/LR specific antibodies and shRNAs as potential therapeutic tools for AD treatment. PMID:24990253
The Indicative and Subjunctive "da"-complements in Serbian A Syntactic-Semantic Approach
ERIC Educational Resources Information Center
Todorovic, Natasa
A syntactic-semantic investigation of subjunctive and indicative "da"-complements in Serbian is conducted in this project. After a careful comparison of Serbian sentence constructions with "da"-complements to the equivalent sentence structures in languages of the Balkans as well as other Slavic languages, it is clearly…
Paśu Ayurvĕda (veterinary medicine) in Garudapurăņa.
Varanasi, Subhose; Narayana, A
The history of veterinary medicine is closely tied to the development of human medicine. Evidence of animal medicine has been found in ancient civilizations, such as those of the Hindu, Babylonians, Hebrews, Arabs, Greeks, and Romans. Ancient Indian literature in the form of the holy Vĕda, Purăna, Brăhmaņa, epics, etc. is flooded with information on animal care. The Purăņa are ancient scriptures discuss varied topics like devotion to God and his various aspects, traditional sciences like Ayurvĕda, Jyŏtişa (Astrology), cosmology, concepts like dharma, karma, reincarnation and many others. The treatment of animal diseases using Ayurvedic medicine has been mentioned in Garudapurăna, Agnipurăņa, Atri-samhită, Matsyapurăņa and many other texts. The Garudapurăņa is one of the important Săttvika purăna, the subject matter is divided into two parts, viz. Pŭrvakhaņda (first part) and an Uttarakhaņda (subsequent part). Gavăyurvĕda, Gajăyurvĕda narrated briefly and Aśvăyurvĕda described detailly in Pŭrvakhaņda. PMID:19580108
A Value-Engaged Approach for Evaluating the Bunche-Da Vinci Learning Academy
ERIC Educational Resources Information Center
Greene, Jennifer C.
In 2001, the Bunche Academy was chosen by its district to join in partnership with the Da Vinci Learning Corporation to embark on an ambitious whole-school reform initiative, especially designed by the corporation for low-performing schools. In this chapter, the author describes how, as illustrated in the Bunche-Da Vinci Learning Academy context,…
Processing and Visualizing Planetary Data Using DaVinci: Updates for Portability and Scriptable Execution
NASA Astrophysics Data System (ADS)
Edwards, C. S.; Anwar, S.; Hagee, W.; Doerres, D.; Dickensheid, S.; Christensen, P. R.
We present updates to the DaVinci software toolkit that enable stand-alone reading of ISIS3 files, a link between DaVinci and JMARS, the ingestion and use of geospatial information, and a series of executable THEMIS processing scripts.
32 CFR Appendix F to Part 623 - Power of Attorney (DA Form 4881-4-R)
Code of Federal Regulations, 2010 CFR
... 32 National Defense 3 2010-07-01 2010-07-01 true Power of Attorney (DA Form 4881-4-R) F Appendix F to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. F Appendix F to Part 623—Power of Attorney (DA Form...
Topological studies of spinach 22 kDa protein of Photosystem II.
Kim, S; Pichersky, E; Yocum, C F
An intrinsic 22 kDa polypeptide is associated with the O2-evolving Photosystem II core complex in a variety of green plants, although it does not appear to be required for O2 evolution. Digestion of thylakoid membranes and isolated Photosystem II preparations with trypsin, followed by immunoblotting using spinach anti-22 kDa antibodies, leads to two observations: (1) the domain between the 2nd and 3rd transmembrane helices of the 22 kDa protein is stromally exposed, and (2) only in a reaction center complex preparation, lacking the chlorophyll a/b-light harvesting complex II, is there extensive proteolytic cleavage of the 22 kDa protein. We also found that after, but not prior to, selective extraction of the 22 and 10 kDa proteins from Photosystem II membranes, the chlorophyll a/b-light harvesting complex II can be separated from the Photosystem II reaction center core by precipitation with MgCl2. This result suggests that the 22 kDa polypeptide is located between the Photosystem II reaction center polypeptides and light-harvesting complex II; it is possible that the protein serves as a link between the two protein complexes. The presence of the 22 kDa protein in several species was also examined by immunoblotting with polyclonal spinach anti-22 kDa antibodies. PMID:7803450
32 CFR Appendix E to Part 623 - Surety Bond (DA Form 4881-3-R)
Code of Federal Regulations, 2010 CFR
... 32 National Defense 3 2010-07-01 2010-07-01 true Surety Bond (DA Form 4881-3-R) E Appendix E to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. E Appendix E to Part 623—Surety Bond (DA Form...
32 CFR Appendix F to Part 623 - Power of Attorney (DA Form 4881-4-R)
Code of Federal Regulations, 2011 CFR
... 32 National Defense 3 2011-07-01 2009-07-01 true Power of Attorney (DA Form 4881-4-R) F Appendix F to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. F Appendix F to Part 623—Power of Attorney (DA Form...
A Creative Approach to the Common Core Standards: The Da Vinci Curriculum
ERIC Educational Resources Information Center
Chaucer, Harry
"A Creative Approach to the Common Core Standards: The Da Vinci Curriculum" challenges educators to design programs that boldly embrace the Common Core State Standards by imaginatively drawing from the genius of great men and women such as Leonardo da Vinci. A central figure in the High Renaissance, Leonardo made extraordinary contributions as a…
The Effect of DA-9701 in Opioid-induced Bowel Dysfunction of Guinea Pig
Hussain, Zahid; Rhee, Kwang Won; Lee, Young Ju; Park, Hyojin
Background/Aims Opioid induced bowel dysfunction (OIBD) is associated with decreased gastrointestinal (GI) propulsive activity due to intake of opioid analgesics. DA-9701, a novel prokinetic agent formulated with Pharbitis Semen and Corydalis Tuber has promising effects on GI motor function. Therefore, we aim to evaluate the prokinetic effects of DA-9701 in an OIBD model of guinea pig. Methods The ileal and distal colon muscle contraction in presence of different doses of DA-9701, morphine, and combination (morphine + DA-9701) was measured by tissue bath study. The prokinetic effect of DA-9701 was assessed by charcoal transit and fecal pellet output assay in an OIBD model of guinea pig. Results DA-9701 significantly increased the amplitude and area under the curve of ileal muscle contraction, while there was insignificant effect on the distal colon compared to the control. The maximal amplitude of ileal muscle contraction was acquired at a concentration of 10 μg/mL of DA-9701. In contrast, morphine significantly decreased the amplitude of ileal and distal colon muscle contraction compared to the control. Morphine delayed both upper (P < 0.01) and lower (P < 0.05) GI transit, and delayed GI transit was restored by the administration of DA-9701. Morphine induced reduction of contractility was significantly ameliorated by addition of DA-9701 in both ileal and distal colon muscles. Conclusions DA-9701 significantly increased the amplitude of contraction of the ileal muscle, however the distal colon muscle contraction was insignificant. Additionally, it restored delayed upper and lower GI transit in an OIBD model of guinea pig, and it might prove to be a useful candidate drug in a clinical trial for OIBD. PMID:26932898
Pharmacological action of DA-9701 on the motility of feline stomach circular smooth muscle.
Nguyen, Thanh Thao; Song, Hyun Ju; Ko, Sung Kwon; Sohn, Uy Dong
DA-9701, a new prokinetic agent for the treatment of functional dyspepsia, is formulated with Pharbitis semen and Corydalis tuber. This study wasconducted to determine the pharmacological action of DA-9701 and to identify the receptors involved in DA-9701 -induced contractile responsesin the feline gastric corporal, fundic and antral circular smooth muscle. Concentration-response curve to DA-9701 was established. The tissue trips were exposed to methylsergide, ketanserin, ondansetron, GR 113808, atropine and dopamine before administration of DA-9701. The contractile force was determined before and after administration of drugs by a polygraph.DA-9701 enhanced the spontaneous contractile amplitude of antrum, corpus and fundus. However, it did not change the spontaneous contractile frequency of antrum and corpus, but concentration-dependently reduced that of fundus. In the fundus, DA-9701 -induced tonic contractions were inhibited by dopamine, methylsergide, ketanserine, ondansetron or GR 113808 respectively, but not by atropine, indicating that the contractile responses are mediated by multiple receptors: 5-HT2, 5-HT3, 5-HT4, and dopamine receptors. In the corpus, DA-9701-induced contractions were blocked by atropine, dopamine or GR 113808, but not by methysergide, ketanserin or ondansetron, indicating that they are involved in receptors on both, smooth muscles and neurons: 5-HT4 and dopamine receptors. However, contractile responses to DA-9701 are mainly mediated by dopamine receptors in the antrum. These results suggest that DA-9701 has important roles in gastric accommodation by enhancing tonic activity of fundus, and in gastric emptying and gastrointestinal transit by phasic contractions of corpus and antrum mediated by multiple receptors. PMID:25980179
A 92-kDa human immunostimulatory protein.
Fontan, E; Briend, E; Saklani-Jusforgues, H; d'Alayer, J; Vandekerckhove, J; Fauve, R M
We purified to apparent homogeneity a human urinary glycoprotein of 92 kDa (HGP.92) that, administered intravenously at 250 micrograms/kg, fully protected mice against a lethal inoculum of Listeria monocytogenes. Since HGP.92 protected scid mice, which lack B and T lymphocytes, this increased resistance to Listeria did not appear to be lymphocyte mediated. Furthermore, inflammatory macrophages incubated with 6 nM HGP.92 inhibited the growth of Lewis carcinoma cells in vitro. These two activities appeared to depend on an oligosaccharide moiety, as they were lost after N-Glycanase treatment of HGP.92. Thus, the biological activity of HGP.92 was in some way related to a glycan moiety. Images PMID:8078887
DBS -- An rlogin multiplexer and output logger for DA systems
Oleynik, G.; Appleton, L.; Udumula, L.; Votava, M.
DART Bootstrap Services (dbs) is the first component of run-control for the DART Data Acquisition system -- the DA for the 96` round of experiments at Fermilab -- though it has potential usefulness as a powerful tool in other distributed applications. dbs is an rlogin session multiplexer. It allows a user, running a single program, to start up any number of remote login sessions, feed shell commands to them, and collect their output into a single (or multiple) log files (a server keeps the sessions open and collects their output). From this program, any session can be attached to interactively so it appears just like an rlogin session -- dbs becomes transparent. When finished with this interactive mode, the user can escape back to dbs and attach to a different session if so desired. Among many other useful features, dbs supplies a mechanism for cleanup (deletion) of all processes created under a session, allowing a fresh start.
VLBI Radar of the 2012 DA14 Asteroid
NASA Astrophysics Data System (ADS)
Nechaeva, M. B.; Dugin, N. A.; Antipenko, A. A.; Bezrukov, D. A.; Bezrukov, V. V.; Voytyuk, V. V.; Dement'ev, A. F.; Jekabsons, N.; Klapers, M.; Konovalenko, A. A.; Kulishenko, V. F.; Nabatov, A. S.; Nesteruk, V. N.; Putillo, D.; Reznichenko, A. M.; Salerno, E.; Snegirev, S. D.; Tikhomirov, Yu. V.; Khutornoy, R. V.; Skirmante, K.; Shmeld, I.; Chagunin, A. K.
An experiment on VLBI radar of the 2012 DA14 asteroid was carried out on February 15-16, 2011 at the time of its closest approach to the Earth. The research teams of Kharkov (Institute of Radio Astronomy of the National Academy of Sciences of Ukraine), Evpatoria (National Space Facilities Control and Test Center), Nizhny Novgorod (Radiophysical Research Institute), Bologna (Istituto di Radioastronomia (INAF)), and Ventspils (Ventspils International Radioastronomy Center) took part in the experiment. The asteroid was irradiated by the RT-70 planetary radar (Evpatoria) at a frequency of 5 GHz. The reflected signal was received using two 32-m radio telescopes in Medicina (Italy) and Irbene (Latvia) in radiointerferometric mode. The Doppler frequency shifts in bi-static radar mode and interference frequency in VLBI mode were measured. Accuracy of the VLBI radar method for determining the radial and angular velocities of the asteroid were estimated.
Zhao, J. K.; Luo, A. L.; Zhao, G.; Oswalt, T. D. E-mail: firstname.lastname@example.com E-mail: email@example.com
We present a spectroscopically identified catalog of 70 DA white dwarfs (WDs) from the LAMOST pilot survey. Thirty-five are found to be new identifications after cross-correlation with the Eisenstein et al. and Villanova catalogs. The effective temperature and gravity of these WDs are estimated by Balmer lines fitting. Most of them are hot WDs. The cooling times and masses of these WDs are estimated by interpolation in theoretical evolution tracks. The peak of the mass distribution is found to be {approx}0.6 M {sub Sun }, which is consistent with prior work in the literature. The distances of these WDs are estimated using the method of synthetic spectral distances. All of these WDs are found to be in the Galactic disk from our analysis of space motions. Our sample supports the expectation that WDs with high mass are concentrated near the plane of the Galactic disk.
DBS - an rlogin multiplexor and output logger for DA systems
Oleynik, G.; Appleton, L.; Udumula, L.; Votava, M.
DART Bootstrap Services (dbs) is the first component of run-control for the DART Data Acquisition system - the DA for the 96{prime} round of experiments at Fermilab - though it has potential usefulness as a powerful tool in other distributed applications, dbs is an rlogin session multiplexer. It allows a user, running a single program, to start up any number of remote login sessions, feed shell commands to them, and collect their output into a single (or multiple) logfiles (a server keeps the sessions open and collects their output). From this program, any session can be attached to interactively so it appears just like an rlogin session - dbs becomes transparent. When finished with this interactive mode, the user can escape back to dbs and attach to a different session if so desired. Among many other useful features, dbs supplies a mechanism for cleanup (deletion) of all processes created under a session, allowing a fresh start.
[Regarding the Manuscript D " Dell' occhio " of Leonardo da Vinci].
Heitz, Robert F
Leonardo da Vinci's Manuscript D consists of five double pages sheets, which, folded in two, comprise ten folios. This document, in the old Tuscan dialect and mirror writing, reveals the ideas of Leonardo on the anatomy of the eye in relation to the formation of images and visual perception. Leonardo explains in particular the behavior of the rays in the eye in terms of refraction and reflection, and is very mechanistic in his conception of the eye and of the visual process. The most significant innovations found in these folios are the concept of the eye as a camera obscura and the intersection of light rays in the interior of the eye. His texts nevertheless show hesitation, doubts and a troubled confusion, reflecting the ideas and uncertainties of his era. He did not share his results in his lifetime, despite both printing and etching being readily available to him. PMID:19852385
40 CFR 60.45Da - Alternative standards for combined nitrogen oxides (NOX) and carbon monoxide (CO).
Code of Federal Regulations, 2012 CFR
... nitrogen oxides (NOX) and carbon monoxide (CO). 60.45Da Section 60.45Da Protection of Environment... SOURCES Standards of Performance for Electric Utility Steam Generating Units § 60.45Da Alternative standards for combined nitrogen oxides (NOX) and carbon monoxide (CO). (a) The owner or operator of...
40 CFR 60.45Da - Alternative standards for combined nitrogen oxides (NOX) and carbon monoxide (CO).
Code of Federal Regulations, 2013 CFR
... nitrogen oxides (NOX) and carbon monoxide (CO). 60.45Da Section 60.45Da Protection of Environment... SOURCES Standards of Performance for Electric Utility Steam Generating Units § 60.45Da Alternative standards for combined nitrogen oxides (NOX) and carbon monoxide (CO). (a) The owner or operator of...
40 CFR 60.45Da - Alternative standards for combined nitrogen oxides (NOX) and carbon monoxide (CO).
Code of Federal Regulations, 2014 CFR
... nitrogen oxides (NOX) and carbon monoxide (CO). 60.45Da Section 60.45Da Protection of Environment... SOURCES Standards of Performance for Electric Utility Steam Generating Units § 60.45Da Alternative standards for combined nitrogen oxides (NOX) and carbon monoxide (CO). (a) The owner or operator of...
Antimicrobial Properties of an Immunomodulator - 15 kDa Human Granulysin
Wei, Hung-Mu; Lin, Li-Chih; Wang, Chiu-Feng; Lee, Yi-Jang; Chen, Yuan-Tsong; Liao, You-Di
Granulysin, a cationic protein expressed by human natural killer cells and cytotoxic T lymphocytes, is a mediator for drug-induced Stevens-Johnson syndrome and graft-versus-host disease. Some 15 kDa granulysin are processed into 9 kDa forms and sequestered in cytolytic granules, while others are constitutively secreted into body fluids. Both 9 and 15 kDa granulysin have been shown to be a serum marker for cell-mediated immunity. Furthermore, 15 kDa is able to activate monocyte differentiation. However, its antimicrobial properties have not been clearly addressed. Here, we report a novel method to prepare both the soluble 9 and 15 kDa granulysin and show that the 15 kDa form is more effective than the 9 kDa form in exerting specific antimicrobial activity against Pseudomonas aeruginosa within a range of few micromolars. We also show that the 15 kDa granulysin is able to hyperpolarize the membrane potential and increase membrane permeability of treated bacteria. Interestingly, the bactericidal activity and membrane permeability of the granulysins were markedly reduced at lower pH (pH 5.4) as a result of probable increase in hydrophobicity of the granulysins. Additionally, we've also shown the granulysin to inhibit biofilm formation by P. aeruginosa. These results suggest that the 15 kDa granulysin exhibits a novel mechanism in bacteria killing in a way that's different from most antimicrobial peptides. Our novel granulysin preparation methodology will be useful for further study of action mechanisms of other antimicrobial, cytotoxic and immunomodulating properties in granulysin-mediated diseases. PMID:27276051
Neospora caninum: identification of 19-, 38-, and 40-kDa surface antigens and a 33-kDa dense granule antigen using monoclonal antibodies.
Schares, G; Dubremetz, J F; Dubey, J P; Bärwald, A; Loyens, A; Conraths, F J
Neospora caninum, a coccidian parasite closely related to Toxoplasma gondii, can infect a broad host range and is regarded as an important cause of bovine abortion worldwide. In the present study, four antigens of N. caninum were partially characterized using monoclonal antibodies. Immunofluorescence of viable tachyzoites as well as the immunoprecipitation of antigens extracted from tachyzoites previously labeled by surface biotinylation revealed that three of these antigens with apparent molecular weights of 40, 38, and 19 kDa are located in the outer surface membrane of this parasite stage. Further evidence for the surface localization of the 38-kDa antigen was obtained by immunoelectron microscopy. In addition to the surface molecules, an antigen located in dense granules and in the tubular network of the parasitophorous vacuole was detected by another monoclonal antibody. When tachyzoite antigens separated under nonreducing conditions were probed on Western blots, this antibody reacted mainly with a 33-kDa antigen. Immunohistochemical analysis of infected tissue sections indicated that the 33-kDa dense granule antigen is present in both tachyzoites and bradyzoites, while the 38-kDa surface antigen from tachyzoites seems to be absent in bradyzoites. PMID:10366536
Respostas religiosas à aids no Brasil: impressões de pesquisa acerca da Pastoral de DST/Aids da Igreja Católica1
Seffner, Fernando; Silva, Cristiane Gonçalves Meireles da; Maksud, Ívia; Garcia, Jonathan; Rios, Luís Felipe; Natividade, Marcelo; Borges, Priscila Rodrigues; Parker, Richard; Terto, Veriano
O texto encontra-se estruturado em quatro partes. Na primeira delas, apresentamos um conjunto de considerações e informações acerca da situação da aids no Brasil, das relações entre religião, sexualidade, aids e estado laico, bem como uma descrição mais clara do Projeto Respostas Religiosas ao HIV/Aids no Brasil, do qual este texto apresenta algumas impressões de pesquisa preliminares. A seguir, dedicamos um item a apresentação da Pastoral de DST/Aids, sua história, estrutura e objetivos. No item seguinte problematizamos diversas questões em particular no âmbito das relações Estado e Igreja, relações entre agentes de pastoral e hierarquia da Igreja, e questões ligadas mais diretamente à sexualidade e aids, todas referenciadas ao trabalho da Pastoral de DST/Aids. Ao final, apresentamos a bibliografia e fontes consultadas. PMID:20442806
PD2P: PanDA Dynamic Data Placement for ATLAS
NASA Astrophysics Data System (ADS)
Maeno, T.; De, K.; Panitkin, S.
The PanDA (Production and Distributed Analysis) system plays a key role in the ATLAS distributed computing infrastructure. PanDA is the ATLAS workload management system for processing all Monte-Carlo (MC) simulation and data reprocessing jobs in addition to user and group analysis jobs. The PanDA Dynamic Data Placement (PD2P) system has been developed to cope with difficulties of data placement for ATLAS. We will describe the design of the new system, its performance during the past year of data taking, dramatic improvements it has brought about in the efficient use of storage and processing resources, and plans for the future.
Stability of new carbapenem DA-1131 to renal dipeptidase (dehydropeptidase I).
Park, Sung Wook; We, Jeoung Soon; Kim, Gye Won; Choi, Seong Hak; Park, Haeng Soon
The stability of DA-1131 to renal dipeptidase (RDPase) (EC 18.104.22.168) was compared with that of imipenem and meropenem by V(max)/K(m) ratios as an index of the enzyme's preference for substrates. Our results showed a decreasing order of imipenem (6.24), meropenem (2.41), and DA-1131 (1.39). The biochemical evaluation of DA-1131 as the least preferred substrate of RDPase suggests its potential use as a novel beta-lactam antibiotic which may be usable without coadministration of RDPase inhibitors once its clinical suitability is proven. PMID:11796382
Arbitrary Amplitude DIA and DA Solitary Waves in Adiabatic Dusty Plasmas
Mamun, A. A.; Jahan, N.; Shukla, P. K.
The dust-ion-acoustic (DIA) as well as the dust-acoustic (DA) solitary waves (SWs) in an adiabatic dusty plasma are investigated by the pseudo-potential approach which is valid for arbitrary amplitude SWs. The role of the adiabaticity of electrons and ions in modifying the basic features (polarity, speed, amplitude and width) of arbitrary amplitude DIA and DA SWs are explicitly examined. It is found that the effects of the adiabaticity of electrons and ions significantly modify the basic features (polarity, speed, amplitude and width) of the DIA and DA SWs. The implications of our results in space and laboratory dusty plasmas are briefly discussed.
2012 DA14 and Chelyabinsk: Same Day Coincidence?
NASA Astrophysics Data System (ADS)
Chapman, Clark R.
A long anticipated near-miss by near-Earth asteroid (NEA) 2012 DA14 on 15 February 2013 was upstaged 16 hours earlier by the impact and atmospheric explosion of a ~20 m NEA over Chelyabinsk, Russia. DA14 was earlier estimated to be 45 m across and passage at a distance under geosynchronous satellites was considered to be a record close approach by an object of that size, a once-in-40-years event. Actual Earth impact by a 20 m NEA had been estimated to be a once-in-200-years event. A simplistic calculation gives a probability of these happening on the same day of 1-in-a-billion. Within hours of the Chelyabinsk impact, it was recognized and widely reported that the two asteroids were in very different orbits and could not, at least in any readily understandable way, be physically related to each other (e.g. fragments of the same precursor body). Also, some early reports suggested that the two asteroids had very different compositions, further undermining the possibility they were pieces of the same body. (It seems unlikely, though certainly conceivable, that the two NEAs could have different compositions yet have some kind of causal connection resulting in the same-day events.) Nevertheless, incredibly tiny probabilities beg for an explanation. Here, with the benefit of hindsight, I re-examine issues that affect the probabilities, such as size, albedo, probable composition, and numbers of NEAs of these sizes. I also consider difficult issues of framing the apparent coincidence, which dominate other uncertainties. Updated information about the physical nature of the two asteroids somewhat modifies the original estimates of probabilities. Moreover, more proper ways of framing elements of the coincidence considerably reduce the improbability of the same-day events, but not nearly to the level (e.g. 1-in-1000 or perhaps 1-in-10,000) where we could rationalize dismissing the events as "just a coincidence." The events of 15 February 2013 deserve more sophisticated
Miltefosine Increases Lipid and Protein Dynamics in Leishmania amazonensis Membranes at Concentrations Similar to Those Needed for Cytotoxicity Activity
Moreira, Rodrigo Alves; Mendanha, Sebastião Antonio; Fernandes, Kelly Souza; Matos, Grazzielle Guimaraes; Alonso, Lais; Dorta, Miriam Leandro
Miltefosine (MT) is a membrane-active alkylphospholipid licensed for the topical treatment of breast cancer skin metastases and the oral treatment of leishmaniasis, although its mechanism of action remains unclear. Electron paramagnetic resonance (EPR) spectroscopy of a spin-labeled lipid and a thiol-specific spin label in the plasma membrane of Leishmania promastigotes showed that MT causes dramatic increases in membrane dynamics. Although these alterations can be detected using a spin-labeled lipid, our experimental results indicated that MT interacts predominantly with the protein component of the membrane. Cell lysis was also detected by analyzing the supernatants of centrifuged samples for the presence of spin-labeled membrane fragments and cytoplasmic proteins. Using a method for the rapid incorporation of MT into the membrane, these effects were measured immediately after treatment under the same range of MT concentrations that cause cell growth inhibition. Cytotoxicity, estimated via microscopic counting of living and dead cells, indicated ∼70% cell death at the concentration of MT at which EPR spectroscopy detected a significant change in membrane dynamics. After this initial impact on the number of viable parasites, the processes of cell death and growth continued during the first 4 h of incubation. The EPR spectra of spin-labeled membrane-bound proteins were consistent with more expanded and solvent-exposed protein conformations, suggesting a detergent-like action. Thus, MT may form micelle-like structures around polypeptide chains, and proteins with a higher hydrophobicity may induce the penetration of hydrophilic groups of MT into the membrane, causing its rupture. PMID:24614380
HIV-1 Tat protein enhances the intracellular growth of Leishmania amazonensis via the ds-RNA induced protein PKR
Vivarini, Áislan de Carvalho; Santos Pereira, Renata de Meirelles; Barreto-de-Souza, Victor; Temerozo, Jairo Ramos; Soares, Deivid C.; Saraiva, Elvira M.; Saliba, Alessandra Mattos; Bou-Habib, Dumith Chequer; Lopes, Ulisses Gazos
HIV-1 co-infection with human parasitic diseases is a growing public health problem worldwide. Leishmania parasites infect and replicate inside macrophages, thereby subverting host signaling pathways, including the response mediated by PKR. The HIV-1 Tat protein interacts with PKR and plays a pivotal role in HIV-1 replication. This study shows that Tat increases both the expression and activation of PKR in Leishmania-infected macrophages. Importantly, the positive effect of Tat addition on parasite growth was dependent on PKR signaling, as demonstrated in PKR-deficient macrophages or macrophages treated with the PKR inhibitor. The effect of HIV-1 Tat on parasite growth was prevented when the supernatant of HIV-1-infected macrophages was treated with neutralizing anti-HIV-1 Tat prior to Leishmania infection. The addition of HIV-1 Tat to Leishmania-infected macrophages led to inhibition of iNOS expression, modulation of NF-kB activation and enhancement of IL-10 expression. Accordingly, the expression of a Tat construct containing mutations in the basic region (49–57aa), which is responsible for the interaction with PKR, favored neither parasite growth nor IL-10 expression in infected macrophages. In summary, we show that Tat enhances Leishmania growth through PKR signaling. PMID:26608746
Near-UV absorption in very cool DA white dwarfs
Saumon, D.; Holberg, J. B.; Kowalski, P. M. E-mail: firstname.lastname@example.com
The atmospheres of very cool, hydrogen-rich white dwarfs (WDs) (T{sub eff} < 6000 K) are challenging to model because of the increased complexity of the equation of state, chemical equilibrium, and opacity sources in a low-temperature, weakly ionized dense gas. In particular, many models that assume relatively simple models for the broadening of atomic levels and mostly ideal gas physics overestimate the flux in the blue part of their spectra. A solution to this problem that has met with some success is that additional opacity at short wavelengths comes for the extreme broadening of the Lyman α line of atomic H by collisions primarily with H{sub 2}. For the purpose of validating this model more rigorously, we acquired Hubble Space Telescope STIS spectra of eight very cool WDs (five DA and three DC stars). Combined with their known parallaxes, BVRIJHK, and Spitzer IRAC photometry, we analyze their entire spectral energy distribution (from 0.24 to 9.3 μm) with a large grid of model atmospheres and synthetic spectra. We find that the red wing of the Lyman α line reproduces the rapidly decreasing near-UV flux of these very cool stars very well. We determine better constrained values of T{sub eff} and gravity as well as upper limits to the helium abundance in their atmospheres.
Granite geomorphology and its geological controls, Serra da Estrela, Portugal
NASA Astrophysics Data System (ADS)
Migoń, Piotr; Vieira, Gonçalo
Serra da Estrela is an elevated granite massif in central Portugal, characterized by extensive plateau surfaces incised by deep valleys affected by Quaternary glaciation, bounded by steep fault-generated escarpments. The presence of seven major textural variants of granite provides an opportunity to study the relationships between lithology and relief, whereas DEM analysis helped to show the relationships between lithology and topography objectively. The higher ground is associated with fine- to medium-grained granites and is typified by planar surfaces of low gradient, with occasional angular tors and rock pedestals. Block fields built by angular material are common in the parts that were not previously glaciated. Less elevated parts of the plateau are supported by medium- to coarse-grained granites and show more varied topography, with an abundance of tors, boulder piles, and depressions. Lithological boundaries locally coincide with slope breaks but this is not the rule. In the northern part of the massif a deep topographic basin has evolved in biotite granite, whereas deeply incised valleys follow major fault lines. Geological controls show a hierarchy, in that gross relief reflects the pattern of tectonic uplift and subsidence, whereas lithology and then fracture patterns become more and more important if one focuses on smaller and smaller landforms.
D.A. Henderson: acting globally, thinking locally.
Henderson, D A
They had said that it couldn't be done-the worldwide eradication of smallpox. To hear D.A. Henderson tell it, the job of leading the World Health Organization's initiative to conquer the disease in the 1960s and 1970s rather fell into his lap. In fact, he describes each of the posts that he has held with great modesty, beginning with his military service at the Centers for Disease Control and Prevention all the way through his assignments as Dean of Public Health at Johns Hopkins, Associate Director of the Office of Science and Technology Policy in the Executive Office of the President, and more recently as Director of the Office of Public Health Preparedness under Secretary Thompson at the Department of Health and Human Services. Confronted with enormous challenges in terms of public health initiatives, Henderson describes each assignment as a matter of communicating with the people he works with and the people that he serves, and drawing on their insights to devise strategies for accomplishing the task at hand. With bioterrorism posing one of the major public health concerns to face the United States and the world, it's gratifying to know that someone with Henderson's track record and wide-ranging expertise is paying attention and making sure that medical and government officials are preparing to respond to the threat. Again and again, Henderson appears to have the knack for showing up in the right place at the right time with just the right idea. PMID:14993437
Robotic Partial Nephrectomy with the Da Vinci Xi
Kallingal, George J. S.; Swain, Sanjaya; Darwiche, Fadi; Punnen, Sanoj; Manoharan, Murugesan; Gonzalgo, Mark L.; Parekh, Dipen J.
Purpose. The surgical expertise to perform robotic partial nephrectomy is heavily dependent on technology. The Da Vinci Xi (XI) is the latest robotic surgical platform with significant advancements compared to its predecessor. We describe our operative technique and experience with the XI system for robotic partial nephrectomy (RPN). Materials and Methods. Patients with clinical T1 renal masses were offered RPN with the XI. We used laser targeting, autopositioning, and a novel "in-line" port placement to perform RPN. Results. 15 patients underwent RPN with the XI. There were no intraoperative complications and no operative conversions. Mean console time was 101.3 minutes (range 44–176 minutes). Mean ischemia time was 17.5 minutes and estimated blood loss was 120 mLs. 12 of 15 patients had renal cell carcinoma. Two patients had oncocytoma and one had benign cystic disease. All patients had negative surgical margins and pathologic T1 disease. Two postoperative complications were encountered, including one patient who developed a pseudoaneurysm and one readmitted for presumed urinary tract infection. Conclusions. RPN with the XI system can be safely performed. Combining our surgical technique with the technological advancements on the XI offers patients acceptable pathologic and perioperative outcomes. PMID:26977144
Audio annotation watermarking with robustness against DA/AD conversion
NASA Astrophysics Data System (ADS)
Qian, Kun; Kraetzer, Christian; Biermann, Michael; Dittmann, Jana
In the paper we present a watermarking scheme developed to meet the specific requirements of audio annotation watermarking robust against DA/AD conversion (watermark detection after playback by loudspeaker and recording with a microphone). Additionally the described approach tries to achieve a comparably low detection complexity, so it could be embedded in the near future in low-end devices (e.g. mobile phones or other portable devices). We assume in the field of annotation watermarking that there is no specific motivation for attackers to the developed scheme. The basic idea for the watermark generation and embedding scheme is to combine traditional frequency domain spread spectrum watermarking with psychoacoustic modeling to guarantee transparency and alphabet substitution to improve the robustness. The synchronization and extraction scheme is designed to be much less computational complex than the embedder. The performance of the scheme is evaluated in the aspects of transparency, robustness, complexity and capacity. The tests reveals that 44% out of 375 tested audio files pass the simulation test for robustness, while the most appropriate category shows even 100% robustness. Additionally the introduced prototype shows an averge transparency of -1.69 in SDG, while at the same time having a capacity satisfactory to the chosen application scenario.
Zhang Yueyang; Deng Licai; Liu Chao; Carrell, Kenneth; Yang Fan; Gao Shuang; Xu Yan; Li Jing; Zhang Haotong; Zhao Yongheng; Luo Ali; Bai Zhongrui; Yuan Hailong; Lepine, Sebastien; Newberg, Heidi Jo; Carlin, Jeffrey L.; Jin Ge
A total of {approx}640, 000 objects from the LAMOST pilot survey have been publicly released. In this work, we present a catalog of DA white dwarfs (DAWDs) from the entire pilot survey. We outline a new algorithm for the selection of white dwarfs (WDs) by fitting Sersic profiles to the Balmer H{beta}, H{gamma}, and H{delta} lines of the spectra, and calculating the equivalent width of the Ca II K line. Two thousand nine hundred sixty-four candidates are selected by constraining the fitting parameters and the equivalent width of the Ca II K line. All the spectra of candidates are visually inspected. We identify 230 DAWDs (59 of which are already included in the Villanova and SDSS WD catalogs), 20 of which are DAWDs with non-degenerate companions. In addition, 128 candidates are classified as DAWDs/subdwarfs, which means the classifications are ambiguous. The result is consistent with the expected DAWD number estimated based on the LEGUE target selection algorithm.
Adenovirus type 2 encoded early 11 kDa protein
Murthy, S.V.K.N.; Kapoor, Q.S.
Several adenovirus type 2 (Ad2) encoded early proteins have been identified in viral infected human KB cells. These proteins are of great interest as they play key roles in cell transformation, viral DNA synthesis and gene expression. They have partially purified an AD2 encoded early polypeptide of an apparent molecular weight of 11 kilodaltons from the nuclei of viral infected cells labelled with /sup 35/S-methionine. After DNA removal from the nuclear extracts, the polypeptide was isolated using DEAE-Sephacel anion exchange and Biogel P-10 gel filtration columns. This simple two step procedure yielded several fold purification of the polypeptide. Antisera raised in mice against an Ad2 transformed rat cell line 8617 was found to immunoprecipitate the 11 kDa polypeptide from the nuclear extract of Ad2 infected KB cells. After relating this protein to an open reading frame of an Ad2 early gene block by matching the amino acid sequences to the nucleotide sequences of early genes, they plan to functionally characterize this protein by using monoclonal antibodies in in vivo and in vitro experiments.
Biogeochemical Indicators of Paleoenvironmental Changes in a Humid Region of Amazonia (Lagoa da Pata, Sao Gabriel da Cachoeira)
NASA Astrophysics Data System (ADS)
Cordeiro, R. C.; Turcq, B.; Mendes, L. G.; Azeredo, J. B.; Moura, M. S.; de Oliveira, A. R.; Monteiro, F.; Santelli, R. E.
Analysis of four sediment core collected in Lagoa da Pata lake show significant changes in the environmental history of Amazonia during the last 40,000 years. The Lagoa da Pata Lake (0°17'S, 66°40'W) is located on the top of the Morro dos Seis Lagos, an isolated hill in the extensive forested plane surface of high Rio Negro basin. The climate in the region presents mean annual precipitation around 3000 mm without dry season. Three distinct sections are clearly identified in the LPT III, LPT IV and LPT V cores. They consist of upper and lower organic-rich layers, separated by a clastic layer which represents a short period of rapid accumulation around 18 ka BP. Analysis of the organic matter composition by d13C and d15N, total Carbon and Nitrogen, sedimentary chlorophyll concentration and organic matter petrography show three different phases related to organic matter deposition. Between, at least 46,000 and 31,000 14C yr B.P., the lake presented a high water level attested by high accumulation rates of organic carbon. Values of d13C (around -32%) associated to high C/N ratio demonstrate a lignocellulosic material contribution in this phase; the second phase between the 31000 yr 14C B.P. and 18000 14C B.P. is characterised by a lower productivity attested by low sedimentary chlorophyll and total organic carbon values. The d13C values increase with medium values around -28% that may correspond to an input of organic matter rich in C4 plants (grasses). The sedimentation rate decreases significantly. High charcoal fluxes were observed in this period. At around 18000 14C yr BP, there appears to have been a sudden input of clastic material. This is represented by a sandy facies, which exhibits lower carbon contents. From 18000 14C yr B.P. to the present time occurred an increase in lacustrine productivity marked by an increase in carbon and chlorophyll derivate accumulation rates. The C/N values dropped indicting an algae organic matter contribution. It probably
Temperature and Gravity Dependence of Trace Element Abundances in Hot DA White Dwarfs (94-EUVE-094)
NASA Technical Reports Server (NTRS)
Finley, David S.
EUV spectroscopy has shown that DA white dwarfs hotter than about 45,000 K may contain trace heavy elements, while those hotter than about 50,000 K almost always have significant abundances of trace heavy elements. One of our continuing challenges is to identify and determine the abundances of these trace constituents, and then to relate the observed abundance patterns to the present conditions and previous evolutionary histories of the hot DA white dwarfs.
The 32 kDa Enamelin Undergoes Conformational Transitions upon Calcium Binding
Fan, Daming; Lakshminarayanan, Rajamani; Moradian-Oldak, Janet
The 32 kDa hydrophilic and acidic enamelin, the most stable cleavage fragment of the enamel specific glycoprotein, is believed to play vital roles in controlling crystal nucleation or growth during enamel biomineralization. Circular dichroism and Fourier transform infrared spectra demonstrate that the secondary structure of the 32 kDa enamelin has a high content of α-helix (81.5%). Quantitative analysis on the circular dichroism data revealed that the 32 kDa enamelin undergoes conformational changes with a structural preference to β-sheet as a function of calcium ions. We suggest that the increase of β-sheet conformation upon presence of Ca2+ may allow preferable interaction of the 32 kDa enamelin with apatite crystal surfaces during enamel biomineralization. The calcium association constant of the 32 kDa enamelin calculated from the fitting curve of ellipticity at 222 nm is Ka = 1.55 (±0.13) × 103 M−1, indicating a relatively low affinity. Our current biophysical studies on the 32 kDa enamelin structure provide novel insights towards understanding the enamelin-mineral interaction and subsequently the functions of enamelin during enamel formation. PMID:18508280
OpenDA: Open Source Generic Data Assimilation Environment and its Application in Geophysical Process Models
NASA Astrophysics Data System (ADS)
Weerts, A.; van Velzen, N.; Verlaan, M.; Sumihar, J.; Hummel, S.; El Serafy, G.; Dhondia, J.; Gerritsen, H.; Vermeer-Ooms, S.; Loots, E.; Markus, A.; Kockx, A.
Data Assimilation techniques are essential elements in state-of-the-art development of models and their use in operational forecasting and real-time control in the fields of groundwater, surface water and soil systems. In meteorological and atmospheric sciences, steady improvements in numerical weather forecasting and climate prediction over the last couple of decades have been enabled to a large degree by the development of community-based models and data assimilation systems. The hydrologic community should learn from the experiences of the meteorological and atmospheric communities by accelerating the transition of hydrologic DA research into operations and developing community-supported, open-source modeling and forecasting systems and data assimilation tools. In 2010, a community based open source initiative named OpenDA was started. The openDA initiative bears similarities with the well-known openMI initiative. OpenDA makes use of a set of interfaces that describe the interaction between models, observations and data assimilation algorithms. It focuses on flexible applications in portable systems for modeling geophysical processes. It provides a generic interfacing protocol that allows combination of the implemented data assimilation techniques with, in principle, any time-stepping model describing a process (atmospheric processes, 3D circulation, 2D water flow, rainfall-runoff, unsaturated flow, groundwater flow, etc.). Presently, openDA features filtering techniques and calibration techniques. The presentation will give an overview of openDA and the results of some of its practical applications.
In vitro degradation of the 32kDa PS II reaction centre protein
Eckenswiller, L.C.; Greenberg, B.M. )
The 32kDa thylakoid membrane protein is an integral component of the PS II reaction centre. The protein, although stable in the dark, undergoes light dependent turnover. Light from the UV, visible and far-red spectral regions induce 32kDa protein degradation. To better understand 32kDa protein metabolism, an in vitro degradation system is being developed. It consists of isolated thylakoid membranes than contain radiolabelled protein. The 32kDa protein is actively and specifically degraded when the thylakoid preparation is exposed to UV or visible radiation. The protein is stable in the dark. The herbicides (atrazine and DCMU) inhibit degradation in the in vitro system as they do in vivo. Additionally, several methods of isolating thylakoids are being compared to optimize the 32kDa protein degradation reaction. The preparations will be evaluated based on their ability to permit light dependent degradation of the 32kDa protein without affecting the other membrane components.
[The behavior of superoxide dismutase (SOD) in serum of cows with abomasal displacement (DA)].
Fürll, M; Dabbagh, M N; Fürll, B; Sattler, T
After surgical reposition of displaced organs (abomasum, uterus, intestines) restoration of blood flow and oxygen supply generates oxygen radicals and other reactive oxygen species. SOD indicates radical stress of the organism. Subject of the study was the question if SOD can be detected in blood serum samples of cows and if there are differences in SOD activity between healthy cows and cows with Dislocatio abomasi (DA). We also wanted to investigate the influence of breed "Schwarzbunte" with DA (16 left/5 rights). The samples were drawn before and 1, 3 and 24 post op. Ten healthy cows of the same breed were also examined (2 weeks and 4-6 weeks after calving). There are no significant differences between the SOD activity of healthy cows and cows with DA, but the SOD activity of cows with left DA is significant lower than the activity of cows with right DA. Post op. SOD activity decreases; 24 h after surgery cows with left but not with right DA show an increase of SOD activity similar to values before surgery. There is a close positive correlation between SOD activity and protein concentration as well as negative correlation to concentration of free fatty acids after surgery. The behaviour of SOD activity shows that the surgical replacement of the displaced abomasum can generate a depression of the antioxidative capacity of the organism. PMID:14983749
The sandfly fauna (Diptera: Psychodidae: Phlebotominae) of the Parque Estadual da Serra da Tiririca, Rio de Janeiro, Brazil
Rodrigues, Andressa Alencastre Fuzari; Barbosa, Vanessa de Araújo; Andrade, José Dilermando; Brazil, Reginaldo Peçanha
Cutaneous leishmaniasis (CL) in the state of Rio de Janeiro is sporadic and can be characterised as a peridomestic transmission that occurs in modified natural environments. The aim of this work was to study the fauna and ecological characteristics of sandflies in an environmentally protected area (the State Park of Serra da Tiririca) within the remnants of the Atlantic Forest in the municipalities of Niterói and Maricá and their possible relationship with leishmaniasis. Captures were performed using light traps during the night once a month for one year in both sylvatic environments and areas surrounding homes near the park. A total of 1,037 sandflies were captured, belonging to nine genera and 12 species: Evandromyia tupynambai (34.1%), Migonemyia migonei (20.6%), Brumptomyia cunhai (13.8%), Micropygomyia schreiberi (9.7%), Psathyromyia lanei (6.5%), Brumptomyia nitzulescui (5.7%), Evandromyia edwardsi (5.4%), Nyssomyia intermedia (2.8%), Evandromyia cortelezzii (0.6%), Pintomyia bianchigalatiae (0.5%), Lutzomyia longipalpis (0.2%) and Sciopemyia microps (0.1%). Both Mg. migonei and Ny. intermedia may be acting as vectors of CL in this area. PMID:24141956
Negative regulation of human immunodeficiency virus type 1 expression in monocytes: role of the 65-kDa plus 50-kDa NF-kappa B dimer.
Raziuddin; Mikovits, J A; Calvert, I; Ghosh, S; Kung, H F; Ruscetti, F W
Although monocytic cells can provide a reservoir for viral production in vivo, their regulation of human immunodeficiency virus type 1 (HIV-1) transcription can be either latent, restricted, or productive. These differences in gene expression have not been molecularly defined. In THP-1 cells with restricted HIV expression, there is an absence of DNA-protein binding complex formation with the HIV-1 promoter-enhancer associated with markedly less viral RNA production. This absence of binding was localized to the NF-kappa B region of the HIV-1 enhancer; the 65-kDa plus 50-kDa NF-kappa B heterodimer was preferentially lost. Adding purified NF-kappa B protein to nuclear extracts from cells with restricted expression overcomes this lack of binding. In addition, treatment of these nuclear extracts with sodium deoxycholate restored their ability to form the heterodimer, suggesting the presence of an inhibitor of NF-kappa B activity. Furthermore, treatment of nuclear extracts from these cells that had restricted expression with lipopolysaccharide increased viral production and NF-kappa B activity. Antiserum specific for NF-kappa B binding proteins, but not c-rel-specific antiserum, disrupted heterodimer complex formation. Thus, both NF-kappa B-binding complexes are needed for optimal viral transcription. Binding of the 65-kDa plus 50-kDa heterodimer to the HIV-1 enhancer can be negatively regulated in monocytes, providing one mechanism restricting HIV-1 gene expression. PMID:1946356
Myelin management by the 18.5–kDa and 21.5–kDa classic myelin basic protein isoforms
Harauz, George; Boggs, Joan M.
The classic myelin basic protein (MBP) splice isoforms range in nominal molecular mass from 14 to 21.5 kDa, and arise from the gene in the oligodendrocyte lineage (Golli) in maturing oligodendrocytes. The 18.5-kDa isoform that predominates in adult myelin adheres the cytosolic surfaces of oligodendrocyte membranes together, and forms a two-dimensional molecular sieve restricting protein diffusion into compact myelin. However, this protein has additional roles including cytoskeletal assembly and membrane extension, binding to SH3-domains, participation in Fyn-mediated signaling pathways, sequestration of phosphoinositides, and maintenance of calcium homeostasis. Of the diverse post-translational modifications of this isoform, phosphorylation is the most dynamic, and modulates 18.5-kDa MBP's protein-membrane and protein-protein interactions, indicative of a rich repertoire of functions. In developing and mature myelin, phosphorylation can result in microdomain or even nuclear targeting of the protein, supporting the conclusion that 18.5-kDa MBP has significant roles beyond membrane adhesion. The full-length, early-developmental 21.5-kDa splice isoform is predominantly karyophilic due to a non-traditional P-Y nuclear localization signal, with effects such as promotion of oligodendrocyte proliferation. We discuss in vitro and recent in vivo evidence for multifunctionality of these classic basic proteins of myelin, and argue for a systematic evaluation of the temporal and spatial distributions of these protein isoforms, and their modified variants, during oligodendrocyte differentiation. PMID:23398367
Characteristics of 26 kDa antigen of H. Pylori by Monoclonal Antibody.
Ghahremani, Hossein; Farshad, Shohreh; Amini Najafabadi, Hossein; Kashanian, Susan; Momeni Moghaddam, Mohammad Amin; Moradi, Nariman; Paknejad, Maliheh
Alkylhydroperoxide reductase (AhpC, the 26 kDa antigen) is one of the abundant antioxidant enzymes in Helicobacter pylori and seems to have a good potential for use in development of immunoassays to detect H. pylori infection in clinical specimens. This study aimed to investigate some properties of this antigen by the produced monoclonal antibodies. Five established hybridoma cell lines secreting monoclonal antibodies (MAbs) against 26 kDa antigen of H. pylori were cultivated and MAbs were purified by affinity chromatography. Subsequently, MAbs were conjugated with biotin, and different combinations of capture and tracer antibodies used in sandwich ELISA. Immunoblotting of bacterial extracts were performed to estimate aggregation status of the antigen. Release of antigen from the cultivated bacteria on solid media was examined by sandwich ELISA, and also, existence of interference in fecal extract was investigated by immunoblotting and sandwich ELISA. Our findings showed that the MAbs against 26 kDa antigen of H. pylori could recognize three bands of nearly 25 kDa, 50 kDa, and 75 kDa in immunoblotting. This study also indicated presence of more antigens in the culture medium around the bacteria than the bacterial extract itself. The results of sandwich ELISA and immunoblotting on fecal extracts suggest the presence of interfering agents that prevent detection of antigen by antibody in ELISA but not in immunoblotting. In this study the oligomerization of the 26 kDa antigen, presence of interfering agents in stool matrix, and release of antigen to outside of bacteria, were demonstrated. PMID:25530147
Naloxone-induced jumping activity in morphine-dependent mice: a pharmacological study on the role of DA2 receptors.
Cerbo, R; Boni, B; De Lena, C; Cesarino, F; Meco, G; Feliciani, M; Formisano, R; Casacchia, M
The morphine withdrawal syndrome is strictly related to modifications on neurotransmittorial systems at central and peripheral levels. Dopamine seems to play an important role. Particularly in the experimental withdrawal syndrome the jumping activity seems to be related to a hypersensitivity of DA receptors. While the role of DA1 receptors has been extensively studied, using agonist and antagonist drugs, the role of DA2 receptors is still little known. In our study we evaluated the changes of the jumping response that was induced by substances active on DA2 receptors (lisuride, tiapride) in mice made acutely dependent on morphine, in which we induced a withdrawal syndrome with naloxone. Haloperidol (DA1-DA2 blocker) did not provoke significant changes. Either tiapride or lisuride determined a sharp increase of jumping. PMID:6712813
11 CFR 111.11 - Written questions under order (2 U.S.C. 437d(a)(1)).
Code of Federal Regulations, 2011 CFR
... 11 Federal Elections 1 2011-01-01 2011-01-01 false Written questions under order (2 U.S.C. 437d(a... PROCEDURE (2 U.S.C. 437g, 437d(a)) Enforcement § 111.11 Written questions under order (2 U.S.C. 437d(a)(1... submit sworn written answers to written questions and may specify a date by which such answers must...
11 CFR 111.11 - Written questions under order (2 U.S.C. 437d(a)(1)).
Code of Federal Regulations, 2010 CFR
... 11 Federal Elections 1 2010-01-01 2010-01-01 false Written questions under order (2 U.S.C. 437d(a... PROCEDURE (2 U.S.C. 437g, 437d(a)) Enforcement § 111.11 Written questions under order (2 U.S.C. 437d(a)(1... submit sworn written answers to written questions and may specify a date by which such answers must...
82-kDa choline acetyltransferase and SATB1 localize to β-amyloid induced matrix attachment regions.
Winick-Ng, Warren; Caetano, Fabiana A; Winick-Ng, Jennifer; Morey, Trevor M; Heit, Bryan; Rylett, R Jane
The M-transcript of human choline acetyltransferase (ChAT) produces an 82-kDa protein (82-kDa ChAT) that concentrates in nuclei of cholinergic neurons. We assessed the effects of acute exposure to oligomeric amyloid-β1-42 (Aβ1-42) on 82-kDa ChAT disposition in SH-SY5Y neural cells, finding that acute exposure to Aβ1-42 results in increased association of 82-kDa ChAT with chromatin and formation of 82-kDa ChAT aggregates in nuclei. When measured by chromatin immunoprecipitation with next-generation sequencing (ChIP-seq), we identified that Aβ1-42-exposure increases 82-kDa ChAT association with gene promoters and introns. The Aβ1-42-induced 82-kDa ChAT aggregates co-localize with special AT-rich binding protein 1 (SATB1), which anchors DNA to scaffolding/matrix attachment regions (S/MARs). SATB1 had a similar genomic association as 82-kDa ChAT, with both proteins associating with synapse and cell stress genes. After Aβ1-42 -exposure, both SATB1 and 82-kDa ChAT are enriched at the same S/MAR on the APP gene, with 82-kDa ChAT expression attenuating an increase in an isoform-specific APP mRNA transcript. Finally, 82-kDa ChAT and SATB1 have patterned genomic association at regions enriched with S/MAR binding motifs. These results demonstrate that 82-kDa ChAT and SATB1 play critical roles in the response of neural cells to acute Aβ-exposure. PMID:27052102
An Introduction to the DA-T Gibbs Sampler for the Two-Parameter Logistic (2PL) Model and beyond
ERIC Educational Resources Information Center
Maris, Gunter; Bechger, Timo M.
The DA-T Gibbs sampler is proposed by Maris and Maris (2002) as a Bayesian estimation method for a wide variety of "Item Response Theory (IRT) models". The present paper provides an expository account of the DA-T Gibbs sampler for the 2PL model. However, the scope is not limited to the 2PL model. It is demonstrated how the DA-T Gibbs sampler for…
82-kDa choline acetyltransferase and SATB1 localize to β-amyloid induced matrix attachment regions
Winick-Ng, Warren; Caetano, Fabiana A.; Winick-Ng, Jennifer; Morey, Trevor M.; Heit, Bryan; Rylett, R. Jane
The M-transcript of human choline acetyltransferase (ChAT) produces an 82-kDa protein (82-kDa ChAT) that concentrates in nuclei of cholinergic neurons. We assessed the effects of acute exposure to oligomeric amyloid-β1–42 (Aβ1–42) on 82-kDa ChAT disposition in SH-SY5Y neural cells, finding that acute exposure to Aβ1–42 results in increased association of 82-kDa ChAT with chromatin and formation of 82-kDa ChAT aggregates in nuclei. When measured by chromatin immunoprecipitation with next-generation sequencing (ChIP-seq), we identified that Aβ1–42 -exposure increases 82-kDa ChAT association with gene promoters and introns. The Aβ1–42 -induced 82-kDa ChAT aggregates co-localize with special AT-rich binding protein 1 (SATB1), which anchors DNA to scaffolding/matrix attachment regions (S/MARs). SATB1 had a similar genomic association as 82-kDa ChAT, with both proteins associating with synapse and cell stress genes. After Aβ1–42 -exposure, both SATB1 and 82-kDa ChAT are enriched at the same S/MAR on the APP gene, with 82-kDa ChAT expression attenuating an increase in an isoform-specific APP mRNA transcript. Finally, 82-kDa ChAT and SATB1 have patterned genomic association at regions enriched with S/MAR binding motifs. These results demonstrate that 82-kDa ChAT and SATB1 play critical roles in the response of neural cells to acute Aβ -exposure. PMID:27052102
High resolution structure of cleaved Serpin 42 Da from Drosophila melanogaster
Background The Drosophila melanogaster Serpin 42 Da gene (previously Serpin 4) encodes a serine protease inhibitor that is capable of remarkable functional diversity through the alternative splicing of four different reactive centre loop exons. Eight protein isoforms of Serpin 42 Da have been identified to date, targeting the protease inhibitor to both different proteases and cellular locations. Biochemical and genetic studies suggest that Serpin 42 Da inhibits target proteases through the classical serpin 'suicide' inhibition mechanism, however the crystal structure of a representative Serpin 42 Da isoform remains to be determined. Results We report two high-resolution crystal structures of Serpin 42 Da representing the A/B isoforms in the cleaved conformation, belonging to two different space-groups and diffracting to 1.7 Å and 1.8 Å. Structural analysis reveals the archetypal serpin fold, with the major elements of secondary structure displaying significant homology to the vertebrate serpin, neuroserpin. Key residues known to have central roles in the serpin inhibitory mechanism are conserved in both the hinge and shutter regions of Serpin 42 Da. Furthermore, these structures identify important conserved interactions that appear to be of crucial importance in allowing the Serpin 42 Da fold to act as a versatile template for multiple reactive centre loops that have different sequences and protease specificities. Conclusions In combination with previous biochemical and genetic studies, these structures confirm for the first time that the Serpin 42 Da isoforms are typical inhibitory serpin family members with the conserved serpin fold and inhibitory mechanism. Additionally, these data reveal the remarkable structural plasticity of serpins, whereby the basic fold is harnessed as a template for inhibition of a large spectrum of proteases by reactive centre loop exon 'switching'. This is the first structure of a Drosophila serpin reported to date
Characterization of the 105-kDa molecular chaperone. Identification, biochemical properties, and localization.
Matsumori, Mika; Itoh, Hideaki; Toyoshima, Itaru; Komatsuda, Atsushi; Sawada, Ken-Ichi; Fukuda, Jun; Tanaka, Toshinobu; Okubo, Atsuya; Kinouchi, Hiroyuki; Mizoi, Kazuo; Hama, Tokiko; Suzuki, Akira; Hamada, Fumio; Otaka, Michiro; Shoji, Yutaka; Takada, Goro
We have characterized the biochemical properties of the testis and brain-specific 105-kDa protein which is cross-reacted with an anti-bovine HSP90 antibody. The protein was induced in germ cells by heat stress, resulting in a protein which is one of the heat shock proteins [Kumagai, J., Fukuda, J., Kodama, H., Murata, M., Kawamura, K., Itoh, H. & Tanaka, T. (2000) Eur. J. Biochem.267, 3073-3078]. In the present study, we characterized the biochemical properties of the protein. The 105-kDa protein inhibited the aggregation of citrate synthase as a molecular chaperone in vitro. ATP/MgCl2 has a slight influence of the suppression of the citrate synthase aggregation by the 105-kDa protein. The protein possessed chaperone activity. The protein was able to bind to ATP-Sepharose like the other molecular chaperone HSP70. A partial amino-acid sequence (24 amino-acid residues) of the protein was determined and coincided with those of the mouse testis- and brain-specific APG-1 and osmotic stress protein 94 (OSP94). The 105-kDa protein was detected only in the medulla of the rat kidney sections similar to OSP94 upon immunoblotting. The purified 105-kDa protein was cross-reacted with an antibody against APG-1. These results suggested that APG-1 and OSP94 are both identical to the 105-kDa protein. There were highly homologous regions between the 105-kDa protein/APG-1/OSP94 and HSP90. The region of HSP90 was also an immunoreactive site. An anti-bovine HSP90 antibody may cross-react with the 105-kDa protein similar to HSP90 in the rat testis and brain. We have investigated the localization and developmental induction of the protein in the rat brain. In the immunohistochemical analysis, the protein was mainly detected in the cytoplasm of the nerve and glial cells of the rat brain. Although the 105-kDa protein was localized in all rat brain segments, the expression pattern was fast in the cerebral cortex and hippocampus and slow in the cerebellum. PMID:12423363
The rare DAT coding variant Val559 perturbs DA neuron function, changes behavior, and alters in vivo responses to psychostimulants.
Mergy, Marc A; Gowrishankar, Raajaram; Gresch, Paul J; Gantz, Stephanie C; Williams, John; Davis, Gwynne L; Wheeler, C Austin; Stanwood, Gregg D; Hahn, Maureen K; Blakely, Randy D
Despite the critical role of the presynaptic dopamine (DA) transporter (DAT, SLC6A3) in DA clearance and psychostimulant responses, evidence that DAT dysfunction supports risk for mental illness is indirect. Recently, we identified a rare, nonsynonymous Slc6a3 variant that produces the DAT substitution Ala559Val in two male siblings who share a diagnosis of attention-deficit hyperactivity disorder (ADHD), with other studies identifying the variant in subjects with bipolar disorder (BPD) and autism spectrum disorder (ASD). Previously, using transfected cell studies, we observed that although DAT Val559 displays normal total and surface DAT protein levels, and normal DA recognition and uptake, the variant transporter exhibits anomalous DA efflux (ADE) and lacks capacity for amphetamine (AMPH)-stimulated DA release. To pursue the significance of these findings in vivo, we engineered DAT Val559 knock-in mice, and here we demonstrate in this model the presence of elevated extracellular DA levels, altered somatodendritic and presynaptic D2 DA receptor (D2R) function, a blunted ability of DA terminals to support depolarization and AMPH-evoked DA release, and disruptions in basal and psychostimulant-evoked locomotor behavior. Together, our studies demonstrate an in vivo functional impact of the DAT Val559 variant, providing support for the ability of DAT dysfunction to impact risk for mental illness. PMID:25331903
The rare DAT coding variant Val559 perturbs DA neuron function, changes behavior, and alters in vivo responses to psychostimulants
Mergy, Marc A.; Gowrishankar, Raajaram; Gresch, Paul J.; Gantz, Stephanie C.; Williams, John; Davis, Gwynne L.; Wheeler, C. Austin; Stanwood, Gregg D.; Hahn, Maureen K.; Blakely, Randy D.
Despite the critical role of the presynaptic dopamine (DA) transporter (DAT, SLC6A3) in DA clearance and psychostimulant responses, evidence that DAT dysfunction supports risk for mental illness is indirect. Recently, we identified a rare, nonsynonymous Slc6a3 variant that produces the DAT substitution Ala559Val in two male siblings who share a diagnosis of attention-deficit hyperactivity disorder (ADHD), with other studies identifying the variant in subjects with bipolar disorder (BPD) and autism spectrum disorder (ASD). Previously, using transfected cell studies, we observed that although DAT Val559 displays normal total and surface DAT protein levels, and normal DA recognition and uptake, the variant transporter exhibits anomalous DA efflux (ADE) and lacks capacity for amphetamine (AMPH)-stimulated DA release. To pursue the significance of these findings in vivo, we engineered DAT Val559 knock-in mice, and here we demonstrate in this model the presence of elevated extracellular DA levels, altered somatodendritic and presynaptic D2 DA receptor (D2R) function, a blunted ability of DA terminals to support depolarization and AMPH-evoked DA release, and disruptions in basal and psychostimulant-evoked locomotor behavior. Together, our studies demonstrate an in vivo functional impact of the DAT Val559 variant, providing support for the ability of DAT dysfunction to impact risk for mental illness. PMID:25331903
A Influência da Escola e da Mídia na Obtenção dos Conhecimentos Astronômicos
NASA Astrophysics Data System (ADS)
da Cunha, W. S.; Voelzke, M. R.; Amaral, L. H.
O universo em que vivemos possui segredos jamais imaginados pelo homem, fruto da pequena proporção que ocupamos diante de um cosmos infinito. A astronomia funciona como um instrumento que sacia a sede humana em desvendar esses segredos, estudando as galáxias, quasares, pulsares além da existência de todos os elementos que preenchem o espaço cósmico, o vácuo, não mais vazio como se pensou em outros tempos. A mídia explora esse campo de forma mística e também científica. A difusão adequada e dentro dos padrões educacionais vigentes, de conceitos astronômicos, se faz necessária, devido sua presença nos conteúdos escolares, centros de ciência e na mídia, de modo geral. A reforma do Currículo Básico da Escola Pública da maioria dos Estados brasileiros introduziu conceitos astronômicos desde a pré-escola até o Ensino Médio. Num estudo realizado com 1.032 alunos do Ensino Médio de seis escolas estaduais, revelou-se através da aplicação de um questionário que abordava conheciment os básicos em astronomia, como estes foram adquiridos e qual era a infra-estrutura oferecida pela escola: para 66,2% dos entrevistados o professor já fez alguma apresentação a respeito de astronomia. O presente estudo revelou também que 44,5% dos alunos entrevistados adquiriram seus conhecimentos astronômicos na escola, 28,1% através da televisão e 27,4% afirmaram que seus conhecimentos foram obtidos através de outros tipos de mídia (internet, filmes e revistas). Embora o estudo revele claramente que há certo equilíbrio na forma em que os alunos obtiveram seus conhecimentos astronômicos, a mídia ainda explora e muitas vezes desperta o interesse pela astronomia com assuntos que não fazem parte de nosso cotidiano como, por exemplo, o homem na Lua, os projetos espaciais e a primeira viagem de um astronauta brasileiro no espaço. Porém, em certos casos, a mídia pode promover conhecimentos incompletos ou mesmo equivocados sobre os conceitos que envolvam
In vitro study on the interaction between the 32 kDa enamelin and amelogenin
Fan, Daming; Du, Chang; Sun, Zhi; Lakshminarayanan, Rajamani; Moradian-Oldak, Janet
Enamel extracelluar matrix components play vital roles in controlling crystal nucleation and growth during enamel formation. We investigated the interaction between the 32 kDa enamelin fragment and amelogenin using immunochemical and biophysical methods. Immunoprecipitation studies revealed that the 32 kDa enamelin and amelogenin eluted together from a Protein A column. Dynamic light scattering results showed that the 32 kDa enamelin had a profound effect on amelogenin assembly at pH 8.0, causing partial dissociation of the nanospheres, in a dose-dependent manner. The appearance of an isodichroic point and the shifting and intensity decrease of the ellipticity minima in the circular dichroism spectra of amelogenin following the addition of the 32 kDa enamelin were indicative of conformational changes in amelogenin and of a direct interaction between the two macromolecules. Our results collectively demonstrate that the 32 kDa enamelin has a direct interaction with amelogenin in vitro. Our current studies provide novel insights into understanding possible cooperation between enamelin and amelogenin in macromolecular self-assembly and in controlling enamel mineral formation. PMID:19263522
Morphogenetic Studies of the Drosophila DA1 Ventral Olfactory Projection Neuron.
Shen, Hung-Chang; Wei, Jia-Yi; Chu, Sao-Yu; Chung, Pei-Chi; Hsu, Tsai-Chi; Yu, Hung-Hsiang
In the Drosophila olfactory system, odorant information is sensed by olfactory sensory neurons and relayed from the primary olfactory center, the antennal lobe (AL), to higher olfactory centers via olfactory projection neurons (PNs). A major portion of the AL is constituted with dendrites of four groups of PNs, anterodorsal PNs (adPNs), lateral PNs (lPNs), lateroventral PNs (lvPNs) and ventral PNs (vPNs). Previous studies have been focused on the development and function of adPNs and lPNs, while the investigation on those of lvPNs and vPNs received less attention. Here, we study the molecular and cellular mechanisms underlying the morphogenesis of a putative male-pheromone responding vPN, the DA1 vPN. Using an intersection strategy to remove background neurons labeled within a DA1 vPN-containing GAL4 line, we depicted morphological changes of the DA1 vPN that occurs at the pupal stage. We then conducted a pilot screen using RNA interference knock-down approach to identify cell surface molecules, including Down syndrome cell adhesion molecule 1 and Semaphorin-1a, that might play essential roles for the DA1 vPN morphogenesis. Taken together, by revealing molecular and cellular basis of the DA1 vPN morphogenesis, we should provide insights into future comprehension of how vPNs are assembled into the olfactory neural circuitry. PMID:27163287
Morphogenetic Studies of the Drosophila DA1 Ventral Olfactory Projection Neuron
Yu, Hung-Hsiang
In the Drosophila olfactory system, odorant information is sensed by olfactory sensory neurons and relayed from the primary olfactory center, the antennal lobe (AL), to higher olfactory centers via olfactory projection neurons (PNs). A major portion of the AL is constituted with dendrites of four groups of PNs, anterodorsal PNs (adPNs), lateral PNs (lPNs), lateroventral PNs (lvPNs) and ventral PNs (vPNs). Previous studies have been focused on the development and function of adPNs and lPNs, while the investigation on those of lvPNs and vPNs received less attention. Here, we study the molecular and cellular mechanisms underlying the morphogenesis of a putative male-pheromone responding vPN, the DA1 vPN. Using an intersection strategy to remove background neurons labeled within a DA1 vPN-containing GAL4 line, we depicted morphological changes of the DA1 vPN that occurs at the pupal stage. We then conducted a pilot screen using RNA interference knock-down approach to identify cell surface molecules, including Down syndrome cell adhesion molecule 1 and Semaphorin-1a, that might play essential roles for the DA1 vPN morphogenesis. Taken together, by revealing molecular and cellular basis of the DA1 vPN morphogenesis, we should provide insights into future comprehension of how vPNs are assembled into the olfactory neural circuitry. PMID:27163287
Low strain, long life creep fatigue of AF2-1DA and INCO 718
Thakker, A.B.; Cowles, B.A.
Two aircraft turbine disk alloys, GATORIZED AF2-DA and INCO 718 were evaluated for their low strain long life creep-fatigue behavior. Static (tensile and creep rupture) and cyclic properties of both alloys were characterized. The cntrolled strain LCF tests were conducted at 760 C (1400 F) and 649 C (1200 F) for AF2-1DA and INCO 718, respectively. Hold times were varied for tensile, compressive and tensile/compressive strain dwell (relaxation) tests. Stress (creep) hold behavior of AF2-1DA was also evaluated. Generally, INCO 718 exhibited more pronounced reduction in cyclic life due to hold than AF2-1DA. The percent reduction in life for both alloys for strain dwell tests was greater at low strain ranges (longer life regime). Changing hold time from 0 to 0.5, 2.0 and 15.0 min. resulted in corresponding reductions in life. The continuous cycle and cyclic/dwell initiation failure mechanism was predominantly transgranular for AF2-1DA and intergranular for INCO 718.
PanDA: Exascale Federation of Resources for the ATLAS Experiment at the LHC
NASA Astrophysics Data System (ADS)
Barreiro Megino, Fernando; Caballero Bejar, Jose; De, Kaushik; Hover, John; Klimentov, Alexei; Maeno, Tadashi; Nilsson, Paul; Oleynik, Danila; Padolski, Siarhei; Panitkin, Sergey; Petrosyan, Artem; Wenaus, Torre
After a scheduled maintenance and upgrade period, the world's largest and most powerful machine - the Large Hadron Collider(LHC) - is about to enter its second run at unprecedented energies. In order to exploit the scientific potential of the machine, the experiments at the LHC face computational challenges with enormous data volumes that need to be analysed by thousand of physics users and compared to simulated data. Given diverse funding constraints, the computational resources for the LHC have been deployed in a worldwide mesh of data centres, connected to each other through Grid technologies. The PanDA (Production and Distributed Analysis) system was developed in 2005 for the ATLAS experiment on top of this heterogeneous infrastructure to seamlessly integrate the computational resources and give the users the feeling of a unique system. Since its origins, PanDA has evolved together with upcoming computing paradigms in and outside HEP, such as changes in the networking model, Cloud Computing and HPC. It is currently running steadily up to 200 thousand simultaneous cores (limited by the available resources for ATLAS), up to two million aggregated jobs per day and processes over an exabyte of data per year. The success of PanDA in ATLAS is triggering the widespread adoption and testing by other experiments. In this contribution we will give an overview of the PanDA components and focus on the new features and upcoming challenges that are relevant to the next decade of distributed computing workload management using PanDA.
Influence of 120 kDa Pyruvate:Ferredoxin Oxidoreductase on Pathogenicity of Trichomonas vaginalis
Song, Hyun-Ouk
Trichomonas vaginalis is a flagellate protozoan parasite and commonly infected the lower genital tract in women and men. Iron is a known nutrient for growth of various pathogens, and also reported to be involved in establishment of trichomoniasis. However, the exact mechanism was not clarified. In this study, the author investigated whether the 120 kDa protein of T. vaginalis may be involved in pathogenicity of trichomonads. Antibodies against 120 kDa protein of T. vaginalis, which was identified as pyruvate:ferredoxin oxidoreductase (PFOR) by peptide analysis of MALDI-TOF-MS, were prepared in rabbits. Pretreatment of T. vaginalis with anti-120 kDa Ab decreased the proliferation and adherence to vaginal epithelial cells (MS74) of T. vaginalis. Subcutaneous tissue abscess in anti-120 kDa Ab-treated T. vaginalis-injected mice was smaller in size than that of untreated T. vaginalis-infected mice. Collectively, the 120 kDa protein expressed by iron may be involved in proliferation, adhesion to host cells, and abscess formation, thereby may influence on the pathogenicity of T. vaginalis. PMID:26951982
Experience with ATLAS MySQL PanDA database service
NASA Astrophysics Data System (ADS)
Smirnov, Y.; Wlodek, T.; De, K.; Hover, J.; Ozturk, N.; Smith, J.; Wenaus, T.; Yu, D.
The PanDA distributed production and analysis system has been in production use for ATLAS data processing and analysis since late 2005 in the US, and globally throughout ATLAS since early 2008. Its core architecture is based on a set of stateless web services served by Apache and backed by a suite of MySQL databases that are the repository for all PanDA information: active and archival job queues, dataset and file catalogs, site configuration information, monitoring information, system control parameters, and so on. This database system is one of the most critical components of PanDA, and has successfully delivered the functional and scaling performance required by PanDA, currently operating at a scale of half a million jobs per week, with much growth still to come. In this paper we describe the design and implementation of the PanDA database system, its architecture of MySQL servers deployed at BNL and CERN, backup strategy and monitoring tools. The system has been developed, thoroughly tested, and brought to production to provide highly reliable, scalable, flexible and available database services for ATLAS Monte Carlo production, reconstruction and physics analysis.
Synergistic effects of melatonin and deprenyl against MPTP-induced mitochondrial damage and DA depletion.
Khaldy, Hoda; Escames, Germaine; León, Josefa; Bikjdaouene, Leila; Acuña-Castroviejo, Darío
Previous studies showed a synergistic effect of melatonin and deprenyl against dopamine (DA) autoxidation in vitro. Since oxidative stress is implicated in Parkinson's disease (PD), we explored the effects of melatonin plus deprenyl administration in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD in C57/Bl6 mice. Melatonin, but not deprenyl prevents the inhibition of mitochondrial complex I and the oxidative damage in nigrostriatal neurons induced by MPTP. With the dose used deprenyl recovers 50% DA levels and tyrosine hydroxylase activity depressed by the neurotoxin, normalizing locomotor activity of mice. Melatonin, which was unable to counteract MPTP-induced DA depletion and inhibition of tyrosine hydroxylase activity, potentiates the effect of deprenyl on catecholamine turnover and mice ambulatory activity. These results suggest a dissociation of complex I inhibition from DA depletion in this model of Parkinson's disease. The data also support that a combination of melatonin, which improves mitochondrial electron transport chain and reduces oxidative damage, and deprenyl, which promotes the specific function of the rescued neurons, i.e. DA turnover, may be a promising strategy for the treatment of PD. PMID:12600724
Low strain, long life creep fatigue of AF2-1DA and INCO 718
NASA Technical Reports Server (NTRS)
Thakker, A. B.; Cowles, B. A.
Two aircraft turbine disk alloys, GATORIZED AF2-DA and INCO 718 were evaluated for their low strain long life creep-fatigue behavior. Static (tensile and creep rupture) and cyclic properties of both alloys were characterized. The cntrolled strain LCF tests were conducted at 760 C (1400 F) and 649 C (1200 F) for AF2-1DA and INCO 718, respectively. Hold times were varied for tensile, compressive and tensile/compressive strain dwell (relaxation) tests. Stress (creep) hold behavior of AF2-1DA was also evaluated. Generally, INCO 718 exhibited more pronounced reduction in cyclic life due to hold than AF2-1DA. The percent reduction in life for both alloys for strain dwell tests was greater at low strain ranges (longer life regime). Changing hold time from 0 to 0.5, 2.0 and 15.0 min. resulted in corresponding reductions in life. The continuous cycle and cyclic/dwell initiation failure mechanism was predominantly transgranular for AF2-1DA and intergranular for INCO 718.
Essential 170-kDa subunit for degradation of crystalline cellulose by Clostridium cellulovorans cellulase
Shoseyov, O.; Doi, R.H. )
The cellulase complex from Clostridium cellulovorans has been purified and its subunit composition determined. The complex exhibits cellulase activity against crystalline cellulose as well as carboxymethylcellulase (CMCase) and cellobiohydrolase activities. Three major subunits are present with molecular masses of 170, 100, and 70 kDa. The 100-kDa subunit is the major CMCase, although at least four other, minor subunits show CMCase activity. The 170-kDa subunit has the highest affinity for cellulose, does not have detectable enzymatic activity, but is necessary for cellulase activity. Immunological studies indicate that the 170-kDa subunit is not required for binding of the catalytic subunits to cellulose and therefore does not function solely as an anchor protein. Thus this core subunit must have multiple functions. The authors propose a working hypothesis that the binding of the 170-kDa subunit converts the crystalline cellulose to a form that is capable of being hydrolyzed in a cooperative fashion by the associated catalytic subunits.
Detection of trace helium in G104 - 27, a 26,000 K DA white dwarf
NASA Technical Reports Server (NTRS)
Holberg, J. B.; Kidder, K. M.; Wesemael, F.
The detection of the He I 4471 A feature in the moderately hot DA white dwarf G104 - 27 places this star within the rather sparse class of DAB white dwarfs. It does not appear possible, on the basis of the present data, to distinguish between a stratified and a homogeneously mixed atmosphere. While a stratified structure would be natural for a DA white dwarf, G104 - 27 resides near the red edge of the DB gap, in a region where DA stars could transform into DB stars through convective mixing in the underlying He envelope and thereby destroy envelope stratification. Both atmospheres are consistent with the existing soft X-ray flux limit for this star.
Fluorescence spectroscopy coupled with PARAFAC and PLS DA for characterization and classification of honey.
Lenhardt, Lea; Bro, Rasmus; Zeković, Ivana; Dramićanin, Tatjana; Dramićanin, Miroslav D
Fluorescence spectroscopy coupled with parallel factor analysis (PARAFAC) and Partial least squares Discriminant Analysis (PLS DA) were used for characterization and classification of honey. Excitation emission spectra were obtained for 95 honey samples of different botanical origin (acacia, sunflower, linden, meadow, and fake honey) by recording emission from 270 to 640 nm with excitation in the range of 240-500 nm. The number of fluorophores present in honey, excitation and emission spectra of each fluorophore, and their relative concentration are determined using a six-component PARAFAC model. Emissions from phenolic compounds and Maillard reaction products exhibited the largest difference among classes of honey of different botanical origin. The PLS DA classification model, constructed from PARAFAC model scores, detected fake honey samples with 100% sensitivity and specificity. Honey samples were also classified using PLS DA with errors of 0.5% for linden, 10% for acacia, and about 20% for both sunflower and meadow mix. PMID:25577082
gLExec Integration with the ATLAS PanDA Workload Management System
NASA Astrophysics Data System (ADS)
Karavakis, E.; Barreiro, F.; Campana, S.; De, K.; Di Girolamo, A.; Litmaath, M.; Maeno, T.; Medrano, R.; Nilsson, P.; Wenaus, T.
ATLAS user jobs are executed on Worker Nodes (WNs) by pilots sent to sites by pilot factories. This paradigm serves to allow a high job reliability and although it has clear advantages, such as making the working environment homogeneous, the approach presents security and traceability challenges. To address these challenges, gLExec can be used to let the payloads for each user be executed under a different UNIX user id that uniquely identifies the ATLAS user. This paper describes the recent improvements and evolution of the security model within the ATLAS PanDA system, including improvements in the PanDA pilot, in the PanDA server and their integration with MyProxy, a credential caching system that entitles a person or a service to act in the name of the issuer of the credential. Finally, it presents results from ATLAS user jobs running with gLExec and describes the deployment campaign within ATLAS.
Chronic 5-HT transporter blockade reduces DA signaling to elicit basal ganglia dysfunction.
Morelli, Emanuela; Moore, Holly; Rebello, Tahilia J; Gray, Neil; Steele, Kelly; Esposito, Ennio; Gingrich, Jay A; Ansorge, Mark S
Serotonin (5-HT)-selective reuptake inhibitors (SSRIs) are widely administered for the treatment of depression, anxiety, and other neuropsychiatric disorders, but response rates are low, and side effects often lead to discontinuation. Side effect profiles suggest that SSRIs inhibit dopaminergic activity, but mechanistic insight remains scarce. Here we show that in mice, chronic 5-HT transporter (5-HTT) blockade during adulthood but not during development impairs basal ganglia-dependent behaviors in a dose-dependent and reversible fashion. Furthermore, chronic 5-HTT blockade reduces striatal dopamine (DA) content and metabolism. A causal relationship between reduced DA signaling and impaired basal ganglia-dependent behavior is indicated by the reversal of behavioral deficits through L-DOPA administration. Our data suggest that augmentation of DA signaling would reduce side effects and increase efficacies of SSRI-based therapy. PMID:22049417
The Da Vinci European BioBank: A Metabolomics-Driven Infrastructure
Carotenuto, Dario; Luchinat, Claudio; Marcon, Giordana; Rosato, Antonio; Turano, Paola
We present here the organization of the recently-constituted da Vinci European BioBank (daVEB, https://www.davincieuropeanbiobank.org/it). The biobank was created as an infrastructure to support the activities of the Fiorgen Foundation (http://www.fiorgen.net/), a nonprofit organization that promotes research in the field of pharmacogenomics and personalized medicine. The way operating procedures concerning samples and data have been developed at daVEB largely stems from the strong metabolomics connotation of Fiorgen and from the involvement of the scientific collaborators of the foundation in international/European projects aimed to tackle the standardization of pre-analytical procedures and the promotion of data standards in metabolomics. PMID:25913579
Evolution of the ATLAS PanDA Production and Distributed Analysis System
NASA Astrophysics Data System (ADS)
Maeno, T.; De, K.; Wenaus, T.; Nilsson, P.; Walker, R.; Stradling, A.; Fine, V.; Potekhin, M.; Panitkin, S.; Compostella, G.
The PanDA (Production and Distributed Analysis) system has been developed to meet ATLAS production and analysis requirements for a data-driven workload management system capable of operating at LHC data processing scale. PanDA has performed well with high reliability and robustness during the two years of LHC data-taking, while being actively evolved to meet the rapidly changing requirements for analysis use cases. We will present an overview of system evolution including automatic rebrokerage and reattempt for analysis jobs, adaptation for the CernVM File System, support for the multi-cloud model through which Tier-2 sites act as members of multiple clouds, pledged resource management and preferential brokerage, and monitoring improvements. We will also describe results from the analysis of two years of PanDA usage statistics, current issues, and plans for the future.
The coolest DA white dwarfs detected at soft X-ray wavelengths
NASA Technical Reports Server (NTRS)
Kidder, K. M.; Holberg, J. B.; Barstow, M. A.; Tweedy, R. W.; Wesemael, F.
New soft X-ray/EUV photometric observations of the DA white dwarfs KPD 0631 + 1043 = WD 0631 + 107 and PG 1113 + 413 = WD 1113 + 413 are analyzed. Previously reported soft X-ray detections of three other DAs and the failure to detect a fourth DA in deep Exosat observations are investigated. New ground-based spectra are presented for all of the objects, with IUE Ly-alpha spectra for some. These data are used to constrain the effective temperatures and surface gravities. The improved estimates of these parameters are employed to refer a photospheric He abundance for the hotter objects and to elucidate an effective observational low-temperature threshold for the detection of pure hydrogen DA white dwarfs at soft X-ray wavelengths.
Integration of PanDA workload management system with Titan supercomputer at OLCF
NASA Astrophysics Data System (ADS)
De, K.; Klimentov, A.; Oleynik, D.; Panitkin, S.; Petrosyan, A.; Schovancova, J.; Vaniachine, A.; Wenaus, T.
The PanDA (Production and Distributed Analysis) workload management system (WMS) was developed to meet the scale and complexity of LHC distributed computing for the ATLAS experiment. While PanDA currently distributes jobs to more than 100,000 cores at well over 100 Grid sites, the future LHC data taking runs will require more resources than Grid computing can possibly provide. To alleviate these challenges, ATLAS is engaged in an ambitious program to expand the current computing model to include additional resources such as the opportunistic use of supercomputers. We will describe a project aimed at integration of PanDA WMS with Titan supercomputer at Oak Ridge Leadership Computing Facility (OLCF). The current approach utilizes a modified PanDA pilot framework for job submission to Titan's batch queues and local data management, with light-weight MPI wrappers to run single threaded workloads in parallel on Titan's multicore worker nodes. It also gives PanDA new capability to collect, in real time, information about unused worker nodes on Titan, which allows precise definition of the size and duration of jobs submitted to Titan according to available free resources. This capability significantly reduces PanDA job wait time while improving Titan's utilization efficiency. This implementation was tested with a variety of Monte-Carlo workloads on Titan and is being tested on several other supercomputing platforms. Notice: This manuscript has been authored, by employees of Brookhaven Science Associates, LLC under Contract No. DE-AC02-98CH10886 with the U.S. Department of Energy. The publisher by accepting the manuscript for publication acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes.
Recurring X-ray outbursts in the supernova impostor SN 2010da in NGC 300
NASA Astrophysics Data System (ADS)
Binder, B.; Williams, B. F.; Kong, A. K. H.; Gaetz, T. J.; Plucinsky, P. P.; Skillman, E. D.; Dolphin, A.
We present new observations of the `supernova impostor' SN 2010da using the Chandra X-ray Observatory and the Hubble Space Telescope. During the initial 2010 outburst, the 0.3-10 keV luminosity was observed by Swift to be ˜5 × 1038 erg s-1 and faded by a factor of ˜25 in a four month period. Our two new Chandra observations show a factor of ˜10 increase in the 0.35-8 keV X-ray luminosity, from ˜4 × 1036 to 4 × 1037 erg s-1 in ˜6 months, and the X-ray spectrum is consistent in both observations with a power-law with a photon index of Γ ˜ 0. We find evidence of X-ray spectral state changes: when SN 2010da is in a high-luminosity state, the X-ray spectrum is harder (Γ ˜0) compared to the low-luminosity state (Γ ˜ 1.2 ± 0.8). Using our Hubble observations, we fit the colour-magnitude diagram of the coeval stellar population to estimate a time since formation of the SN 2010da progenitor system of ≲5 Myr. Our observations are consistent with SN 2010da being a high-mass X-ray binary (HMXB) composed of a neutron star and a luminous blue variable-like companion, although we cannot rule out the possibility that SN 2010da is an unusually X-ray bright massive star. The ≲5 Myr age is consistent with the theoretically predicted delay time between the formation of a massive binary and the onset of the HMXB phase. It is possible that the initial 2010 outburst marked the beginning of X-ray production in the system, making SN 2010da possibly the first massive progenitor binary ever observed to evolve into an HMXB.
Characterization of the 70 kDa polypeptide of the Na/Ca exchanger.
Saba, R I; Bollen, A; Herchuelz, A
The Na/Ca exchanger is associated with 160, 120 and 70 kDa polypeptides whose nature is poorly understood. We have purified and characterized the Na/Ca exchanger from bovine cardiac sarcolemmal vesicles (SLVs) by using ion-exchange and affinity chromatographies. The Na/Ca exchanger-enriched fraction was reconstituted into asolectin liposomes [lipid to protein ratio 10:1 (w/w)] that showed Na/Ca exchange activity. Under non-reducing conditions, SDS/PAGE showed a single 70 kDa polypeptide, which was further characterized by immunoblots with different antibodies: SWant, raised against the purified exchanger protein; NH2-terminus, residues 1-21; NCX1, residues 393-406; and Exon F, residues 622-644. Immunoblots under reducing conditions with SWant, NH2-terminus and NCX1 showed three bands migrating at 160, 120 and 70 kDa for SLV preparations, whereas Exon F reacted only with the 160 and 120 kDa bands. Under non-reducing conditions, immunoblots with purified reconstituted Na/Ca exchanger showed a single band at 70 kDa reacting with SWant, NH2-terminus and NCX1 but not with Exon F. We conclude that the 70 kDa protein is associated with Na/Ca exchange activity, has the same N-terminal sequence as the cloned bovine cardiac exchanger, and has its length decreased by at least 35% from its C-terminal portion as compared with that of the wild-type exchanger. PMID:9931309
Solubilization of insoluble inorganic phosphate by Burkholderia cepacia DA23 isolated from cultivated soil
Song, Ok-Ryul; Lee, Seung-Jin; Lee, Yong-Seok; Lee, Sang-Cheol; Kim, Keun-Ki; Choi, Yong-Lark
A mineral phosphate solubilizing bacterium, Burkholderia cepacia DA23 has been isolated from cultivated soils. Phosphate-solubilizing activities of the strain against three types of insoluble phosphate were quantitatively determined. When 3% of glucose concentration was used for carbon source, the strain had a marked mineral phosphate-solubilizing activity. Mineral phosphate solubilization was directly related to the pH drop by the strain. Analysis of the culture medium by high pressure liquid chromatography identified gluconic acid as the main organic acid released by Burkholderia cepacia DA23. Gluconic acid production was apparently the result of the glucose dehydrogenase activity and glucose dehydrogenase was affected by phosphate regulation. PMID:24031195
Spectroscopy of the DA white dwarfs - Automatic atmospheric parameterization and mass distribution
NASA Technical Reports Server (NTRS)
Mcmahan, Robert K.
A method for the automatic calculation of the atmospheric parameters (Teff and log g) of hydrogen-rich degenerate stars from low-resolution spectra is described, and then applied to the spectra of 53 DA white dwarfs. A value for the width of the DA mass distribution of sigma M/solar-M not greater than +0.10 is obtained using the proposed approach. The data indicate that the distribution is asymmetrically skewed to low masses; however, there is also evidence of a high-mass non-Gaussian tail.
[Euclides da Cunha in the Amazon: historical discontinuities in how the forest is seen and narrated].
Guimarães, Leandro Belinaso
The article examines how the discursive, social, and political processes that nationalized the Amazon gained firm footing during Brazil's First Republic. It is argued that Euclides da Cunha's texts on the Amazon see the forest and narrate its story in a way that differs from the travel accounts of nineteenth-century naturalists, above all foreign ones. Drawing inspiration from Latin American cultural studies, the article reinforces the argument that there is a historical discontinuity between the romantic foreign travel literature of the nineteenth century and the early twentieth-century writings of Euclides da Cunha. PMID:21461535
Aerodynamic design of the Cal Poly Da Vinci Human-Powered Helicopter
NASA Technical Reports Server (NTRS)
Larwood, Scott; Saiki, Neal
This paper will discuss the methodology used in designing the rotor and drive propellers for the third generation Cal Poly Da Vinci Human-Powered Helicopter. The rotor was designed using a lifting surface, uniform inflow hover analysis code and the propeller was designed using a minimum induced-loss method. Construction, geometry, and operating considerations are discussed as they impact the designs. Optimization of the design performance is also explained. The propellers were tested in a wind tunnel and results are compared with theoretical data. Successful flight tests of the Da Vinci III are discussed.
The prototypical 4.1R-10-kDa domain and the 4.1g-10-kDa paralog mediate fodrin-actin complex formation.
Kontrogianni-Konstantopoulos, A; Frye, C S; Benz, E J; Huang, S C
A complex family of 4.1R isoforms has been identified in non-erythroid tissues. In this study we characterized the exonic composition of brain 4.1R-10-kDa or spectrin/actin binding (SAB) domain and identified the minimal sequences required to stimulate fodrin/F-actin association. Adult rat brain expresses predominantly 4.1R mRNAs that carry an extended SAB, consisting of the alternative exons 14/15/16 and part of the constitutive exon 17. Exon 16 along with sequences carried by exon 17 is necessary and sufficient to induce formation of fodrin-actin-4.1R ternary complexes. The ability of the respective SAB domains of 4.1 homologs to sediment fodrin/actin was also investigated. 4.1G-SAB stimulates association of fodrin/actin, although with an approximately 2-fold reduced efficiency compared with 4.1R-10-kDa, whereas 4.1N and 4.1B do not. Sequencing of the corresponding domains revealed that 4.1G-SAB carries a cassette that shares significant homology with 4.1R exon 16, whereas the respective sequence is divergent in 4.1N and absent from brain 4.1B. An approximately 150-kDa 4.1R and an approximately 160-kDa 4.1G isoforms are present in PC12 lysates that occur in vivo in a supramolecular complex with fodrin and F-actin. Moreover, proteins 4.1R and 4.1G are distributed underneath the plasma membrane in PC12 cells. Collectively, these observations suggest that brain 4.1R and 4.1G may modulate the membrane mechanical properties of neuronal cells by promoting fodrin/actin association. PMID:11274145
Análise da aplicação e dos resultados do modelo OPM3® para a área da saúde
Augusto dos Santos, Luis; de Fátima Marin, Heimar
Esta pesquisa procurou analisar se um modelo de questionário criado por uma comunidade internacional de gerenciamento de projetos e se é aplicavél a organizações voltadas a área de saúde. O modelo OPM3® (Organizational Project Management Maturity Model) foi criado para que organizações de qualquer área ou porte pudessem identificar a presença, ou ausência, de boas práticas de gerenciamento. O objetivo da aplicação desse modelo é avaliar sempre a organização e não o entrevistado. No presente artigo, são apresentados os resultados da aplicação desse modelo em uma organização que possuía produtos e serviços de tecnologia da informação aplicados à área de saúde. Este estudo verificou que o modelo é aplicável de forma rápida e que a organização analisada possuía um número expressivo de boas práticas. PMID:26924862
Detection and differentiation of 22 kDa and 20 kDa Growth Hormone proteoforms in human plasma by LC-MS/MS.
Such-Sanmartín, Gerard; Bache, Nicolai; Bosch, Jaume; Gutiérrez-Gallego, Ricardo; Segura, Jordi; Jensen, Ole N
Human growth hormone (GH) is suspected to be widely and illegally used in sport to improve athletes' performance. For the detection of GH abuse, blood samples are screened for abnormal ratios between the 22 and 20 kDa GH proteoforms that demonstrate the administration of the synthetic hormone. Current detection methods are based on classical immunoassays as they provide sufficient sensitivity for the detection of GH proteoforms. These antibody based methods, however, suffer from unclear selectivity and potential cross-reactivity towards similar proteins. For unambiguous GH detection, we report a Mass Spectrometry ImmunoAssay (MSIA) that first enriches GH from plasma with an antibody of relatively low specificity, and subsequently quantifies the 22 and 20 kDa proteoforms by Selected Reaction Monitoring (SRM) LC-MS/MS analysis. This method proved superior to an antibody-free strategy based on GH purification by protein precipitation. Using GH-MSIA we successfully quantified the 22/20 kDa GH ratio in post-exercise capillary plasma extracted from two individuals. This GH-MSIA is applicable to anti-doping and GH-related disease analysis. PMID:25556001
76 FR 56455 - DaVita, Inc.; Analysis of Agreement Containing Consent Orders To Aid Public Comment
Federal Register 2010, 2011, 2012, 2013, 2014
... terms of the Consent Agreement, DaVita is required to divest 28 dialysis clinics and terminate one... Commission Act, as amended, 15 U.S.C. 45, by lessening competition for the provision of outpatient dialysis... provider of outpatient dialysis services in the United States. DaVita operates 1,612 outpatient...
The 37/67kDa laminin receptor (LR) inhibitor, NSC47924, affects 37/67kDa LR cell surface localization and interaction with the cellular prion protein
Sarnataro, Daniela; Pepe, Anna; Altamura, Gennaro; De Simone, Imma; Pesapane, Ada; Nitsch, Lucio; Montuori, Nunzia; Lavecchia, Antonio; Zurzolo, Chiara
The 37/67 kDa laminin receptor (LR) is a non-integrin protein, which binds both laminin-1 of the extracellular matrix and prion proteins, that hold a central role in prion diseases. The 37/67 kDa LR has been identified as interactor for the prion protein (PrPC) and to be required for pathological PrP (PrPSc) propagation in scrapie-infected neuronal cells, leading to the possibility that 37/67 kDa LR-PrPC interaction is related to the pathogenesis of prion diseases. A relationship between 37/67 kDa LR and PrPC in the presence of specific LR inhibitor compounds has not been investigated yet. We have characterized the trafficking of 37/67 kDa LR in both neuronal and non-neuronal cells, finding the receptor on the cell surface and nuclei, and identified the 67 kDa LR as the almost exclusive isoform interacting with PrPC. Here, we show that the treatment with the 37/67 kDa LR inhibitor, NSC47924, affects both the direct 37/67 kDa LR-PrPC interaction in vitro and the formation of the immunocomplex in live cells, inducing a progressive internalization of 37/67 kDa LR and stabilization of PrPC on the cell surface. These data reveal NSC47924 as a useful tool to regulate PrPC and 37/67 kDa LR trafficking and degradation, representing a novel small molecule to be tested against prion diseases. PMID:27071549
Effect of DA-9701 on the Normal Motility and Clonidine-induced Hypomotility of the Gastric Antrum in Rats
Kang, Je Wook; Han, Dae Kyeong; Kim, Ock Nyun; Lee, Kwang Jae
Background/Aims DA-9701 is a novel prokinetic agent. In the present study, we investigated the effect of DA-9701 on the motility of the gastric antrum in the normal and clonidine-induced hypomotility in an in vivo animal model. Methods A strain gauge force transducer was sutured on the gastric antrum to measure the contractile activity in rats. A total of 28 rats were subclassified into the 4 groups: (1) the placebo group, (2) the DA-9701 group, (3) the placebo group in the clonidine-pretreated rats, and (4) the DA-9701 group in the clonidine-pretreated rats. After the basal recording, either placebo (3% [w/v] hydroxypropylmethyl cellulose) or DA-9701 was administered. Contractile signals were measured after the administration and after a meal. In the clonidine-pretreated rats, either placebo or DA-9701 was administered. Contractile signals were measured after the administration and after a meal. Results Oral administration of DA-9701 did not significantly alter the motility index of the gastric antrum in the preprandial and postprandial periods, compared with the placebo group. The administration of clonidine decreased the motility index of the gastric antrum in the preprandial and postprandial periods, compared with the administration of placebo. This reduction of the antral motility by the administration of clonidine was not observed in the clonidine-pretreated DA-9701 group. The percentage of the motility index in the postprandial period was significantly greater in the clonidine-pretreated DA-9701 group, compared with the clonidine-pretreated placebo group. Conclusions DA-9701 improves the hypomotility of the gastric antrum induced by clonidine, suggesting its gastroprokinetic effect in the pathologic condition. PMID:26755679
32 CFR 516.14 - Service of process on DA or Secretary of Army.
Code of Federal Regulations, 2011 CFR
... 32 National Defense 3 2011-07-01 2009-07-01 true Service of process on DA or Secretary of Army. 516.14 Section 516.14 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY AID OF CIVIL AUTHORITIES AND PUBLIC RELATIONS LITIGATION Service of Process § 516.14 Service of process on...
32 CFR 644.512 - DA-SBA joint set-aside determination.
Code of Federal Regulations, 2011 CFR
... 32 National Defense 4 2011-07-01 2011-07-01 false DA-SBA joint set-aside determination. 644.512 Section 644.512 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL PROPERTY REAL ESTATE HANDBOOK Disposal Disposal of Standing Timber, Crops, and Embedded Gravel, Sand...
32 CFR 644.512 - DA-SBA joint set-aside determination.
Code of Federal Regulations, 2013 CFR
... 32 National Defense 4 2013-07-01 2013-07-01 false DA-SBA joint set-aside determination. 644.512 Section 644.512 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL PROPERTY REAL ESTATE HANDBOOK Disposal Disposal of Standing Timber, Crops, and Embedded Gravel, Sand...
32 CFR 644.512 - DA-SBA joint set-aside determination.
Code of Federal Regulations, 2012 CFR
... 32 National Defense 4 2012-07-01 2011-07-01 true DA-SBA joint set-aside determination. 644.512 Section 644.512 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL PROPERTY REAL ESTATE HANDBOOK Disposal Disposal of Standing Timber, Crops, and Embedded Gravel, Sand...
32 CFR 644.512 - DA-SBA joint set-aside determination.
Code of Federal Regulations, 2010 CFR
... 32 National Defense 4 2010-07-01 2010-07-01 true DA-SBA joint set-aside determination. 644.512 Section 644.512 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL PROPERTY REAL ESTATE HANDBOOK Disposal Disposal of Standing Timber, Crops, and Embedded Gravel, Sand...
32 CFR 644.512 - DA-SBA joint set-aside determination.
Code of Federal Regulations, 2014 CFR
... 32 National Defense 4 2014-07-01 2013-07-01 true DA-SBA joint set-aside determination. 644.512 Section 644.512 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL PROPERTY REAL ESTATE HANDBOOK Disposal Disposal of Standing Timber, Crops, and Embedded Gravel, Sand...
Purification and characterization of the acid soluble 26-kDa polypeptide from soybean seeds.
Momma, M; Haraguchi, K; Saito, M; Chikuni, K; Harada, K
Whey proteins from soybean seeds of Japanese varieties were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Among 11 varieties of soybean, three green and one black soybeans lacked a 26-kDa band that was found in all yellow soybeans. In this paper, the 26-kDa protein was named AS26k (acid soluble 26-kDa protein) temporarily. The AS26k protein was purified from Glycine max cv. Nattosyoryu, which is yellow soybean, through four purification steps: 30-35% saturated ammonium sulfate fractionation, ion exchange chromatography on S Sepharose Fast Flow, gel filtration on Sephadex G-100, and hydrophobic chromatography on phenyl Sepharose CL-4B. Purified AS26k was cleaved with V8 proteinase from Staphylococcus aureus or CNBr. The cleaved polypeptide contained two typical dehydrin motif sequences: DEYGNPV and (M)DKIKEKLPG, and a 19 amino acids sequence similar to a pea dehydrin. Native AS26k had a molecular mass of 32 kDa on gel filtration and a pl of 7.2 on two-dimensional PAGE. Similarly to other dehydrins and late embryogenesis abundant (LEA) proteins, AS26k was rich in hydrophilic amino acids, and highly heat stable. These results showed that AS26k was a dehydrin, a group II LEA protein in soybean seeds. PMID:9301109
Assessments of the economic impacts of natural hazards in Albania: application of DaLA methodology.
NASA Astrophysics Data System (ADS)
Massabo, Marco; Toto, Emanuela; Deda, Miranda; Prenci, Shemsi; Dhima, Maksimiljan
Disaster loss datasets are built using economic assessments done by different parties and using different approaches which are in many cases inconsistent. This is observed in more than 75% of the Alabanian records with missing economic evaluations. To assess the economic impact of disasters at all scales in Global Assessment Report on Disaster Risk Reduction 2011 and 2013 has used the DaLA methodology that takes advantage of a common and homogeneous set of quantitative physical damage indicators contained in the disaster databases. These have to be taken as the lower bound of the damage, which in general is higher. The UN-ECLAC DaLA Methodology takes into account in its assessments the overall status of economy of the affected country. It uses the national accounts and statistics of the country government as baseline data to assess damage and loss. It also factors in the impact of disasters on individual livelihoods and incomes to fully define the needs for recovery and reconstruction. Similarly to national disaster loss databases DaLA assessments have a national level of observation and data produced at sub-national resolution, resulting in highly detailed and accurate data sets. In 2013 in Albania was implemented a collection of disaster losses based on Desinventar. We applied the DaLa methodology to the Albanian database of impacts of natural disasters.
40 CFR 60.40Da - Applicability and designation of affected facility.
Code of Federal Regulations, 2014 CFR
... combination with any other fuel) in the combustion turbine engine and associated heat recovery steam generator... Performance for Electric Utility Steam Generating Units § 60.40Da Applicability and designation of affected... subpart applies is each electric utility steam generating unit: (1) That is capable of combusting...
40 CFR 60.40Da - Applicability and designation of affected facility.
Code of Federal Regulations, 2012 CFR
... combination with any other fuel) in the combustion turbine engine and associated heat recovery steam generator... Performance for Electric Utility Steam Generating Units § 60.40Da Applicability and designation of affected... subpart applies is each electric utility steam generating unit: (1) That is capable of combusting...
40 CFR 60.40Da - Applicability and designation of affected facility.
Code of Federal Regulations, 2013 CFR
... combination with any other fuel) in the combustion turbine engine and associated heat recovery steam generator... Performance for Electric Utility Steam Generating Units § 60.40Da Applicability and designation of affected... subpart applies is each electric utility steam generating unit: (1) That is capable of combusting...
Long-term evolution of 1991 DA: A dynamically evolved extinct Halley-type comet
NASA Technical Reports Server (NTRS)
Hahn, Gerhard; Bailey, M. E.
The long-term dynamical evolution of 21 variational orbits for the intermediate-period asteroid 1991 DA was followed for up to +/-10(exp 5) years from the present. 1991 DA is close to the 2:7 resonance with Jupiter; it has avoided close encounters, within 1 AU, with this planet for at least the past 30,000 years, even at the node crossing. The future evolution typically shows no close encounters with Jupiter within at least 50,000 years. This corresponds to the mean time between node crossings with either Jupiter or Saturn. Close encounters with Saturn and Jupiter lead to a chaotic evolution for the whole ensemble, while secular perturbations cause large-amplitude swings in eccentricity and inclination (the latter covering the range 15 deg approximately less than i approximately less than 85 deg) which correlate with deep excursions of the perihelion distance to values much less than 1 AU. These variations are similar to those found in P/Machholz and a variety of other high-inclination orbits, e.g., P/Hartley-IRAS. We emphasize the connection between the orbital evolution of 1991 DA and that of Halley-type comets. If 1991 DA was once a comet, it is not surprising that it is now extinct.
Computerized Dynamic Assessment (C-DA): Diagnosing L2 Development According to Learner Responsiveness to Mediation
ERIC Educational Resources Information Center
Poehner, Matthew E.; Zhang, Jie; Lu, Xiaofei
Dynamic assessment (DA) derives from the sociocultural theory of mind as elaborated by Russian psychologist L. S. Vygotsky. By offering mediation when individuals experience difficulties and carefully tracing their responsiveness, Vygotsky (1998) proposed that diagnoses may uncover abilities that have fully formed as well as those still in the…
GS32, a novel Golgi SNARE of 32 kDa, interacts preferentially with syntaxin 6.
Wong, S H; Xu, Y; Zhang, T; Griffiths, G; Lowe, S L; Subramaniam, V N; Seow, K T; Hong, W
Syntaxin 1, synaptobrevins or vesicle-associated membrane proteins, and the synaptosome-associated protein of 25 kDa (SNAP-25) are key molecules involved in the docking and fusion of synaptic vesicles with the presynaptic membrane. We report here the molecular, cell biological, and biochemical characterization of a 32-kDa protein homologous to both SNAP-25 (20% amino acid sequence identity) and the recently identified SNAP-23 (19% amino acid sequence identity). Northern blot analysis shows that the mRNA for this protein is widely expressed. Polyclonal antibodies against this protein detect a 32-kDa protein present in both cytosol and membrane fractions. The membrane-bound form of this protein is revealed to be primarily localized to the Golgi apparatus by indirect immunofluorescence microscopy, a finding that is further established by electron microscopy immunogold labeling showing that this protein is present in tubular-vesicular structures of the Golgi apparatus. Biochemical characterizations establish that this protein behaves like a SNAP receptor and is thus named Golgi SNARE of 32 kDa (GS32). GS32 in the Golgi extract is preferentially retained by the immobilized GST-syntaxin 6 fusion protein. The coimmunoprecipitation of syntaxin 6 but not syntaxin 5 or GS28 from the Golgi extract by antibodies against GS32 further sustains the preferential interaction of GS32 with Golgi syntaxin 6. PMID:9880331
Tumour-infiltrating lymphocytes bear the 75 kDa tumour necrosis factor receptor.
Trentin, L.; Zambello, R.; Bulian, P.; Cerutti, A.; Enthammer, C.; Cassatella, M.; Nitti, D.; Lise, M.; Agostini, C.; Semenzato, G.
Tumour necrosis factor alpha (TNF-alpha) is a cytokine with a variety of immunological properties. The identification of two receptors for this molecule, i.e. the 75 kDa and the 55 kDa TNF receptors (TNF-R), recently clarified the mechanisms through which this cytokine provides its wide range of immunomodulatory activities. In this study we have investigated the expression and the functional properties of these receptors on tumour-infiltrating lymphocytes (TILs) recovered from 17 patients with solid cancers (melanoma, colorectal carcinoma and lung cancer). To this end, TIL lines and freshly isolated TILs were evaluated for (a) the expression and the functional role of TNF receptors following culture in the presence of interleukin 2 (IL-2) and (b) the production of TNF-alpha following culture with IL-2 and the role of this cytokine in IL-2-driven TIL proliferation. Flow cytometry analysis demonstrated that TILs bear the 75 kDa TNF-R. Moreover, TIL lines express detectable messages for TNF-alpha and release this cytokine. Functional in vitro studies have shown that anti-TNF-alpha, as well as anti-75 kDa TNF-R antibodies, are able to inhibit the IL-2-induced TIL proliferation. These data demonstrate that TILs are equipped with a fully functional TNF-R system and suggest a putative role for this receptor and its ligand in the activation and expression of TILs following immunotherapy with IL-2. Images Figure 2 PMID:7841036
Translocation of an 89-kDa periplasmic protein is associated with Holospora infection
Iwatani, Koichi; Dohra, Hideo; Lang, B. Franz; Burger, Gertraud; Hori, Manabu; Fujishima, Masahiro . E-mail: email@example.com
The symbiotic bacterium Holospora obtusa infects the macronucleus of the ciliate Paramecium caudatum. After ingestion by its host, an infectious form of Holospora with an electron-translucent tip passes through the host digestive vacuole and penetrates the macronuclear envelope with this tip. To investigate the underlying molecular mechanism of this process, we raised a monoclonal antibody against the tip-specific 89-kDa protein, sequenced this partially, and identified the corresponding complete gene. The deduced protein sequence carries two actin-binding motifs. Indirect immunofluorescence microscopy shows that during escape from the host digestive vacuole, the 89-kDa proteins translocates from the inside to the outside of the tip. When the bacterium invades the macronucleus, the 89-kDa protein is left behind at the entry point of the nuclear envelope. Transmission electron microscopy shows the formation of fine fibrous structures that co-localize with the antibody-labeled regions of the bacterium. Our findings suggest that the 89-kDa protein plays a role in Holospora's escape from the host digestive vacuole, the migration through the host cytoplasm, and the invasion into the macronucleus.
40 CFR 60.50Da - Compliance determination procedures and methods.
Code of Federal Regulations, 2013 CFR
... Electric Utility Steam Generating Units § 60.50Da Compliance determination procedures and methods. (a) In... operator of an electric utility combined cycle gas turbine that does not meet the definition of an IGCC... of this part. The SO2 and NOX emission rates calculations from the gas turbine used in Method 19...
40 CFR 60.50Da - Compliance determination procedures and methods.
Code of Federal Regulations, 2012 CFR
... Electric Utility Steam Generating Units § 60.50Da Compliance determination procedures and methods. (a) In... calculations from the gas turbine used in Method 19 of appendix A-7 of this part are determined when the gas turbine is performance tested under subpart GG of this part. The potential uncontrolled PM emission...
40 CFR 60.50Da - Compliance determination procedures and methods.
Code of Federal Regulations, 2014 CFR
... Electric Utility Steam Generating Units § 60.50Da Compliance determination procedures and methods. (a) In... operator of an electric utility combined cycle gas turbine that does not meet the definition of an IGCC... of this part. The SO2 and NOX emission rates calculations from the gas turbine used in Method 19...
A Integracao de Ensino das Ciencias da Saude (An Integrated Medical Education Program [in Brazil]).
ERIC Educational Resources Information Center
Pourchet-Campos, M. A.; Guimaraes Junior, Paulino
At the Sixth Annual Reunion of the Brazilian Association of Medical Schools (VI Reuniao Anual da Associacao Brasileira de Escolas Medicas) leaders in the Brazilian medical profession proposed an integrated educational program for training students in the fields of medicine and public health. Under Brazil's present system of education, all…
40 CFR 60.44Da - Standards for nitrogen oxides (NOX).
Code of Federal Regulations, 2012 CFR
... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Standards for nitrogen oxides (NOX). 60... Steam Generating Units § 60.44Da Standards for nitrogen oxides (NOX). (a) Except as provided in....20 Liquid fuels: Coal-derived fuels 210 0.50 Shale oil 210 0.50 All other fuels 130 0.30 Solid...
40 CFR 60.44Da - Standards for nitrogen oxides (NOX).
Code of Federal Regulations, 2014 CFR
... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Standards for nitrogen oxides (NOX). 60... Steam Generating Units § 60.44Da Standards for nitrogen oxides (NOX). (a) Except as provided in....20 Liquid fuels: Coal-derived fuels 210 0.50 Shale oil 210 0.50 All other fuels 130 0.30 Solid...
40 CFR 60.44Da - Standards for nitrogen oxides (NOX).
Code of Federal Regulations, 2013 CFR
... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Standards for nitrogen oxides (NOX). 60... Steam Generating Units § 60.44Da Standards for nitrogen oxides (NOX). (a) Except as provided in....20 Liquid fuels: Coal-derived fuels 210 0.50 Shale oil 210 0.50 All other fuels 130 0.30 Solid...
32 CFR 635.23 - DA Form 4833 (Commander's Report of Disciplinary or Administrative Action).
Code of Federal Regulations, 2010 CFR
... 32 National Defense 4 2010-07-01 2010-07-01 true DA Form 4833 (Commander's Report of Disciplinary or Administrative Action). 635.23 Section 635.23 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) LAW ENFORCEMENT AND CRIMINAL INVESTIGATIONS LAW ENFORCEMENT...
Brominated and radioiodinated derivatives of methylphenidate (MP): Potential imaging agents for the dopamine (DA) transporter
Pan, D.; Gatley, S.J.; Dewey, S.L.
MP (Ritalin) is a psychomotor stimulant used in the treatment of attention-deficit hyperactivity disorder. The therapeutic properties of MP are thought to be mediated by its binding to a site on the DA transporter, resulting in inhibition of DA reuptake and enhanced levels of synaptic dopamine. MP also inhibits reuptake of norepinephrine (NE) in vitro. MP has two chiral centers, but its pharmacological activity is believed due solely to the d-threo isomer. We have found that d,l-threo-C-11 MP has favorable properties for PET studies, and therefore examined the effects of incorporating halogen atoms into the phenyl ring of MP, with a view to preparing C-11 and I-123 MP analogs as potential PET/SPECT tracers. We synthesized the 2-, 3- and 4-bromo MP analogs from the corresponding bromophenylacetonitriles by modification of the original synthesis of MP. In in vitro binding assays all three d,l-threo bromo compounds had higher affinities than MP for DA transporter sites labeled with tritiated WIN 35,428 (3->4-, 2->MP). They also showed high activity with NE reuptake sites labeled with tritiated nisoxetine. They were active in vivo as demonstrated by inhibition of heart uptake of tritiated NE in the mouse, and elevation of striatal extracellular DA (microdialysis) and stimulation of locomotor activity in the rat.
Next Generation PanDA Pilot for ATLAS and Other Experiments
NASA Astrophysics Data System (ADS)
Nilsson, P.; Barreiro Megino, F.; Caballero Bejar, J.; De, K.; Hover, J.; Love, P.; Maeno, T.; Medrano Llamas, R.; Walker, R.; Wenaus, T.; Atlas Collaboration
The Production and Distributed Analysis system (PanDA) has been in use in the ATLAS Experiment since 2005. It uses a sophisticated pilot system to execute submitted jobs on the worker nodes. While originally designed for ATLAS, the PanDA Pilot has recently been refactored to facilitate use outside of ATLAS. Experiments are now handled as plug-ins such that a new PanDA Pilot user only has to implement a set of prototyped methods in the plug-in classes, and provide a script that configures and runs the experiment-specific payload. We will give an overview of the Next Generation PanDA Pilot system and will present major features and recent improvements including live user payload debugging, data access via the Federated XRootD system, stage-out to alternative storage elements, support for the new ATLAS DDM system (Rucio), and an improved integration with glExec, as well as a description of the experiment-specific plug-in classes. The performance of the pilot system in processing LHC data on the OSG, LCG and Nordugrid infrastructures used by ATLAS will also be presented. We will describe plans for future development on the time scale of the next few years.
32 CFR 516.14 - Service of process on DA or Secretary of Army.
Code of Federal Regulations, 2010 CFR
... 32 National Defense 3 2010-07-01 2010-07-01 true Service of process on DA or Secretary of Army. 516.14 Section 516.14 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY AID OF CIVIL AUTHORITIES AND PUBLIC RELATIONS LITIGATION Service of Process § 516.14 Service of process on...
76 FR 55293 - Special Conditions: Diamond Aircraft Industries, Model DA-40NG; Electronic Engine Control (EEC...
Federal Register 2010, 2011, 2012, 2013, 2014
... cantilever low wing, T-tail airplane with the Austro Engine GmbH model E4 diesel engine and an increased..., Model DA-40NG; Electronic Engine Control (EEC) System AGENCY: Federal Aviation Administration (FAA), DOT... design feature(s) associated with an electronic engine control (EEC), also known as a Full...
76 FR 72087 - Special Conditions: Diamond Aircraft Industries, Model DA-40NG; Electronic Engine Control (EEC...
Federal Register 2010, 2011, 2012, 2013, 2014
.... A47CE to include the new model DA- 40NG with the Austro Engine GmbH model E4 Aircraft Diesel Engine (ADE...-tail airplane with the Austro Engine GmbH model E4 diesel engine and an increased maximum takeoff gross... Engine GmbH model E4 aircraft diesel engine. 1. Electronic Engine Control a. For electronic...
Views and Voices of the Working Class: Aída and Rosario
ERIC Educational Resources Information Center
Pao, Maria T.
In 2005, Spanish television audiences saw the debut of the nation's first spinoff, the sitcom "Aída." The show featured the tribulations of its title character and her working-class family in their struggle to "llegar a fin de mes." It seemed to promise a sensibility enacted in the US series "Roseanne," where…
Understanding and Interpreting Japanese NP1 "wa" NP2 "da" Sentences: Mechanism and Contextual Factors
ERIC Educational Resources Information Center
Yoshida, Megumi
This dissertation investigates the contextual factors that affect the understanding and interpretation of one Japanese topicalized construction, NP[subscript 1] wa NP[subscript 2] da sentences, by native speakers of Japanese. The construction allows two possibilities in the relation between the NP[subscript 1] and the NP[subscript 2]. When the two…
In response to Dr. Jose AP da Silva: fall prevention with vitamin D clarifications needed
Technology Transfer Automated Retrieval System (TEKTRAN)
We agree with Dr. da Silva, and wish to clarify three issues raised by the recent IOM report concerning our 2009 meta-analysis on vitamin D and fall prevention. 1. The IOM questioned the inclusion of Broe et al., which did not pre-specify falls as a primary or secondary outcome. While this did viol...
40 CFR 60.43Da - Standards for sulfur dioxide (SO2).
Code of Federal Regulations, 2013 CFR
... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Standards for sulfur dioxide (SO2). 60... Steam Generating Units § 60.43Da Standards for sulfur dioxide (SO2). (a) On and after the date on which... the percent reduction requirement is determined on a 24-hour basis. (d) Sulfur dioxide emissions...
40 CFR 60.43Da - Standards for sulfur dioxide (SO2).
Code of Federal Regulations, 2014 CFR
... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Standards for sulfur dioxide (SO2). 60... Steam Generating Units § 60.43Da Standards for sulfur dioxide (SO2). (a) On and after the date on which... the percent reduction requirement is determined on a 24-hour basis. (d) Sulfur dioxide emissions...
40 CFR 60.43Da - Standards for sulfur dioxide (SO2).
Code of Federal Regulations, 2012 CFR
... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Standards for sulfur dioxide (SO2). 60... Steam Generating Units § 60.43Da Standards for sulfur dioxide (SO2). (a) On and after the date on which... the percent reduction requirement is determined on a 24-hour basis. (d) Sulfur dioxide emissions...
Maternal Atrazine (ATR) alters hypothalamic dopamine (HYP-DA) and serum prolactin (sPRL) in male pups. 1Christopher Langdale, 2Tammy Stoker and 2Ralph Cooper. 1 Dept. of Cell Biology, North Carolina State University College of Veterinary Medicine, Raleigh, NC. 2 Endocrinology ...
Leonardo da Vinci, One Year on...a Different Look at Vocational Training in Europe.
ERIC Educational Resources Information Center
Le Magazine, 1996
Discusses the success of the Leonardo da Vinci program, a European laboratory of innovation in vocational training, a priority focus of investment in human resources and intelligence, and a way to mobilize innovative forces beyond national boundaries. Trends identified by the program focus on new information and communication technologies. (JOW)
Transparency of Vocational Qualifications: The Leonardo da Vinci Approach. CEDEFOP Panorama Series.
ERIC Educational Resources Information Center
Bjornavold, Jens; Pettersson, Sten
This report gives an overview of the situation of transparency of vocational qualifications by presenting measures introduced at the European Community level and by drawing attention to projects within the Leonardo da Vinci Program dealing with the issue. A 16-page executive summary appears first. Chapter 1 provides general background and aims.…
Sobre a largura da última superfície de espalhamento
NASA Astrophysics Data System (ADS)
Nobre, M. A. S.; Pires, N.; Lima, J. A. S.
De acordo com o modelo do "Big-Bang", no universo primordial a matéria estava em equilíbrio térmico com a radiação. Com a expansão a temperatura da radiação cai. Quando a temperatura chega em torno dos 4.000K, os espalhamentos diminuem, começando a recombinação dos prótons e elétrons em Hidrogênio neutro (era conhecida como da recombinação). Ao final da recombinação, os fótons se propagam livremente sofrendo, em princípio, somente os efeitos do "redshift" cosmológico. Esses fótons nos alcançam hoje como a radiação cósmica de fundo (RCF), e parecem vir de uma superfície esférica ao nosso redor, tal que o raio dela é a distância que ele viajou desde seu último espalhamento na época da recombinação. Naturalmente, esse processo não ocorreu abruptamente, implicando na existência de uma largura no espaço dos "redshifts" que deve depender do modelo cosmológico específico e dos processos físicos considerados. Neste trabalho analisamos os efeitos de diferentes modelos - a saber, aqueles com decaimento do vácuo L(t), criação de matéria, quintessência e gás de Chaplygin - sobre a última superfície de espalhamento da RCF, em particular sua largura e a função visibilidade, que determina a probabilidade de um fóton ter tido seu último espalhamento num "redshift" z. No caso particular dos modelos com decaimento do vácuo, existe uma forte dependência da função visibilidade com L(t). Tais efeitos poderão ser testados através da análise dos resultados de experimentos mais precisos que estão atualmente em andamento, como por exemplo, o WMAP.
PanDA Pilot Submission using Condor-G: Experience and Improvements
Zhao X.; Hover John; Wlodek Tomasz; Wenaus Torre; Frey Jaime; Tannenbaum Todd; Livny Miron
PanDA (Production and Distributed Analysis) is the workload management system of the ATLAS experiment, used to run managed production and user analysis jobs on the grid. As a late-binding, pilot-based system, the maintenance of a smooth and steady stream of pilot jobs to all grid sites is critical for PanDA operation. The ATLAS Computing Facility (ACF) at BNL, as the ATLAS Tier1 center in the US, operates the pilot submission systems for the US. This is done using the PanDA 'AutoPilot' scheduler component which submits pilot jobs via Condor-G, a grid job scheduling system developed at the University of Wisconsin-Madison. In this paper, we discuss the operation and performance of the Condor-G pilot submission at BNL, with emphasis on the challenges and issues encountered in the real grid production environment. With the close collaboration of Condor and PanDA teams, the scalability and stability of the overall system has been greatly improved over the last year. We review improvements made to Condor-G resulting from this collaboration, including isolation of site-based issues by running a separate Gridmanager for each remote site, introduction of the 'Nonessential' job attribute to allow Condor to optimize its behavior for the specific character of pilot jobs, better understanding and handling of the Gridmonitor process, as well as better scheduling in the PanDA pilot scheduler component. We will also cover the monitoring of the health of the system.
PanDA Pilot Submission using Condor-G: Experience and Improvements
NASA Astrophysics Data System (ADS)
Zhao, Xin; Hover, John; Wlodek, Tomasz; Wenaus, Torre; Frey, Jaime; Tannenbaum, Todd; Livny, Miron; ATLAS Collaboration
PanDA (Production and Distributed Analysis) is the workload management system of the ATLAS experiment, used to run managed production and user analysis jobs on the grid. As a late-binding, pilot-based system, the maintenance of a smooth and steady stream of pilot jobs to all grid sites is critical for PanDA operation. The ATLAS Computing Facility (ACF) at BNL, as the ATLAS Tier1 center in the US, operates the pilot submission systems for the US. This is done using the PanDA "AutoPilot" scheduler component which submits pilot jobs via Condor-G, a grid job scheduling system developed at the University of Wisconsin-Madison. In this paper, we discuss the operation and performance of the Condor-G pilot submission at BNL, with emphasis on the challenges and issues encountered in the real grid production environment. With the close collaboration of Condor and PanDA teams, the scalability and stability of the overall system has been greatly improved over the last year. We review improvements made to Condor-G resulting from this collaboration, including isolation of site-based issues by running a separate Gridmanager for each remote site, introduction of the 'Nonessential' job attribute to allow Condor to optimize its behavior for the specific character of pilot jobs, better understanding and handling of the Gridmonitor process, as well as better scheduling in the PanDA pilot scheduler component. We will also cover the monitoring of the health of the system.
Deconstructing honeybee vitellogenin: novel 40 kDa fragment assigned to its N terminus
Havukainen, Heli; Halskau, Øyvind; Skjaerven, Lars; Smedal, Bente; Amdam, Gro V.
Vitellogenin, an egg-yolk protein precursor common to oviparous animals, is found abundantly in honeybee workers – a caste of helpers that do not usually lay eggs. Instead, honeybee vitellogenin (180 kDa) participates in processes other than reproduction: it influences hormone signaling, food-related behavior, immunity, stress resistance and longevity. The molecular basis of these functions is largely unknown. Here, we establish and compare the molecular properties of vitellogenin from honeybee hemolymph (blood) and abdominal fat body, two compartments that are linked to vitellogenin functions. Our results reveal a novel 40 kDa vitellogenin fragment in abdominal fat body tissue, the main site for vitellogenin synthesis and storage. Using MALDI-TOF combined with MS/MS mass-spectroscopy, we assign the 40 kDa fragment to the N terminus of vitellogenin, whereas a previously observed 150 kDa fragment corresponded to the remainder of the protein. We show that both protein units are N glycosylated and phosphorylated. Focusing on the novel 40 kDa fragment, we present a homology model based on the structure of lamprey lipovitellin that includes a conserved β-barrel-like shape, with a lipophilic cavity in the interior and two insect-specific loops that have not been described before. Our data indicate that the honeybee fat body vitellogenin experiences cleavage unlike hemolymph vitellogenin, a pattern that can suggest a tissue-specific role. Our experiments advance the molecular understanding of vitellogenin, of which the multiple physiological and behavioral effects in honeybees are well established. PMID:21270306
Site effects inferred from HVSR methodology in Praia da Vitória city (Portugal)
NASA Astrophysics Data System (ADS)
Fontiela, João; Borges, José; Bezzeghoud, Mourad; Rosset, Philippe; Rodrigues, Francisco
Terceira island (Azores) have experienced several strong earthquakes, especially on the eastern and south-eastern part of the island with maximum MM intensity of VIII in 1800, 1801 and 1912 and IX in 1614 and 1841. Seismicity of the eastern part of the island is largely dominated by the tectonic of the NW-SE Lajes graben where are located Praia da Vitória and other setlements. Praia da Vitória is settled partially over pyroclastic material(pumice and ignimbrite) deposited around 20'000 years ago and forming part of the UpperTerceira Group. Its thickness is 15m on the north but in Praia da Vitoria has less than one meter.. It is sited on by basaltic flow of the upper Basaltic Terceira Group (2,000y BP) and a thick andesitic lava flow. A narrow band with sand dunes and beach crossed this deposits along Praia da Vitória and behind it appear a flat area with 7-10m of round cobbles that is interpreted as an old boulders'beach. Thickness of these different deposits are tricky to estimate in urban areas since very few outcrops are present. In order to assess site amplification of seismic waves due to the soil conditions, we perform horizontal-to-vertical spectral ratio (HVSR) method from broad band ambient noise measurements in 51 different sites within Praia da Vitoria. The first results show that the frequencies of the study area are variable, lying between 0.6-12 Hz. ACKNOWLEDGMENTS: João Fontiela is supported by grant M3.1.2/F/060/2011 provided by the Regional Science Fund of the Azores Regional Government. This study was partially funded by the Institute of Earth Sciences, Portugal, under the contract with the FCT, ICT-UID/GEO/04683/2013 (Portugal).
Intestinal mucins from normal subjects and patients with cystic fibrosis. Variable contents of the disulphide-bound 118 kDa glycoprotein and different reactivities with an anti-(118 kDa glycoprotein) antibody.
Mantle, M; Stewart, G
1. A specific antibody was developed against the disulphide-bound 118 kDa glycoprotein of human intestinal mucin and used to establish an e.l.i.s.a. Fourteen purified mucins [eight normal (N) and six cystic fibrosis (CF)] had the same affinity for the antibody in the e.l.i.s.a., but their relative immunoreactivities varied widely (approx. 100,000-fold). In general, CF mucins were more antigenic than N mucins. 2. Variations (approx. 10-fold) were detected in the 118 kDa glycoprotein content of both N and CF mucins (assessed from Coomassie Blue-stained polyacrylamide gels), but these did not appear to be responsible for the differences in mucin immunoreactivity. 3. Variations (approx. 6-fold) were also observed in the size of the 118 kDa peak produced by N and CF mucins on Western blots. These were mostly due to differences in the 118 kDa glycoprotein content of mucins, although a small proportion resulted from changes in the number of antigenic determinants within individual 118 kDa glycoproteins. 4. After concanavalin A affinity chromatography of four reduced mucins (two N and two CF), purified 118 kDa glycoprotein was recovered in the bound fractions from the column, specifically eluted by methyl alpha-mannoside. 5. The amounts of 118 kDa glycoprotein isolated from the four mucins varied as predicted from the size of their 118 kDa bands on Coomassie Blue-stained gels. 6. Three 118 kDa glycoproteins (one N and two CF) showed almost identical reactivity in the e.l.i.s.a.; the fourth had fewer antigenic determinants. 7. Since differences in 118 kDa glycoprotein content and in the number of antigenic determinants within the 118 kDa glycoprotein did not account for variations in the reactivity of native mucins in the e.l.i.s.a., it appeared that accessibility of the 118 kDa glycoprotein to antibody binding may be critical in determining mucin immunoreactivity. This suggests that the three-dimensional conformation of CF mucins may differ from that of N mucins, leading
Qual o Papel da Escola e da Mídia na Obtenção dos Conhecimentos Astronômicos?
NASA Astrophysics Data System (ADS)
da Cunha, W. S.; Voelzke, M. R.; Amaral, L. H.
É amplamente reconhecido que as pessoas em geral têm grande fascínio e interesse pela Astronomia. Por outro lado, o conteúdo dessa área incluído no ensino formal está longe de ser abrangente e suficiente para suprir a demanda. Esse interesse permite aperfeiçoar a divulgação e o ensino não formal de Astronomia através da mídia aliando a adequação do conteúdo à expectativa das pessoas. A reforma do Currículo Básico da Escola Pública da maioria dos Estados brasileiros tem introduzido Astronomia desde a pré-escola até o Ensino Médio. Num estudo realizado com 1180 alunos do Ensino Médio de seis escolas estaduais, revelou-se através da aplicação de um formulário contendo questões que abordavam conhecimentos básicos em Astronomia, como foram adquiridos tais conhecimentos e à infra-estrutura da escola. Para 66,1% dos entrevistados o professor já fez alguma apresentação a respeito de Astronomia. O presente estudo revelou também que 54,4% dos alunos entrevistados adquiriram seus conhecimentos astronômicos na escola, 18,4% através da televisão e 27,1% afirmaram que seus conhecimentos foram obtidos através de outros tipos de mídia (internet, livros didáticos, filmes e revistas). Embora o estudo revele claramente que há certo equilíbrio no meio em que os alunos obtiveram seus conhecimentos astronômicos, a mídia busca não somente sua difusão, mas a disponibilização desse conhecimento de maneira correta, que às vezes, não é atingido, apresentando conceitos sem fundamentação e incorretos. Por outro lado, no dia-a-dia escolar, é comum encontrar professores que vêm para as aulas com concepções prévias, que podem diferir substancialmente das idéias a serem ensinadas, dificultando o aprendizado de futuros conceitos científicos. Uma avaliação criteriosa, avaliando as dificuldades dos professores em determinar o sentido correto dos conceitos e observando a metodologia que utilizam no ensino, poderá proporcionar uma melhor
Dioscorea Extract (DA-9801) Modulates Markers of Peripheral Neuropathy in Type 2 Diabetic db/db Mice
Moon, Eunjung; Lee, Sung Ok; Kang, Tong Ho; Kim, Hye Ju; Choi, Sang Zin; Son, Mi-Won; Kim, Sun Yeou
The purpose of this study was to investigate the therapeutic effects of DA-9801, an optimized extract of Dioscorea species, on diabetic peripheral neuropathy in a type 2 diabetic animal model. In this study, db/db mice were treated with DA-9801 (30 and 100 mg/kg, daily, p.o.) for 12 weeks. DA-9801 reduced the blood glucose levels and increased the withdrawal latencies in hot plate tests. Moreover, it prevented nerve damage based on increased nerve conduction velocity and ultrastructural changes. Decrease of nerve growth factor (NGF) may have a detrimental effect on diabetic neuropathy. We previously reported NGF regulatory properties of the Dioscorea genus. In this study, DA-9801 induced NGF production in rat primary astrocytes. In addition, it increased NGF levels in the sciatic nerve and the plasma of type 2 diabetic animals. DA-9801 also increased neurite outgrowth and mRNA expression of Tieg1/Klf10, an NGF target gene, in PC12 cells. These results demonstrated the attenuation of diabetic peripheral neuropathy by oral treatment with DA-9801 via NGF regulation. DA-9801 is currently being evaluated in a phase II clinical study. PMID:25414776
The Effect of DA-6034 on Intestinal Permeability in an Indomethacin-Induced Small Intestinal Injury Model
Kwak, Dong Shin; Lee, Oh Young; Lee, Kang Nyeong; Jun, Dae Won; Lee, Hang Lak; Yoon, Byung Chul; Choi, Ho Soon
Background/Aims DA-6034 has anti-inflammatory activities and exhibits cytoprotective effects in acute gastric injury models. However, explanations for the protective effects of DA-6034 on intestinal permeability are limited. This study sought to investigate the effect of DA-6034 on intestinal permeability in an indomethacin-induced small intestinal injury model and its protective effect against small intestinal injury. Methods Rats in the treatment group received DA-6034 from days 0 to 2 and indomethacin from days 1 to 2. Rats in the control group received indomethacin from days 1 to 2. On the fourth day, the small intestines were examined to compare the severity of inflammation. Intestinal permeability was evaluated by using fluorescein isothiocyanate-labeled dextran. Western blotting was performed to confirm the association between DA-6034 and the extracellular signal-regulated kinase (ERK) pathway. Results The inflammation scores in the treatment group were lower than those in the control group, but the difference was statistically insignificant. Hemorrhagic lesions in the treatment group were broader than those in the control group, but the difference was statistically insignificant. Intestinal permeability was lower in the treatment group than in the control group. DA-6034 enhanced extracellular signal-regulated kinase expression, and intestinal permeability was negatively correlated with ERK expression. Conclusions DA-6034 may decrease intestinal permeability in an indomethacin-induced intestinal injury model via the ERK pathway. PMID:27114435
Lack of Apoptosis of Infiltrating Cells as the Mechanism of High Susceptibility to EAE in DA Rats
Mensah-Brown, Eric; Galadari, Sehamuddin; Shahin, Allen
Dark Agouti (DA) rats are highly susceptible to induction of Th-l-mediated autoimmunity disease, including experimental allergic encephalomyelitis (EAE). In contrast to other susceptible rat strains in which disease is induced only with encephalitogen emulsified in complete Freund's adjuvants (CFA), in DA rats EAE develops after injection of encephalitogen in incomplete Freund's adjuvants (IFA) or Titermax, putative Th-2 directed adjuvant. Lymph node cells derived from immunized DA rats and stimulated in vitro produce significantly more Interferon-γ (IFN-γ) than resistant Albino Oxford (AO) rats. However, cells derived from both strains produce large amounts of IL-10 but not IL-4. Immunized lymph node cells derived from EAE susceptible (AO × DA) F1rats induce clinical signs of disease in sublethally irradiated parental DA but not AO rats. The pathohistology of the target tissue in these recipients clearly demonstrated infiltration of mononuclear cells in both parental strains. However, the number of CD4+ cells was significantly higher and number of apoptotic cells significantly lower in DA rats sacrificed 8 days after passive transfer. We postulate that in addition to higher IFN-γ and TNF-α production, resistance to early apoptosis of the invading cells in the target tissue possibly due to lack of downregulation by TGF-β leads to exceptional susceptibility to EAE in DA rats. PMID:11785669
Loss of 45 Da from a2 Ions and Preferential Loss of 48 Da from a2 Ions Containing Methionine in Peptide Ion Tandem Mass Spectra
NASA Astrophysics Data System (ADS)
Simón-Manso, Yamil; Neta, Pedatsur; Yang, Xiaoyu; Stein, Stephen E.
While analyzing tandem mass spectra of tryptic tripeptides, intense unassigned peaks were observed, corresponding to neutral loss of 45 Da from a2 ions. This process was confirmed by MS3 experiments. Based on exact mass analysis, the loss was ascribed to (NH3 + CO) or formamide. The proposed mechanism involves a cyclic form of the a2 ions. The structure of the a2 - 45 ions was confirmed by their fragmentation in MS3 experiments. Loss of (NH3 + CO) from the a2 ions occurs in competition with other paths, such as the loss of H2O or the formation of immonium ions. However, if the a2 ion contains methionine, a neutral loss of 48 Da (ascribed to CH3SH) predominates, and is followed by the loss of (NH3 + CO). These processes were confirmed by MS3 experiments. The intensity of the a2 - 48 peak formed from XaaMet has a maximum value of 42% (of the total intensity of all ions) for Xaa=Gly, varies between 15% and 40% for most other Xaa residues, is lower for residues that can undergo loss of water or ammonia, and is very low for Lys or Arg. When the order of the residues is reversed to MetXaa, the loss of 48 Da is much smaller. This effect can be used to determine the sequence of b2 ions containing Met in proteomic studies. Considerable loss of CH3SH is observed from doubly protonated tryptic tripeptides with N-terminal Met, but the loss is much less when they are singly protonated or when Met is in the center position.
(3H)domperidone binding to the kidney inner medullary collecting duct dopamine-2K (DA2K) receptor
Huo, T.; Healy, D.P. )
Previous studies by the authors laboratory have indicated that inner medullary collecting ducts (IMCDs) express a novel dopamine (DA) receptor, designated DA2K, that is linked to stimulation of prostaglandin E2 production. This receptor has a distinct pharmacological profile and is similar in size, but not homologous to, the brain D2 receptor. Because the DA2-selective antagonist domperidone blocks DA-mediated stimulation of prostaglandin E2 production in IMCD cells, we utilized (3H)domperidone to study the binding characteristics of the DA2K receptor in IMCD cells. (3H)Domperidone binding was saturable and best fit to a single high density site (KD, 57.6 {plus minus} 10.5 nM; Bmax, 14.9 {plus minus} 2.7 pmol/mg protein). The specificity of (3H)domperidone binding in IMCD cells was verified by competition analysis with a variety of dopaminergic and nondopaminergic agents. Dopaminergic drugs were less potent competitors for (3H)domperidone binding in IMCD cells than previously reported for brain DA receptors, but the rank order was consistent with the labeling of a DA receptor (antagonists: domperidone greater than spiperone greater than haloperidol greater than Sch 23390 much greater than (-)-sulpiride; agonists: norapomorphine greater than fenoldopam much greater than dopamine = quinpirole), and was better correlated with the pharmacological profile for the brain D2 receptor than the brain D3 receptor. In addition, quinpirole, the most D3-selective ligand currently available, did not compete for (3H)domperidone binding in IMCD cells. These results add further support to the existence of a novel high density DA receptor, DA2K, expressed in IMCD cells.
[The nuclear matrix proteins (mol. mass 38 and 50 kDa) are transported by chromosomes in mitosis].
Murasheva, M I; Chentsov, Iu S
It was shown by immunofluorescence method that serum M68 and serum K43 from patients with autoimmune disease stain interphase nuclei and periphery of mitotic chromosomes of pig kidney cells. Western blotting reveals the polypeptide with mol. mass of 50 kDa in serum M68, and the polypeptide with mol. mass of 38 kDa in serum K43. In the nuclear protein matrix, the antibodies to protein with mol. mass of 38 kDa stained only nucleolar periphery, while the antibodies to the protein with mol. mass of 50 kDa stained both the nucleolar periphery and all the interphase nucleus. It shows that among all components of nuclear protein matrix (lamina, internuclear network, residual nucleoli) only nucleolar periphery contains the 38 kDa protein, while the 50 kDa protein is a part of residual nucleolar periphery and takes part in nuclear protein network formation. In the interphase cells, both proteins were in situ localized in the nuclei, but one of them with mol. mass of 50 kDa was in the form of small clearly outlined granules, while the other (38 kDa) was in the form of small bright granules against the background of diffusely stained nuclei. Both proteins were also revealed as continuous ring around nucleolar periphery. During all mitotic stages, the 50 kDa protein was seen on the chromosomal periphery as a cover, and the 38 kDa protein formed separate fragments and granules around them. After nuclear and chromosome decondensation induced by hypotonic treatment, both antibodies stain interphase nuclei in diffuse manner, but in mitotic cells they stained the surface of the swollen chromosomes. The polypeptide with mol. mass of 50 kDa maintained strong connection with chromosome periphery both in norm and under condition of decondensation induced by hypotonic treatment and at subsequent recondensation in isotonic medium. In contrast, the protein with mol. mass of 38 kDa partially lost the contact with a chromosome during recondensation appearing also in the form of granules in
Approaches to Sample Size Determination for Multivariate Data: Applications to PCA and PLS-DA of Omics Data.
Saccenti, Edoardo; Timmerman, Marieke E
Sample size determination is a fundamental step in the design of experiments. Methods for sample size determination are abundant for univariate analysis methods, but scarce in the multivariate case. Omics data are multivariate in nature and are commonly investigated using multivariate statistical methods, such as principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA). No simple approaches to sample size determination exist for PCA and PLS-DA. In this paper we will introduce important concepts and offer strategies for (minimally) required sample size estimation when planning experiments to be analyzed using PCA and/or PLS-DA. PMID:27322847
Hunting for the Tristan plume - An upper mantle tomography around the volcanic island Tristan da Cunha
NASA Astrophysics Data System (ADS)
Schlömer, Antje; Geissler, Wolfram H.; Jokat, Wilfried; Jegen, Marion
Tristan da Cunha is a volcanic island in the South Atlantic close to the Mid-Atlantic Ridge. It is part of an area consisting of widely scattered seamounts and small islands at the western and youngest end of the aseismic Walvis Ridge. Tristan da Cunha together with the Walvis Ridge represents the classical example of a mantle plume track, because of the connection to the Cretaceous Etendeka flood basalt province in NW Namibia. The genesis of the island has so far remained enigmatic. It is hotly debated, if Tristan da Cunha sits actually above a deep mantle plume or if it is only originated by upwelling material from weak (leaky) fracture zones. It also has to be clarified if there are any indications for a plume-ridge interaction. Geochemical investigations have shown complex compositions of magmatic samples from Tristan da Cunha, which could be interpreted as a mixing of plume-derived melts and depleted upper mantle sources. To improve our understanding about the origin of Tristan and to test the mantle plume hypothesis, we deployed 24 broadband ocean-bottom seismometers and 2 seismological land stations around and on the island during an expedition in January 2012 with the German research vessel Maria S. Merian. After acquiring continuous seismological data for almost one year, the seismometers were recovered in early January 2013. We cross-correlated the arrival times of teleseismic P and PKP phases to perform a finite-frequency tomography of the upper mantle beneath the study area. Here we show the 3D mantle structure in terms of velocity variations: We do not image a "classical" plume-like structure directly beneath Tristan da Cunha, but we observe regions of low velocities at the edges of our array that we relate to local mantle upwelling from potentially deeper sources. Additionally we discuss local seismicity within the Tristan da Cunha region, which show processes along the nearby mid-ocean ridge and transform faults. Furthermore, the local seismicity
Estudo de não gaussianidade nas anisotropias da RCF medidas Wmap
NASA Astrophysics Data System (ADS)
Andrade, A. P. A.; Wuensche, C. A.; Ribeiro, A. L. B.
A investigação do campo de flutuações da Radiação Cósmica de Fundo (RCF) pode oferecer um importante teste para os modelos cosmológicos que descrevem a origem e a evolução das flutuações primordiais. De um lado, apresenta-se o modelo inflacionário que prevê um espectro de flutuações adiabáticas distribuídas segundo uma gaussiana e, de outro, os modelos de defeitos topológicos (dentre outros) que descrevem um mecanismo para a geração de flutuações de isocurvatura que obedecem a uma distribuição não gaussiana. Este trabalho tem como objetivo caracterizar traços do modelo não gaussiano de campo misto (entre flutuações adiabáticas e de isocurvatura) nos mapas do Wilkinson Microwave Anisotropy Probe (WMAP). Simulações das anisotropias da RCF no contexto de mistura indicam traços marcantes na distribuição das flutuações de temperatura, mesmo quando consideradas pequenas contribuições do campo de isocurvatura (da ordem de 0.001). O efeito da mistura entre os campos resulta na transferência de potência de flutuações em escalas angulares intermediárias para flutuações em pequenas escalas angulares. Este efeito pode ser caracterizado pela relação entre as amplitudes dos primeiros picos acústicos no espectro de potência da RCF. Neste trabalho, investigamos a contribuição do campo de isocurvatura, no contexto de mistura, sobre as observações recentes da RCF realizadas pelo WMAP. As previsões do modelo de campo misto, uma vez confrontadas com as observações em pequenas escalas angulares, podem ajudar a revelar a natureza das flutuações primordiais.
O Imperio como Argumento: Um Contraponto entre Joaquim Nabuco e o Bispo D. Jose Mauricio da Rocha (The Empire as Argument: A Counterpoint between Joaquim Nabuco and the Bishop Dom Jose Mauricio da Rocha).
ERIC Educational Resources Information Center
Marson, Izabel Andrade
Dialogs with the previous article establishing a comparative analysis between the political acceptance present in the reactionary speech of Dom Jose Mauricio da Rocha and in the monarchist words of Joaquim Nabuco. (PA)
Iron abundance in the hot DA white dwarfs Feige 24 and G191 B2B
NASA Technical Reports Server (NTRS)
Vennes, Stephane; Chayer, Pierre; Thorstensen, John R.; Bowyer, Stuart; Shipman, Harry L.
Attention is given to model calculations of the far- and extreme-UV line spectra of highly ionized Fe species (Fe IV, Fe V, and Fe VI) for hot high-gravity H-rich stars. A spectral analysis of 31 hr of exposure of the DA white dwarf Feige 24 with IUE in the echelle mode reveals the presence of Fe with an abundance relative to H by number of (5-10) x 10 exp -6 with an uncertainty dominated by the determination of stellar parameters. An analysis of IUE data from the white dwarf G191 B2B results in a similar Fe abundance if this star shares similar atmospheric parameters (Teff, g) with Feige 24. Fe is thus the second most abundant photospheric element in hot DA white dwarfs.
Crosslinking of hemin to a specific site on the 90-kDa ferritin repressor protein
Lin, Jihjing; Thach, R.E. ); Patino, M.M.; Gaffield, L.; Walden. W.E. ); Smith, A. )
Incubation of a 90-kDa ferritin repressor protein (FRP) with small amounts of radiolabeled hemin resulted in the formation of a strong interaction between the two that was stable to SDS/PAGE. Of seven other proteins tested individually, only apohemopexin and bovine serum albumin showed similar crosslinking ability, albeit to a much lower extent. ({sup 14}C)Hemin specifically crosslinked to FRP in the presence of a 50-fold excess of total wheat germ proteins. Inclusion of catalase did not prevent the reaction of hemin with FRP, suggesting that H{sub 2}O{sub 2} is not involved. The subsequent addition of a stoichiometric amount of apohemopexin did not reverse the reaction. Exhaustive digestion of the complex with Staphylococcus aureus V8 protease produced a major labeled peptide of 17 kDa. These results show the existence of a highly specific, uniquely reactive hemin binding site on FRP.
[Freud's identification with men who had 2 mothers: Oedipus, Leonardo da Vinci, Michealangelo and Moses].
Harsch, H E
In view of the fact that as a child Sigmund Freud was looked after by two mothers--his actual mother and a nursemaid--it is hardly surprising that traces of this pre-oedipal situation, fraught as it was with traumatisation and loss, should be discernible in the works of the creator of psychoanalysis. Freud's continued preoccupation with the Oedipus myth, his interest in "great men" like da Vinci and Michelangelo, and finally his identification with the figure of Moses are pointers not only to the paternal dimension (as long suggested by Freud's biographers) but also to the maternal dimension and its significance for Freud's life and work. The author demonstrates that those mythical and historical figures which Freud identified with--Oedipus, da Vinci, Michelangelo, Moses--themselves all had two mothers and sublimated this traumatic experience into outstanding achievements, the same being true of Freud himself "who solved the famous riddle and was a most powerful man". PMID:8153361
Towards the Implementation of an Autonomous Camera Algorithm on the da Vinci Platform.
Eslamian, Shahab; Reisner, Luke A; King, Brady W; Pandya, Abhilash K
Camera positioning is critical for all telerobotic surgical systems. Inadequate visualization of the remote site can lead to serious errors that can jeopardize the patient. An autonomous camera algorithm has been developed on a medical robot (da Vinci) simulator. It is found to be robust in key scenarios of operation. This system behaves with predictable and expected actions for the camera arm with respect to the tool positions. The implementation of this system is described herein. The simulation closely models the methodology needed to implement autonomous camera control in a real hardware system. The camera control algorithm follows three rules: (1) keep the view centered on the tools, (2) keep the zoom level optimized such that the tools never leave the field of view, and (3) avoid unnecessary movement of the camera that may distract/disorient the surgeon. Our future work will apply this algorithm to the real da Vinci hardware. PMID:27046563
PSI-O, a new 10-kDa subunit of eukaryotic photosystem I.
Knoetzel, Jürgen; Mant, Alexandra; Haldrup, Anna; Jensen, Poul Erik; Scheller, Henrik Vibe
A novel polypeptide with an apparent molecular mass of 9 kDa was detected after sodium dodecyl sulphate-polyacrylamide gel electrophoresis of Arabidopsis photosystem I (PSI) and was N-terminally sequenced. Corresponding cDNA clones encode a precursor protein of 140 amino acid residues which was imported into isolated intact chloroplasts and processed to the mature protein, designated PSI-O. The mature protein has two transmembrane helices and a calculated mass of 10104 Da. The PSI-O protein was also shown to be present in PSI isolated from barley and spinach, and was essentially absent in chloroplast grana. Expressed sequences encoding similar proteins are available from many species of plants and green algae. PMID:11801243
A 32-kDa protein associated with phospholipase A2-inhibitory activity from human placenta.
Hayashi, H; Owada, M K; Sonobe, S; Kakunaga, T; Kawakatsu, H; Yano, J
Two monomeric 32-kDa proteins, termed 32K-I (pI 5.8) and 32K-II (pI 5.1), were isolated from human placenta, which was solubilized by a Ca2+-chelator. Only 32K-I was associated with PLA2-inhibitory activity. CNBr peptide mapping indicated that 32K-I was distinct from 32K-II and two 36-kDa proteins, called calpactin I and II or lipocortin II and I, which have been shown to possess PLA2-inhibitory activity. 32K-I bound to PS in a Ca2+-dependent manner. 32K-I was detected in many tissues except brain, cardiac and skeletal muscle. PMID:3666152
Solid state of a new PDE-5 inhibitor DA-8159: characterization, dissolution, transformation.
Ryu, Ji Yoon; Sohn, Young-Taek
The polymorphic forms of a new PDE-5 inhibitor DA-8159 were prepared and characterized by differential scanning calorimetry (DSC), powder X-ray diffractometry (PXRD) and thermogravimetric analysis (TG). Two crystal forms and one amorphous form of DA-8159 have been isolated by recrystallization and characterized by DSC, TG and PXRD. From the TG data it was confirmed that two crystal forms are neither solvates nor hydrates. The PXRD patterns of the two crystal forms were different. In the dissolution studies in simulated intestinal fluid at 37 ± 0.5°C, the solubility decreased in the order of amorphous form > Form 1 > Form 2. After storage of 60 days, Form 1 was transformed to Form 2. Form 2 was not transformed. The amorphous form was transformed to Form 2 at 52% R.H. and 95% R.H., but it did not transform at 0% R.H. PMID:22644853
Maximizing use of robot-arm no. 3 in daVinci-assisted thoracic surgery.
Kajiwara, Naohiro; Maeda, Junichi; Yoshida, Koichi; Kato, Yasufumi; Hagiwara, Masaru; Kakihana, Masatoshi; Ohira, Tatsuo; Kawate, Norihiko; Ikeda, Norihiko
We have previously reported on the importance of appropriate robot-arm settings and replacement of instrument ports in robot-assisted thoracic surgery, because the thoracic cavity requires a large space to access all lesions in various areas of the thoracic cavity from the apex to the diaphragm and mediastinum and the chest wall. (1 - 3) Moreover, it can be difficult to manipulate the da Vinci Surgical System using only arms No. 1 and No. 2 depending on the tumor location. However, arm No. 3 is usually positioned on the same side as arm No. 2, and sometimes it is only used as an assisting-arm to avoid conflict with other arms ( Fig. 1 ). In this report, we show how robot-arm No. 3 can be used with maximum effectiveness in da Vinci-assisted thoracic surgery. [Figure: see text]. PMID:26011219
[Da Vinci robot-assisted thoracoscopy for primary hyperparathyroidism: a new application in endocrine surgery].
Brunaud, L; Ayav, A; Bresler, L; Schjött, B
Primary hyperparathyroidism is a relatively frequent disease whose incidence is often underestimated. It is caused by one or more hyperfunctioning parathyroid glands. Almost all pathologic glands (hyperplasia or adenoma) are located in the neck but 1-2% may be located in the mediastinum and may require a sternotomy or thoracotomy approach for resection. The thoracoscopic approach was proposed in 1994 and the DaVinci robotic system allows performance of this thoracoscopic procedure with the benefits of 3-dimensional vision, and improved surgical dexterity and ergonomics. We report a case of a patient with a parathyroid adenoma located in the aorto-pulmonary window which was resected using a left thoracoscopic approach aided by the Da Vinci robotic system. PMID:18645560
Visual degradation in Leonardo da Vinci's iconic self-portrait: A nanoscale study
NASA Astrophysics Data System (ADS)
Conte, A. Mosca; Pulci, O.; Misiti, M. C.; Lojewska, J.; Teodonio, L.; Violante, C.; Missori, M.
The discoloration of ancient paper, due to the development of oxidized groups acting as chromophores in its chief component, cellulose, is responsible for severe visual degradation in ancient artifacts. By adopting a non-destructive approach based on the combination of optical reflectance measurements and time-dependent density functional theory ab-initio calculations, we describe and quantify the chromophores affecting Leonardo da Vinci's iconic self-portrait. Their relative concentrations are very similar to those measured in modern and ancient samples aged in humid environments. This analysis quantifies the present level of optical degradation of the Leonardo da Vinci's self-portrait which, compared with future measurements, will assess its degradation rate. This is a fundamental information in order to plan appropriate conservation strategies.
First experience DaTSCAN imaging using cadmium-zinc-telluride gamma camera SPECT.
Farid, Karim; Queneau, Mathieu; Guernou, Mohamed; Lussato, David; Poullias, Xavier; Petras, Slavomir; Caillat-Vigneron, Nadine; Songy, Bernard
We report our first experience of brain DaTSCAN SPECT imaging using cadmium-zinc-telluride gamma camera (CZT-GC) in 2 cases: a 64-year-old patient suffering from essential tremor and a 73-year-old patient presenting with atypical bilateral extrapyramidal syndrome. In both cases, 2 different acquisitions were performed and compared, using a double-head Anger-GC, followed immediately by a second acquisition on CZT-GC. There were no significant visual differences between images generated by different GC. Our first result suggests that DaTSCAN SPECT is feasible on CZT-GC, allowing both injected dose and acquisition time reductions without compromising image quality. This experience needs to be evaluated in larger series. PMID:22785531
Development of noSQL data storage for the ATLAS PanDA Monitoring System
NASA Astrophysics Data System (ADS)
Potekhin, M.; ATLAS Collaboration
For several years the PanDA Workload Management System has been the basis for distributed production and analysis for the ATLAS experiment at the LHC. Since the start of data taking PanDA usage has ramped up steadily, typically exceeding 500k completed jobs/day by June 2011. The associated monitoring data volume has been rising as well, to levels that present a new set of challenges in the areas of database scalability and monitoring system performance and efficiency. These challenges are being met with a R&D effort aimed at implementing a scalable and efficient monitoring data storage based on a noSQL solution (Cassandra). We present our motivations for using this technology, as well as data design and the techniques used for efficient indexing of the data. We also discuss the hardware requirements as they were determined by testing with actual data and realistic loads.
On the interpretation of the Exosat photometry of two hot DA white dwarfs
NASA Technical Reports Server (NTRS)
Vennes, Stephane; Shipman, Harry L.; Petre, Robert
Exosat photometric observations of the hot DA white dwarfs GD 2 and EG 70 are presented and analyzed using model atmosphere and synthetic spectrum calculations. The numerical accuracy of the calculations and their impact on the reliability of the results are addressed. The optimum atmospheric parameters that explain the complete spectral coverage are derived. The consequences of these conjectures for the spectral evolution theory of white dwarf stars are considered.
76 FR 19903 - Special Conditions: Diamond Aircraft Industry Model DA-40NG; Diesel Cycle Engine
Federal Register 2010, 2011, 2012, 2013, 2014
...These special conditions are issued for the Diamond Aircraft Industry (DAI) GmbH model DA-40NG the Austro Engine GmbH model E4 aircraft diesel engine (ADE) using turbine (jet) fuel. This airplane will have a novel or unusual design feature(s) associated with the installation of a diesel cycle engine utilizing turbine (jet) fuel. The applicable airworthiness regulations do not contain adequate......
Hirabayashi, Masatoshi; Scheeres, Daniel J.
Rozitis et al. recently reported that near-Earth asteroid (29075) 1950 DA, whose bulk density ranges from 1.0 g cm{sup –3} to 2.4 g cm{sup –3}, is a rubble pile and requires a cohesive strength of at least 44-76 Pa to keep from failing due to its fast spin period. Since their technique for giving failure conditions required the averaged stress over the whole volume, it discarded information about the asteroid's failure mode and internal stress condition. This paper develops a finite element model and revisits the stress and failure analysis of 1950 DA. For the modeling, we do not consider material hardening and softening. Under the assumption of an associated flow rule and uniform material distribution, we identify the deformation process of 1950 DA when its constant cohesion reaches the lowest value that keeps its current shape. The results show that to avoid structural failure the internal core requires a cohesive strength of at least 75-85 Pa. It suggests that for the failure mode of this body, the internal core first fails structurally, followed by the surface region. This implies that if cohesion is constant over the whole volume, the equatorial ridge of 1950 DA results from a material flow going outward along the equatorial plane in the internal core, but not from a landslide as has been hypothesized. This has additional implications for the likely density of the interior of the body.
Purification and properties of an oestrogen-stimulated mouse uterine glycoprotein (approx. 70 kDa).
Teng, C T; Walker, M P; Bhattacharyya, S N; Klapper, D G; DiAugustine, R P; McLachlan, J A
An oestrogen-induced secretory protein from mouse uterine luminal fluid was purified by CM-Affi-Gel Blue chromatography and reverse-phase h.p.l.c. This protein has an apparent molecular mass of approx. 70 kDa both by SDS/polyacrylamide-gel electrophoresis (with or without 2-mercaptoethanol) and by gel-filtration column chromatography, indicating that it exists as a single-chain polypeptide. Further analysis of the protein revealed that it is highly basic (pI greater than or equal to 10) and is a glycoprotein. The N-terminus appears to be blocked to Edman degradation. The partial amino acid sequence of a fragment was obtained by cleavage with CNBr; no sequence homology was apparent between the analysed fragment and other known sequences. The incorporation of [35S]methionine into uterine proteins in vitro revealed that oestrogen treatment of immature mice stimulates both synthesis and secretion of the 70 kDa protein. An enzyme-linked immunosorbent assay with polyclonal antibody was used to determine the tissue distribution of the protein. Tissues such as lung, brain, spleen, muscle, intestine, liver, kidney and ovary of oestrogen-treated mice did not have detectable amounts of the 70 kDa protein. Immunoreactivity was present in uterine and vaginal tissues from oestrogen-treated animals. The 70 kDa protein was not induced by testosterone or progesterone. Although the function of this protein is unknown, it is useful as a marker for the study of oestrogen action in the mammalian uterus as well as regulation of gene expression at the molecular level. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 6. PMID:3814091
The Handedness of Leonardo da Vinci: A Tale of the Complexities of Lateralisation
ERIC Educational Resources Information Center
McManus, I. C.; Drury, Helena
The handedness of Leonardo da Vinci is controversial. Although there is little doubt that many of his well-attributed drawings were drawn with the left hand, the hatch marks of the shading going downwards from left to right, it is not clear that he was a natural left-hander, there being some suggestion that he may have become left-handed as the…
Diagnoses behind patients with hard-to-classify tremor and normal DaT-SPECT: a clinical follow up study
Menéndez-González, Manuel; Tavares, Francisco; Zeidan, Nahla; Salas-Pacheco, José M.; Arias-Carrión, Oscar
The [123I]ioflupane—a dopamine transporter radioligand—SPECT (DaT-SPECT) has proven to be useful in the differential diagnosis of tremor. Here, we investigate the diagnoses behind patients with hard-to-classify tremor and normal DaT-SPECT. Therefore, 30 patients with tremor and normal DaT-SPECT were followed up for 2 years. In 18 cases we were able to make a diagnosis. The residual 12 patients underwent a second DaT-SPECT, were then followed for additional 12 months and thereafter the diagnosis was reconsidered again. The final diagnoses included cases of essential tremor, dystonic tremor, multisystem atrophy, vascular parkinsonism, progressive supranuclear palsy, corticobasal degeneration, fragile X–associated tremor ataxia syndrome, psychogenic parkinsonism, iatrogenic parkinsonism and Parkinson's disease. However, for 6 patients the diagnosis remained uncertain. Larger series are needed to better establish the relative frequency of the different conditions behind these cases. PMID:24744729
Laparoscopically assisted total daVinci aorto bifemoral graft bypass with a unique system of graft delivery.
Martinez, B D; George, P M; Ameer, A; Gerhardinger, A M; Moses, J T; Hansen, J M
We report the first known example of a fully robotic aortobifemoral graft and aortic endarterectomy using the daVinci Surgical System and a unique method for graft delivery using the Endo-Vein Harvester. PMID:18657387
Diagnoses behind patients with hard-to-classify tremor and normal DaT-SPECT: a clinical follow up study.
Menéndez-González, Manuel; Tavares, Francisco; Zeidan, Nahla; Salas-Pacheco, José M; Arias-Carrión, Oscar
The [(123)I]ioflupane-a dopamine transporter radioligand-SPECT (DaT-SPECT) has proven to be useful in the differential diagnosis of tremor. Here, we investigate the diagnoses behind patients with hard-to-classify tremor and normal DaT-SPECT. Therefore, 30 patients with tremor and normal DaT-SPECT were followed up for 2 years. In 18 cases we were able to make a diagnosis. The residual 12 patients underwent a second DaT-SPECT, were then followed for additional 12 months and thereafter the diagnosis was reconsidered again. The final diagnoses included cases of essential tremor, dystonic tremor, multisystem atrophy, vascular parkinsonism, progressive supranuclear palsy, corticobasal degeneration, fragile X-associated tremor ataxia syndrome, psychogenic parkinsonism, iatrogenic parkinsonism and Parkinson's disease. However, for 6 patients the diagnosis remained uncertain. Larger series are needed to better establish the relative frequency of the different conditions behind these cases. PMID:24744729
Fully digital controlled A.C. servo engraving machine based on DEC4DA
NASA Astrophysics Data System (ADS)
Shu, Zhibing; Chen, Xianfeng; Zhang, Hairong; Huang, Yiqun; Yan, Caizhong
A novel engraving machine (NUT-1A) is presented, in which fully digital controlled AC system based on DEC4DA was used to improve the machining precision and sensitivity. This engraving machine was constructed around AC servo motor with encoder, controlled by a servo motor control card - DEC4DA. As the upper unit of AC servo motor, DEC4DA was a numerical control generator, which received pulses form CPU by ISA bus, and these pulses were amplified and converted to drive AC servo actuator. This novel engraving machine can achieve a higher positioning accuracy of +/-0.01mm and positioning repetition of +/-0.005mm, and its resolution is 0.001mm/0.0001mm. Moreover, because of multi-closed loops were used in the system, the steady and transient performances are more excellent. This system ensures a much quicker current regulation in closed-loop operation, of acceleration and braking in both directions, as well as stable speed characteristics. Amplifier boards are protected against excessive current, excessive temperature and short circuiting of the motor supply cables.
A 115 kDa calmodulin-binding protein is located in rat liver endosome fractions.
Enrich, C; Bachs, O; Evans, W H
The distribution of calmodulin-binding polypeptides in various rat liver subcellular fractions was investigated. Plasma-membrane, endosome, Golgi and lysosome fractions were prepared by established procedures. The calmodulin-binding polypeptides present in the subcellular fractions were identified by using an overlay technique after transfer from gels to nitrocellulose sheets. Distinctive populations of calmodulin-binding polypeptides were present in all the fractions examined except lysosomes. A major 115 kDa calmodulin-binding polypeptide of pI 4.3 was located to the endosome subfractions, and it emerges as a candidate endosome-specific protein. Partitioning of endosome fractions between aqueous and Triton X-114 phases indicated that the calmodulin-binding polypeptide was hydrophobic. Major calmodulin-binding polypeptides of 140 and 240 kDa and minor polypeptides of 40-60 kDa were present in plasma membranes. The distribution of calmodulin in the various endosome and plasma-membrane fractions was also analysed, and the results indicated that the amounts were high compared with those in the cytosol. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. PMID:3214436
The validity of the autism spectrum disorders-diagnosis for intellectually disabled adults (ASD-DA).
Matson, Johnny L; Wilkins, Jonathan; Boisjoli, Jessica A; Smith, Kimberly R
Psychologists interviewed direct-care staff using a battery of assessment measures including the autism spectrum disorders-diagnosis for intellectually disabled adults (ASD-DA), the Diagnostic Assessment for the Severely Handicapped-II (DASH-II), the Matson Evaluation of Social Skills for Individuals with Severe Retardation (MESSIER), the Socialization domain of the Vineland Adaptive Behavior Scales (VABS), and a checklist containing criteria for autism and PDD-NOS from the DSM-IV-TR and ICD-10. Three hundred and seven intellectually disabled (ID) adolescents and adults ranging in age from 16 to 88 were assessed. Participants were diagnosed with either ID and ASD (autism or PDD-NOS; n=156) or ID and no Axis I diagnosis (n=151). A modification of the multitrait-multimethod approach was used to establish the convergent and discriminant validity of the ASD-DA. The scale proved to have robust convergent validity when correlated with the DSM-IV-TR/ICD-10 checklist, MESSIER, and Socialization domain of the VABS. Additionally, discriminant validity was demonstrated by comparing the ASD-DA to items from the DASH-II (measure of general psychopathology). The implications of these data are discussed. PMID:17981435
Petrology of potassic alkaline ultramafic and carbonatitic rocks from Planalto da Serra (Mato Grosso State), Brazil
NASA Astrophysics Data System (ADS)
Comin-Chiaramonti, Piero; Gomes, Celso B.; De Min, Angelo; Ruberti, Excelso; Girardi, Vicente A. V.; Slejko, Francesca; Neder, Renato D.; Pinho, Francisco E. C.
The Planalto da Serra igneous rocks form plugs, necks and dykes of carbonate-rich ultramafic lamprophyres (aillikites and glimmerites with kamafugitic affinity) and carbonatites (alvikites and beforsites). Phlogopite and/or tetraphlogopite, diopside and melanitic garnet are restricted to aillikitic rock-types, whereas pyroclore occurs only in carbonatites. Aillikites and carbonatites are altered to hydrotermalites, having chlorite and serpentine as dominant minerals. Planalto da Serra igneous rock association has kamafugitic affinity (i.e. effusive, ultrapotassic. High LREE/HREE fractionation, incompatible elements data and Sr-Nd isotopes, suggest that the K-ultramafic alkaline and carbonatite rocks originated from a variably metasomatized mantle source enriched in radiogenic Sr. Crustal contamination is negligible or absent. Age values of 600 Ma rule out the geochronological relationship between the investigated intrusions and the Mesozoic alkaline bodies from the Azimuth 125° lineament. The TDM model ages allow to conclude that Planalto da Serra magma is derived from the partial melting of a mantle source metasomatised by K-rich carbonatated melt during the Early to Late Neoproterozoic. On the basis of alkaline magmatism repetitions at 600 Ma and 90-80 Ma we question the subsistence of a stationary mantle plume for so long time.
da Vinci robot-assisted keyhole neurosurgery: a cadaver study on feasibility and safety.
Marcus, Hani J; Hughes-Hallett, Archie; Cundy, Thomas P; Yang, Guang-Zhong; Darzi, Ara; Nandi, Dipankar
The goal of this cadaver study was to evaluate the feasibility and safety of da Vinci robot-assisted keyhole neurosurgery. Several keyhole craniotomies were fashioned including supraorbital subfrontal, retrosigmoid and supracerebellar infratentorial. In each case, a simple durotomy was performed, and the flap was retracted. The da Vinci surgical system was then used to perform arachnoid dissection towards the deep-seated intracranial cisterns. It was not possible to simultaneously pass the 12-mm endoscope and instruments through the keyhole craniotomy in any of the approaches performed, limiting visualization. The articulated instruments provided greater dexterity than existing tools, but the instrument arms could not be placed in parallel through the keyhole craniotomy and, therefore, could not be advanced to the deep cisterns without significant clashing. The da Vinci console offered considerable ergonomic advantages over the existing operating room arrangement, allowing the operating surgeon to remain non-sterile and seated comfortably throughout the procedure. However, the lack of haptic feedback was a notable limitation. In conclusion, while robotic platforms have the potential to greatly enhance the performance of transcranial approaches, there is strong justification for research into next-generation robots, better suited to keyhole neurosurgery. PMID:25516094
New Insights into the Problem of the Surface Gravity Distribution of Cool DA White Dwarfs
NASA Astrophysics Data System (ADS)
Tremblay, P.-E.; Bergeron, P.; Kalirai, J. S.; Gianninas, A.
We review at length the longstanding problem in the spectroscopic analysis of cool hydrogen-line (DA) white dwarfs (T eff< 13,000 K) where gravities are significantly higher than those found in hotter DA stars. The first solution that has been proposed for this problem is a mild and systematic helium contamination from convective mixing that would mimic the high gravities. We constrain this scenario by determining the helium abundances in six cool DA white dwarfs using high-resolution spectra from the Keck I 10 m telescope. We obtain no detections, with upper limits as low as He/H = 0.04 in some cases. This allows us to put this scenario to rest for good. We also extend our model grid to lower temperatures using improved Stark profiles with non-ideal gas effects from Tremblay & Bergeron and find that the gravity distribution of cool objects remains suspiciously high. Finally, we find that photometric masses are, on average, in agreement with expected values, and that the high-log g problem is so far unique to the spectroscopic approach.
Tremblay, P.-E.; Bergeron, P.; Gianninas, A.; Kalirai, J. S. E-mail: firstname.lastname@example.com E-mail: firstname.lastname@example.com
We review at length the longstanding problem in the spectroscopic analysis of cool hydrogen-line (DA) white dwarfs (T{sub eff}< 13,000 K) where gravities are significantly higher than those found in hotter DA stars. The first solution that has been proposed for this problem is a mild and systematic helium contamination from convective mixing that would mimic the high gravities. We constrain this scenario by determining the helium abundances in six cool DA white dwarfs using high-resolution spectra from the Keck I 10 m telescope. We obtain no detections, with upper limits as low as He/H = 0.04 in some cases. This allows us to put this scenario to rest for good. We also extend our model grid to lower temperatures using improved Stark profiles with non-ideal gas effects from Tremblay and Bergeron and find that the gravity distribution of cool objects remains suspiciously high. Finally, we find that photometric masses are, on average, in agreement with expected values, and that the high-log g problem is so far unique to the spectroscopic approach.
Shark cartilage 14 kDa protein as a dendritic cells activator.
Safari, Elahe; Hassan, Zuhair M; Moazzeni, Seyed Mohammad
Low molecular weight components of shark cartilage are reported to have anti-tumor as well as immuno-stimulating effects. Dendritic cells (DCs) are potent antigen-presenting cells (APCs) that have a key role in establishment of anti-cancer immune response. In this study, the effect of 14 kDa protein from shark cartilage was investigated on stimulation and maturation of dendritic cells. The isolated 14 kDa protein from shark cartilage extract was added to DCs medium during overnight culture and their maturation and T cells stimulation potential was investigated. The majority of shark-cartilage-treated DCs expressed higher levels of maturation markers and were more effective in stimulation of allogenic T cells compared with non-treated DCs (p < 0.05). Our results showed that shark cartilage 14 kDa protein can potentially be used in DC-mediated T-cells stimulation and induction of desirable immune responses in clinical trials such as cancer immunotherapy. However, further studies are required to examine this proposal. PMID:25669314
OnDA: online data analysis and feedback for serial X-ray imaging1
Mariani, Valerio; Morgan, Andrew; Yoon, Chun Hong; Lane, Thomas J.; White, Thomas A.; O'Grady, Christopher; Kuhn, Manuela; Aplin, Steve; Koglin, Jason; Barty, Anton; Chapman, Henry N.
This article describes a free and open-source data analysis utility designed for fast online feedback during serial X-ray diffraction and scattering experiments: OnDA (online data analysis). Three complete real-time monitors for common types of serial X-ray imaging experiments are presented. These monitors are capable of providing the essential information required for quick decision making in the face of extreme rates of data collection. In addition, a set of modules, functions and algorithms that allow developers to modify the provided monitors or develop new ones are provided. The emphasis here is on simple, modular and scalable code that is based on open-source libraries and protocols. OnDA monitors have already proven to be invaluable tools in several experiments, especially for scoring and monitoring of diffraction data during serial crystallography experiments at both free-electron laser and synchrotron facilities. It is felt that in the future the kind of fast feedback that OnDA monitors provide will help researchers to deal with the expected very high throughput data flow at next-generation facilities such as the European X-ray free-electron laser. PMID:27275150
Albumin interacts specifically with a 60-kDa microvascular endothelial glycoprotein.
Schnitzer, J E; Carley, W W; Palade, G E
Confluent monolayers of microvascular endothelial cells, derived from the rat epididymal fat pad and grown in culture, were radioiodinated by using the lactoper-oxidase method. Their radioiodinated surface polypeptides were detected by NaDodSO4/PAGE (followed by autoradiography) and were characterized by both lectin affinity chromatography and protease digestion to identify the proteins involved in albumin binding. All detected polypeptides were sensitive to Pronase digestion, whereas several polypeptides were resistant to trypsin. Pronase treatment of the cell monolayer significantly reduced the specific binding of radioiodinated rat serum albumin, but trypsin digestion did not. Limax flavus, Ricinus communis, and Triticum vulgaris agglutinins competed significantly with radioiodinated rat serum albumin binding, whereas other lectins did not. A single 60-kDa glyco-protein was precipitated in common by these three lectins and was trypsin-resistant and Pronase-sensitive. Rat serum albumin affinity chromatography columns weakly but specifically bound a 60-kDa polypeptide from cell lysates derived from radioiodinated cell monolayers. These findings indicate that the 60-kDa glycoprotein is directly involved in a specific interaction of albumin with the cultured microvascular endothelial cells used in these experiments. Images PMID:3413125
32 CFR Appendix C to Part 623 - Agreement for Loan of US Army Materiel (DA Form 4881-R)
Code of Federal Regulations, 2011 CFR
... 32 National Defense 3 2011-07-01 2009-07-01 true Agreement for Loan of US Army Materiel (DA Form... ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. C Appendix C to Part 623—Agreement for Loan of US Army Materiel (DA Form 4881-R) EC24OC91.013 EC24OC91.014 EC24OC91.015 EC24OC91.016...
32 CFR Appendix C to Part 623 - Agreement for Loan of US Army Materiel (DA Form 4881-R)
Code of Federal Regulations, 2010 CFR
... 32 National Defense 3 2010-07-01 2010-07-01 true Agreement for Loan of US Army Materiel (DA Form... ARMY SUPPLIES AND EQUIPMENT LOAN OF ARMY MATERIEL Pt. 623, App. C Appendix C to Part 623—Agreement for Loan of US Army Materiel (DA Form 4881-R) EC24OC91.013 EC24OC91.014 EC24OC91.015 EC24OC91.016...
Evaluation of the immunomodulatory effect of the 14 kDa protein isolated from aged garlic extract on dendritic cells.
Ahmadabad, Hasan Namdar; Hassan, Zuhair Mohammad; Safari, Elahe; Bozorgmehr, Mahmood; Ghazanfari, Tooba; Moazzeni, Seyed Mohammad
Garlic is used all over the world for treatment of different diseases. A wide range of biological activities of garlic has been verified in vitro and in vivo. One of major proteins of garlic which has been isolated and purified is the 14 kDa protein. This protein has been shown to have immunomodulatory effects. In this study, the effect of the 14 kDa protein isolated from aged garlic extract (AGE) was investigated on maturation and immunomodulatory activity of dendritic cells (DC). Proteins were purified from AGE by biochemical method; the semi-purified 14 kDa protein was run on gel filtration Sephadex G50 and its purity was checked by SDS-PAGE. DC were isolated from spleen of BALB/c mice by Nycodenz centrifugation and their adhesiveness to plastic dish. 14 kDa protein from AGE was added to overnight culture of DC medium and the expression percentage of CD40, CD86, and MHC-II was evaluated by flowcytometric analysis. Also, proliferation of T-cells was measured by allogenic mixed lymphocyte reaction (MLR) test. The purified 14 kDa protein isolated from AGE increased the expression of CD40 molecule on DC, but it did not influence CD86 and MHCII molecules. Furthermore, no significant differences were noticed in the pulsed-DC with 14 kDa protein and non-pulsed DC on the MLR. PMID:21570063
Histopathological effects and determination of the putative receptor of Bacillus thuringiensis Cry1Da toxin in Spodoptera littoralis midgut.
BenFarhat-Touzri, Dalel; Saadaoui, Marwa; Abdelkefi-Mesrati, Lobna; Saadaoui, Imen; Azzouz, Hichem; Tounsi, Slim
Bacillus thuringiensis subsp. aizawai strain HD133, known by its effectiveness against Spodoptera species, produces many insecticidal proteins including Cry1Ab, Cry1Ca and Cry1Da. In the present study, the insecticidal activity of Cry1Da against Spodoptera littoralis was investigated. It showed toxicity with an LC(50) of 224.4 ng/cm(2) with 95% confidence limits of (178.61-270.19) and an LC(90) of 467.77 ng/cm(2) with 95% confidence limits of (392.89-542.65). The midgut histopathology of Cry1Da fed larvae showed vesicle formation in the apical region, vacuolization and destruction of epithelial cells. Biotinylated-activated Cry1Da toxin bound protein of about 65 kDa on blots of S. littoralis brush border membrane preparations. This putative receptor differs in molecular size from those recognized by Cry1C and Vip3A which are active against this polyphagous insect. This difference in midgut receptors strongly supports the use of Cry1Da as insecticidal agent, particularly in case of Cry and/or Vip-resistance management. PMID:23220238
Tremblay, P.-E.; Bergeron, P.; Gianninas, A. E-mail: firstname.lastname@example.com
We present an improved spectroscopic and photometric analysis of hydrogen-line DA white dwarfs from the Sloan Digital Sky Survey Data Release 4 (SDSS DR4) based on model atmospheres that include improved Stark broadening profiles with non-ideal gas effects. We also perform a careful visual inspection of all spectroscopic fits with high signal-to-noise ratios (S/Ns > 12) and present improved atmospheric parameters (T{sub eff} and log g) for each white dwarf. Through a comparison of spectroscopic and photometric temperatures, we report the discovery of 35 DA+DB/DC double degenerate candidates and two helium-rich DA stars. We also determine that a cutoff at S/N = 15 optimizes the size and quality of the sample for computing the mean mass of DA white dwarfs, for which we report a value of 0.613 M{sub sun}. We compare our results to previous analyses of the SDSS DR4 and find a good agreement if we account for the shift produced by the improved Stark profiles. Finally, the properties of DA white dwarfs in the SDSS are weighed against those of the Villanova White Dwarf Catalog sample of Gianninas et al. We find systematically lower masses (by about 3% on average), a difference that we trace back to the data reduction procedure of the SDSS. We conclude that a better understanding of these differences will be important to determine the absolute temperature scale and mean mass of DA white dwarfs.
Ubiquitous expression of the 43- and 44-kDa forms of transcription factor USF in mammalian cells.
Sirito, M; Lin, Q; Maity, T; Sawadogo, M
USF is a helix-loop-helix transcription factor that, like Myc, recognizes the DNA binding motif CACGTG. Two different forms of USF, characterized by apparent molecular weights of 43,000 and 44,000, were originally identified in HeLa cells by biochemical analysis. Clones for the 43-kDa USF were first characterized, but only partial clones for the human 44-kDa USF (USF2, or FIP) have been reported. Here we describe a complete cDNA for the 44-kDa USF from murine cells. Analysis of this clone has revealed that the various USF family members are quite divergent in their N-terminal amino acid sequences, while a high degree of conservation characterizes their dimerization and DNA-binding domains. Interestingly, the 3' noncoding region of the 44-kDa USF cDNAs displayed an unusual degree of conservation between human and mouse. In vitro transcription/translation experiments indicated a possible role for this region in translation regulation. Alternative splicing forms of the 44-kDa USF messages exist in both mouse and human. Examination of the tissue and cell-type distribution of USF by Northern blot and gel retardation assays revealed that while expression of both the 43- and 44-kDa USF species is ubiquitous, different ratios of USF homo- and heterodimers are found in different cells. Images PMID:8127680
The human 64-kDa polyadenylylation factor contains a ribonucleoprotein-type RNA binding domain and unusual auxiliary motifs.
Takagaki, Y; MacDonald, C C; Shenk, T; Manley, J L
Cleavage stimulation factor is one of the multiple factors required for 3'-end cleavage of mammalian pre-mRNAs. We have shown previously that this factor is composed of three subunits with estimated molecular masses of 77, 64, and 50 kDa and that the 64-kDa subunit can be UV-crosslinked to RNA in a polyadenylylation signal (AAUAAA)-dependent manner. We have now isolated cDNAs encoding the 64-kDa subunit of human cleavage stimulation factor. The 64-kDa subunit contains a ribonucleoprotein-type RNA binding domain in the N-terminal region and a repeat structure in the C-terminal region in which a pentapeptide sequence (consensus MEARA/G) is repeated 12 times and the formation of a long alpha-helix stabilized by salt bridges is predicted. An approximately 270-amino acid segment surrounding this repeat structure is highly enriched in proline and glycine residues (approximately 20% for each). When cloned 64-kDa subunit was expressed in Escherichia coli, an N-terminal fragment containing the RNA binding domain bound to RNAs in a polyadenylylation-signal-independent manner, suggesting that the RNA binding domain is directly involved in the binding of the 64-kDa subunit to pre-mRNAs. Images PMID:1741396
Identification and purification of a novel 120-kDa protein that recognizes the cAMP-responsive element
Andrisani, O.; Dixon, J.E. )
The TGACGTCA (CRE) motif required for function by a number of cellular (somatostatin, enkephalin, alpha-human chorionic gonadotropin) and viral (Ad5 E1A-inducible, HTLV-1 TAX-inducible) genes is the site of interaction of multiple sequence-specific complexes. A protocol has been developed for the fractionation and purification of these activities. We report here the purification from HeLa nuclear extracts of a novel 120-kDa polypeptide which by Southwestern blots, gel retardation, and UV cross-linking assays displays CRE-specific binding. The CRE-affinity purified 120-kDa protein displays properties distinct from those of the 43-kDa CREB/ATF polypeptide. The 120-kDa protein is readily phosphorylated in vitro by protein kinase C but not by protein kinase A, suggesting that this molecule may mediate cellular signals distinct from the cAMP-responsive pathway. In vitro transcription-complementation assays utilizing the purified 120-kDa protein failed to transactivate the cAMP-responsive somatostatin promoter suggesting that the mode of action of this 120-kDa polypeptide may require an activation step distinct from the cAMP-signaling pathway.
Atmospheric parameters of a soft X ray selected set of hot DA white dwarfs
NASA Astrophysics Data System (ADS)
Kidder, Kenneth Mark
Soft x-ray/EUV observations provide a sensitive means of probing the structure and composition of the atmospheres of hot hydrogen-rich (DA) white dwarf stars. This is due to the fact that soft x-rays originate from hotter, deeper layers of the photosphere than do longer wavelengths. A primary aim of this research is to expand the sample of DA's observed with soft x-ray instrumentation through a search for serendipitous observations of catalogued hot DA white dwarfs in existing soft x-ray databases. The positional coincidences of 31 catalogued DA's (McCook and Sion 1987) were examined in Einstein IPC and EXOSAT LE fields. As a result, three soft x-ray sources have been identified corresponding to the white dwarfs (WD0631 + 107, WD1113 + 413 and WD0425 + 168) and a fourth source which is probably not due to photospheric thermal emission from the coincident white dwarf. The three detected DA's have relatively low effective temperatures (27,200 +/- 400 K, 26,200 +/- 1100 K and 24,000 +/- 500 K, respectively), as determined independently using complementary optical and UV spectroscopy. Applying these temperature constraints to the soft x-ray photometric data, the photospheres of these stars must be composed of effectively pure hydrogen (n(He)/n(H) less than 10 exp -5), although the hottest star, WD0631 + 107, can have trace levels of homogeneously mixed helium less than 10 exp -4.2. The analysis of the soft x-ray observations are supplemented with independent determinations of temperature, gravity, and V magnitude. The method of determination employed is to fit the well-sampled wings of the broad hydrogen absorption profiles with self-consistent model atmosphere predictions. Precise temperatures and gravities are obtained by fitting the observed profiles independently, then jointly determining a consistent solution for each object. In this effort, a ground-based observation program was established to obtain high quality spectra of the Balmer delta and gamma lines and in
OpenDA Open Source Generic Data Assimilation Environment and its Application in Process Models
NASA Astrophysics Data System (ADS)
El Serafy, Ghada; Verlaan, Martin; Hummel, Stef; Weerts, Albrecht; Dhondia, Juzer
Data Assimilation techniques are essential elements in state-of-the-art development of models and their optimization with data in the field of groundwater, surface water and soil systems. They are essential tools in calibration of complex modelling systems and improvement of model forecasts. The OpenDA is a new and generic open source data assimilation environment for application to a choice of physical process models, applied to case dependent domains. OpenDA was introduced recently when the developers of Costa, an open-source TU Delft project [http://www.costapse.org; Van Velzen and Verlaan; 2007] and those of the DATools from the former WL|Delft Hydraulics [El Serafy et al 2007; Weerts et al. 2009] decided to join forces. OpenDA makes use of a set of interfaces that describe the interaction between models, observations and data assimilation algorithms. It focuses on flexible applications in portable systems for modelling geophysical processes. It provides a generic interfacing protocol that allows combination of the implemented data assimilation techniques with, in principle, any time-stepping model duscribing a process(atmospheric processes, 3D circulation, 2D water level, sea surface temperature, soil systems, groundwater etc.). Presently, OpenDA features filtering techniques and calibration techniques. The presentation will give an overview of the OpenDA and the results of some of its practical applications. Application of data assimilation in portable operational forecasting systems—the DATools assimilation environment, El Serafy G.Y., H. Gerritsen, S. Hummel, A. H. Weerts, A.E. Mynett and M. Tanaka (2007), Journal of Ocean Dynamics, DOI 10.1007/s10236-007-0124-3, pp.485-499. COSTA a problem solving environment for data assimilation applied for hydrodynamical modelling, Van Velzen and Verlaan (2007), Meteorologische Zeitschrift, Volume 16, Number 6, December 2007 , pp. 777-793(17). Application of generic data assimilation tools (DATools) for flood
The Near-Earth Flyby of Asteroid 2012 DA14 (Invited)
NASA Astrophysics Data System (ADS)
Moskovitz, N. A.; Endicott, T.; Lister, T.; Ryan, B.; Ryan, E. V.; Willman, M.; Hergenrother, C.; Binzel, R. P.; Polishook, D.; DeMeo, F.; Bennechi, S.; Sheppard, S.; Marchis, F.; Angusteijn, T.; Birthwhistle, P.; Verveer, A.; Gulbis, A.; Nagayama, T.; Gilmore, A.; Kilmartin, P.
On UT 2013 February 15, the elongated 20x40 meter near-Earth asteroid 2012 DA14 made a close flyby of the Earth at a distance of approximately 27,000 km or 4 Earth-radii. This passage was inside the orbital distance of geosynchronous satellites and close to the Earth's Roche limit. This close encounter was unprecedented in our advance knowledge of the incoming trajectory, thereby allowing us to forge a detailed coordinated plan for observation and testing of physical models predicting tidal effects near our planet's Roche limit. Predicted outcomes of such close encounters include tidal disruption, changes in rotation state, and mass loss or mass redistribution due to seismic shaking. Each of these effects sensitively depend on how tidal energy is dissipated in the interior of the asteroid: a rubble pile will dissipate tidal energy differently than a solid or partially fractured monolith. The internal properties of asteroids in general and particularly those in the size range of 2012 DA14 are largely unknown. This encounter represents one of the first ever opportunities to observationally test models of the interior structure of small planetary bodies. We will present the results of a large multi-observatory campaign focused on 2012 DA14. This campaign includes data from shortly after discovery in 2012 and surrounding the close approach of the asteroid in February of 2013. These data were collected with over a dozen telescopes worldwide, ranging in aperture from 1m to 8m and include major facilities in Chile, Australia, New Zealand, South Africa, Israel, Hawaii, Arizona, New Mexico, and the Canary Islands. We will highlight the outcome of pre- and post-flyby visible-wavelength photometric and spectroscopic observations. This will include clear evidence that 2012 DA14 is now in a non-principal axis rotation state, requiring a multi-periodic tumbling model to fit its post-flyby rotational light curve. Such a complex rotation state is consistent with models for the
The Greek Database of Seismogenic Sources (GreDaSS): the new version.
NASA Astrophysics Data System (ADS)
Caputo, Riccardo; Pavlides, Spyros; GreDaSS Working Group
The Greek Database of Seismogenic Sources (GreDaSS) is a repository of geological, tectonic and active fault data for the Greek territory and its surroundings including sectors of western Anatolia, southern Bulgaria, FYR of Macedonia and Albania. The principal aim of this international project is to create a homogenized framework of all data relevant to the seismotectonics, and especially the seismic hazard assessment, of Greece and its surroundings as well as a common research platform for performing seismic hazard analyses, modelling and scenarios from specific seismogenic structures. GreDaSS represents an on-going project based on the results of decades of investigations by the GreDaSS Working Group* and by numerous other researchers working on the active tectonics of the broader Aegean Region. Members of the WG have contributed in several national and European projects like FAUST (Faults as a seismological tool, 1998-2000) and SHARE (Seismic Hazard and Armonization in Europe, 2009-2012). The database is continuously updated by the WG, with the aim of improving its completeness in terms of seismogenic sources analysed and included, but also of enriching and better defining all parametric and ancillary information on each source. Preliminary and partial versions of GreDaSS have been already presented at previous congresses (Pavlides et al., 2010; Caputo et al., 2012; Sboras et al., 2013), but at next EGU the whole area and the most recent version of the database will be displayed. *) the principal scientific contributors belonging to the GreDaSS WG are: Sboras. S., Chatzipetros A., Koukouvelas I., Michailidou A., Valkaniotis S., Tarabusi G. Caputo R., Chatzipetros A., Pavlides S. and Sboras S. (2012): Ann. Geophysics, 55(5), 859-894, doi: 10.4401/ag-5168 Pavlides S., Caputo R., Sboras S., Chatzipetros A., Papathanasiou G. & Valkaniotis S. (2010): Bull. Geol. Soc. Greece, 43(1), 486-494 Sboras S., Pavlides S., Caputo R., Chatzipetros A., Michailidou A
Rapid purification and direct microassay of calbindin9kDa utilizing its solubility in perchloric acid.
Hubbard, M J
The 9 kDa calcium-binding protein, calbindin9kDa, was found to be soluble in 7% (v/v) perchloric acid. Calbindin9kDa was easily purified from rat duodenum in 1 day with perchloric acid precipitation followed by reverse-phase h.p.l.c. The yield was 21.4 +/- 2.3 nmol/g wet weight of tissue (mean +/- S.E.M.; n = 3) from normally fed 7-8-week-old rats (approx. 70% recovery). The purification was also effective with rabbit duodenum calbindin9kDa, but not with various other EF-hand calcium-binding proteins tested in the rat. Several criteria (h.p.l.c., u.v. spectrum, denaturing two-dimensional PAGE, N-terminal sequencing) indicated that the rat calbindin9kDa was purified to homogeneity and was not affected by proteolysis. High-affinity calcium-binding properties were retained and no evidence of isoforms or charge modification was observed. Residue 59, identified as Asn (not Asp as previously reported), was fully amidated. When adopted as a microassay with isocratic h.p.l.c., the perchloric acid procedure enabled rapid (less than 6 min) and direct (peptide bond absorbance) quantification of less than 1 pmol of calbindin9kDa. This new approach to purification and assay will be of particular utility for investigations of calbindin9kDa in previously intractable low-abundance sources (e.g. cultured cells). Images Figure 1 Figure 2 PMID:8392333
15 kDa Granulysin versus GM-CSF for monocytes differentiation: analogies and differences at the transcriptome level
Background Granulysin is an antimicrobial and proinflammatory protein with several isoforms. While the 9 kDa isoform is a well described cytolytic molecule with pro-inflammatory activity, the functions of the 15 kDa isoform is less well understood. Recently it was shown that 15 kDa Granulysin can act as an alarmin that is able to activate monocytes and immature dendritic cells. Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) is a growth factor widely used in immunotherapy both for in vivo and ex vivo applications, especially for its proliferative effects. Methods We analyzed gene expression profiles of monocytes cultured with 15 kDa Granulysin or GM-CSF for 4, 12, 24 and 48 hours to unravel both similarities and differences between the effects of these stimulators. Results The analysis revealed a common signature induced by both factors at each time point, but over time, a more specific signature for each factor became evident. At all time points, 15 kDa Granulysin induced immune response, chemotaxis and cell adhesion genes. In addition, only 15 kDa Granulsyin induced the activation of pathways related to fundamental dendritic cell functions, such as co-stimulation of T-cell activation and Th1 development. GM-CSF specifically down-regulated genes related to cell cycle arrest and the immune response. More specifically, cytokine production, lymphocyte mediated immunity and humoral immune response were down-regulated at late time points. Conclusion This study provides important insights on the effects of a novel agent, 15 kDa granulysin, that holds promise for therapeutic applications aimed at the activation of the immune response. PMID:21501511
Synergetic effects of DA-6/GA₃ with EDTA on plant growth, extraction and detoxification of Cd by Lolium perenne.
He, Shanying; Wu, Qiuling; He, Zhenli
Research is needed to improve efficiency of phytoextraction of heavy metals from contaminated soils. A pot experiment was carried out to study the effects of plant growth regulators (PGRs) (diethyl aminoethyl hexanoate (C18H33NO8, DA-6) and gibberellic acid 3 (C19H22O6, GA3)) and/or EDTA on Cd extraction, subcellular distribution and chemical forms in Lolium perenne. The addition of EDTA or PGRs significantly enhanced Cd extraction efficiency (P<0.05), with the decreasing order of: 1 μM DA-6>10 μM DA-6>10 μM GA3>2.5 mmol kg(-1) EDTA>other treatments of PGR alone. PGRs+EDTA resulted in a further increase in Cd extraction efficiency, with EDTA+1 μM DA-6 being the most efficient. At the subcellular level, about 44-57% of Cd was soluble fraction, 18-44% in cell walls, and 12-25% in cellular organelles fraction. Chemical speciation analysis showed that 40-54% of Cd was NaCl extractable, 7-23% HAc extractable, followed by other fractions. EDTA increased the proportions of Cd in soluble and cellular organelles fraction, as well as the metal migration in shoot; therefore, the toxicity to plant increased and plant growth was inhibited. Conversely, PGRs fixed more Cd in cell walls and reduced Cd migration in shoot; thus, metal toxicity was reduced. In addition, PGRs promoted plant biomass growth significantly (P<0.05), with 1 μM DA-6 being the most effective. A combination of DA-6/GA3 with EDTA can alleviate the adverse effect of EDTA on plant growth, and the treatment of EDTA+1 μM DA-6 appears to be optimal for improving the remediation efficiency of L. perenne for Cd contaminated soil. PMID:24999226
Sequence and expression of an Eisenia-fetida-derived cDNA clone that encodes the 40-kDa fetidin antibacterial protein.
Lassegues, M; Milochau, A; Doignon, F; Du Pasquier, L; Valembois, P
Fetidins are 40-kDa and 45-kDa hemolytic and antibacterial glycoproteins present in the coelomic fluid of the earthworm Eisenia fetida andrei. By screening a cDNA library with a polyclonal antifetidin serum, we have cloned a cDNA that encoded the 40-kDa fetidin. The clone contains an insert of 1.44 kb encoding a protein of 34 kDa, which corresponds to the size of deglycosylated fetidins. The recombinant protein inhibits Bacillus megaterium growth. Restriction fragment polymorphisms were observed on Southern blots and correspond to a known protein polymorphism. The sequence of the cDNA contains a peroxidase signature and fetidins from earthworm coelomic fluid have peroxidase activity. The 40-kDa and 45-kDa fetidins therefore represent two related polymorphic defence factors in invertebrates. PMID:9219536
11 CFR 111.15 - Motions to quash or modify a subpoena (2 U.S.C. 437d(a) (3), (4)).
Code of Federal Regulations, 2012 CFR
... 11 Federal Elections 1 2012-01-01 2012-01-01 false Motions to quash or modify a subpoena (2 U.S.C. 437d(a) (3), (4)). 111.15 Section 111.15 Federal Elections FEDERAL ELECTION COMMISSION GENERAL COMPLIANCE PROCEDURE (2 U.S.C. 437g, 437d(a)) Enforcement § 111.15 Motions to quash or modify a subpoena (2 U.S.C. 437d(a) (3), (4)). (a) Any person...
11 CFR 111.15 - Motions to quash or modify a subpoena (2 U.S.C. 437d(a) (3), (4)).
Code of Federal Regulations, 2013 CFR
... 11 Federal Elections 1 2013-01-01 2012-01-01 true Motions to quash or modify a subpoena (2 U.S.C. 437d(a) (3), (4)). 111.15 Section 111.15 Federal Elections FEDERAL ELECTION COMMISSION GENERAL COMPLIANCE PROCEDURE (2 U.S.C. 437g, 437d(a)) Enforcement § 111.15 Motions to quash or modify a subpoena (2 U.S.C. 437d(a) (3), (4)). (a) Any person...
Evolution of DA white dwarfs in the context of a new theory of convection
NASA Astrophysics Data System (ADS)
Althaus, L. G.; Benvenuto, O. G.
In this study we compute the structure and evolution of carbon-oxygen DA (hydrogen-rich envelope) white dwarf models by means of a detailed and updated evolutionary code. We consider models with masses from 0.5 to 1.0Msolar and we vary the hydrogen layer mass in the interval 10^-13<=M_H/M≮10^-4. In particular, we treat the energy transport by convection within the formalism of the full-spectrum turbulence theory, as given by the Canuto, Goldman & Mazzitelli (CGM) model. We explore the effect of various hydrogen layer masses on both the surface gravity and the hydrogen burning. Convective mixing at low luminosities is also considered. One of our main interests in this work has been to study the evolution of ZZ Ceti models, with the aim of comparing the CGM and mixing-length theory (MLT) predictions. In this connection, we find that the temperature profile given by the CGM model is markedly different from that of the ML1 and ML2 versions of the MLT. In addition, the evolving outer convection zone behaves differently in both theories. We have also computed approximate effective temperatures for the theoretical blue edge of the DA instability strip by using thermal time-scale arguments for our evolving DA models. In this context, we found that the CGM theory leads to blue edges that are cooler than the observed ones. However, because the determination of atmospheric parameters of ZZ Ceti stars is dependent on the assumed convection description in model atmosphere calculations, observed blue edges based on model atmospheres computed considering the CGM theory are required in order to perform a self-consistent comparison of our results with observations. Finally, detailed non-adiabatic pulsational computations of ZZ Ceti models considering the CGM convection would be necessary to place the results found in this paper on a firmer basis.
[The Vitruvian Man: an anatomical drawing for proportions by Leonardo Da Vinci].
Le Floch-Prigent, P
The aim of the study was to find out and to analyse the text by Vitruvius which inspired the famous drawing by Leonardo da Vinci (circa 1490) kept in the Galleria dell'Accademia, in Venezia, Italy: the man inscribed in one circle and in one square. The book "de Architectura" by Vitruvius Marcus Pollio was printed several times since the Renaissance when both the roman architecture of antiquity and this text became very popular. From a French translation by Claude Perrault in 1864, it became easy to find a French translation with the original text in Latin (Paris, 2003, Les Belles Lettres, French text by Pierre Gros). The drawing by Leonardo da Vinci illustrates with great accuracy and fidelity the quotation of Vitruvius (with the exception of two of the 12 main relationships). The genius of Leonardo da Vinci was to keep only one trunk, head and neck for two pairs of limbs: scapular and pelvic; to make the circle tangent to the lower edge of the square; to adjust a few features of the quotation for the equilibrium of the whole figure; and of course to bring his incredible skill as a drawer (one of the best of his century). The drawing was made on a sheet of paper 344x245mm, in black ink which became dark brown with time; several lines complete the figure above and below; a short caption and a horizontal scale appear just under the drawing. The celebrity of the drawing, a symbol of the Renaissance, of the equilibrium of man and mankind, of the universality of the artists and intellectuals of the time (Humanism) made it iconic and it has been constantly reproduced and adapted especially for advertisement and logos, not only in the medical field. PMID:18951824
Structural requirements to obtain highly potent and selective 18 kDa Translocator Protein (TSPO) Ligands.
Taliani, Sabrina; Pugliesi, Isabella; Da Settimo, Federico
The (18 kDa) Translocator Protein (TSPO), was initially identified in 1977 as peripheral binding site for the benzodiazepine diazepam and named "Peripheral-type benzodiazepine receptor (PBR)". It is an evolutionarily well-conserved protein particularly located at the outer/inner mitochondrial membrane contact sites, in closely association with the 32 kDa voltage-dependent anion channel (VDAC) and the 30 kDa adenine nucleotide translocase (ANT), thus forming the mitochondrial permeability transition pore (MPTP). TSPO is ubiquitary expressed in peripheral tissues (steroid producing tissues, liver, heart, kidney, lung, immune system) and in lower levels in the central nervous system, where it is mainly located in glial cells, and in neurons. TSPO is involved in a variety of biological processes such as cholesterol transport, steroidogenesis, calcium homeostasis, lipid metabolism, mitochondrial oxidation, cell growth and differentiation, apoptosis induction, and regulation of immune functions. In the last decade, many studies have reported that TSPO basal expression is altered in a number of human pathologies, such as cancer and neurodegenerative disorders (Huntington's and Alzheimer's diseases), as well as in various forms of brain injury and inflammation and anxiety. Consequently, TSPO has not only been suggested as a promising drug target for a number of therapeutic applications (anticonvulsant, anxiolytic, immunomodulating, etc.), but also as valid diagnostic marker for related-disease state and progression, prompting the development of specific labelled ligands as powerful tools for imaging techniques. A number of structurally different classes of ligands have been reported, showing high affinity and selectivity towards TSPO. Indeed, most of these ligands have been designed starting from selective CBR ligands which were structurally modified in order to shift their affinity towards TSPO. Extensive structure-activity relationship studies were performed allowing to
Análise da medição do raio solar em ultravioleta
NASA Astrophysics Data System (ADS)
Saraiva, A. C. V.; Giménez de Castro, C. G.; Costa, J. E. R.; Selhorst, C. L.; Simões, P. J. A.
A medição acurada do raio solar em qualquer banda do espectro eletromagnético é de relevância na formulação e calibração de modelos da estrutura e atmosfera solar. Esses modelos atribuem emissão do contínuo do Sol calmo em microondas à mesma região da linha Ha do Hell. Apresentamos a medição do raio solar em UV com imagens do EIT (Extreme Ultraviolet Image Telescope) entre 1996 e 2002, no comprimento de onda 30,9 nm (Ha do Hell), que se forma na região de transição/cromosfera solar. A técnica utilizada para o cálculo do raio UV foi baseada na transformada Wavelet B3spline. Fizemos um banco de dados com 1 imagem por dia durante o período citado. Obtivemos como resultado o raio médio da ordem de 975.61" e uma diminuição do mesmo para o período citado variando em média -0,45" /ano. Comparamos estes dados com os valores obtidos pelo ROI (Radio Observatório de Itapetinga) em 22/48 GHz e Nobeyama Radio Heliograph em 17 GHz mostrando que os raios médios são muito próximos o que indica que a região de formação nessas freqüências é a mesma conforme os modelos. Comparamos os resultados também com outros índices de atividade solar.
Crystal structure of the 500-kDa yeast acetyl-CoA carboxylase holoenzyme dimer.
Wei, Jia; Tong, Liang
Acetyl-CoA carboxylase (ACC) has crucial roles in fatty acid metabolism and is an attractive target for drug discovery against diabetes, cancer and other diseases. Saccharomyces cerevisiae ACC (ScACC) is crucial for the production of very-long-chain fatty acids and the maintenance of the nuclear envelope. ACC contains biotin carboxylase (BC) and carboxyltransferase (CT) activities, and its biotin is linked covalently to the biotin carboxyl carrier protein (BCCP). Most eukaryotic ACCs are 250-kilodalton (kDa), multi-domain enzymes and function as homodimers and higher oligomers. They contain a unique, 80-kDa central region that shares no homology with other proteins. Although the structures of the BC, CT and BCCP domains and other biotin-dependent carboxylase holoenzymes are known, there is currently no structural information on the ACC holoenzyme. Here we report the crystal structure of the full-length, 500-kDa holoenzyme dimer of ScACC. The structure is remarkably different from that of the other biotin-dependent carboxylases. The central region contains five domains and is important for positioning the BC and CT domains for catalysis. The structure unexpectedly reveals a dimer of the BC domain and extensive conformational differences compared to the structure of the BC domain alone, which is a monomer. These structural changes reveal why the BC domain alone is catalytically inactive and define the molecular mechanism for the inhibition of eukaryotic ACC by the natural product soraphen A and by phosphorylation of a Ser residue just before the BC domain core in mammalian ACC. The BC and CT active sites are separated by 80 Å, and the entire BCCP domain must translocate during catalysis. PMID:26458104
Quadrupole Ion Mass Spectrometer for Masses of 2 to 50 Da
NASA Technical Reports Server (NTRS)
Helms, William; Griffin, Timothy P.; Ottens, Andrew; Harrison, Willard
A customized quadrupole ion-trap mass spectrometer (QITMS) has been built to satisfy a need for a compact, rugged instrument for measuring small concentrations of hydrogen, helium, oxygen, and argon in a nitrogen atmosphere. This QITMS can also be used to perform quantitative analyses of other gases within its molecular-mass range, which is 2 to 50 daltons (Da). (More precisely, it can be used to perform quantitative analysis of gases that, when ionized, are characterized by m/Z ratios between 2 and 50, where m is the mass of an ion in daltons and Z is the number of fundamental electric charges on the ion.
The strainrange partitioning behavior of an advanced gas turbine disk alloy, AF2-1DA
NASA Technical Reports Server (NTRS)
Halford, G. R.; Nachtigall, A. J.
The low-cycle, creep-fatigue characteristics of the advanced gas turbine disk alloy, AF2-1DA have been determined at 1400 F and are presented in terms of the method of strainrange partitioning (SRP). The mean stresses which develop in the PC and CP type SRP cycles at the lowest inelastic strainrange were observed to influence the cyclic lives to a greater extent than the creep effects and hence interfered with a conventional interpretation of the results by SRP. A procedure is proposed for dealing with the mean stress effects on life which is compatible with SRP.
Study of the hadronic interactions of kaons in light nuclei at DA{Phi}ne
Vazquez Doce, O.
The AMADEUS experiment at the Da{Phi}ne accelerator of the Frascati National Laboratories (Italy) of INFN, will perform, for the first time, full-acceptance studies of antikaon interaction in light nuclei, with a complete experimental program for the case of the kaonic clusters. Studying the absorption of antikaon by the nucleus will provide information concerning the K-barN interaction and the modification of the kaon mass in the nuclear medium.A preliminar study of these kind of hadronic interactions is being done by the AMADEUS collaboration by analyzing the existent KLOE data.
The magmatic source of the Tristan da Cunha hotspot: Implication from electrical conductivity
NASA Astrophysics Data System (ADS)
Chen, Jin; Baba, Kiyoshi; Jegen, Marion; Utada, Hisashi; Geissler, Wolfram; Jokat, Wilfried
Tristan da Cunha Island is one of the classical hot spots in the Atlantic Ocean, situated at the western end of the aseismic Walvis Ridge which forms a connection to the Cretaceous Etendeka flood basalt province in northwestern Namibia. The discussion about its source (in shallow asthenosphere or deeper mantle) have not reached consensus yet because of lack of the geophysical observations in the area. A marine magnetotelluric (MT) experiment was conducted together with seismological observations in the area in 2012-2013 through a German-Japanese collaboration with the goal to constrain the physical state of the mantle beneath the area. A total of 26 MT seafloor stations were deployed around the Tristan da Cunha Islands and available data were retrieved and processed from 24 stations. We applied iterative topographic effect correction and one-dimensional (1-D) conductivity structure inversion to the data. Then, three-dimensional (3-D) inversion analysis incorporating the topographic effect was carried out, using the 1-D model as the initial model. The local small-scale topography and the far continental coast effects are incorporated as the distortion term in the 3-D inversion. The preliminary result of our analysis shows no evidence of a significant conductive anomaly arising from the mantle transition zone, suggesting that the current magmatic source (major place of melting) of the hotspot activity is in the shallow upper mantle. This is in contrast to results from geochemical analysis, in which samples along the Tristan track exhibit an ocean-island-basalt-type incompatible element pattern pointing to a deep mantle source of the melt. Our findings therefore might indicate that the deep mantle up-welling underneath Tristan da Cunha Islands may be almost dead. A conductive anomaly at approx. 100 km depth in our derived conductivity model to the southwest of Tristan da Cunha Islands suggests an interaction between the mid-ocean ridge and/or up-welling further south
Middle Paleolithic human remains from the Gruta da Oliveira (Torres Novas), Portugal.
Trinkaus, Erik; Maki, Julia; Zilhão, João
Ongoing excavations in the Middle Paleolithic levels at the Gruta da Oliveira, Portugal have yielded four fragmentary human remains, a manual phalanx and an ulna from levels 9 and 10, and a humerus and a tibia from levels 18 and 19. The first two remains date to approximately 39 ka 14C BP ( approximately 43.5 ka cal BP), whereas the latter two derive from earlier in oxygen isotope stage 3. The preserved portions of the phalanx, humerus, and tibia align them morphologically with the Neandertals. In addition, the Oliveira 4 tibial diaphysis shows evidence of carnivore (probably canid) gnawing. PMID:17632802
Implementação de um algoritmo para a limpeza de mapas da RCFM
NASA Astrophysics Data System (ADS)
Souza, C. L.; Wuensche, C. A.
A Radiação Cósmica de Fundo em Microondas (RCFM), descoberta por Penzias e Wilson em 1965, é uma das ferramentas mais poderosas para o estudo da cosmologia. Com a descoberta de flutuações de temperatura na RCFM, da ordem de uma parte em 105, pelo COBE (1992), uma nova era teve início. Nos últimos onze anos, diversos instrumentos fizeram novas medidas de alta precisão, refinando os resultados apresentados pelo COBE, culminando com os resultados recentes do satélite WMAP. A análise de dados da RCFM, especialmente no caso de experimentos com pequena cobertura do céu, apresenta uma série de dificuldades devido a emissões de contaminantes externos, tais como a emissão da Galáxia e de fontes pontuais, e de ruídos intrínsecos tanto ao sistema de detecção quanto à estratégia de observação do céu. Uma das soluções típicas para a filtragem de dados brutos de um experimento para medir flutuações de temperatura é aplicar um gabarito (template) e um filtro passa alta ao produzir mapas simplificados (sem considerar matrizes de correlação ou covariância). No caso de experimentos que utilizam detectores HEMT, essa combinação de filtros remove, satisfatoriamente, ruídos do tipo 1/f gerados pela instabilidade no ganho do detector acoplado ao movimento do instrumento, definido pela estratégia de observação. Entretanto, o sinal resultante medido, tanto em simulações quanto em séries temporais reais, sugere que parte do sinal cosmológico pode estar sendo removido junto com o ruído dos detectores. Este trabalho descreve as etapas para a produção de um mapa típico (simulado) e os testes preliminares de um algoritmo para remover ruídos do tipo 1/f introduzidos pela estratégia de observação sem prejudicar a qualidade do sinal cosmológico presente no mapa.
[Examples of textual research on gynecological literature included in Yong le da dian (Yongle's Great Canon)].
Zhang, Xue-dan; Zhang, Ru-qing
Yongle's Great Canon is a series of books to be read exclusively by the emperors of the Ming Dynasty, so all of the books included are from many rare books collected in Wen Yuan Ge of early Ming Dynasty. These rare books, ranging from Song, Jin, and Yuan Dynasties or from folk sources preserve the original appearance of the ancient books almost without any changes. From a philological viewpoint, the gynecological books included in Yong le da dian are studied, proof-read and checked for its lost texts and different texts. This can serve as a basis for the reprinting and re-publishing of ancient TCM books. PMID:21122346
Prospective EUV observations of hot DA white dwarfs with the EUV Explorer
NASA Technical Reports Server (NTRS)
Finley, David S.; Malina, Roger F.; Bowyer, Stuart
The Extreme Ultraviolet Explorer (EUVE) will perform a high sensitivity EUV all-sky survey. A major category of sources which will be detected with the EUVE instruments consists of hot white dwarfs. Detailed preliminary studies of synthetic EUV observations of white dwarfs have been carried out using the predicted EUVE instrumental response functions. Using available information regarding space densities of white dwarfs and the distribution of neutral hydrogen in the interstellar medium, the numbers of DA white dwarfs which will be detectable in the different EUV bandpasses have been estimated.
Molluscan mega-hemocyanin: an ancient oxygen carrier tuned by a ~550 kDa polypeptide
Background The allosteric respiratory protein hemocyanin occurs in gastropods as tubular di-, tri- and multimers of a 35 × 18 nm, ring-like decamer with a collar complex at one opening. The decamer comprises five subunit dimers. The subunit, a 400 kDa polypeptide, is a concatenation of eight paralogous functional units. Their exact topology within the quaternary structure has recently been solved by 3D electron microscopy, providing a molecular model of an entire didecamer (two conjoined decamers). Here we study keyhole limpet hemocyanin (KLH2) tridecamers to unravel the exact association mode of the third decamer. Moreover, we introduce and describe a more complex type of hemocyanin tridecamer discovered in fresh/brackish-water cerithioid snails (Leptoxis, Melanoides, Terebralia). Results The "typical" KLH2 tridecamer is partially hollow, whereas the cerithioid tridecamer is almost completely filled with material; it was therefore termed "mega-hemocyanin". In both types, the staggering angle between adjoining decamers is 36°. The cerithioid tridecamer comprises two typical decamers based on the canonical 400 kDa subunit, flanking a central "mega-decamer" composed of ten unique ~550 kDa subunits. The additional ~150 kDa per subunit substantially enlarge the internal collar complex. Preliminary oxygen binding measurements indicate a moderate hemocyanin oxygen affinity in Leptoxis (p50 ~9 mmHg), and a very high affinity in Melanoides (~3 mmHg) and Terebralia (~2 mmHg). Species-specific and individual variation in the proportions of the two subunit types was also observed, leading to differences in the oligomeric states found in the hemolymph. Conclusions In cerithioid hemocyanin tridecamers ("mega-hemocyanin") the collar complex of the central decamer is substantially enlarged and modified. The preliminary O2 binding curves indicate that there are species-specific functional differences in the cerithioid mega-hemocyanins which might reflect different physiological
[Science and nation: romanticism and natural history in the works of E. J. da Silva Maia].
Kury, L
The works of physician and naturalist Emílio Joaquim da Silva Maia (1808-59) can be viewed as a scientific project that discovers Brazil and its inhabitants. Maia's nationalism and his romantic view of nature formed the underpinnings of his scientific theories, especially his studies on zoological geography. He subordinated the issue of the biological specificity of different regions of the world to his era's debates on the construction of Brazil as an independent nation. In his interpretations of European natural history, Maia endeavored to understand Brazilian nature as a specific achievement of the Cosmos, in keeping with Alexander von Humboldt's approach. PMID:16676427
The discontinuity near 1600 A in the spectra of DA white dwarfs
NASA Technical Reports Server (NTRS)
Wegner, G.
Ultraviolet spectroscopic observations of two relatively cool DA white dwarfs, L481 - 60 (= WD 1544 - 37) and BPM 1266 ( = WD 2105 - 82), with the International Ultraviolet Explorer (IUE) satellite show a strong drop in their spectral energy distributions below 1600 A. Published model atmospheres and thier visual spectra suggest that these two stars have effective temperatures in the vicinity of 9,000-10,000 K, and it is proposed that the 1600 A feature could be due to the 342(1S) 3s2(1S) photoionization edge of Mg I.
OCT structural examination of Madonna dei Fusi by Leonardo da Vinci
NASA Astrophysics Data System (ADS)
Targowski, Piotr; Iwanicka, Magdalena; Sylwestrzak, Marcin; Kaszewska, Ewa A.; Frosinini, Cecilia
Madonna dei Fusi (`Madonna of the Yarnwider') is a spectacular example of Italian Renaissance painting, attributed to Leonardo da Vinci. The aim of this study is to give an account of past restoration procedures. The evidence of a former retouching campaign will be presented with cross-sectional images obtained non-invasively with Optical Coherence Tomography (OCT). Specifically, the locations of overpaintings/retouchings with respect to the original paint layer and secondary varnishes will be given. Additionally, the evidence of a former transfer of the pictorial layer to the new canvas support by detecting the presence of its structure incised into paint layer will be shown.
Pietro Anzolino da Eboli and the thermal therapy of renal pathologies.
Mezzogiorno, Antonio; Caruso, Arturo Armone; Lorio, Luigi; Papa, Michele; Esposito, Vincenzo
In the Campi Flegrei area of Campania, a region of Southern Italy, some thermal baths still exist, known for the therapeutical properties of their waters. Utilized since remote ages for the treatment of a variety of pathologies. In "De Balneis Terrae Laboris", an Italian poet of the Middle Ages and scientist, Pietro Anzolino da Eboli, reports every detail about more than thirty different thermal sites in the Campi Flegrei, including all therapeutic effects of their waters. Based on Pietro's manuscript, we report in the current paper all the sites whose waters were - among other indications - also recommended for the treatment of some urinary diseases. PMID:15293539
Purification and characterization of a novel 88 kDa protein from serum and vitreous of patients with Eales' disease.
Sulochana, K N; Rajesh, M; Ramakrishnan, S
Eales' disease is a perivasculitis that affects the peripheral retina of young adults and results in recurrent vitreous hemorrhage. Although increased oxidative stress and decreased antioxidant defense have been reported to be associated with Eales' disease, the exact cause for the disease and its pathogenesis are not known. Here is reported the identification, purification and characterization of a new protein from the serum and vitreous of patients with Eales' disease. This protein was purified using preparative electrophoresis and HPLC. The purified protein had a retention time of 9.2 min in RP HPLC. Its molecular weight as determined by gel permeation chromatography was 88 kDa hence, it was termed as 88 kDa protein. Alcian blue and Schiffs staining revealed 88 kDa protein to be a glycoprotein. Proteins purified from both serum and vitreous exhibited anti lipid peroxidation effect on erythrocyte when added during in vitro assay of thiobarbuteric acid reactive substances (TBARS). In addition to this property the protein also has Fe(2+)sequestering effect. The anti TBARS activity of 88 kDa protein was completely inhibited by 0.1 m M concentration of parachlromercuric benzoate (PCMB) and 5,5' dithiobis(2-nitrobenzoic acid) DTNB. The total thiol content (cysteine) of the purified 88 kDa protein was found to be 8% by mass. Eighty eight kDa protein from both the sources namely vitreous and serum are immunologically identical when studied using polyclonal antibodies raised in goat against purified serum protein. The N terminal sequence of 88 kDa protein by automated Edman's degradation chemistry is A D D P N S L S P S A F A E A L A L L R D S X L A R F V. The protein and DNA data base search revealed no match to 88 kDa protein and hence this was considered as unique protein. Further knowledge on the in vivo function of 88 kDa protein is very important to understand its role in the pathogenesis of Eales' disease. PMID:11825025
Circulating non-22 kDa growth hormone isoforms after a repeated GHRH stimulus in normal subjects.
Coya, R; Algorta, J; Boguszewski, C L; Vela, A; Carlsson, L M S; Aniel-Quiroga, A; Busturia, M A; Martul, P
The aim of this study was to evaluate the proportion of non-22 kDa GH isoforms in relation to total GH concentration after a repeated GHRH stimulus in healthy subjects. We studied 25 normal volunteers (12 males and 13 females, mean age 13.1 years, range 6-35), who received two GHRH bolus (1.5 mug/kg body weight, i.v.) administered separately by an interval of 120 minutes. The proportion of non-22 kDa GH was determined by the 22 kDa GH exclusion assay (GHEA), which is based on immunomagnetic extraction of monomeric and dimeric 22 kDa GH from serum, and quantitation of non-22 kDa GH isoforms using a polyclonal GH assay. Samples were collected at baseline and at 15-30 min intervals up to 240 min for total GH concentration. Non-22 kDa GH isoforms were measured in samples where peak GH after GHRH was observed. Total GH peaked after the first GHRH bolus in all subjects (median 37.2 ng/ml; range: 10.4-94.6). According to GH response to the second GHRH stimulus, the study group was divided in "non-responders" (n=7; 28%), with GH peak levels lower than 10 ng/ml (median GH: 8.7 ng/ml; range 7.3-9.6) and "responders" (n=18; 72%), who showed a GH response greater than 10 ng/ml (median 17 ng/ml; range 10.1-47.0). The median proportion of non-22 kDa GH on the peak of GH secretion after the first GHRH administration was similar in both groups ("responders" median: 8.6%, range 7-10.9%; "non-responders" median: 8.7%, range 6.7-10.3%), independently of the type of response after the second GHRH. In contrast, the median proportion of non-22 kDa GH was greater at time of GH peak after the second GHRH bolus in the "non-responders" (median 11.4%; range 9.1-14.3%) in comparison with the "responders" (median 9.1%; range 6.7-11.9%; p=0.003). A significant negative correlation between the total GH secreted and the percentage of non-22 kDa isoforms was seen in the "non-responders" (p=0.003). These differences in GH response to repeated GHRH stimulation and in the pattern of GH isoforms at GH
Da0324, an inhibitor of nuclear factor-κB activation, demonstrates selective antitumor activity on human gastric cancer cells
Jin, Rong; Xia, Yiqun; Chen, Qiuxiang; Li, Wulan; Chen, Dahui; Ye, Hui; Zhao, Chengguang; Du, Xiaojing; Shi, Dengjian; Wu, Jianzhang; Liang, Guang
Background The transcription factor nuclear factor-κB (NF-κB) is constitutively activated in a variety of human cancers, including gastric cancer. NF-κB inhibitors that selectively kill cancer cells are urgently needed for cancer treatment. Curcumin is a potent inhibitor of NF-κB activation. Unfortunately, the therapeutic potential of curcumin is limited by its relatively low potency and poor cellular bioavailability. In this study, we presented a novel NF-κB inhibitor named Da0324, a synthetic asymmetric mono-carbonyl analog of curcumin. The purpose of this study is to research the expression of NF-κB in gastric cancer and the antitumor activity and mechanism of Da0324 on human gastric cancer cells. Methods The expressions between gastric cancer tissues/cells and normal gastric tissues/cells of NF-κB were evaluated by Western blot. The inhibition viability of compounds on human gastric cancer cell lines SGC-7901, BGC-823, MGC-803, and normal gastric mucosa epithelial cell line GES-1 was assessed with the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. Absorption spectrum method and high-performance liquid chromatography method detected the stability of the compound in vitro. The compound-induced changes of inducible NF-κB activation in the SGC-7901 and BGC-823 cells were examined by Western blot analysis and immunofluorescence methods. The antitumor activity of compound was performed by clonogenic assay, matrigel invasion assay, flow cytometric analysis, Western blot analysis, and Hoechst 33258 staining assay. Results High levels of p65 were found in gastric cancer tissues and cells. Da0324 displayed higher growth inhibition against several types of gastric cancer cell lines and showed relatively low toxicity to GES-1. Moreover, Da0324 was more stable than curcumin in vitro. Western blot analysis and immunofluorescence methods showed that Da0324 blocked NF-κB activation. In addition, Da0324 significantly inhibited tumor proliferation
The Ubiquitin Receptor DA1 Regulates Seed and Organ Size by Modulating the Stability of the Ubiquitin-Specific Protease UBP15/SOD2 in Arabidopsis[W
Du, Liang; Li, Na; Chen, Liangliang; Xu, Yingxiu; Li, Yu; Zhang, Yueying; Li, Chuanyou; Li, Yunhai
Although the control of organ size is a fundamental question in developmental biology, little is known about the genetic and molecular mechanisms that determine the final size of seeds in plants. We previously demonstrated that the ubiquitin receptor DA1 acts synergistically with the E3 ubiquitin ligases DA2 and ENHANCER1 OF DA1 (EOD1)/BIG BROTHER to restrict seed growth in Arabidopsis thaliana. Here, we describe UBIQUITIN-SPECIFIC PROTEASE15 (UBP15), encoded by SUPPRESSOR2 OF DA1 (SOD2), which acts maternally to regulate seed size by promoting cell proliferation in the integuments of ovules and developing seeds. The sod2/ubp15 mutants form small seeds, while overexpression of UBP15 increases seed size of wild-type plants. Genetic analyses indicate that UBP15 functions antagonistically in a common pathway with DA1 to influence seed size, but does so independently of DA2 and EOD1. Further results reveal that DA1 physically associates with UBP15 in vitro and in vivo and modulates the stability of UBP15. Therefore, our findings establish a genetic and molecular framework for the regulation of seed size by four ubiquitin-related proteins DA1, DA2, EOD1, and UBP15 and suggest that they are promising targets for increasing seed size in crops. PMID:24585836
Increased susceptibility of ethanol-treated gastric mucosa to naproxen and its inhibition by DA-9601, an Artemisia asiatica extract
Oh, Tae Young; Ahn, Gook Jun; Choi, Seul Min; Ahn, Byoung Ok; Kim, Won Bae
AIM: To examine the effect of DA-9601, a new gastroprotective agent, on the vulnerability of ethanol-treated rat's stomach to naproxen (NAP). METHODS: Male Sprague-Dawley rats were pretreated with 1 mL of 50% ethanol twice a day for 5 d and then NAP (50 mg/kg) was administered. DA-9601 was administered 1 h before NAP. Four hours after NAP, the rats were killed to examine gross injury index (mm2), histologic change and to determine mucosal levels of malondialdehyde (MDA), prostaglandin E2 (PGE2), glutathione (GSH) and myeloperoxidase (MPO). RESULTS: Pretreatment of ethanol significantly increased NAP-induced gastric lesions, as well as an increase in MDA and MPO. On the contrary, mucosal PGE2 and GSH contents were decreased dramatically by ethanol pretreatment, which were aggravated by NAP. DA-9601 significantly reduced NAP-induced gastric injury grossly and microscopically, regardless of pretreatment with ethanol. DA-9601 preserved, or rather, increased mucosal PGE2 and GSH in NAP-treated rats (P<0.05), with reduction in mucosal MDA and MPO levels. CONCLUSION: These results suggest that repeated alcohol consumption renders gastric mucosa more susceptible to NSAIDs though, at least in part, reduction of endogenous cytoprotectants including PGE2 and GSH, and increase in MPO activation, and that DA-9601, a new gastroprotectant, can reduce the increased vulnerability of ethanol consumers to NSAIDs-induced gastric damage via the mechanism in which PGE2 and GSH are involved. PMID:16437715
Evaluating the potential of marine Bacteriovorax sp. DA5 as a biocontrol agent against vibriosis in Litopenaeus vannamei larvae.
Wen, Chongqing; Xue, Ming; Liang, Huafang; Zhou, Shining
The potential application of Bdellovibrio-and-like organisms (BALOs) for the biocontrol of bacterial diseases has been widely recognized. However, no marine BALOs have been reported for Vibrio-related infections in penaeid shrimp. In the present study, the bacteriolytic ability of the marine Bacteriovorax strain DA5 against Vibrio alginolyticus zouA was examined by cocultivation and electron microscopy, and optimal lysis was observed at 30-35°C and 20-30‰ salinity along with a high multiplicity of infection. Then, we showed that experimentally infected Litopenaeus vannamei larvae exhibited significantly higher survival with incremental DA5 levels. Finally, variation in the bacterial counts and the bacterial community in larval rearing water was investigated after prophylactic application of DA5. The elimination effect of DA5 on vibrios was visible at early time points, whereas only a few non-dominant bacteria, rather than the predominant populations, were affected through analysis of denaturing gradient gel electrophoresis of the 16S rDNA V3 region. Accordingly, the prophylactic and therapeutic efficacies of DA5 on vibriosis associated with L. vannamei could markedly enhance larval survivability, and these results will facilitate the application of marine Bacteriovorax to control vibriosis in shrimp larviculture. PMID:25139659
Autoradiographic localization of dopamine DA1 receptors in rat kidney with ( sup 3 H)Sch 23390
Huo, T.; Healy, D.P. )
Dopamine receptors in the rat kidney were characterized by homogenate binding and in vitro autoradiography using the dopamine 1 (DA1)-selective antagonist ({sup 3}H)Sch 23390. ({sup 3}H)Sch 23390 binding in cortical membrane preparations was saturable, stereoselective, and competed for by DA agonists and antagonists with a rank order of potency consistent with the labeling of the DA1 receptor. ({sup 3}H)Sch 22390 binding was best fit to a two-site model: a high affinity-low density site (KD1 4.9 +/- 1.4 nM, Bmax 1 31.4 +/- 13.8 fmol/mg protein) and a low affinity-high density site (KD2 86.4 +/- 23.9 nM, Bmax 2 848.0 +/- 227.4 fmol/mg protein). In vitro autoradiography indicated that ({sup 3}H)Sch 22390 binding sites were restricted to the cortex. High-resolution autoradiography further indicated that ({sup 3}H)Sch 22390 binding sites were localized primarily on proximal tubules. Glomeruli and other vascular elements were devoid of ({sup 3}H)Sch 23390 binding sites. These results suggest that DA and DA1 agonists may affect sodium excretion by a direct action on proximal tubule sodium reabsorption.
Agarose and Polyacrylamide Gel Electrophoresis Methods for Molecular Mass Analysis of 5–500 kDa Hyaluronan
Bhilocha, Shardul; Amin, Ripal; Pandya, Monika; Yuan, Han; Tank, Mihir; LoBello, Jaclyn; Shytuhina, Anastasia; Wang, Wenlan; Wisniewski, Hans-Georg; de la Motte, Carol; Cowman, Mary K.
Agarose and polyacrylamide gel electrophoresis systems for the molecular mass-dependent separation of hyaluronan (HA) in the size range of approximately 5–500 kDa have been investigated. For agarose-based systems, the suitability of different agarose types, agarose concentrations, and buffers systems were determined. Using chemoenzymatically synthesized HA standards of low polydispersity, the molecular mass range was determined for each gel composition, over which the relationship between HA mobility and logarithm of the molecular mass was linear. Excellent linear calibration was obtained for HA molecular mass as low as approximately 9 kDa in agarose gels. For higher resolution separation, and for extension to molecular masses as low as approximately 5 kDa, gradient polyacrylamide gels were superior. Densitometric scanning of stained gels allowed analysis of the range of molecular masses present in a sample, and calculation of weight-average and number-average values. The methods were validated for polydisperse HA samples with viscosity-average molecular masses of 112, 59, 37, and 22 kDa, at sample loads of 0.5 µg (for polyacrylamide) to 2.5 µg (for agarose). Use of the methods for electrophoretic mobility shift assays was demonstrated for binding of the HA-binding region of aggrecan (recombinant human aggrecan G1-IGD-G2 domains) to a 150 kDa HA standard. PMID:21684248
A estabilidade dos PAHS em função da energia da radiação interestelar nas faixas UV e raios-X
NASA Astrophysics Data System (ADS)
Pinotti, R.; Costa, R. K.; Boechat-Roberty, H. M.; Lago, A.; Souza, G. B.
A nebulosa CRL 618, uma proto-nebulosa planetária cuja nuvem molecular espessa envolve uma estrela B0, contém uma grande quantidade de C2H2 e CH4. Estas moléculas são consideradas os tijolos da criação de grandes moléculas carbonadas como os Hidrocarbonetos Policíclicos Aromáticos (PAHs). Esta nebulosa, por estar exposta a intensos campos de UV e Raios-X, é uma região de fotodissociação molecular que propicia a formação de novas moléculas, confirmada pela presença de C4H2 e C6H6 (Benzeno), que é a unidade básica dos PAHs. Atribui-se a esta família de moléculas orgânicas duas propriedades fundamentais, a resistência para sobreviver ao campo de radiação UV interestelar e a geração das bandas de emissão não identificadas (UIR) observadas no infravermelho. No entanto, alguns autores questionam a resistência dos PAHs ao campo de radiação UV interestelar. Empregando a técnica de Espectrometria de Massas por Tempo de Vôo, no modo de coincidência fotoelétron-fotoíon, estudamos a ionização e fragmentação das seguintes moléculas: Benzeno, Benzeno deuterado, Naftaleno, Antraceno e Fenantreno. Utilizamos uma fonte de Hélio monocromática em 21,21 eV (584,5 Å) e a radiação Síncroton do Laboratório Nacional de Luz Síncroton (LNLS) em diferentes energias nas proximidades da borda do C 1s ( 290 eV). Comprovamos a estabilidade dos PAHs sob ação de UV (21,21 eV), onde eles apresentam um baixo nível de fotodissociação, produzindo fragmentos ionizados com rendimento total na ordem de 5 por cento em relação ao íon molecular pai. Entretanto, em altas energias, na faixa de Raios-X, a quebra destas moléculas torna-se mais intensa, com a produção de muitos fragmentos. Como uma das rotas de fragmentação do Naftaleno é [(C10H8) = > (C6H6+) + (C4H2) + (e-)], e como temos as evidências observacionais da existência do C4H2 e C6H6 na nebulosa CRL 618, sugerimos que este ambiente também possui o Naftaleno.
Divergent regulation by growth factors and cytokines of 95 kDa and 72 kDa gelatinases and tissue inhibitors or metalloproteinases-1, -2, and -3 in rabbit aortic smooth muscle cells.
Fabunmi, R P; Baker, A H; Murray, E J; Booth, R F; Newby, A C
The migration and proliferation of vascular smooth muscle cells (SMCs) during neointima formation in atherosclerosis and angioplasty restenosis is mediated by certain growth factors and cytokines, one action of which may be to promote basement-membrane degradation. To test this hypothesis further, the effects of such growth factors and cytokines on the synthesis of two basement-membrane-degrading metalloproteinases, namely the 72 kDa gelatinase (MMP-2, gelatinase A) and the 95 kDa gelatinase (MMP-9, gelatinase B) and three tissue inhibitors of metalloproteinases (TIMPs) was studied in primary cultured rabbit aortic SMCs. Expression of the 95 kDa gelatinase was increased by phorbol myristate acetate, foetal calf serum, thrombin and interleukin-1alpha (IL-1alpha); platelet-derived growth factor (PDGF) BB alone had no effect but acted synergistically with IL-1alpha. A selective protein kinase C inhibitor, Ro 31-8220, abolished induction of the 95 kDa gelatinase. In contrast, none of the agents tested modulated the synthesis of the 72 kDa gelatinase. We conclude that maximal up-regulation of 95 kDa gelatinase expression requires the concerted action of growth factors and inflammatory cytokines mediated, in part, by a protein kinase C-dependent pathway. TIMP-1 and TIMP-2 were highly expressed, and their synthesis was not affected by growth factors or cytokines. Expression of TIMP-3 mRNAs was, however, increased by PDGF and transforming growth factor beta, especially in combination. Divergent regulation of gelatinase and TIMP expression implies that either net synthesis or net degradation of basement membrane can be mediated by appropriate combinations of growth factors and cytokines. PMID:8670128
Divergent regulation by growth factors and cytokines of 95 kDa and 72 kDa gelatinases and tissue inhibitors or metalloproteinases-1, -2, and -3 in rabbit aortic smooth muscle cells.
Fabunmi, R P; Baker, A H; Murray, E J; Booth, R F; Newby, A C
The migration and proliferation of vascular smooth muscle cells (SMCs) during neointima formation in atherosclerosis and angioplasty restenosis is mediated by certain growth factors and cytokines, one action of which may be to promote basement-membrane degradation. To test this hypothesis further, the effects of such growth factors and cytokines on the synthesis of two basement-membrane-degrading metalloproteinases, namely the 72 kDa gelatinase (MMP-2, gelatinase A) and the 95 kDa gelatinase (MMP-9, gelatinase B) and three tissue inhibitors of metalloproteinases (TIMPs) was studied in primary cultured rabbit aortic SMCs. Expression of the 95 kDa gelatinase was increased by phorbol myristate acetate, foetal calf serum, thrombin and interleukin-1alpha (IL-1alpha); platelet-derived growth factor (PDGF) BB alone had no effect but acted synergistically with IL-1alpha. A selective protein kinase C inhibitor, Ro 31-8220, abolished induction of the 95 kDa gelatinase. In contrast, none of the agents tested modulated the synthesis of the 72 kDa gelatinase. We conclude that maximal up-regulation of 95 kDa gelatinase expression requires the concerted action of growth factors and inflammatory cytokines mediated, in part, by a protein kinase C-dependent pathway. TIMP-1 and TIMP-2 were highly expressed, and their synthesis was not affected by growth factors or cytokines. Expression of TIMP-3 mRNAs was, however, increased by PDGF and transforming growth factor beta, especially in combination. Divergent regulation of gelatinase and TIMP expression implies that either net synthesis or net degradation of basement membrane can be mediated by appropriate combinations of growth factors and cytokines. PMID:8670128
Genes influencing spinal bone mineral density in inbred F344, LEW, COP, and DA rats
Alam, Imranul; Sun, Qiwei; Koller, Daniel L.; Liu, Lixiang; Liu, Yunlong; Edenberg, Howard J.; Foroud, Tatiana
Previously, we identified the regions of chromosomes 10q12–q31 and 15p16–q21 harbor quantitative trait loci (QTLs) for lumbar volumetric bone mineral density (vBMD) in female F2 rats derived from Fischer 344 (F344) × Lewis (LEW) and Copenhagen 2331 (COP) × Dark Agouti (DA) crosses. The purpose of this study is to identify the candidate genes within these QTL regions contributing to the variation in lumbar vBMD. RNA was extracted from bone tissue of F344, LEW, COP, and DA rats. Microarray analysis was performed using Affymetrix Rat Genome 230 2.0 Arrays. Genes differentially expressed among the rat strains were then ranked based on the strength of the correlation with lumbar vBMD in F2 animals derived from these rats. Quantitative PCR (qPCR) analysis was performed to confirm the prioritized candidate genes. A total of 285 genes were differentially expressed among all strains of rats with a false discovery rate less than 10%. Among these genes, 18 candidate genes were prioritized based on their strong correlation (r2 > 0.90) with lumbar vBMD. Of these, 14 genes (Akap1, Asgr2, Esd, Fam101b, Irf1, Lcp1, Ltc4s, Mdp-1, Pdhb, Plxdc1, Rabep1, Rhot1, Slc2a4, Xpo4) were confirmed by qPCR. We identified several novel candidate genes influencing spinal vBMD in rats. PMID:19841953
Studies in pig heart tissue on various 60,000 Da phosphoproteins.
Guesdon, F; David-Pfeuty, T
Pig heart tissue have been shown to contain 3 different 60,000 Da phosphoproteins. Different purification procedures were used in order to separate them, suggesting that the 3 phosphoproteins differ in their environmental parameters. The 2 major ones appear essentially as peripheral phosphoproteins that are associated with cellular membranes through ionic forces, whereas the third minor phosphoprotein behaves as an integral plasma membrane protein. The three phosphoproteins also differ in their relative amount of phosphorylated serine, threonine and tyrosine residues after in vitro protein kinase assay. Evidence that the 3 phosphoproteins are related arises from the similarity between their respective phosphopeptide maps after partial hydrolysis with proteases, an experiment that also points out relatedness in primary structure between them and the transforming protein of Rous sarcoma virus, pp60v-src. The 3 phosphoproteins, however, do not appear to be immunologically related to pp60v-src since none of them is immunoprecipitated by sera that precipitate pp60v-src. The possibility that the three 60,000 Da phosphoproteins under study represent 3 differentially localized and phosphorylated products of c-src and/or c-src related genes is discussed. PMID:2472841
3D-printing of transparent bio-microfluidic devices in PEG-DA.
Urrios, Arturo; Parra-Cabrera, Cesar; Bhattacharjee, Nirveek; Gonzalez-Suarez, Alan M; Rigat-Brugarolas, Luis G; Nallapatti, Umashree; Samitier, Josep; DeForest, Cole A; Posas, Francesc; Garcia-Cordero, José L; Folch, Albert
The vast majority of microfluidic systems are molded in poly(dimethylsiloxane) (PDMS) by soft lithography due to the favorable properties of PDMS: biocompatible, elastomeric, transparent, gas-permeable, inexpensive, and copyright-free. However, PDMS molding involves tedious manual labor, which makes PDMS devices prone to assembly failures and difficult to disseminate to research and clinical settings. Furthermore, the fabrication procedures limit the 3D complexity of the devices to layered designs. Stereolithography (SL), a form of 3D-printing, has recently attracted attention as a way to customize the fabrication of biomedical devices due to its automated, assembly-free 3D fabrication, rapidly decreasing costs, and fast-improving resolution and throughput. However, existing SL resins are not biocompatible and patterning transparent resins at high resolution remains difficult. Here we report procedures for the preparation and patterning of a transparent resin based on low-MW poly(ethylene glycol) diacrylate (MW 250) (PEG-DA-250). The 3D-printed devices are highly transparent and cells can be cultured on PEG-DA-250 prints for several days. This biocompatible SL resin and printing process solves some of the main drawbacks of 3D-printed microfluidic devices: biocompatibility and transparency. In addition, it should also enable the production of non-microfluidic biomedical devices. PMID:27217203
Integration of Globus Online with the ATLAS PanDA Workload Management System
NASA Astrophysics Data System (ADS)
Contreras, C.; Deng, W.; Maeno, T.; Nilsson, P.; Potekhin, M.
The PanDA Workload Management System is the basis for distributed production and analysis for the ATLAS experiment at the LHC. In this role, it relies on sophisticated dynamic data movement facilities developed in ATLAS. In certain scenarios, such as small research teams in ATLAS Tier-3 sites and non-ATLAS Virtual Organizations, the overhead of installation and operation of these components makes their use not very cost effective. Globus Online is an emerging new tool from the Globus Alliance, which already proved popular within the research community. It provides the users with fast and robust file transfer capabilities that can also be managed from a Web interface, and in addition to grid sites, can have individual workstations and laptops serving as data transmission endpoints. We will describe the integration of the Globus Online functionality into the PanDA suite of software, in order to give more flexibility in choosing the method of data transfer to ATLAS Tier-3 and Open Science Grid (OSG) users.
VaccineDA: Prediction, design and genome-wide screening of oligodeoxynucleotide-based vaccine adjuvants
Nagpal, Gandharva; Gupta, Sudheer; Chaudhary, Kumardeep; Kumar Dhanda, Sandeep; Prakash, Satya; Raghava, Gajendra P. S.
Immunomodulatory oligodeoxynucleotides (IMODNs) are the short DNA sequences that activate the innate immune system via toll-like receptor 9. These sequences predominantly contain unmethylated CpG motifs. In this work, we describe VaccineDA (Vaccine DNA adjuvants), a web-based resource developed to design IMODN-based vaccine adjuvants. We collected and analyzed 2193 experimentally validated IMODNs obtained from the literature. Certain types of nucleotides (e.g., T, GT, TC, TT, CGT, TCG, TTT) are dominant in IMODNs. Based on these observations, we developed support vector machine-based models to predict IMODNs using various compositions. The developed models achieved the maximum Matthews Correlation Coefficient (MCC) of 0.75 with an accuracy of 87.57% using the pentanucleotide composition. The integration of motif information further improved the performance of our model from the MCC of 0.75 to 0.77. Similarly, models were developed to predict palindromic IMODNs and attained a maximum MCC of 0.84 with the accuracy of 91.94%. These models were evaluated using a five-fold cross-validation technique as well as validated on an independent dataset. The models developed in this study were integrated into VaccineDA to provide a wide range of services that facilitate the design of DNA-based vaccine adjuvants (http://crdd.osdd.net/raghava/vaccineda/). PMID:26212482
Crystal Structure of the 25 kDa Subunit of Human Cleavage Factor I{m}
Coseno,M.; Martin, G.; Berger, C.; Gilmartin, G.; Keller, W.; Doublie, S.
Cleavage factor Im is an essential component of the pre-messenger RNA 3'-end processing machinery in higher eukaryotes, participating in both the polyadenylation and cleavage steps. Cleavage factor Im is an oligomer composed of a small 25 kDa subunit (CF Im25) and a variable larger subunit of either 59, 68 or 72 kDa. The small subunit also interacts with RNA, poly(A) polymerase, and the nuclear poly(A)-binding protein. These protein-protein interactions are thought to be facilitated by the Nudix domain of CF Im25, a hydrolase motif with a characteristic {alpha}/{beta}/{alpha} fold and a conserved catalytic sequence or Nudix box. We present here the crystal structures of human CF Im25 in its free and diadenosine tetraphosphate (Ap4A) bound forms at 1.85 and 1.80 Angstroms, respectively. CF Im25 crystallizes as a dimer and presents the classical Nudix fold. Results from crystallographic and biochemical experiments suggest that CF Im25 makes use of its Nudix fold to bind but not hydrolyze ATP and Ap4A. The complex and apo protein structures provide insight into the active oligomeric state of CF Im and suggest a possible role of nucleotide binding in either the polyadenylation and/or cleavage steps of pre-messenger RNA 3'-end processing.
Leonardo da Vinci and Andreas Vesalius; the shoulder girdle and the spine, a comparison.
Ganseman, Y; Broos, P
Leonardo Da Vinci and Andreas Vesalius were two important renaissance persons; Vesalius was a surgeon-anatomist who delivered innovative work on the study of the human body, Leonardo da Vinci was an artist who delivered strikingly accurate and beautiful drawings on the human body. Below we compare both masters with regard to their knowledge of the working of the muscles, their method and system of dissection and their system and presentation of the drawings. The investigation consisted of a comparison between both anatomists, in particular concerning their study on the shoulder girdle and spine, by reviewing their original work as well as already existing literature on this subject. The investigation led to the conclusion that the drawings mentioned meant a change in history, and were of high quality, centuries ahead of their time. Both were anatomists, both were revolutionary, only one changed history at the moment itself, while the other changed history centuries later. Leonardo has made beautiful drawings that are at a match with the drawings of today or are even better. Vesalius set the start for medicine as a science as it is until this day. Their lives differed as strongly as their impact. In the light of their time, the achievement they made was extraordinary. PMID:18807610
UniDA: Uniform Device Access Framework for Human Interaction Environments
Varela, Gervasio; Paz-Lopez, Alejandro; Becerra, Jose Antonio; Vazquez-Rodriguez, Santiago; Duro, Richard José
Human interaction environments (HIE) must be understood as any place where people carry out their daily life, including their work, family life, leisure and social life, interacting with technology to enhance or facilitate the experience. The integration of technology in these environments has been achieved in a disorderly and incompatible way, with devices operating in isolated islands with artificial edges delimited by the manufacturers. In this paper we are presenting the UniDA framework, an integral solution for the development of systems that require the integration and interoperation of devices and technologies in HIEs. It provides developers and installers with a uniform conceptual framework capable of modelling an HIE, together with a set of libraries, tools and devices to build distributed instrumentation networks with support for transparent integration of other technologies. A series of use case examples and a comparison to many of the existing technologies in the field has been included in order to show the benefits of using UniDA. PMID:22163700
Blood species identification using Near-Infrared diffuse transmitted spectra and PLS-DA method
NASA Astrophysics Data System (ADS)
Zhang, Linna; Zhang, Shengzhao; Sun, Meixiu; Wang, Zhennan; Li, Hongxiao; Li, Yingxin; Li, Gang; Lin, Ling
Blood species identification is of great significance for blood supervision and wildlife investigations. The current methods used to identify the blood species are destructive. Near-Infrared spectroscopy method is known for its non-invasive properties. In this research, we combined Near-Infrared diffuse transmitted spectra and Partial Least Square Discrimination Analysis (PLS-DA) to identify three blood species, including macaque, human and mouse. Blind test and external test were used to assess the PLS-DA model. The model performed 100% accuracy in its identification between three blood species. This approach does not require a specific knowledge of biochemical features for each individual class but relies on a spectroscopic statistical differentiation of the whole components. This is the first time to demonstrate Near-Infrared diffuse transmitted spectra have the ability to identify the species of origin of blood samples. The results also support a good potential of absorption and scattering spectroscopy for species identification in practical applications for real-time detection.
Interactions of the 67 kDa laminin receptor and its precursor with laminin
Fatehullah, Aliya; Doherty, Caroline; Pivato, Géraldine; Allen, George; Devine, Lynda; Nelson, John; Timson, David J.
The 67LR (67 kDa laminin receptor) enables cells to interact with components of the extracellular matrix. The molecule is derived from the 37LRP (37 kDa laminin receptor precursor); however, the precise molecular mechanism of this conversion is unknown. Recombinant 37LRP, expressed in and purified from Escherichia coli, bound to human laminin in a SPR (surface plasmon resonance) experiment. 67LR isolated from human breast-cancer-derived cells in culture was also shown to bind to laminin by SPR. However, the kinetics of association are qualitatively different. 37LRP, but not 67LR, binds to heparan sulfate. The binding of 37LRP to heparan sulfate did not affect the interaction of 37LRP with laminin. In contrast, heparan sulfate reduces the extent of binding of laminin to 67LR. Taken together, these results show that 37LRP has some of the biological activities of 67LR, even prior to the conversion event. However, the conversion affects the sites of interaction with both laminin and heparan sulfate. PMID:19691449
The uncatchable smile in Leonardo da Vinci's La Bella Principessa portrait.
Soranzo, Alessandro; Newberry, Michelle
A portrait of uncertain origin recently came to light which, after extensive research and examination, was shown to be that rarest of things: a newly discovered Leonardo da Vinci painting entitled La Bella Principessa. This research presents a new illusion which is similar to that identified in the Mona Lisa; La Bella Principessa's mouth appears to change slant depending on both the Viewing Distance and the Level of Blur applied to a digital version of the portrait. Through a series of psychophysics experiments, it was found that a perceived change in the slant of the La Bella Principessa's mouth influences her expression of contentment thus generating an illusion that we have coined the "uncatchable smile". The elusive quality of the Mona Lisa's smile has been previously reported (Science, 290 (2000) 1299) and so the existence of a similar illusion in a portrait painted prior to the Mona Lisa becomes more interesting. The question remains whether Leonardo da Vinci intended this illusion. In any case, it can be argued that the ambiguity created adds to the portrait's allure. PMID:26049039
Gentile da Foligno, a pioneer of cardionephrology: commentary on Carmina de urinarum iudiciis and De pulsibus.
Timio, M
Gentile da Foligno was an influential professor and teacher of medicine at the Universities of Bologna, Padua, Siena and Perugia in the first part of the 14th century. He was one of the first physicians to perform human dissection. He authored many important texts and commentaries, among which is the Consilla, a collection of therapeutic advices for many diseases. He made a commentary on Carmina de urinarum iudiciis (Songs of urinary judgements) and to De pulsibus (About pulses) composed by Egidius Corbaliensis. This work is more than a commentary: it is a text attempting to conceptualise the physiology of urine formation. According to his writing, urine associated to the blood passes 'per poros euritides' (through the porous tubules) of the kidney and is then delivered to the bladder. Commenting and explaining the writing on De pulsibus, he stressed the importance of heart disease on modulating the colour and output of urine and the relationship between fast pulse rate and urine output. For the originality of his thought and for the conceptualization of the relationship between pulse rate and urine characteristic, Gentile da Foligno could be indicated as the 'first' cardionephrologist in the history of medicine. PMID:10213818
Magnetisches Tracking für die Navigation mit dem da Vinci® Surgical System
NASA Astrophysics Data System (ADS)
Nickel, Felix; Wegner, Ingmar; Kenngott, Hannes; Neuhaus, Jochen; Müller-Stich, Beat P.; Meinzer, Hans-Peter; Gutt, Carsten N.
In dieser Studie wurde untersucht ob in einem typischen OP-Aufbau mit dem da Vinci® Telemanipulator elektromagnetisches Tracking für die Realisation eines Navigationssystems eingesetzt werden kann. Hierfür wurde in einem realen OP-Aufbau untersucht, wie stark metallische und ferromagnetisch wirksame Objekte wie Operationstisch und Telemanipulator das elektromagnetische Feld des Trackingsystems beeinflussen. Die Ergebnisse zeigen, dass der Telemanipulator nur unwesentlich die Störung des Magnetfeldes durch den OP-Tisch verstärkt. Insbesondere die Bewegung der Instrumente im Trackingvolumen verursachte keine zusätzliche relevante Störung des Magnetfeldes. Bei Begrenzung des Trackingvolumens auf eine Länge von 190 mm, Höhe von 200mm und Breite von 400 mm war der maximale Fehler in diesem Bereich an allen Messpunkten kleiner 10 mm. Der Einsatz von elektromagnetischem Tracking für die Navigation mit dem da Vinci® Surgical System ist somit in einem begrenzten Arbeitsvolumen mit hinreichender Genauigkeit möglich.
Time Resolved Detection of Infrared Synchrotron Radiation at DA{phi}NE
Bocci, A.; Marcelli, A.; Drago, A.; Guidi, M. Cestelli; Pace, E.; Piccinini, M.; Sali, D.; Morini, P.
Synchrotron radiation is characterized by a very wide spectral emission from IR to X-ray wavelengths and a pulsed structure that is a function of the source time structure. In a storage ring, the typical temporal distance between two bunches, whose duration is a few hundreds of picoseconds, is on the nanosecond scale. Therefore, synchrotron radiation sources are a very powerful tools to perform time-resolved experiments that however need extremely fast detectors. Uncooled IR devices optimized for the mid-IR range with sub-nanosecond response time, are now available and can be used for fast detection of intense IR sources such as synchrotron radiation storage rings. We present here different measurements of the pulsed synchrotron radiation emission at DA{phi}NE (Double Annular {phi}-factory for Nice Experiments), the collider of the Laboratori Nazionali of Frascati (LNF) of the Istituto Nazionale di Fisica Nucleare (INFN), performed with very fast uncooled infrared detectors with a time resolution of a few hundreds of picoseconds. We resolved the emission time structure of the electron bunches of the DA{phi}NE collider when it works in a normal condition for high energy physics experiments with both photovoltaic and photoconductive detectors. Such a technology should pave the way to new diagnostic methods in storage rings, monitoring also source instabilities and bunch dynamics.
[History of robotics: from Archytas of Tarentum until da Vinci robot. (Part I)].
Sánchez Martín, F M; Millán Rodríguez, F; Salvador Bayarri, J; Palou Redorta, J; Rodríguez Escovar, F; Esquena Fernández, S; Villavicencio Mavrich, H
Robotic surgery is the newst technologic option in urology. To understand how new robots work is interesting to know their history. The desire to design machines imitating humans continued for more than 4000 years. There are references to King-su Tse (clasic China) making up automaton at 500 a. C. Archytas of Tarentum (at around 400 a.C.) is considered the father of mechanical engineering, and one of the occidental robotics classic referents. Heron of Alexandria, Hsieh-Fec, Al-Jazari, Roger Bacon, Juanelo Turriano, Leonardo da Vinci, Vaucanson o von Kempelen were robot inventors in the middle age, renaissance and classicism. At the XIXth century, automaton production underwent a peak and all engineering branches suffered a great development. At 1942 Asimov published the three robotics laws, based on mechanics, electronics and informatics advances. At XXth century robots able to do very complex self governing works were developed, like da Vinci Surgical System (Intuitive Surgical Inc, Sunnyvale, CA, USA), a very sophisticated robot to assist surgeons. PMID:17645084
Development of noSQL data storage for the ATLAS PanDA Monitoring System
NASA Astrophysics Data System (ADS)
Ito, H.; Potekhin, M.; Wenaus, T.
For several years the PanDA Workload Management System has been the basis for distributed production and analysis for the ATLAS experiment at the LHC. Since the start of data taking PanDA usage has ramped up steadily, typically exceeding 500k completed jobs/day by June 2011. The associated monitoring data volume has been rising as well, to levels that present a new set of challenges in the areas of database scalability and monitoring system performance and efficiency. These challenges are being met with an R&D effort aimed at implementing a scalable and efficient monitoring data storage based on a noSQL solution (Cassandra). We present our motivations for using this technology, as well as data design and the techniques used for efficient indexing of the data. We also discuss the hardware requirements as they were determined by testing with actual data and realistic rate of queries. In conclusion, we present our experience with operating a Cassandra cluster over an extended period of time and with data load adequate for planned application.
Food residue recycling by swine breeders in a developing economy: a case study in Da Nang, Viet Nam.
Kato, Takaaki; Pham, Dung Thi Xuan; Hoang, Hai; Xue, Yonghai; Tran, Quang Van
This study provides a detailed description of food residue collection by swine breeders in Da Nang, Viet Nam. In January 2011, the study surveyed 30 swine breeders in two villages with respect to locations, methods, prices, quantities, and prospects for food residue collection. The sampled swine breeders regularly visited 55 locations in central Da Nang to collect raw food residue. They then transferred the food residue to their piggeries, boiled it, and fed it to their swine. A regression analysis revealed that the total amount of food residue collected by a farm depends on the number of swine in the farm and the number of collections made per day. Swine breeders in Da Nang were estimated to collect 26.3 metric tons of organic waste per day, which amounted to 4.1% of domestic waste collected by the local government. Among the sampled swine breeders, 93% answered that they would continue using food residue for the next five years. PMID:22883688
Análise Temporal da Coma de CO+ no Cometa P/Halley
NASA Astrophysics Data System (ADS)
Voelzke, M. R.; Schlosser, W.; Schmidt-Kaler, Th.
Observações fotográficas e fotoelétricas da coma de gás ionizado do cometa P/Halley a nível de CO+ em 4250 angstroms fizeram parte do programa de Monitoramento do Halley desenvolvido pela Universidade de Bochum (Ruhr-Universitaet Bochum) na Alemanha, realizado de 17 de fevereiro a 17 de abril de 1986 no European Southern Observatory (ESO) em La Silla (Chile). Nesta faixa espectral é possível observar a contínua formação, bem como o movimento e expansão das estruturas de plasma. Para observar a morfologia destas estruturas foram analisadas 32 placas fotográficas de CO+ (placas de vidro) do cometa P/Halley. Tais placas possuem um campo de visão de 28,6 por 28,6 graus sendo obtidas entre 29 de março e 17 de abril de 1986 com tempos de exposição entre 20 e 120 minutos. Todas as placas foram digitadas com o auxílio de um microdensitômetro PDS 2020 GM (Photometric Data System) do Instituto Astronómico da Westfaelischen Wilhelms-Universitaet em Muenster, Alemanha (um pixel = 25 por 25 micrômetros correspondendo aproximadamente a 46,88 por 46,88 segundos de arco). Após a digitação os dados foram reduzidos à intensidades relativas, sendo que os posssíveis de calibração também foram reduzidos à intensidades absolutas, expressas em termos de densidade colunar utilizando-se dos sistemas de tratamento de imagens MIDAS (Munich Image Data Analysis System; ESO - Image Processing Group, 1988) e IHAP (Image Handling And Processing; Middleburg, 1983). Com o auxílio do método de teta mínimo de Stellingwerf (Stellingwerf, 1978) obteve-se um período de 2,22 +/- 0,09 dias a partir da análise de estruturas na coma de plasma através da subtração de imagens subsequentes. Este método foi comparado com o método de Fourier. Provavelmente exista um segundo ciclo com período aproximado de 3,6 dias. A idéia de subtrair imagens subsequentes é devido ao fato de que os efeitos de rotação são apenas 10% dos fenômenos de distribuição gasosa. Portanto as
Expression, purification, and characterization of a bifunctional 99-kDa peptidoglycan hydrolase from Pediococcus acidilactici ATCC 8042.
García-Cano, Israel; Campos-Gómez, Manuel; Contreras-Cruz, Mariana; Serrano-Maldonado, Carlos Eduardo; González-Canto, Augusto; Peña-Montes, Carolina; Rodríguez-Sanoja, Romina; Sánchez, Sergio; Farrés, Amelia
Pediococcus acidilactici ATCC 8042 is a lactic acid bacteria that inhibits pathogenic microorganisms such as Staphylococcus aureus through the production of two proteins with lytic activity, one of 110 kDa and the other of 99 kDa. The 99-kDa one has high homology to a putative peptidoglycan hydrolase (PGH) enzyme reported in the genome of P. acidilactici 7_4, where two different lytic domains have been identified but not characterized. The aim of this work was the biochemical characterization of the recombinant enzyme of 99 kDa. The enzyme was cloned and expressed successfully and retains its activity against Micrococcus lysodeikticus. It has a higher N-acetylglucosaminidase activity, but the N-acetylmuramoyl-L-alanine amidase can also be detected spectrophotometrically. The protein was then purified using gel filtration chromatography. Antibacterial activity showed an optimal pH of 6.0 and was stable between 5.0 and 7.0. The optimal temperature for activity was 60 °C, and all activity was lost after 1 h of incubation at 70 °C. The number of strains susceptible to the recombinant 99-kDa enzyme was lower than that susceptible to the mixture of the 110- and 99-kDa PGHs of P. acidilactici, a result that suggests synergy between these two enzymes. This is the first PGH from LAB that has been shown to possess two lytic sites. The results of this study will aid in the design of new antibacterial agents from natural origin that can combat foodborne disease and improve hygienic practices in the industrial sector. PMID:25940238
Culture and Dehydration: A Comparative Study of Caída de la Mollera (Fallen Fontanel) in Three Latino Populations.
Pachter, Lee M; Weller, Susan C; Baer, Roberta D; de Alba Garcia, Javier E Garcia; Glazer, Mark; Trotter, Robert; Klein, Robert E; Gonzalez, Eduardo
A sunken soft-spot or fontanel is a sign for dehydration in infants. Around the world, folk illnesses, such as caída de la mollera in some Latin American cultures, often incorporate this sign as a hallmark of illness, but may or may not incorporate re-hydration therapies in treatment strategies. This report describes a study of lay descriptions of causes, symptoms, and treatments for caída de la mollera in three diverse Latin American populations. A mixed-methods approach was used. Representative community-based samples were interviewed in rural Guatemala, Guadalajara, Mexico, and Edinburgh, Texas, with a 132 item questionnaire on the causes, susceptibility, symptoms, and therapies for caída de la mollera. Cultural consensus analysis was used to estimate community beliefs about caída. Interviews conducted in rural Guatemala (n = 60), urban Mexico (n = 62), and rural Texas on the Mexican border (n = 61) indicated consistency in thematic elements within and among these three diverse communities. The high degree of consistency in the illness explanatory models indicated shared beliefs about caída de la mollera in each of the communities and a core model shared across communities. However, an important aspect of the community beliefs was that rehydration therapies were not widely endorsed. The consistency in explanatory models in such diverse communities, as well as the high degree of recognition and experience with this illness, may facilitate communication between community members, and health care providers/public health intervention planners to increase use of rehydration therapies for caída de la mollera. Recommendations for culturally informed and respectful approaches to clinical communication are provided. PMID:26239172
Membrane-associated 41-kDa GTP-binding protein in collagen-induced platelet activation
Walker, G.; Bourguignon, L.Y. )
Initially we established that the binding of collagen to human blood platelets stimulates both the rapid loss of PIP2 and the generation of inositol-4,5-bisphosphate (IP2) and inositol-1,4,5-triphosphate (IP3). These results indicate that the binding of collagen stimulates inositol phospholipid-specific phospholipase C during platelet activation. The fact that GTP or GTP-gamma-S augments, and pertussis toxin inhibits, collagen-induced IP3 formation suggests that a GTP-binding protein or (or proteins) may be directly involved in the regulation of phospholipase C-mediated phosphoinositide turnover in human platelets. We have used several complementary techniques to isolate and characterize a platelet 41-kDa polypeptide (or polypeptides) that has a number of structural and functional similarities to the regulatory alpha i subunit of the GTP-binding proteins isolated from bovine brain. This 41-kDa polypeptide (or polypeptides) is found to be closely associated with at least four membrane glycoproteins (e.g., gp180, gp110, gp95, and gp75) in a 330-kDa complex that can be dissociated by treatment with high salt plus urea. Most important, we have demonstrated that antilymphoma 41-kDa (alpha i subunit of GTP-binding proteins) antibody cross-reacts with the platelet 41-kDa protein (or proteins) and the alpha i subunit of bovine brain Gi alpha proteins, and blocks GTP/collagen-induced IP3 formation. These data provide strong evidence that the 41-kDa platelet GTP-binding protein (or proteins) is directly involved in collagen-induced signal transduction during platelet activation.
Purification and characteristics of a 30 kDa Cd-binding protein from rat testis cytosol
Kinoshita, C.M.
The main objective of this dissertation was to purify the 30 kDa Cd-binding protein from rat testis cytosol (Cd-testin) in a form retaining 1 or more moles of Cd per mole of protein, following the addition of /sup 109/CdCl/sub 2/ to cytosol in amounts just sufficient to saturate the 30 kDa peak. Two successful procedures were developed based on (1) ammonium sulfate fractionation, hydrophobic interaction chromatography, and gel filtration; or (2) heat treatment (90/sup 0/C, 5 min) and gel filtration. Application of Method 1 to cytosol labeled with /sup 65/ZnCL/sub 2/ showed that Zn formed a complex analogous to Cd-testin. The Cd content (moles/mole protein) in two preparations of 30 kDa protein was 2.1 to 2.6 (Method 1); Method 2 gave more variable results (1.1 to 3.4). In neither case was a 30 kDa polypeptide detected by 20 SDS PAGE, but a major subunit (22 kDa/pl 4.6) was observed in both preparations, along with smaller subunits. Method 1 gave higher purity (3 spots) but lower yield (about 4% of cytosolic 30 kDa Cd) whereas Method 2 gave a much higher yield (about 45%) but somewhat lower purity based on 2D SDS PAGE. On the basis of these results, Cd-testin consists of subunits rather than a single polypeptide chain. It is concluded that rat testis contains novel proteins binding Cd or Zn, and can be isolated in forms that retain a high Cd content. The manner in which the Cd is bound, and the subunits involved, have not been determined.
gLExec and MyProxy integration in the ATLAS/OSG PanDA workload management system
NASA Astrophysics Data System (ADS)
Caballero, J.; Hover, J.; Litmaath, M.; Maeno, T.; Nilsson, P.; Potekhin, M.; Wenaus, T.; Zhao, X.
Worker nodes on the grid exhibit great diversity, making it difficult to offer uniform processing resources. A pilot job architecture, which probes the environment on the remote worker node before pulling down a payload job, can help. Pilot jobs become smart wrappers, preparing an appropriate environment for job execution and providing logging and monitoring capabilities. PanDA (Production and Distributed Analysis), an ATLAS and OSG workload management system, follows this design. However, in the simplest (and most efficient) pilot submission approach of identical pilots carrying the same identifying grid proxy, end-user accounting by the site can only be done with application-level information (PanDA maintains its own end-user accounting), and end-user jobs run with the identity and privileges of the proxy carried by the pilots, which may be seen as a security risk. To address these issues, we have enabled PanDA to use gLExec, a tool provided by EGEE which runs payload jobs under an end-user's identity. End-user proxies are pre-staged in a credential caching service, MyProxy, and the information needed by the pilots to access them is stored in the PanDA DB. gLExec then extracts from the user's proxy the proper identity under which to run. We describe the deployment, installation, and configuration of gLExec, and how PanDA components have been augmented to use it. We describe how difficulties were overcome, and how security risks have been mitigated. Results are presented from OSG and EGEE Grid environments performing ATLAS analysis using PanDA and gLExec.
Ontogeny of DA1 receptor-mediated natriuresis in the rat: in vivo and in vitro correlations.
Kaneko, S; Albrecht, F; Asico, L D; Eisner, G M; Robillard, J E; Jose, P A
The natriuretic and diuretic effects of dopamine are attenuated in the young. Because dopamine has actions on receptors (e.g., adrenergic, serotonin) other than dopamine, we studied a novel dopamine agonist, pramipexole, which has a selectivity to both DA1 and DA2-receptor subtypes. Intravenous administration of pramipexole resulted in a dose-related (1, 10, and 100 micrograms.kg-1.min-1) increase in urine flow and absolute and fractional sodium excretion and a decrease in mean arterial pressure (MAP) in three groups of rats studied. Pramipexole induced a greater decrease in MAP in 6- to 7- (n = 5) and 9- to 16- (n = 6) than in 3- to 4-wk-old (n = 8) rats; the natriuresis and diuresis were greatest in 12- to 16- and least in 3- to 4-wk-old rats. The renal effects of pramipexole were mainly due to actions at the DA1 receptor, since these effects were completely blocked by the coinfusion of a DA1 antagonist, SKF 83742. To explore further a cause of the attenuated natriuretic effect of pramipexole in the young, we studied the effect of a selective DA1-receptor agonist, fenoldopam, on amiloride-sensitive 22Na+ uptake in renal brush-border membrane vesicles. The 3-s amiloride-sensitive uptake was inhibited (45%) by fenoldopam (5 x 10(-5)M) in 9- to 16- (n = 6) but not in 3- to 4-wk-old (n = 5) rats. These studies suggest that the attenuated natriuretic effect of dopamine in the young is in part due to decreased DA1 action on the brush-border membrane Na(+)-H+ exchanger. PMID:1357983
Hopf-Ranãda linked and knotted light beam solution viewed as a null electromagnetic field.
Besieris, Ioannis M; Shaarawi, Amr M
The Hopf-Ranãda linked and knotted light beam solution, which has been interpreted physically and extended analytically by Irvine and Bouwmeester recently, is viewed in this Letter as a null electromagnetic field. It is shown, in particular, that the Hopf-Ranãda solution is a variant of a luminal null electromagnetic wave due originally to Robinson and Troutman and reported by Bialynicki-Birula recently. This analogy is motivated by means of a method due to Whittaker and Bateman, and a relationship to well-known scalar luminal localized waves is examined. PMID:20016647
Genome Sequence of the Psychrophilic Bacterium Tenacibaculum ovolyticum Strain da5A-8 Isolated from Deep Seawater
Zhai, Zhenyu; Komatsu, Ayumi; Shibayama, Keigo
Some bacterial species of the genus Tenacibaculum, including Tenacibaculum ovolyticum, have been known as fish pathogens in the sea. So far, the only published genome sequence for this genus is for Tenacibaculum dicentrarchi, which could also be a fish pathogen. Strain da5A-8, showing 100% identity to the 16S rRNA gene sequence of T. ovolyticum DSM 18103T, was isolated from seawater at a depth of 344 m in Kochi, Japan, and grew optimally at 10 to 20°C. The genome sequence of strain da5A-8 revealed the possible virulence genes commonly observed in the genus Tenacibaculum. PMID:27365358
L151-81A/B - A unique white dwarf binary with DB and DA components
NASA Technical Reports Server (NTRS)
Oswalt, Terry D.; Hintzen, Paul M.; Liebert, James W.; Sion, Edward M.
Spectroscopic observations of the wide binary L151-81A/B reveal that both components are white dwarfs with spectral types DB3 and DA4, the first such binary identified in a comprehensive survey of Luyten and Giclas common proper motion pairs. Assuming log g = 8, measurements of the helium line profiles in the DB primary yield Teff = 16,000 + or - 2,000, while the hydrogen profiles for the DA companion yield Teff =12,000 + or - 2,000. The existence of a helium-rich/hydrogen-rich degenerate pair offers several interesting tests of theoretical white dwarf formation channels and surface abundance evolution.
Purification, characterization and allergenicity assessment of 26kDa protein, a major allergen from Cicer arietinum.
Verma, Alok Kumar; Sharma, Akanksha; Kumar, Sandeep; Gupta, Rinkesh Kumar; Kumar, Dinesh; Gupta, Kriti; Giridhar, B H; Das, Mukul; Dwivedi, Premendra D
Chickpea (CP), a legume of the family Fabaceae, is an important nutrient-rich food providing protein, essential amino acids, vitamins, dietary fibre, and minerals. Unfortunately, several IgE-binding proteins in CP have been detected that are responsible for allergic manifestations in sensitized population. Therefore, the prevalence of CP induced allergy prompted us towards purification, characterization and allergenicity assessment of a major ∼26kDa protein from chickpea crude protein extract (CP-CPE). Purification of CP 26kDa protein was done using a combination of fractionation and anion exchange chromatography. This protein was further characterized as "Chain A, crystal structure of a plant albumin" from Cicer arietinum with Mol wt 25.8kDa by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Further, allergenic potential of purified 25.8kDa protein was assessed using in vivo and in vitro model. Purified protein showed IgE-binding capacity with sensitized BALB/c mice and CP allergic patient's sera. Enhanced levels of specific and total IgE, MCP-1, MCPT-1, myeloperoxidase, histamine, prostaglandin D2, and cysteinyl leukotriene were found in sera of mice treated with CP ∼26kDa protein. Further, expressions of Th2 cytokines (i.e. IL-4, IL-5, IL-13), transcription factors (i.e. GATA-3, STAT-6, SOCS-3) and mast cell signaling proteins (Lyn, cFgr, Syk, PLC-γ2, PI-3K, PKC) were also found increased at mRNA and protein levels in the intestines of mice treated with CP ∼26kDa protein. In addition, enhanced release of β-hexosaminidase, histamine, cysteinyl leukotriene and prostaglandin D2 were observed in RBL2H3 cell line when treated (125μg) with CP 26kDa protein. Conclusively, in vivo and in vitro studies revealed the allergenic potential of purified CP 26kDa protein. Being a potential allergen, plant albumin may play a pivotal role in CP induced allergenicity. Current study will be helpful for better development of therapeutic approaches to
Genome Sequence of the Psychrophilic Bacterium Tenacibaculum ovolyticum Strain da5A-8 Isolated from Deep Seawater.
Teramoto, Maki; Zhai, Zhenyu; Komatsu, Ayumi; Shibayama, Keigo; Suzuki, Masato
Some bacterial species of the genus Tenacibaculum, including Tenacibaculum ovolyticum, have been known as fish pathogens in the sea. So far, the only published genome sequence for this genus is for Tenacibaculum dicentrarchi, which could also be a fish pathogen. Strain da5A-8, showing 100% identity to the 16S rRNA gene sequence of T. ovolyticum DSM 18103(T), was isolated from seawater at a depth of 344 m in Kochi, Japan, and grew optimally at 10 to 20°C. The genome sequence of strain da5A-8 revealed the possible virulence genes commonly observed in the genus Tenacibaculum. PMID:27365358
Eugenia uniflora L. Essential Oil as a Potential Anti-Leishmania Agent: Effects on Leishmania amazonensis and Possible Mechanisms of Action
Amorim, Layane Valéria; de Oliveira, Jamylla Mirck Guerra; Dias, Clarice Noleto; Moraes, Denise Fernandes Coutinho; Andrade, Eloisa Helena de Aguiar; Maia, Jose Guilherme Soares; Carneiro, Sabrina Maria Portela; Carvalho, Fernando Aécio de Amorim
Eugenia uniflora L. is a member of the Myrtaceae family and is commonly known as Brazilian cherry tree. In this study, we evaluated the chemical composition of Eugenia uniflora L. essential oil (EuEO) by using gas chromatography-mass spectrometry (GC-MS) and assessed its anti-Leishmania activity. We also explored the potential mechanisms of action and cytotoxicity of EuEO. Thirty-two compounds were identified, which constituted 92.65% of the total oil composition. The most abundant components were sesquiterpenes (91.92%), with curzerene (47.3%), γ-elemene (14.25%), and trans-β-elemenone (10.4%) being the major constituents. The bioactivity shown by EuEO against promastigotes (IC50, 3.04 μg·mL−1) and amastigotes (IC50, 1.92 μg·mL−1) suggested significant anti-Leishmania activity. In the cytotoxicity determination, EuEO was 20 times more toxic to amastigotes than to macrophages. Hemolytic activity was 63.22% at the highest concentration tested (400 μg·mL−1); however, there appeared to be no toxicity at 50 μg·mL−1. While the data show that EuEO activity is not mediated by nitric oxide production, they do suggest that macrophage activation may be involved in EuEO anti-Leishmania activity, as evidenced by increases in both the phagocytic capacity and the lysosomal activity. More studies are needed to determine in vivo activity as well as additional mechanisms of the anti-Leishmania activity. PMID:23533469
The integrated endoplasmic reticulum stress response in Leishmania amazonensis macrophage infection: the role of X-box binding protein 1 transcription factor.
Dias-Teixeira, Karina Luiza; Calegari-Silva, Teresa Cristina; dos Santos, Guilherme R R M; Vitorino Dos Santos, José; Lima, Carolina; Medina, Jorge Mansur; Aktas, Bertal Huseyin; Lopes, Ulisses G
Endoplasmic reticulum (ER) stress triggers the integrated ER-stress response (IERSR) that ensures cellular survival of ER stress and represents a primordial form of innate immunity. We investigated the role of IERSR duringLeishmania amazonensisinfection. Treatment of RAW 264.7 infected macrophages with the ER stress-inducing agent thapsigargin (TG; 1 μM) increasedL. amazonensisinfectivity in an IFN1-α receptor (IFNAR)-dependent manner. In Western blot assays, we showed thatL. amazonensisactivates the inositol-requiring enzyme (IRE1)/ X-box binding protein (XBP)-1-splicing arms of the IERSR in host cells. In chromatin immunoprecipitation (ChIP) assays, we showed an increased occupancy of enhancer and promoter sequences for theIfnbgene by XBP1 in infected RAW 264.7 cells. Knocking down XBP1 expression by transducing RAW 264.7 cells with the short hairpin XBP1 lentiviral vector significantly reduced the parasite proliferation associated with impaired translocation of phosphorylated IFN regulatory transcription factor (IRF)-3 to the nucleus and a decrease in IFN1-β expression. Knocking down XBP1 expression also increased NO concentration, as determined by Griess reaction and reduced the expression of antioxidant genes, such as heme oxygenase (HO)-1, that protect parasites from oxidative stress. We conclude thatL. amazonensisactivation of XBP1 plays a critical role in infection by protecting the parasites from oxidative stress and increasing IFN1-β expression.-Dias-Teixeira, K. L., Calegari-Silva, T. C., Dos Santos, G. R. R. M., Vitorino dos Santos, J., Lima, C., Medina, J. M., Aktas, B. H., Lopes, U. G. The integrated endoplasmic reticulum stress response inLeishmania amazonensismacrophage infection: the role of X-box binding protein 1 transcription factor. PMID:26678450
Eugenia uniflora L. Essential Oil as a Potential Anti-Leishmania Agent: Effects on Leishmania amazonensis and Possible Mechanisms of Action.
Rodrigues, Klinger Antonio da Franca; Amorim, Layane Valéria; de Oliveira, Jamylla Mirck Guerra; Dias, Clarice Noleto; Moraes, Denise Fernandes Coutinho; Andrade, Eloisa Helena de Aguiar; Maia, Jose Guilherme Soares; Carneiro, Sabrina Maria Portela; Carvalho, Fernando Aécio de Amorim
Eugenia uniflora L. is a member of the Myrtaceae family and is commonly known as Brazilian cherry tree. In this study, we evaluated the chemical composition of Eugenia uniflora L. essential oil (EuEO) by using gas chromatography-mass spectrometry (GC-MS) and assessed its anti-Leishmania activity. We also explored the potential mechanisms of action and cytotoxicity of EuEO. Thirty-two compounds were identified, which constituted 92.65% of the total oil composition. The most abundant components were sesquiterpenes (91.92%), with curzerene (47.3%), γ -elemene (14.25%), and trans- β -elemenone (10.4%) being the major constituents. The bioactivity shown by EuEO against promastigotes (IC50, 3.04 μ g·mL(-1)) and amastigotes (IC50, 1.92 μ g·mL(-1)) suggested significant anti-Leishmania activity. In the cytotoxicity determination, EuEO was 20 times more toxic to amastigotes than to macrophages. Hemolytic activity was 63.22% at the highest concentration tested (400 μ g·mL(-1)); however, there appeared to be no toxicity at 50 μ g·mL(-1). While the data show that EuEO activity is not mediated by nitric oxide production, they do suggest that macrophage activation may be involved in EuEO anti-Leishmania activity, as evidenced by increases in both the phagocytic capacity and the lysosomal activity. More studies are needed to determine in vivo activity as well as additional mechanisms of the anti-Leishmania activity. PMID:23533469
In para totale...una cosa da panico...sulla lingua dei giovani in Italia (In para totale...una cosa da panico...The Language of Young People in Italy).
ERIC Educational Resources Information Center
Marcato, Carla
Describes and analyzes the language of young people in Italy today. Particular focus is on the expressions using "para" (e.g., "in para totale" = to be very bored or worried) and the phrase "una cosa da panico" (something terrible or its opposite something wonderful). (CFM)
DaVIE: Database for the Visualization and Integration of Epigenetic data
Fejes, Anthony P.; Jones, Meaghan J.; Kobor, Michael S.
One of the challenges in the analysis of large data sets, particularly in a population-based setting, is the ability to perform comparisons across projects. This has to be done in such a way that the integrity of each individual project is maintained, while ensuring that the data are comparable across projects. These issues are beginning to be observed in human DNA methylation studies, as the Illumina 450k platform and next generation sequencing-based assays grow in popularity and decrease in price. This increase in productivity is enabling new insights into epigenetics, but also requires the development of pipelines and software capable of handling the large volumes of data. The specific problems inherent in creating a platform for the storage, comparison, integration, and visualization of DNA methylation data include data storage, algorithm efficiency and ability to interpret the results to derive biological meaning from them. Databases provide a ready-made solution to these issues, but as yet no tools exist that that leverage these advantages while providing an intuitive user interface for interpreting results in a genomic context. We have addressed this void by integrating a database to store DNA methylation data with a web interface to query and visualize the database and a set of libraries for more complex analysis. The resulting platform is called DaVIE: Database for the Visualization and Integration of Epigenetics data. DaVIE can use data culled from a variety of sources, and the web interface includes the ability to group samples by sub-type, compare multiple projects and visualize genomic features in relation to sites of interest. We have used DaVIE to identify patterns of DNA methylation in specific projects and across different projects, identify outlier samples, and cross-check differentially methylated CpG sites identified in specific projects across large numbers of samples. A demonstration server has been setup using GEO data at http
Micromorphology of selected relict slope deposits from Serra da Estrela, Portugal
NASA Astrophysics Data System (ADS)
Nieuwendam, Alexandre; Vieira, Gonçalo; Schaefer, Carlos
Serra da Estrela is the highest mountain in Portugal (1,993 m ASL) and part of the Iberian Central Cordillera. The mountain has a strong relief and a lithological diversity with several types of granitoids and metasediments. Most of the western plateau area was glaciated during the Last Glacial Maximum and its morphology is dominated by glacial landforms. Vieira (2004) produced a detailed geomorphological map of Serra da Estrela and described several sites showing stratified slope, head and debris-flow deposits. Based on the geomorphological analysis of the relationships between glacial and periglacial evidence, a first relative chronology was presented. However, a detailed and systematical sedimentological analysis has not been conducted before and absolute ages are also lacking. Micromorphology analysis has proven to be of considerable value in the interpretation of mountain soils and sediments. Such interpretation depends on identifying diagnostic features, indicating factors as the presence or absence of permafrost, thickness of the active layer, ice segregation and the operation of processes of mass-wasting. In this study, micromorphology was used to answer questions concerning the composition, structure, origin and depositional processes of relict slope deposits. Micromorphology allowed a systematic description of the physical characteristics of the sediments. Lamination and sorting, when preserved, are good evidence for overland flow. Features due to deformation (folds, boudins, coatings and tails due to the rotation of clasts) are associated with sliding. Other mass-movements such as debris flows, earth flows, and to a certain extent, dry grain flows may be characterized by similar microscopic facies, typically a poorly sorted, porphyric material. Porosity gives evidence for both liquefaction (debris flows) and frost-induced mass-movement (solifluction). The relict slope deposits of the Serra da Estrela show an increase in cryogenic micromorphological
Detection of cohesive forces in the rubble-pile asteroid (29075) 1950 DA
NASA Astrophysics Data System (ADS)
Rozitis, Benjamin; MacLennan, Eric; Emery, Joshua P.
The physical behavior of rubble-pile asteroids has traditionally been described using only gravitational and frictional forces within a granular material. Cohesive forces in the form of small van der Waals forces between constituent grains have recently been predicted to be important for small rubble-pile asteroids (less than 10 kilometers in size), and could potentially explain how small fast spinning asteroids remain intact. It is possible to infer the existence of cohesive forces within a rubble-pile asteroid by determining if it has insufficient self-gravity, dictated by its bulk density, to prevent rotational breakup by centrifugal forces. The kilometer-sized and potentially-hazardous asteroid (29075) 1950 DA is one of the largest known candidates for being held together by cohesive forces, as it has a rotation period of 2.1216 h that is just beyond the critical spin limit of 2.2 h estimated for a cohesionless asteroid. Using the Advanced Thermophysical Model (or ATPM), in combination with the radar shape model, WISE thermal-infrared data, and Yarkovsky orbital drift measurement, we determined the thermal inertia, bulk density, and cohesive strength of (29075) 1950 DA (Rozitis et al., 2014, Nature, 512, 174-176). The thermal inertia value is remarkably low at 24 +20/-14 SI units, which gives a corresponding bulk density of 1.7 ± 0.7 g/cm^3 in the Yarkovsky orbital drift analysis. This bulk density is typical of a rubble-pile asteroid, and a minimum cohesive strength of 64 +12/-20 Pa is therefore required to prevent surface mass shedding and structural failure by centrifugal forces. This strength is comparable to, though somewhat less than, the cohesive forces found between the grains of lunar regolith. Finally, as (29075) 1950 DA has a 1 in 19,800 chance of impacting the Earth in 2880, and has the potential to disrupt like main-belt comet P/2013 R3, it raises new implications for impact mitigation against fast spinning rubble-pile asteroids.
Toxicokinetics of the phytoestrogen daidzein in female DA/Han rats.
Janning, P; Schuhmacher, U S; Upmeier, A; Diel, P; Michna, H; Degen, G H; Bolt, H M
Female DA/Han rats were given the phytoestrogen daidzein, either intravenously (10 mg/kg b.w.) or orally by gavage (10 or 100 mg/kg b.w.). The plasma concentration-time curve determined after i.v. administration of daidzein was fitted to a triexponential model, resulting in a final half-life (gamma-phase) of approximately 4 h. The oral bioavailability of 10 mg daidzein/kg was 9.7%, while that of 100 mg/kg was 2.2%; the higher dose (100 mg/kg) was apparently absorbed to a four- to fivefold lower extent than the smaller dose. The plasma concentration time curves after oral administration of daidzein to female DA/Han rats revealed pronounced interindividual differences and multiple peaks, pointing to extensive enterohepatic circulation and/or protracted absorption from the gastrointestinal tract. As shown in a separate experiment with bile duct-cannulated rats, daidzein (i.p. 10 mg/kg b.w.) is efficiently excreted with bile: glucuronide/sulfate metabolites amounting to approximately 30% of the dose in 8 h. Conjugates were also the main circulating metabolites upon i.v. or gavage administration of daidzein, indicating efficient phase II metabolism in female DA/Han rats. Since only few data have been published on tissue levels of isoflavones, their concentrations were measured in various organs and compared to plasma levels determined at the time the animals were killed, with one exception 32 or 48 h after rats had received a single dose of daidzein (i.v. or per os). As expected, the daidzein concentrations depended upon dose and administration route. Despite notable differences in the absolute amounts of total daidzein (free plus hydrolyzed conjugates), the levels were usually three- to fivefold higher in liver and kidney than in plasma; in most samples of uteri, the concentrations were similar, or up to twofold higher, than the respective plasma levels. These data point to an uptake and storage of isoflavones and metabolites in tissues. Experimental toxicokinetics
Chlorophyll antenna proteins of photosystem I: topology, synthesis, and regulation of the 20-kDa subunit of Chlamydomonas light-harvesting complex of photosystem I
Herrin, D.L.; Plumley, F.G.; Ikeuchi, M.; Michaels, A.S.; Schmidt, G.W.
The light-harvesting complex of photosystem I (LHCI) was isolated from wild-type cells of Chlamydomonas reinhardtii; the Chl a/b-protein complex contains four major polypeptides of approximately 27, 26, 24, and 20 kDa (polypeptides 14, 15, 17.2, and 22, respectively, in the nomenclature for Chlamydomonas thylakoid proteins). Antiserum against the 20-kDa subunit of LHCI was prepared and used to determine the membrane topology, subcellular site of synthesis, and cell-cycle regulation of this polypeptide. The results indicate that the 20-kDa subunit as well as the other major LHCI polypeptides are integral membrane proteins. Moreover, protease digestion experiments reveal that the 20-kDa polypeptide is completely protected by the membrane bilayer but the 27- and 26-kDa LHCI polypeptides are exposed at the membrane surface. In vivo synthesis of the 20-kDa polypeptide is sensitive to cycloheximide but not to chloramphenicol; the form of the polypeptide recovered from in vitro translations of polyadenylated RNA is approximately 24 kDa, 4 kDa larger than the mature polypeptide. It is concluded that this LHCI polypeptide is nuclear encoded and synthesized in the cytoplasm as a higher molecular weight precursor. Synthesis of the 20-kDa polypeptide is restricted to the light period in light-dark synchronized cells. Translatable mRNA for this polypeptide accumulates during the light but levels are dramatically reduced during the dark period. Thus, synthesis of the 20-kDa subunit of LHCI appears to be transcriptionally regulated during the cell cycle.
Molecular cloning, tissue distribution, and expression of a 14-kDa bile acid-binding protein from rat ileal cytosol.
Gong, Y Z; Everett, E T; Schwartz, D A; Norris, J S; Wilson, F A
A cDNA clone encoding the major intestinal cytosolic 14-kDa bile acid-binding protein (14-kDa I-BABP) was isolated from a rat ileal lambda gt22A library following immunoscreening using a monospecific antiserum raised against a 14-kDa polypeptide found in the rat ileal cytosol. One clone of 516 bp encoded a 128-amino acid protein with a predicted molecular mass of 14,544 Da. The deduced amino acid sequence of 14-kDa I-BABP showed 100% homology to rat intestinal 15-kDa protein (I-15P) and 72% homology to porcine 15-kDa gastrotropin, whereas comparison of I-BABP to rat 14-kDa fatty acid-binding proteins of liver, intestine, and heart revealed homologies of 44%, 25%, and 28%, respectively. Northern blot analysis revealed a single transcript of approximately 0.5 kb in ileum and ovary; however, the abundance of I-BABP mRNA was much greater in ileum than in ovary. No transcript was seen in RNA extracted from stomach, jejunum, colon, liver, adrenal, brain, heart, kidney, or testis. Transfection of the I-BABP cDNA into COS-7 cells resulted in the expression of a 14-kDa protein that was identical to the ileal cytosolic I-BABP as determined by immunoblotting. Photoaffinity labeling of expressed 14-kDa protein was saturable with respect to increasing concentrations of 7,7-azo[3H]taurocholate (Km, 83.3 microM; Vmax, 6.7 pmol/mg per 5 min). Taurocholate inhibited 7,7-azotaurocholate labeling by > 96% with lesser inhibition by taurochenodeoxycholate (83.1%), chenodeoxycholate (74.6%), cholate (50.5%), and progesterone (38.5%), whereas oleic acid and estradiol did not inhibit binding. Images PMID:8197128
Chlorophyll antenna proteins of photosystem I: topology, synthesis, and regulation of the 20-kDa subunit of Chlamydomonas light-harvesting complex of photosystem I.
Herrin, D L; Plumley, F G; Ikeuchi, M; Michaels, A S; Schmidt, G W
The light-harvesting complex of photosystem I (LHCI) was isolated from wild-type cells of Chlamydomonas reinhardtii; the Chl a/b-protein complex contains four major polypeptides of approximately 27, 26, 24, and 20 kDa (polypeptides 14, 15, 17.2, and 22, respectively, in the nomenclature for Chlamydomonas thylakoid proteins). Antiserum against the 20-kDa subunit of LHCI was prepared and used to determine the membrane topology, subcellular site of synthesis, and cell-cycle regulation of this polypeptide. The results indicate that the 20-kDa subunit as well as the other major LHCI polypeptides are integral membrane proteins. Moreover, protease digestion experiments reveal that the 20-kDa polypeptide is completely protected by the membrane bilayer but the 27- and 26-kDa LHCI polypeptides are exposed at the membrane surface. In vivo synthesis of the 20-kDa polypeptide is sensitive to cycloheximide but not to chloramphenicol; the form of the polypeptide recovered from in vitro translations of polyadenylated RNA is approximately 24 kDa, 4 kDa larger than the mature polypeptide. It is concluded that this LHCI polypeptide is nuclear encoded and synthesized in the cytoplasm as a higher molecular weight precursor. Synthesis of the 20-kDa polypeptide is restricted to the light period in light-dark synchronized cells. Translatable mRNA for this polypeptide accumulates during the light but levels are dramatically reduced during the dark period. Thus, synthesis of the 20-kDa subunit of LHCI appears to be transcriptionally regulated during the cell cycle. PMID:3555343
Constitutive expression of DaCBF7, an Antarctic vascular plant Deschampsia antarctica CBF homolog, resulted in improved cold tolerance in transgenic rice plants.
Byun, Mi Young; Lee, Jungeun; Cui, Li Hua; Kang, Yoonjee; Oh, Tae Kyung; Park, Hyun; Lee, Hyoungseok; Kim, Woo Taek
Deschampsia antarctica is an Antarctic hairgrass that grows on the west coast of the Antarctic peninsula. In this report, we have identified and characterized a transcription factor, D. antarctica C-repeat binding factor 7 (DaCBF7), that is a member of the monocot group V CBF homologs. The protein contains a single AP2 domain, a putative nuclear localization signal, and the typical CBF signature. DaCBF7, like other monocot group V homologs, contains a distinct polypeptide stretch composed of 43 amino acids in front of the AP2 motif. DaCBF7 was predominantly localized to nuclei and interacted with the C-repeat/dehydration responsive element (CRT/DRE) core sequence (ACCGAC) in vitro. DaCBF7 was induced by abiotic stresses, including drought, cold, and salinity. To investigate its possible cellular role in cold tolerance, a transgenic rice system was employed. DaCBF7-overexpressing transgenic rice plants (Ubi:DaCBF7) exhibited markedly increased tolerance to cold stress compared to wild-type plants without growth defects; however, overexpression of DaCBF7 exerted little effect on tolerance to drought or salt stress. Transcriptome analysis of a Ubi:DaCBF7 transgenic line revealed 13 genes that were up-regulated in DaCBF7-overexpressing plants compared to wild-type plants in the absence of cold stress and in short- or long-term cold stress. Five of these genes, dehydrin, remorin, Os03g63870, Os11g34790, and Os10g22630, contained putative CRT/DRE or low-temperature responsive elements in their promoter regions. These results suggest that overexpression of DaCBF7 directly and indirectly induces diverse genes in transgenic rice plants and confers enhanced tolerance to cold stress. PMID:26025521
Test of ''Crab-Waist'' Collisions at the DA{Phi}NE {Phi} Factory
Zobov, M.; Alesini, D.; Biagini, M. E.; Biscari, C.; Bocci, A.; Boni, R.; Boscolo, M.; Bossi, F.; Buonomo, B.; Clozza, A.; Delle Monache, G. O.; Demma, T.; Di Pasquale, E.; Di Pirro, G.; Drago, A.; Gallo, A.; Ghigo, A.; Guiducci, S.; Ligi, C.; Marcellini, F.
The electron-positron collider DA{Phi}NE, the Italian {Phi} factory, has been recently upgraded in order to implement an innovative collision scheme based on large crossing angle, small beam sizes at the crossing point, and compensation of beam-beam interaction by means of sextupole pairs creating a ''crab-waist'' configuration in the interaction region. Experimental tests of the novel scheme exhibited an increase by a factor of 3 in the peak luminosity of the collider with respect to the performances reached before the upgrade. In this Letter we present the new collision scheme, discuss its advantages, describe the hardware modifications realized for the upgrade, and report the results of the experimental tests carried out during commissioning of the machine in the new configuration and standard operation for the users.
Toward a Network of Faint DA White Dwarfs as High-precision Spectrophotometric Standards
NASA Astrophysics Data System (ADS)
Narayan, G.; Axelrod, T.; Holberg, J. B.; Matheson, T.; Saha, A.; Olszewski, E.; Claver, J.; Stubbs, C. W.; Bohlin, R. C.; Deustua, S.; Rest, A.
We present the initial results from a program aimed at establishing a network of hot DA white dwarfs to serve as spectrophotometric standards for present and future wide-field surveys. These stars span the equatorial zone and are faint enough to be conveniently observed throughout the year with large-aperture telescopes. The spectra of these white dwarfs are analyzed in order to generate a non-local-thermodynamic-equilibrium model atmosphere normalized to Hubble Space Telescope colors, including adjustments for wavelength-dependent interstellar extinction. Once established, this standard star network will serve ground-based observatories in both hemispheres as well as space-based instrumentation from the UV to the near IR. We demonstrate the effectiveness of this concept and show how two different approaches to the problem using somewhat different assumptions produce equivalent results. We discuss the lessons learned and the resulting corrective actions applied to our program.
The binary Feige 24 - The mass, radius, and gravitational redshift of the DA white dwarf
NASA Technical Reports Server (NTRS)
Vennes, Stephane; Shipman, Harry L.; Thorstensen, John R.; Thejll, Peter
Observations are reported which refine the binary ephemeris of the Feige 24 system, which contains a peculiar hot DA white dwarf and an M dwarf with an atmosphere illuminated by extreme ultraviolet radiation from the white dwarf. With the new ephemeris and a set of IUE high-dispersion spectra, showing phase-dependent redshifted C IV, N V, and Si IV resonance lines, the orbital velocity, and hence the mass (0.54 + or - 0.20 solar masses), and the gravitational redshift of the white dwarf (14.1 + or - 5.2 km/s) are determined independently. It is shown that the measured Einstein redshift is consistent with an estimated radius for the white dwarf obtained from a model atmosphere solid angle and a parallax measurement. This radius is twice the Hamada-Salpeter radius for the given mass and offers a prospect to investigate the presence of a massive hydrogen envelope in that white dwarf star.
Holberg, J. B.; Howell, Steve B. E-mail: email@example.com
The faint (g = 16.9) hot white dwarf BOKS 53856 was observed by the Kepler Mission in short cadence mode during mid-2009. Analysis of these observations reveals a highly stable modulation with a period of 6.1375 hr and a 2.46% half-amplitude. The folded light curve has an unusual shape that is difficult to explain in terms of a binary system containing an unseen companion more luminous than an L0 brown dwarf. Optical spectra of BOKS 53856 show a T{sub eff} = 34,000 K, log g = 8.0 DA white dwarf. There are few, if any, known white dwarfs in this temperature range exhibiting photometric variations similar to those we describe. A magnetic spin-modulated white dwarf model can in principle explain the light curve, an interpretation supported by spectral observations of the H{alpha} line showing evidence of Zeeman splitting.
Sensitivity of EIA in the diagnosis of tuberculosis using 38-kDa antigen.
Chander, J; Subrahmanyan, S; Gupta, R
Patients, who reported to Government Medical College Hospital, Chandigarh with suspected signs and symptoms of pulmonary tuberculosis, were examined. Their sputum was investigated for the presence of acid-fast bacilli, using the routine Ziehl-Neelsen staining, for 3 consecutive days. The positive cases were referred for treatment and the rest were referred for supplementary tests like x-ray, Mantoux test, FNAC and the presence of IgG antibodies in their serum against recombinant 38-kDa antigen derived from Mycobacterium tuberculosis, using micro-ELISA plates. It was found that 37.8% of the cases who were negative for acid-fast bacilli found to be positive for the presence of antimycobacterial IgG antibodies with an antibody titre greater than the cut off value in addition to one of the above mentioned positive supplementary finding. Hence, EIA for tuberculosis is more sensitive as compared to the routine acid-fast bacilli staining. PMID:9141876
First Results of the VLBI Experiment on Radar Location of the Asteroid 2012 DA14
NASA Astrophysics Data System (ADS)
Nechaeva, M.; Antipenko, A.; Bezrukov, D.; Bezrukovs, Vl.; Dementjev, A.; Dugin, N.; Jekabsons, N.; Khutornoy, R.; Klapers, M.; Konovalenko, A.; Kulishenko, V.; Nabatov, A.; Nesteruk, V.; Pupillo, G.; Reznichenko, A.; Salerno, E.; Shmeld, I.; Skirmante, K.; Tikhomirov, Yu.; Voytyuk, V.
An international VLBI experiment on radio location of the asteroid 2012 DA14 was organized on 2013 February 15--16, during its flyby close to Earth. The purpose of observations was to investigate and specify orbital parameters of the asteroid, as well as to evaluate its rotation period and other characteristics. The irradiation of the asteroid was performed by the RT-70 transmitter at Evpatoria (Crimea, Ukraine), while the reflected signals were successfully accepted by the two 32 m radio telescopes at Medicina (Bologna, Italy) and Irbene (Ventspils, Latvia). Processing and interpretation of the data were performed both in the Radiophysical Research Institute and in the Ventspils International Radio Astronomy Center. The first results of this experiment are presented and discussed.
Shadowing and absorption effects on J/psi production in dA collisions
Vogt, R.
The authors study medium modifications of J/{psi} production in cold nuclear media in deuterium-nucleus collisions. They discuss several parameterizations of the modifications of the parton densities in the nucleus, known as shadowing, an initial-state effect. They also include absorption of the produced J/{psi} by nucleons, a final-state effect. Both spatially homogeneous and inhomogeneous shadowing and absorption are considered. They use the number of binary nucleon-nucleon collisions as a centrality measure. Results are presented for d+Au collisions at {radical}S{sub NN} = 200 GeV and for d+Pb collisions at {radical}S{sub NN} = 6.2 TeV. To contrast the centrality dependence in pA and dA collisions, they also present pPb results at {radical}S{sub NN} = 8.8 TeV.
A 138-kDa glycoprotein from Dictyostelium membranes with folate deaminase and folate binding activity.
Greiner, R A; Jacobs-Krahnen, D; Mutzel, R; Malchow, D; Wurster, B
A 138-kDa glycoprotein comprising folate deaminase activity was purified to apparent homogeneity from membranes of Dictyostelium discoideum. Deaminase activity could be effectively inhibited by p-chloromercuriphenylsulfonate. This treatment protected folate from deamination and thus allowed investigation of folate binding to deaminase fractions. Two types of folate binding sites, differing in affinity and specificity, were detected on the folate deaminase glycoprotein. One type displays high affinity and binds folate stronger than N10-methylfolate. This binding site appears to be identical with the catalytic site of folate deaminase. The other type of binding site shows lower affinity but prefers N10-methylfolate relative to folate. A similar preference for N10-methylfolate was observed in chemotaxis tests pointing to the possibility that the second type of binding site is involved in chemotactic perception of folate compounds. Folate perception and deamination could thus be performed by activities residing on the same polypeptide. PMID:1544893
Preliminary XCOV26 Results For The Da White Dwarf EC14012-1446
NASA Astrophysics Data System (ADS)
Provencal, Judith L.; Thompson, S.; Montgomery, M.; Shipman, H.; Dalessio, J.; Whole Earth Telescope Team
EC14012-1446 is a hydrogen atmosphere (DA) white dwarf pulsator. Its rich pulsation spectrum displays a range of excited modes with complex multiplet structure, in addition to numerous combination frequencies. In April 2008, EC14012-1446 was the primary target of XCOV26. We obtained over 300 hrs of nearly continuous high speed photometry with the goal of using the nonlinear pulse shapes to empirically determine the parameters of the convection zone. The Fourier transform (FT) of the light curve contains power between 1000 to 4000 microHz, with the dominant peak at 1234 microHz. We find 13 independent frequencies distributed in 8 modes, as well as a myriad of combination frequencies. We present preliminary results of our asteroseismological investigation and lay the groundwork for future investigation leading to light curve fitting of EC14012-1446. This research is funded by Mt. Cuba Observatory (Greenville, DE) and the Crystal Trust Foundation.
Middle Paleolithic human remains from the Gruta Da Oliveira (Torres Novas), Portugal.
Willman, John C; Maki, Julia; Bayle, Priscilla; Trinkaus, Erik; Zilhão, João
Additional Middle Paleolithic human remains from layers 17, 18, and 22 of the Gruta da Oliveira, Portugal consist of a proximal manual phalanx 2 (Oliveira 5), a partial postcanine tooth (Oliveira 6), a humeral diaphysis (Oliveira 7), a distal mandibular molar (Oliveira 8), and a mandibular premolar (P(3) ) (Oliveira 9). Oliveira 5, 6, and 8 are unremarkable for Late Pleistocene humans. The Oliveira 7 right humerus is moderately robust or the individual had the stocky body proportions of other European (including Iberian) Neandertals. The Oliveira 9 P(3) has a large and symmetrical crown and lacks a distal accessory ridge and accessory lingual cusps, overlapping both Neandertal and recent human ranges of variation. It contrasts with at least recent human P(3) s in having relatively thin enamel. These join the Oliveira 1 to 4 remains in further documenting early MIS 3 Neandertal morphology in western Iberia. PMID:22610966
Hermes, J. J.; Montgomery, M. H.; Winget, D. E.; Mullally, Fergal; Bischoff-Kim, A.
We report the most rapid rate of period change measured to date for a pulsating DA (hydrogen atmosphere) white dwarf (WD), observed in the 292.9 s mode of WD 0111+0018. The observed period change, faster than 10{sup -12} s s{sup -1}, exceeds by more than two orders of magnitude the expected rate from cooling alone for this class of slow and simply evolving pulsating WDs. This result indicates the presence of an additional timescale for period evolution in these pulsating objects. We also measure the rates of period change of nonlinear combination frequencies and show that they share the evolutionary characteristics of their parent modes, confirming that these combination frequencies are not independent modes but rather artifacts of some nonlinear distortion in the outer layers of the star.
Basil as functional and preserving ingredient in "Serra da Estrela" cheese.
Carocho, Márcio; Barros, Lillian; Barreira, João C M; Calhelha, Ricardo C; Soković, Marina; Fernández-Ruiz, Virginia; Buelga, Celestino Santos; Morales, Patricia; Ferreira, Isabel C F R
Antitumor, antimicrobial and antioxidant activities of basil were studied, along with its characterization in phenolic compounds, organic acids and soluble sugars. The results placed basil as a valuable candidate for functionalization and conservation of food products, maintaining their nutritional properties, while increasing their shelf life and potential health effects. The basil leaves were then incorporated in "Serra da Estrela Cheese", either in its dehydrated form or as a decoction. The cheeses were then subject to a nutritional evaluation, being characterized for their fatty acids, minerals and CIE color parameters. To assess the combined effects of plant incorporation and storage time, a 2-way ANOVA was used to process the results, further analysed through a linear discriminant analysis. Overall, basil leaves provided antioxidant activity to the cheeses, reduced the moisture, and preserved the unsaturated fatty acids and proteins. Comparing both incorporation types, the decoctions had a higher functionalizing and conservative effect. PMID:27080879
Application of da Vinci® Robot in simple or radical hysterectomy: Tips and tricks
Iavazzo, Christos; Gkegkes, Ioannis D.
The first robotic simple hysterectomy was performed more than 10 years ago. These days, robotic-assisted hysterectomy is accepted as an alternative surgical approach and is applied both in benign and malignant surgical entities. The two important points that should be taken into account to optimize postoperative outcomes in the early period of a surgeon's training are how to achieve optimal oncological and functional results. Overcoming any technical challenge, as with any innovative surgical method, leads to an improved surgical operation timewise as well as for patients' safety. The standardization of the technique and recognition of critical anatomical landmarks are essential for optimal oncological and clinical outcomes on both simple and radical robotic-assisted hysterectomy. Based on our experience, our intention is to present user-friendly tips and tricks to optimize the application of a da Vinci® robot in simple or radical hysterectomies. PMID:27403078
Microbiological Analysis of Surfaces of Leonardo Da Vinci's Atlantic Codex: Biodeterioration Risk
Moroni, Catia; Pasquariello, Giovanna; Maggi, Oriana
Following the discovery of discoloration on some pages of the Atlantic Codex (AC) of Leonardo da Vinci kept in the Biblioteca Ambrosiana in Milan, some investigations have been carried out to verify the presence of microorganisms, such as bacteria and fungi. To verify the presence of microorganisms a noninvasive method of sampling has been used that was efficient and allowed us to highlight the microbial facies of the material that was examined using conventional microbiological techniques. The microclimatic conditions in the storage room as well as the water content of the volume were also assessed. The combined observations allowed the conclusion that the discoloration of suspected biological origin on some pages of AC is not related to the presence or current attack of microbial agents. PMID:25574171
The "tobacco mosaic virus" 126-kDa protein associated with virus replication and movement suppresses RNA silencing
Technology Transfer Automated Retrieval System (TEKTRAN)
Systemic symptoms induced on "Nicotiana tabacum" cv. Xanthi by "Tobacco mosaic virus" (TMV) are modulated by one or both amino-coterminal viral 126- and 183-kDa proteins, proteins involved in virus replication and cell-to-cell movement. Here we compare the systemic accumulation and gene silencing c...
Lemon oil vapor causes an anti-stress effect via modulating the 5-HT and DA activities in mice.
Komiya, Migiwa; Takeuchi, Takashi; Harada, Etsumori
We examined the anti-stress action of the essential oils of lavender, rose, and lemon using an elevated plus-maze task (EPM), a forced swimming task (FST), and an open field task (OFT) in mice. Lemon oil had the strongest anti-stress effect in all three behavioral tasks. We further investigated a regulatory mechanism of the lemon oil by pre-treatments with agonists or antagonists to benzodiazepine, 5-HT, DA, and adrenaline receptors by the EPM and the FST. The anti-stress effect of lemon oil was significantly blocked by pre-treatment with frumazenil, benzodiazepine receptor antagonist, or apomorphine, a nonselective DA receptor agonist. In contrast, agonists or antagonists to the 5-HT receptor and the alpha-2 adrenaline receptor did not affect the anti-stress effect of lemon oil. Buspirone, DOI, and mianserine blocked the antidepressant-like effect of lemon oil in the FST, but WAY100,635 did not. These findings suggest that the antidepressant-like effect of lemon oil is closely related with the 5-HTnergic pathway, especially via 5-HT(1A) receptor. Moreover, the lemon oil significantly accelerated the metabolic turnover of DA in the hippocampus and of 5-HT in the prefrontal cortex and striatum. These results suggest that lemon oil possesses anxiolytic, antidepressant-like effects via the suppression of DA activity related to enhanced 5-HTnergic neurons. PMID:16780969
Bringing the ZPD into the Equation: Capturing L2 Development during Computerized Dynamic Assessment (C-DA)
ERIC Educational Resources Information Center
Poehner, Matthew E.; Lantolf, James P.
Dynamic Assessment (DA) derives from Vygotsky's (1987) insight that the use of mediation, attuned to learner needs, enables learners to perform beyond their current level of functioning, thereby providing insights into emerging capabilities. The instructional quality of mediation means that this process simultaneously creates possibilities…
AVALIAÇÃO DA PRESENÇA DE ENDOSSIMBIONTES Cardinium em DIFERENTES ESPÉCIES DE ARTRÓPODES.
Technology Transfer Automated Retrieval System (TEKTRAN)
A presença de endossimbiontes do gênero Cardinium em alguns grupos de artrópodes foi recentemente relatada e relacionada com diversas alterações reprodutivas em seus hospedeiros, tais como feminilização de ácaros, partenogênese em parasitóides, incompatibilidade citoplasmática e aumento da fecundida...
Simultaneous voltammetric determination for DA, AA and NO₂⁻ based on graphene/poly-cyclodextrin/MWCNTs nanocomposite platform.
Zhang, Yu; Yuan, Ruo; Chai, Yaqin; Li, Wenjuan; Zhong, Xia; Zhong, Huaan
In the present work, graphene sheets (GS) and multiwall carbon nanotubes (MWCNTs) were dispersed in the mixed solution of cyclodextrin (CD) and cyclodextrin prepolymer (pre-CD) and were used as modifier to fabricate chemical modified electrode to simultaneous detect dopamine (DA), ascorbic acid (AA) and nitrite (NO2(-)). CD cross-linked pre-CD (CDP) displays excellent film forming ability, which made the electrode stable. Comparing with CDP-GS, CDP-MWCNTs and CDP-GS-MWCNTs modified electrodes, the CDP-GS-MWCNTs displays higher catalytic activity and selectivity toward the oxidation of DA, AA and NO2(-), revealing that MWCNTs effectively inhibited the stacking of individual GS and enhanced the utilization of GS based composites. The host-guest chemical reaction ability of CD and π-π stacking interaction between detected molecules and GS-MWCNTs surface were considered as the main reasons of the successfully simultaneous detection of DA, AA and NO2(-). Cyclic voltammetry (CV), scanning electron microscopy (SEM) and different pulse voltammetry (DPV) were employed to characterize the biosensor. The linear response range for AA, DA and NO2(-) were 5 μM-0.48 mM, 0.15-21.65 μM and 5 μM-6.75 mM, respectively and the detection limits were 1.65 μM, 0.05 μM and 1.65 μM. PMID:21497078
Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori
Son, Mina; Lee, June Yong
Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm. PMID:26770033
Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori.
Jeong, Kyoung Yong; Son, Mina; Lee, June Yong; Park, Kyung Hee; Lee, Jae-Hyun; Park, Jung-Won
Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm. PMID:26770033
An 11-kDa form of human immunodeficiency virus protease expressed in Escherichia coli is sufficient for enzymatic activity.
Graves, M C; Lim, J J; Heimer, E P; Kramer, R A
In order to define the protease domain of human immunodeficiency virus 1, various regions of the pol open reading frame were cloned and expressed in Escherichia coli. Antiserum directed against the conserved retroviral protease active site was used to identify pol precursor and processed species containing the presumed protease domain. The smallest product that accumulates is about 11 kDa as measured by NaDodSO4/PAGE. This size agrees with that predicted from the presence in this region of two Phe-Pro sequences, which is one of the cleavage sites recognized by HIV protease. DNA encoding only the predicted 11-kDa protein was cloned, bypassing the need for autoprocessing, and the protein was expressed to a high level in E. coli. This form is active as demonstrated by its ability to specifically cleave protease-deficient pol protein in vivo in E. coli. Extracts of E. coli containing the 11-kDa protease also process human immunodeficiency virus gag substrates in vitro. These results demonstrate that the 11-kDa protease is sufficient for enzymatic activity and are consistent with a major role for this form in virus maturation. Images PMID:3282230

subset: commoncrawl