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I mentioned a couple of posts ago that I was going to dust off my trusty two-wheeler and start pedaling to get my cardio fix.
Well, the best laid plans...
A surge of work came in nearly two weeks ago that has allowed little free time. Cycling takes more planning than most other forms of exercise, and to get a decent workout, I feel like I need to ride for at least an hour. So I haven't gone for a ride yet since we moved here over a month ago.
However, I've made time to lift weights three days per week. Last week, I had an overwhelming urge to do a short "cooldown" run after each workout. So, last Monday, Wednesday and Friday, I ran two loops around my neighborhood each time for a total of 6.3 miles for the week. There was a time not long ago that I would average that per day, but I'm happy to have "jogged" (that's what it felt like) even that paltry mileage. I did feel some aches and pains, but nothing too extreme.
I'll probably continue with this program for a bit just to see how my body will respond. I still have plans to get out and ride, too.
Even though I intentionally bulked up when I switched to weight lifting back in March, I do need to drop a few of those pounds. The extra 17 lbs. are definitely noticeable during these runs.
So, although it's not much to report, it IS running, even if just a little.


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Eric Allen
Research in our laboratory centers upon the use of environmentally-derived genome sequence information to explore the genetic potential, ecology, and evolution of environmental microbial populations. The nature of this work relies equally upon field-based collections, bioinformatics (genome assembly, annotation, and comparative analyses) and the tools of molecular ecology and genetics. Together, these approaches enable us to utilize environmental genome sequence data to understand natural microbial phenomena including environmental adaptation, evolutionary processes, lateral gene transfer events, biogeographical patterning, biogeochemical cycling, microbial interactions, and in situ metabolic activity.
Current investigations include exploring the genetic composition of microbial populations (Archaea & Bacteria) inhabiting hypersaline environments via cultivation- independent genomic approaches ("metagenomics"). Our primary study site for this project is a hypersaline lake system in NW Victoria, Australia (near the town of Sea Lake approx. 370 km NW of Melbourne). With over 1 billion bases (>1 Gbp) of environmental genome sequence data generated, near complete and complete genome sequences for resident microbial populations can be analyzed in the absence of cultivation requirements. Such a data set uniquely allows analysis of population structure (allelic variation) thus providing insight into natural mechanisms of genomic heterogeneity, diversification, and environmental selection. Additional projects are aiding in the analysis of various marine microbial metagenomic projects including the J. Craig Venter Institute's Global Ocean Sampling (GOS) expedition.
An omnipresent facet of the research in our laboratory is the use of genome sequence information (environmental or isolate-derived) to expand our understanding of microbial physiology and its relationship to environmental adapation and ecosystem function. Abiding by this paradigm, additional projects underway in our laboratory include:
- Diversity and distribution of microbial "secondary lipid" biosynthesis
- Ecology & evolution of microbial polyunsaturated fatty acid biosynthesis
- Biological controls on Hg methylation in marine & estuarine sediments
- Analysis of microbial & meiofaunal diversity at the Land/ Sea Interface
Select Publications
- Zeigler-Allen L, Allen EE, Badger J, McCrow JP, Paulson I, Elbourne LDH, Thiagrijan M, Nealson KH, Williamson SJ, Venter JC, Allen AE. (2012). Influence of nutrients and currents on the genomic composition of microbes across an upwelling mosaic. ISME J in press.
- Ugalde JA, Podell S, Narasingarao P, Allen EE. (2011). Xenorhodopsins, an enigmatic new class of microbial rhodopsins horizontally transferred between Archaea and Bacteria. Biol Direct 6:52.
- Narasingarao P, Podell S, Ugalde JA, Brochier-Armanet C, Emerson J, Brocks JJ, Heidelberg, KB, Banfield JF, Allen EE. (2011). De novo metagenomic assembly reveals abundant novel major lineage of Archaea in hypersaline microbial communities. ISME J Advanced Online Publication, 30 June 2011; doi:10.1038/ismej.2011.78.
- Shulse CN, Allen EE. (2011). Widespread occurence of secondary lipid biosynthesis potential in microbial lineages. PLoS One 6:e20146.
- Eloe EA, Fadrosh DW, Novotny M, Zeigler Allen L, Kim M, Lombardo M-J, Yee-Greenbaum J, Yooseph S, Allen EE, Lasken R, Williamson SJ, Bartlett DH. (2011). Going deeper: Metagenome of a hadopelagic microbial community. PLoS One 6:e20388.
- Jones A, Monroe E, Podell S, Hess W, Klages S, Esquenazi E, Niessen S, Hoover H, Rothmann M, Lasken R, Yates III J, Reinhardt R, Kube M, Burkart M, Allen EE, Gerwick W, Gerwick L. (2011). Genomic insights into the physiology and ecology of the marine filamentous cyanbacterium Lyngbya majuscule. Proc Natl Acad Sci USA 108:8815-8820.
- Shulse CN, Allen EE. (2011). Diversity and distribution of microbial long-chain fatty acid biosynthetic genes in the marine environment. Environ Microbiol 13:684-695.
- Eloe EA, Shulse CN, Fadrosh DW, Williamson SJ, Allen EE, Bartlett DH. (2010). Compositional differences in particle-associated and free-living microbial assemblages from an extreme deep ocean environment. Environ Microbiol Reports (first published online : 24 NOV 2010), DOI: 10.1111/j.1758-2229.2010.00223.
- Sun S, Chen J, Li W, Altintas I, Lin A, Peltier S, Stocks K, Allen EE, Ellisman M, Grethe J, Wooley J. (2010). Community cyberinfrastructure for Advanced Microbial Ecology Research and Analysis: the CAMERA resource. Nucl Acids Res 39:D546-551.
- Han S, Narasingarao P, Obraztsova A, Gieskes J, Hartmann AC, Tebo BM, Allen EE, Deheyn DD. (2010). Mercury speciation in marine sediments under sulfate-limited conditions. Env Sci Technol 44:3752-3757.
- Morin PA, Archer FI, Foote AD, Vilstrup J, Allen EE, Wade P, Durban J, Parsons K, Pitman R, Li L, Bouffard P, Abel Nielsen SC, Rasmussen M, Willerslev E, Gilbert MTP, Harkins T. (2010). Complete mitochondrial genome phylogeographic analysis of killer whales ( Orcinus orca) indicates multiple species. Genome Res 20:908-916.
- Penn, K, Jenkins C, Nett M, Udwary DW, Gontang EA, McGlinchey RP, Foster B, Lapidus A, Podell S, Allen EE, Moore BS, Jensen PR. (2009). Genomic islands link secondary metabolism to functional adaptation in marine Actinobacteria. ISME J 3:1193-1203.
- Podell S, Gaasterland T, Allen EE. (2008). A database of phylogenetically atypical genes in archaeal and bacterial genomes, identified using the DarkHorse algorithm. BMC Bioinformatics 9:419.
- Robidart JC, Bench SR, Feldman RA, Novoradovsky A, Podell SB, Gaasterland T, Allen EE, Felbeck H. (2008). Metabolic versatility of the Riftia pachyptila endosymtiont revealed through metagenomics. Environ Microbiol 10:727-737.
- Lo I, Denef V, VerBerkmoes NC, Shah M, Goltsman D, DiBartolo G, Tyson GW, Allen EE, Ram RJ, Detter C, Richardson P, Thelen MP, Hettich RL, Banfield JF. (2007). Strain-resolved community proteomics reveals that recombination shapes the genomes of acidophilic bacteria. Nature 446:537-541.
- Allen EE, Tyson GW, Whitaker RJ, Detter JC, Richardson PM, Banfield JF. (2007). Genome dynamics in a natural archaeal population. Proc Natl Acad Sci USA 104:1883-1888.
- Baker BJ, Tyson GW, Webb RI, Flanagan J, Hugenholtz P, Allen EE, Banfield JF. (2006). Lineages of acidophilic archaea revealed by community genomic analysis. Science 314:1933-1935.
- Banfield JF, Tyson GW, Allen EE, Whitaker RJ. (2005). The search for a molecular-level understanding of the processes that underpin the Earth's biogeochemical cycles. Rev Mineral Geochem 59:1-7.
- Tyson GW, Lo I, Baker BJ, Allen EE, Hugenholtz P, Banfield JF. (2005). Genome-directed isolation of the key nitrogen fixer, Leptospirillum ferrodiazotrophum sp. nov., from an acidophilic microbial community. Appl Environ Microbiol 71:6319-6324.
- Allen EE, Banfield JF. (2005). Community genomics in microbial ecology and evolution. Nature Rev Microbiol 3:489-498.
- Tyson GW, Chapman J, Hugenholtz P, Allen EE, Ram R, Richardson P, Solovyev V, Rokhsar D, Banfield JF. (2004). Community structure and metabolism through reconstruction of microbial genomes from the environment. Nature 428:37-43.
- Palenik B, Brahamsha B, Larimer FW, Land L, Hauser L, Chain P, Lamerdin J, Regala W, Allen EE, McCarren J, Paulsen I, Dufresne A, Partensky F, Webb EA, Waterbury J. (2003). The genome of a motile marine Synechococcus. Nature 424:1037-1042.
- Allen EE, Bartlett DH. (2002). Structure and regulation of the omega-3 polyunsaturated fatty acid synthase genes from the deep-sea bacterium Photobacterium profundum strain SS9. Microbiology 148:1903-1914.
- Allen EE, Bartlett DH. (2002). Piezophiles: microbial adaptation to the deep-sea environment. In 'Extremophiles', UNESCO Encyclopedia for Life Support Systems. EOLSS Publishers, Oxford, UK.
- Allen EE, Bartlett DH. (2000). FabF is required for piezoregulation of cis-vaccenic acid levels and piezophilic growth of the deep-sea bacterium, Photobacterium profundum strain SS9. J Bacteriol 182:1264-1271.
- Allen EE, Facciotti D, Bartlett DH. (1999). Monounsaturated but not polyunsaturated fatty acids are required for growth of the deep-sea bacterium Photobacterium profundum SS9 at high pressure and low temperature. Appl. Environ Microbiol 65:1710-1720.
Eric Allen received his B.S. from the University of Oregon and his Ph.D. from the Scripps Institution of Oceanography at UC San Diego. He performed postdoctoral research at the University of California, Berkeley as an NSF Postdoctoral Fellow in Microbial Biology and joined the faculty of Biological Sciences and the Scripps Institution of Oceangraphy at UCSD in 2006.

subset: commoncrawl


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Bonaparte, Charles Lucian, 1803-1857 / American ornithology, or, The natural history of birds inhabiting the United States, not given by Wilson : with figures drawn, engraved, and coloured, from nature
Vol. I (1825)
Rocky-Mountain antcatcher. Myiothera obsoleta. Plate I. Fig. 2, pp. 6-11
Page 6
Based on date of publication, this material is presumed to be in the public domain.| For information on re-use, see http://digital.library.wisc.edu/1711.dl/Copyright


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- Research article
- Open Access
Differential development of oil granulomas induced by pristane injection in galectin-3 deficient mice
© Brand et al. 2015
- Received: 4 September 2015
- Accepted: 6 November 2015
- Published: 14 November 2015
Galectin-3 is known to be a lectin that plays an important role in inflammatory processes, acting as pro-inflammatory mediator in activation and migration of neutrophils and macrophages, as well as in the phagocytic function of these cells. The injection of mineral oils into the peritoneal cavity of mice, such as 2, 6, 10, 14-tetramethylpentadecane (pristane), induce a chronic granulomatous inflammatory reaction which is rich in macrophages, B cells and peritoneal plasma cells known as oil granuloma. In addition, this inflammatory microenvironment provided by oil granulomas is also an important site of plasmacytoma induction, which are dependent on cytokine production and cellular mobilization.
Here, we have analyzed the role of galectin-3 in inflammatory cells mobilization and organization after pristane injection characterizing granulomatous reaction through the formation of oil granulomas.
In galectin-3 deficient mice (gal-3−/−), the mobilization of inflammatory cells, between peritoneal cavity and bone marrow, was responsible for the formation of disorganized oil granulomas, which presented scattered cells, large necrotic areas and low amounts of extracellular matrix. The production of inflammatory cytokines partially explained the distribution of cells through peritoneal cavity, since high levels of IL-6 in gal-3−/− mice led to drastically reduction of B1 cells. The previous pro-inflammatory status of these animals also explains the excess of cell death and disruption of oil granulomas architecture.
Our data indicate, for the first time, that the disruption in the inflammatory cells migration in the absence of galectin-3 is a crucial event in the formation and organization of oil granulomas.
- Pristane
- Oil granuloma
- Bone marrow
- Peritoneal cavity
- Inflammation
Chronic inflammation represents a heterogeneous group of physiological conditions that may affect virtually any organ or system. Histologically, the chronic inflammation exhibits a predominance of mononuclear cells, such as macrophages, lymphocytes and plasma cells. In contrast, neutrophils are relevant for the initial phases of the inflammatory process [1, 2]. The stromal milieu (cellular and matrix components) regulates leukocyte production and recruitment from the bone marrow to the site of the inflammatory stimulus, determining the outcome of the process. However, the production of bone marrow cells, including neutrophils, is also largely regulated by the rate of apoptosis of hematopoietic cells in the site of inflammation in a T cell dependent-manner [3, 4].
In this context, continuous and unbalanced recruitment of inflammatory cells such as neutrophils, monocytes, lymphocytes and dendritic cells, from the bone marrow or adjacent tissues, toward the chronic site of stimulus can be determinant to the development of several immune responses [5].
Chronic inflammation can be triggered by long-lasting exposure to naturally occurring hydrocarbon oils such as pristane (2, 6, 10, 14-tetramethylpentadecane, TMPD) [6], which is not metabolized and causes severe physiological consequences to mammals [7]. Although, the clinical significance of these lesions in the development of an infection or neoplasia remains unknown. Epidemiologic studies suggest that occupational exposure to hydrocarbon oils or petroleum waste is associated to rheumatoid arthritis and systemic lupus erythematous [8, 9]. In experimental mouse models, pristane induces a continuous intraperitoneal chronic inflammation followed by oil-granuloma formation and plasma cell tumors, known as plasmacytoma [10–12]. These granulomas are frequently adhered to peritoneal structures correlated to vigorous immune response, autoantibody production and clinical manifestations resembling systemic lupus erythematous [13].
Pristane-induced granulomas are characterized by individual and clustered cells adhered to the mesentery [14] and other peritoneal tissues often covered by mesothelial cells. Angiogenesis in these structures is also established by terminal mesenteric vessels. Moreover, the structure of oil granulomas is regulated by several cytokines, such as IL-6, IFNs, IL-2 and TNF-α [7, 15–17]. These oil granulomas are considered tertiary lymphoid tissues and are enriched by terminally differentiated plasma cells, macrophages and resident self-renewing peritoneal B-1 lymphocytes. In addition, they may be putative targets to understand plasmacytoma development [11, 18]. Although oil granulomas have been well characterized, the role of the first recruited cells to the peritoneal microenvironment and their participation in the organization of the granuloma are not well understood.
Galectin-3, a β-galactoside binding protein, modulates both chronic inflammation and fibrogranulomatous reaction [19], regulating cell-cell and cell-extracellular matrix interactions in healthy and pathological conditions [20–23]. Galectin-3 deficient mice (Gal-3−/− mice) have a delayed monocyte differentiation into macrophages and neutrophil mobilization [24]. On the other hand, these mice have a significant plasmacytogenesis in secondary lymphoid tissues. In addition, the mesenteric membranes histologically correlated with disturbances in B lymphocyte and plasma cell niches [25–27].
In this study, we investigated the histological organization of pristane-induced oil granulomas in the absence of galectin-3. The kinect of cell mobilization and oil granuloma formation were analyzed in different time points of the inflammatory process. In the absence of galectin-3, the organization of oil granulomas was dependent on both bone marrow and peritoneal cavity cells, leading to a necrotic tissue, extracellular matrix disruption and disorganized cellular distribution. On the other hand, oil granulomas formed in the presence of galectin-3, did not present intense cell death, favoring maintenance and amplification of hematopoietic cells, mainly granulocytes.
Cellularity in galectin-3−/− mice is influenced by pristane injection and affects lymphocyte subpopulations
Lymphoid compartment along pristane injection presented differences in CD4+ and CD8+ T lymphocyte populations, both in bone marrow and peritoneal cavity. In the absence of gal-3, CD5 + CD4+ T cells were drastically reduced in bone marrow at 60 days after pristane injection (Fig. 1c and d). However, this cell population was not observed in peritoneal cavity, where CD5 + CD4+ T lymphocytes presented an initial decrease at day 7, but recovered at day 60 to initial numbers compared to gal-3−/− control mice (Fig. 1d). On the other hand, CD5 + CD8+ T lymphocytes presented an opposite behavior, with initial increase, and then, returning to initial total values after 60 days when compared to control group (Fig. 1e). In contrast to gal-3−/−, WT animals presented an increase in CD5 + CD4+ T lymphocytes in bone marrow (Fig. 1c), accompanied by the increase of this population in the peritoneal cavity (Fig. 1d). However, no significant difference was observed in bone marrow CD5 + CD8+ T cells (Fig. 1e). In peritoneal cavity, CD5 + CD8+ T cells behavior was similar to that observed in gal-3−/− mice (Fig. 1f).
The pattern of granulocyte response is modified in the absence of galectin-3 and influences the oil granuloma composition
As described above, the increased number of GrlowCD11bhi myeloid progenitor cells and GrhiCD11bhi mature myeloid cells in the peritoneal cavity of WT mice (Fig. 3b and d, respectivelly) could reflect the cellular composition of oil granulomas in these mice. Indeed, Fig. 3e shows an intense presence of polymorphonuclear cells foci (arrows and insert), indicating a peripheral amplification inside the inflammatory site, in a time independent manner from the bone marrow production. In gal-3−/− mice, oil granulomas were composed mainly by mononuclear cells, without the organization of polymorphonuclear foci (Fig. 3f, arrowheads and insert).
Galectin-3 in granulomas induced by pristane differentially organizes cellular and extracellular components
Oil granulomas in WT animals, macroscopically examined at 60 days after pristane injection (Fig. 4a, white arrows), exhibited two distinct regions when histologically observed: a highly cellular cortical region (Fig. 4c, above dotted line) and an inner region less cellular, but rich in lipid material. The granulomas presented several blood vessels (Fig. 4c, red circles, and e), in many instances structured with a muscular tunica (Fig. 4e, insert). Macroscopic structures of gal-3−/− mice oil granulomas were also obtained (Fig. 4b, white arrows). However, we did not observe a clear compartmentalization of the oil granulomas formed in gal-3−/− mice. Instead, cells were randomly distributed, with blood vessels surrounded by inflammatory cells (Fig. 4d and f, arrowheads). Presence of blood vessels was confirmed by CD31 immunolocalization, as demonstrated in Fig. 4g and h. In WT mice, we observed a higher quantity of blood vessels, as observed in Fig. 4i, when compared to gal-3−/− animals. These results suggest a favoring of cell mobilization process from both bone marrow and peritoneal cavity.
We took advantage of the higher resolution of TEM to study the ultrastructure of granulomas. As previously observed, WT mice oil granulomas presented a large amount of randomly distributed collagen fibrils (Fig. 5c and e, black arrows). On the other hand, we observed by TEM the presence of large amounts of necrotic tissues and dispersed ECM in oil granulomas from gal-3−/−animals (Fig. 5d, white arrows). Furthermore, a drastic reduction in the amount of collagen fibrils was observed. They were dispersed in small quantities in the necrotic tissue, as demonstrated by trichrome staining (Fig. 5f, black arrows).
These data indicate that cellular and extracellular matrix composition inside inflammatory site of oil granulomas could negatively regulate bone marrow response, favoring a peripheral cellular amplification that may lead to different consequences to the outcome of the inflammatory process.
The initial profile of cytokines produced by adherent peritoneal cells is the major responsible for the input of the inflammatory response
As we observed a drastic reduction of B lymphocytes in the absence of galectin-3, both in bone marrow and peritoneal cavity, and a differential oil granuloma organization between WT and gal-3−/− mice, we investigated the expression of major pro-inflammatory cytokines by adherent peritoneal cavity cells. For this, we injected pristane in WT and gal-3−/− mice and at 48 h or at 7 days we quantified cytokine mRNA levels expressed by the cells in the peritoneal cavity.
Macrophage inflammatory protein 2 (MIP-2), a cytokine related to neutrophil recruitment, was also analyzed. As we can observe in Fig. 7d, WT mice presented a down regulation of the mRNA levels at 48 h after pristane inoculation. In contrast, there was no significant difference in the mRNA levels in gal-3−/−. Curiously, when we compared the expression levels of mRNA of MIP-2 between WT and gal-3−/− mice, this cytokine was more expressed in the absence of galectin-3 (Fig. 7d).
Expression of cytokines related to macrophage polarization, arginase and inducible nitric oxide synthase (iNOS), were analyzed in peritoneal adherent cells (Fig. 7e and f, respectivelly). Arginase expression presented the same pattern in WT and gal-3−/− mice, increasing expression after 48 h of pristane stimulus, maintaining these levels increased until 7 days. However, this increase was significantly greater in the absence of galectin-3 when compared to WT animals (Fig. 7e). On the other hand, iNOS expression was not altered both in WT and gal-3−/− mice during treatment. However, expression levels of iNOS in the absence of galectin-3 were increased in all time points analyzed when compared to WT animals (Fig. 7f).
In this study, our results indicate, for the first time, that differential granulomatous reaction and its components, including the presence of cellular debris, regulate mobilization and amplification of inflammatory cells from the bone marrow to the initial site of inflammation.
As observed in our data, oil granulomas formed in WT mice presented greater amounts of neutrophils distributed though parenchyma, followed by a very well organized structure, rich in blood vessels and a scaffold of connective tissue formed by collagen. As described in the literature [7, 28] these structures are formed by large amounts of lipid material, surrounded by inflammatory cells organized as lymphoid follicles, e.g., B lymphocytes nodules surrounded by T and dendritic cells. However, our data showed an intense infiltration of granulocytic cells in WT mice. These conflicting data may be due to the time frame after pristane stimulus chosen to be analyzed. Indeed, Nacionales' group performed their analysis at least 12 weeks after pristane stimulus [7], whereas we analyzed after 8 weeks, using the same mice strain, eliminating possible genetic background influence. However, Chen et al. demonstrated that genetic background contributes to organization and composition of oil granulomas, raising conflicting data in the literature [14, 28]. Thus, both time frame and genetic background can influence separately or synergistically the development of oil granulomas.
On the other hand, oil granulomas formed in gal-3−/− animals were mainly composed by mononuclear cells, including B and T lymphocytes, and reduced amounts of neutrophils. These structures presented low vascularization and ECM, which likely promoted intense cell death. The reduced amounts of neutrophils in peritoneal cavity of gal-3−/− mice could reflect the delay in myeloid cell maturation [27] or a delayed migration from the bone marrow [24], as described in the literature. Schauer and Gresnigt groups have described an important role of neutrophils in resolution of the inflammatory process by reducing proinflammatory cytokines production [29, 30]. In view of these results, it is important to analyze in our model the role of neutrophils in late stages of pristane-induced inflammation and their outcome to plasmacytoma and/or autoimmune diseases development.
The observation of an increase in the expression levels of MIP-2 in gal-3−/− mice, an important cytokine involved in neutrophils recruitment [31, 32], suggests that peritoneal macrophages upregulate MIP-2 in an attempt to compensate the reduced recruitment of neutrophils to inflammatory site or the excess of cell death observed in these animals.
Galectin-3 plays an important role as a negative regulator in apoptotic events [26, 33], so that oil granulomas deficient in this lectin presented a great number of cells positively stained for annexin-V, apoptotic cells and great amounts of cell debris. The persistence of apoptotic cells and cell debris in situ could also be a result of low phagocytic state of macrophages, as our group previously described in gal-3−/− mice [26].
The increase in the expression levels of IL-12 p40 and IL-6 expression is already described in the literature as negative regulator of B1 B lymphocytes [34]. As we observed in our data, in the absence of galectin-3, IL-6 expression in adherent cells was significantly upregulated 7 days after pristane injection. In addition, there was a drastic reduction of B1 lymphocytes in the peritoneal cavity. Interestingly, these same parameters were observed in WT mice only 60 at days after pristane injection. Both IL-6 and IL-12p40 upregulation were observed in WT and gal-3−/− mice. On the other hand, TNF-α down modulation was also observed in WT mice, corroborating data described by Chae and Shin [35], however, this reduction was more drastic in the absence of galectin-3. Adherent cells of WT mice expressed high levels of arginase and presented no differences in iNOS expression after pristane injection, suggesting polarization of macrophages to an anti-inflammatory M2 phenotype. In the gal-3−/− mice, we also observed polarization to M2 phenotype coupled to unchanged levels of no iNOS expression. However, since these animals have high expression of iNOS in the absence of inflammatory stimulus, we suggest that this initial pro-inflammatory M1 background could disturb the response, maintaining the inflammatory status and promoting a rise in cell death within the oil granulomas.
Bone marrow compartment is a productive site of hematopoietic cells, which are continuously mobilized to the primary site of inflammation, and contribute to the development and organization of oil granulomas. In our model, as expected, bone marrow cellularity clearly increased in WT mice after 60 days of pristane injection. An increased cellularity was also observed in the peritoneal cavity fluid, providing evidence that the bone marrow cells are migrating to the initial site of inflammation. However, galectin-3 deficient mice seem not to respond in the same way to the pristane injection, since the cellularity in both bone marrow and peritoneal cavity of these animals was not altered during the experiments. Despite the increase in the total number of mature neutrophils, both in bone marrow and peritoneal cavity, at 60 days after pristane injection, the total number of granulocytic precursors (GrlowCD11bhi) remained unchanged in bone marrow. On the other hand, these precursors were significantly increased in the peritoneal cavity, suggesting a local amplification of these cells. It likely occurred in oil granulomas and in large scale in WT mice, since we observed greater quantities of mature neutrophils in these animals, and more myeloid clusters inside the oil granulomas.
As related in the literature, pristane can be toxic to cells inducing apoptosis and necrosis [17, 36]. In addition, it can accumulate in hematopoietic organs, like spleen and bone marrow [37]. Our group has previously demonstrated that in the absence of galectin-3, lymph node cells are more susceptible to apoptosis [26]. Bone marrow cells of galectin-3 deficient mice might be more sensitive to pristane toxicity, explaining why cellularity in this organ did not increase.
Our data indicated that the first steps of oil granuloma development, with a local amplification of inflammatory myeloid cells, led to a gradual reduction of bone marrow cells mobilization, turning the local responses more autonomous. In contrast, the altered milieu of oil granulomas (low amount of ECM and high of cell debris) could disturb the bone marrow physiology in an attempt to compensate the absence of peripheral amplification. This altered microenvironment of gal-3−/− oil granulomas could be partially explained by an unbalance of pro and anti-inflammatory cytokines produced by macrophages.
Our next steps are to evaluate the consequences of normal and necrotic oil granulomas in the development of lymphoproliferative diseases such as plasmacytoma, which is classically associated with long lasting chronic inflammatory processes.
Galectin-3 deficient mice (gal-3−/−) were generated as described [24] and backcrossed to BALB/cJ mice for nine generations. Age and sex matched wild type (WT) mice on BALB/cJ background were used as controls and were obtained from a colony bred at Federal University of São Paulo (Brazil). All mice procedures were performed at Federal University of Rio de Janeiro in accordance with institutional guidelines (protocol number DAHEICB071, Federal University of Rio de Janeiro). To induce inflammatory process, 8–12 weeks-old mice received a single intraperitoneal injection of 500 μL of 2,6,10,14 tetrametyl-pentadecane (pristane) (95 % pure, Sigma-Aldrich, St Louis, MO, USA). After 7 or 60 days, mice were euthanized using a carbon dioxide chamber. Non injected mice were used as controls.
Histological preparations
For histological analysis, WT and galectin-3−/− mice were sacrificed 60 days after pristane injection. Several oil granulomas were excised, washed in cold saline, fixed in buffered 10 % formalin for 24 h at 4 °C, embedded in paraffin and sectioned into 4-m thick slices. The slices were dewaxed using xylene, hydrated in a graded ethanol, and then washed with distilled water. Lastly, the slices were processed routinely with hematoxylin & eosin (H&E) or with Masson's Trichrome stain. For Masson's Trichrome stain, briefly, slides were placed in Bouin solution for 1 h at 56 °C, then stained with Weigert hematoxylin (10 min; Sigma-Aldrich, St. Louis, MO), followed by Biebrich scarlet acid fuchsin (5 min; Sigma-Aldrich), phosphomolybdic/phosphotungstic acid solution (10 min; Sigma-Aldrich), and aniline blue (5 min; Sigma-Aldrich).
Transmission electron microscopy (TEM)
Oil granulomas were fixed in a solution containing 2.5 % glutaraldehyde, 4 % formaldehyde and 0.1 M sodium phosphate buffer for 4 h at room temperature. Part of the granulomas was cut in two halves to allow the fixative to diffuse faster throughout the granulomas. After rinsing in the same buffer (3 × 10 min), the samples were dehydrated in graded series of acetone till absolute and embedded in epoxy resin (Embed 812). Ultrathin-sections (70 nm) were obtained using a ultramicrotome (Leica) and a diamond knife (Diatome), collected in 300 mesh copper grids and stained with 5 % uranyl acetate for 30 min and 1 % lead citrate for 3 min. The sections were observed in a Zeiss 900 TEM operated at 80 kV and images (2048 × 2048 pixels) acquired using a CCD camera (Olympus).
Paraffin-embedded sections were de-waxed and hydrated. After inhibition of endogenous peroxidase, sections were incubated for 1 h with 0.01 M PBS containing 5 % BSA, 4 % skim milk, 0.1 % Triton X-100 (Sigma Aldrich, USA), 0.05 % Tween-20, and 10 % normal goat serum followed by incubation with purified antibodies overnight at 4 °C in a humid chamber. Primary antibodies were revealed with a biotinylated anti-rat IgG (BA-4001, Vector Laboratories, Burlingame, CA, USA) and developed with avidin-peroxidase (1:200 in PBS) (Sigma Aldrich, USA), using diaminobenzidyne as the chromogen. Sections were counterstained with Harris' hematoxylin.
Cell suspensions
Bone marrow cells were obtained ex vivo by flushing the femoral cavity of WT and gal-3−/− mice with phosphate buffered saline (PBS), pH 7.2, with 3 % fetal bovine serum (FBS, LGC; Cotia, São Paulo, Brazil). Peritoneal cells were harvested on cold PBS with 3 % FBS. A pool of oil granulomas was obtained from each animal, and cells were recovered by digesting the tissues with 2 mg/mL type I-A collagenase and 1 mg/mL dispase in DMEM (Sigma-Aldrich) with constant stirring for 1 h at 37 °C. The resulting cell suspension was removed and filtered through fine nylon mesh [7]. Cells were washed, counted and prepared for further experimentation. When indicated, red blood cell lyses were done with hypotonic ACK (ammonium chloride potassium) solution.
Flow cytometry
In order to saturate Fc receptors, 1 × 106 cells/mL bone marrow, peritoneal and oil granuloma-derived cells were incubated with the Fc blocker antibody, produced by clone 2.4G2 (obtained from the Rio de Janeiro Cell Bank PABCAM, Federal University, Rio de Janeiro, Brazil), for 10 min before adding specific monoclonal antibodies (mAbs). Cells were incubated with mAbs for 30 min; unbound mAbs were washed out with PBS. The following mAbs were used: for T lymphocytes quantification, triple staining was used (FITC-labeled CD8, PE-labeled CD4 and PerCP-labeled CD5); for peritoneal cavity B lymphocytes quantification, double staining was used (FitC-labeled CB11b and APC-labeled B220); for bone marrow B lymphocytes analysis a single staining was used (APC-labeled B220), since there was not observed any double positive (B220+ CD11b+) B1 cells; and for mature and precursor granulocytes analysis a triple staining was used (FitC-labeled CD11b, PerCP-labeled Gr-1 and APC-labeled Ly-6G), where double high positivity for Gr-1 and CD11b was characterized as mature granulocytes, as emphasized by Ly-6G staining, and Gr-1 low and CD11b high cells were characterized as myeloid progenitors. All gating strategies used in this paper are demonstrated in Additional file 1: Figure S1. All antibodies used were obtained from BD Bioscience, San Jose, CA, USA. For cell death analysis, cells were incubated with Annexin-V FITC, according to manufacturer informations (BD Bioscience). Samples were acquired in a flow cytometer (FACScalibur; BD Bioscience) using Cell Quest software and analyzed using WinMDI 2.9 software.
Isolation of adherent peritoneal cells
Animals were divided into four groups with five animals each: (1) WT control group: this group was injected with 500 μL of PBS; (2) WT pristane-treated group: received a single intraperitoneal injection of 500 μL of pristane; (3) gal-3−/− control group: this group was injected with 500 μL of PBS; (4) gal-3−/−pristane-treated group: received a single intraperitoneal injection of 500 μL of pristine. At the end of the experimental period (48 h and 7 days), animals were sacrificed and peritoneal lavage samples were collected by injecting 3 mL of phosphate-buffered saline (PBS). Individual lavage samples were cultured in a 6 well plate at 37 °C with 5 % CO2. After 2 h, peritoneal adherent cells were rinsed with PBS to remove non-adherent cells and subsequently, total RNA was isolated using Tri-Reagent (Sigma).
Total RNA extraction, reverse transcription and quantitative PCR
Primer sequences
Primer sequence (5'-3')
Forward: AGCTGCGTTTTACACCCTTT
Reverse: AAGCCATGCCAATGTTGTCT
Forward: AACCATCTCCTGGTTTGCCA
Reverse: CGGGAGTCCAGTCCACCTC
Forward: CCGAAGCAAACATCACATTCA
Reverse: GGTCTAAAGGCTCCGGGCT
Forward: CAAAAGGACAGCCTCGAGGAG
Reverse: CCCGTGGTCTCTCACGTCAT
Forward: TCAATTCCAGAAACCGCTATGA
Reverse: GAAGTAGGGAAGGCCGTGGT
Forward: GACGTGGAACTGGCAGAAGAG
Reverse: GCCACAAGCAGGAATGAGAAG
Forward: ATCCAGAGCTTGAGTGTGACGC
Reverse: AAGGCAAACTTTTTGACCGCC
Forward: AAAAGGTGTCCCCAAGTA
Reverse: AAGCAGAACTGAACTACCATCG
Statistical analysis
The statistical tests were applied using Tukey's multiple comparison test (t test) and the significance threshold was fixed for α = 0.05. Therefore, P values ≤0.05 were considered statistically significant.
FAPERJ (E-26/111.411/2013) and CNPq (141799/20129) for financial support.
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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text: From the U.S. Department of Education (ED), "Innovative Pathways to School Leadership," which provides examples of successful strategies to prepare candidates for school leadership, is the latest–and last–in a six-booklet series promoting best practices in schools. Published by ED's Office of Innovation and Improvement, these free online resources detail how school systems across the country are working to implement the No Child Left Behind Act (NCLB). Previously released titles include "Creating Strong District School Choice Programs," a compilation of real-life "lessons learned" in implementing NCLB's school-choice provisions; "Creating Strong Supplemental Educational Services Programs," a profile of how five school districts provided low-income children with free tutoring and academic assistance; "Successful Charter Schools," which highlights the successes of eight charter schools nationwide; "Creating Successful Magnet Schools Programs," which includes a guide for starting a magnet program and suggests how to improve existing programs and evaluate their progress; and "Alternative Routes to Teacher Certification," which looks at six programs that prepare people who already have content knowledge, such as mid-career professionals, retired military personnel, and other college graduates, to teach.


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As part of our ongoing water testing, we have included our transportation department water supply, and the test came back positive for PFOA. It is important to note that our transportation department has tested positive, NOT the school campus. The bus garage location is 1.2 miles from our main campus. Our water at the school is safe, and we continue to test our school campus water to ensure that we are providing a safe water supply to our kids.
Once the results were received, we immediately informed the employees at the bus garage. Additionally, we have taken immediate steps to protect the safety of our employees. PFOA was detected in the bus garage water sample at 412 ppt.
The water at the bus garage has not been used for drinking or human consumption since 1997. We have always provided potable water at the bus garage due to other factors. We will continue to provide potable water for our employees.
We have submitted the proposed carbon filtration system plans to the Rensselaer County Department of Health and are waiting for their approval.
We are exploring other options for washing our buses. Bus visibility is part of safety, and to protect our employees from the water spray while washing, we are investigating alternate locations for cleaning the bus fleet.
Again, it is important to reiterate that the water supply for our school campus is safe and free of PFOA. We are, in the spirit of full disclosure and transparency, sharing with our school community that the water at the Transportation Department (bus garage) has tested positive. This positive test at the bus garage does not impact our students or our operations at the school in any way and we are taking steps to work through the situation.


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"Frickin' Damn Cool!"
Fitzgerald Accomplishes the Impossible in Seven Days
At GATS in Dallas during August 2017, Alex and Kenny Laughlin first approached the team from Fitzgerald Glider Kits about the possibility of building a kick-ass truck to transport Alex Laughlin's Gas Monkey Energy Drink sponsored 2018 NHRA Pro Stock Camaro. For good measure, installing a new trailer wrap was an added challenge. Initially, it was thought that having the truck ready before the February 8-11 season opener at Pomona was the target date. But a more urgent December deadline was required as a result of TV production schedules.
Meeting the Challenge
Of the approximately 1,500 trucks in stock, only one was a perfect match. It happened to be a 2017 Peterbilt 389 rolling glider with a 58-in. flattop sleeper on a 312-in. wheelbase. Power is provided by a 60 Series Detroit running through an 18-speed Eaton. The off-white color with red frame and fenders was the only major problem, which turned out to be no problem for the multi-talented crew at Fitzgerald Collision Repair (FCR). A seven-day, seven-night plan even provided a few hours on the last day to carefully assess the week's work against a detailed checklist.
Starting at 8:00 a.m. on Monday, December 11, the 389 was disassembled, the frame was prepped and painted green. The crew worked until 3:00 a.m. on Tuesday and was back in the saddle at 8:00 a.m. This early pace set the tone for the rest of the week as "no way" quickly morphed into "can do." Drawing on years of experience and having successfully completed many rush jobs, the FCR crew marshalled the necessary resources and confidently went about their business.
Simultaneously, a crew of five from SignSmith in Crossville, TN arrived for the multiday process of removing the race trailer wrap and installing the new 2018 version. Shortly after the newly painted frame exited the paint booth, the Fitzgerald team started installing the major pieces on Tuesday morning: cab, sleeper, hood and engine. The build took a good bit of planning as all of the custom parts and components were gathered and staged on the floor adjacent to the 389 Pete. Many parts needed painting or polishing. A crew of five from Almost Chrome out of Nashville came in and polished the trailer, wheels, grille, step boxes, fuel tanks and many other small parts in preparation for reassembly. They worked vigorously to finish everything in only one day.
Arriving from Indiana on Wednesday morning, Brian Davis and his crew from Davis Bros. came down to help layout the paint scheme designed by Brian Bourke. By Wednesday night, the truck was prepped and rolled into the paint booth, awaiting its bright green and black accents. With detailed planning, no wasted motion and great coordination with reliable specialty suppliers, the work progressed smoothly.
On Thursday morning, final reassembly commenced. With a combined team of 10 from Fitzgerald Glider Kits and FCR, the truck took its final form over the next few days and rolled out just in time for the trip to Dallas for its debut appearance at Gas Monkey Garage. When the truck rolled in, the guys from Fast N' Loud along with Alex and Kenny Laughlin laid eyes on this beauty for the first time.
"Most of drag racing is cheatin', and the other half is showin' off. We're going to try covering both, but this truck is definitely showin' off. The Fitzgerald boys killed it on this one! Not only was there a short timeframe like there always is with us; but when we finally pulled the trigger, they were able to get it done. It's frickin' damn cool! They had this thing in a million pieces. It really impressed me, 'cause we do things really, really fast at Gas Monkey, but this is a giant truck. In seven days, they pulled it all apart, rolled the frame in, got it all painted, and put it all back together. Everything is super brand new and ready to rock. We're good, but I don't think we could do that at Gas Monkey."
– Richard Rawlings, Owner of Gas Monkey Garage
Lucky 7
The truck name is derived from all the lights that are in 7s including the T-bars, visor, breather, etc. Also, Alex Laughlin's birthday is on the 7th, and Lucky No. 7 just fit well with the fact this rig is hauling an NHRA Pro Stock race car, plus the seven-day build. Fitzgerald Glider Kits will be an associate sponsor for 23 of the 24 NHRA races during 2018, and will be the primary sponsor when the traveling show visits Bristol, TN for the June 15-17 NHRA Thunder Valley Nationals.


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Anyway, onward and upward. Something else will come along.
It will...and I honestly do think that sometimes things happen to steer us in a direction we might not have chosen. So who knows what might come your way in the future. Good luck!
Yes, things do happen for a reason. The right one is on it's way.
Yes, everything happens for a reason. Perhaps you should consider moving to PEI. Think of it as a retirement parting.


text:
"I Caught a Fish Alive"
One, two, three, four, five,
Once I caught a fish alive,
Six, seven, eight, nine, ten,
But I let it go again.
Why did you let it go?
Because it bit my finger so.
Which finger did it bite?
The little finger on the right.
You must be logged in to post a comment.


text:
Issue: 153
Sent: 31-03-2009 13:42:02
In this issue:
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Small business vs. Government
A US right wing journalist, Jonah Goldberg, wrote this week about the cozy relationship between big business and big government. Here is his main argument:
"... one needs to remember that the New Deal was not the assault on big business that its fans claim. FDR may have talked a good game about going after "economic royalists," and he did love confiscatory personal income taxes. But he and his Brain Trust also loved cartels, big businesses, and other "big units" of society. The notion that big business and big government are at war with one another is one of the great enduring myths of the 20th century. The truth is that ever since Teddy Roosevelt abandoned his love of trust-busting, progressives have liked big businesses big, really big. The bigger the business, the more reliable the partner for big government.
"That's why any huge corporation that plays ball on health care, or "green jobs," or countless other initiatives, is hailed as a "forward-thinking" or "progressive" company. Companies such as GE, which stands to make billions from Obama's energy proposals, are vital sidekicks in the new era of public-private partnerships. Why is Obama working tirelessly to save Detroit automakers? Because GM is a wonderful poster boy for peddling nationalized health care, and UAW is an indispensable cog in the Democratic Party.
"Hillary Clinton's health-care plan required working with large corporations and other firms. It was little guys for whom she had nothing but contempt. When warned her plan would crush smaller businesses, she shrugged, 'I can't go out and save every undercapitalized entrepreneur in America.'
"Again, this is hardly a new story. Chiefly under the auspices of the National Recovery Administration, the New Dealers sought to create huge cartels and trade associations that could work side by side with economic planners. Small and independent firms, from movie theaters [US spelling] to dry cleaners to poultry distributors, were hounded and harassed by a government determined to 'rationalize' the economy by sweeping away all those pesky-but-innovative competitors. Would Barney Frank [a US Congressman] rather work with one giant Fannie Mae that will always take his phone calls and do his bidding, or a thousand smaller firms that would need to be herded like cats? I think we already know the answer.
"Everyone agrees that we are spending trillions of dollars on firms "too big to fail." Many of these firms got so big because politicians in both parties liked to have important businessmen take their phone calls, do their bidding, and fund their campaigns. And maybe, just maybe, the lesson from the financial crisis isn't to get big business and big government even more involved with each other, but to finally bust the trust between them."
Not everyone is going to agree with Goldberg's argument. Some will no doubt have some serious arguments against his thesis. Some will disagree simply because of his politics.
Personally I think he's about on the mark. Think about the practicality of government regulation in Australia. It's a lot easier to regulate a few large super funds than many thousands of small super funds. The ATO find it easier to deal with 11,000 or so small super fund auditors than the 800,000 or so trustees. It's a lot easier for ASIC to deal with a few thousand financial services licensees than the tens of thousands of authorised representatives.
It is easier to regulate a few hundred buses or trains than thousands of cars or motor cycles.
Anyone with experience of dealing with politicians and government departments will know that they prefer to deal with big business. It would be silly to pretend otherwise.
But sometimes small business can make itself powerful by accident. We're seeing that with small super funds. As a whole they are too large and too numerous to attack. This is good for everyone.
This email is general in nature only and does not constitute or convey specific or professional advice. Legislation changes may occur quickly. Formal advice should be sought before acting in any of the areas discussed. Be aware that the information in these articles may become innaccurate with time. Responsibility is disclaimed for any inaccuracies, errors or omissions. Particular investments are neither invited nor recommended and hence this publication is not "financial product advice" as defined in Section 766B of the above legislation. All expressions of opinion by contributors are published on the basis that they are not to be regarded as expressing the official opinion of any other person or entity unless expressly stated. No responsibility for the accuracy of the opinions or information contained in the contributor's articles is accepted by any other person or entity. Copyright: This publication is copyright. If you wish to reproduce this article you require a license, which can be purchased here, to do so.


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D.W. Bradley, the man who made the Wizardry games, was making RPGs when Bioware was but a gleam in a Canadian doctor's eye. But his stature in the industry isn't what it used to be. His last game, Wizards & Warriors, went through a long and troubled development process. It was quietly released to lukewarm reviews and disappointing sales. The man who was once a godfather of the genre seems to have been left behind as the RPG renaissance marches on.
Bradley is nearly naked right now, wearing only a pair of tight boxers that seem to be made of thin leather. Well, okay, not really D.W. Bradley, but his avatar in Dungeon Lords. Courtesy of a demo built with all kinds of developer cheats, he's pretty much carrying one of everything. He opens an inventory window and puts on a colorful suit of armor and wanders out into a dark forest. Things start coming up to him -- rats, skeletons, bandits with bows, that sort of stuff -- and he swings away with a sword while he explains the game.
Based on Bradley's demonstration, Dungeon Lords seems to be a movement towards the Diablo crowd, with a premium on simplicity and cinematic flair. A left-click attacks and a right-click blocks. There are almost no interface elements on the screen; there's only a health bar in the upper left corner. During combat, the amount of damage inflicted flashes briefly at the bottom of the screen. "We keep the interface completely clean," Bradley says. "In other games, players aren't looking at what's going on. Instead, they're clicking icons and watching numbers."
But a minimal interface has it pitfalls. For instance, as he smacks away at the gathering hordes, there's no indication how close a particular opponent is to dying. Is he hurt a little? A lot? Not at all? Bradley is reluctant at this point to use the conventional health bar hovering over a monster's head. "For every element we put on the screen, it draws focus. The last thing I want to do is take away from the player's experience that he's actually in the world."
This is at the heart of what he's trying to accomplish with Dungeon Lords. "It's the same vision I had twenty years ago, but we just didn't have the hardware back then," says Bradley. "That's why role-playing games started as a turn-based system, which let us do party-based games." He says these are both elements that can detract from the player's perception that he's inside another world. Hence Dungeon Lords' single character and more visceral real time focus on what he calls "the heat of the battle."
"After Wizards & Warriors, I took some time off to reevaluate what I learned from that game," Bradley says. "I was studying other games and talking to players. But all along, I've wanted to create for players the experience of a live role-playing setting rather than just sitting around a round table. Thankfully, these days, we have the hardware to do it justice."
Dungeon Lords
Page 2
Once you get past Dungeon Lords' beginning town, you'll pretty much be free to explore a wide-open world with two main plotlines and multiple side quests. You can go through the game with other players, moving your character back and forth between single and multiplayer games and saving your character's progress as you go along. This freedom will also apply to character development. Any character is free to pursue any skill or use any item. Specialization is encouraged by the skills you buy with your experience points, some of which will be cheaper for certain classes and races.
Although Dungeon Lords includes a lot of typical fantasy RPG conventions, it has a unique magic system. "It's not like you have a level one fireball and a level two fireball," Bradley says. "Every spell is different and every school is different."
There are four schools of magic, each with different mechanics. Arcane magic is offensive spells like fireballs and magic missiles, cast from spell books, which are used up as you read from them. Nether magic consists of curses. It works with a component-based system, in which you have to use charms like shrunken heads and bat wings. Celestial magic lets you cast spells like healing and a time warp that makes the action run in slow motion. For celestial spells, you have to draw from crystals that store up mana, but the mana replenishes over time. Rune magic, which consists of buffs, is similar to celestial magic in that you have to use glyphs that store mana and replenish it over time. But each rune magic spell requires different glyph combos.
Ironically enough, Bradley says he's not very good at the game, mainly because he's spent more time creating it than playing it. As he's demoing it, he keeps dying because he'll stop fighting to explain things like the unique magic system. So he'll quickly resurrect himself, courtesy of a developer cheat, and run away, only to have the monsters catch up with him when he stops again to talk. He's got quite a train of baddies following him by now, but he blames me. "It's something about you," he says. "I think you're attracting these bigger monsters. I tell you what, when you play this game, you better be somebody with heavy armor and a lot of combat skills." A big demon dog runs up and breathes poison on him, killing him again.
Dungeon Lords is currently slated for a 2004 release.


text:
Please contact me if you'd like to discuss booking a session. I no long offer set pricing packages. Instead, here's a list of my base pricing.
Rehearsal / Concert Photos: $200
Headshots: $100 - $300
Beauty / Creative: $300
On Location Creative Shoot: $500
Boudoir: $500 - $1,000
Weddings: $1,000 - $3,000
My studio shots are taken in my home, but I often bring my studio lights on location.
I also book spaces for boudoir and creative shoots (the cost of those spaces are included in the package cost).


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Font-size Measurement
The standard unit to measure font height in the print world is the point. In the web world, in addition to the point (pt), there are many other ways to measure font sizes. These can be categorized into two groups: fixed-height measurements and relative font measurements.
Fixed-height Measurement
- points (pt)
- picas (pc)
- inches (in)
- centimeters (cm)
- millimeters (mm)
1 pt = 1/72 of an inch
1 pc = 12 pts [fig.1]
Two other fixed-height measurements are available for the web only:
- X-height (ex) Based on the height of the lowercase x character [fig.2].
- Pixels (px) Based on the resolution of the screen.
Using fixed-height measurement on the web is not recommended because the web usually is viewed on screen, and different screens have different resolutions. When you specify 14-point type, it might display much larger or smaller than you expect on different screens. This is most commonly noticed when moving between Windows and Macintosh computers: type typically displays almost 25% smaller on the Mac.
Thus, a better way to set font-size on the web is to use relative font measurement.
Relative Font Measurement
- Ems (em) Based on the default font size in the viewer's browser.
- Percentages (%) Like ems, based on the default preferences of the browser, and calculated relative to the surrounding text.
1 em = 16 px (the default font-size of most browsers)*
Think in Pixels, use Ems for the measure.
Font sizes are inherited within the DOM by children from their parent elements. This can make em sizing calculations for nested elements difficult. A good idea is to size everything in pixels first and then convert the measurements over to ems.
Composing to a Scale
A good rule of thumb is to select a comfortable size for type: 14pixels and up. Use scales [fig.3] to differentiate headlines, body copy, footers, side notes, and other text-based information.
Once the body text size is determined, the sizes of the other type can be selected within the scale to create hierarchy. The em relationships among different type sizes are also based on where they are on a scale.
*More about Em
Em measures relative sizing and can be specified to three decimal places—for example, 1.375em. It is calculated based on the font-size of the parent element.
Here is the formula to convert pixels to em:
required pixel value ÷ parent font-size = em value
For example, the h1 font-size is 18 pixels. To convert it into em, 18 ÷ 16 = 1.125em (16 pixel is the default body font-size).


text:
contributed by
Dr Andrew Dixon
on January 27, 2017
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A case of gastric band erosion diagnosed on CT. This is a relatively uncommon complication of this form of bariatric surgery. The erosion was conf..."
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Telephoto zoom lens incorporating Sigma's original Optical Stabiliser
Optical Stabiliser
This telephoto zoom lens incorporates Sigma's original OS (Optical Stabiliser) system. It offers the equivalent of using a shutter speed approximately 4 stops faster.
Incorporated with SLD Glass
Three SLD (Special Low Dispersion) glass elements provide excellent correction for chromatic aberration. This lens is equipped with the rear focus system that minimises fluctuation of aberrations caused by focusing. It provides optimum image quality throughout the entire zoom range. The super multi-layer lens coating reduces flare and ghosting. High image quality is assured throughout the entire zoom range.
Minimum focusing distance of 150cm
The minimum focusing distance of 150cm throughout the entire zoom range and has a maximum magnification of 1:4.2 make it useful for close-up photography.
This lens incorporates HSM (Hyper Sonic Motor), which ensures a quiet and high-speed AF as well as full-time manual focus override.
Lens Construction
21 Elements in 15 Groups
Angle of View
20.4 - 6.2 degrees
Number of Diaphragm Blades
9 Blades
Minimum Aperture
Minimum Focusing Distance
Maximum Magnification
Filter Size
Diameter 92mm X Length 203mm
To get exact pricing for a particular product or selection of products simply add the products you desire to your shopping basket and our system will calculate the delivery cost automatically.
We aim to deliver all items within 5 working days of your order.
|United Kingdom
|Smaller items
(eg. lens caps, filters)
|Medium items
(eg. compact cameras, binoculars)
|Larger items
(eg. lighting kits, etc.)


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English and maths matters
Who is this course for?
If you are interested in developing your English or maths skills then this course is for you.
Course content
You will be working, at your own pace, on the aspects of English and maths that you want to improve.
Support and feedback
Throughout the course your tutor will give verbal and written support to help you achieve your personal goals.
Additional costs and requirements
You do not need to bring anything with you and there are no entry requirements.
Will I have to do anything outside of the session?
Sometimes it may be helpful to practise some of the work in your own time.
About the tutor
Barbara Kernick is a tutor working with independent living groups for Learn Devon teaching Drama, Shopping and Independence. Prior to this she worked as a literacy and numeracy tutor and a Skills for Life Coordinator. She has been teaching adults for over 20 years and is a senior practitioner and Internal Verifier.
Before teaching she ran her own business and has been involved in various voluntary roles but now her six grandchildren take up most of her spare time.
"I enjoy encouraging learners to achieve more than they thought they were able to and to surprise themselves".
Barbara Kernick, Independent Living Tutor, Learn Devon


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The fourth of July has come and passed, leaving us all with full stomachs and a general sense of patriotism. You may still be hungry though – hungry for esports. If you didn't catch Immortals' last week of games, you missed out on some great moments. We had some ups and downs; we had a rough week in League, but Immortals' Overwatch squad scored a big win, and our CS:GO team fulfilled a long-standing dream. If you didn't get a chance to catch last week's rollercoaster ride, the Saga will get you back up to date.
Tl;dr: Immortals win Overwatch Contenders Season Zero, and qualify for Season One!
Recommended Match: Immortals v Team Liquid (Grand Finals)
Overwatch Contenders Season Zero is the one of the biggest North American Overwatch tournament to date, and Immortals managed to take it all. After a shaky group phase where we found our footing with KariV and Fate, we came back hard and only dropped one game during the playoffs stages. (Does that still count if it was a draw?) Team Liquid and Immortals met in the finals, and we give them a lot of credit for putting up a tough fight, but Immortals were on fire. We took the prize for Season Zero, and we're automatically qualified for Contenders Season One. GG to everyone who we faced during the course of Season Zero, and we can't wait to come back for Season One!
Tl;dr: Immortals qualify for our first major, and we're heading to Krakow!
Recommended Match: Immortals v Dignitas
It's been a wild year for Immortals' CS:GO team. We've had our eyes on going to a Major, and last year we fell just short in a heartbreaking loss to G2. Since then, we've undergone a few changes, including picking up our newest member kNg. We've been working hard, and while we've slipped along the way, the entire time we kept the goal to make a Major. We've finally succeeded, and the team is now in Europe. We're going to be doubling down and working even harder to make the most of this opportunity!
Tl;dr: Oops! Immortals goes 0-2 against Team SoloMid and Phoenix1
Recommended Match: Immortals v Phoenix1 (Game One)
After a strong start to the split, we didn't have a great couple of matches this week. Unfortunately, we dropped two matches to TSM and Phoenix1, just before they head off to Rift Rivals. We'll definitely be working hard during the break to fix our mistakes and come back stronger. In the meantime, check out a successful game against Phoenix1. We're still making those aggressive plays, we just went a little overboard this weekend. We're working on it!
Want to stay up to date on all things IMT? Make sure you're following us on Twitter, and the Discord is a fantastic place to watch games with other fans.


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Being away at college every year for St. Patty's Day really gets me missing my dad's corned beef and cabbage. Rather than trying to recreate this expensive and time-consuming Irish dish, I created a budget-friendly "Irish-Hungarian fusion", which embraces both my dad's Irish heritage and also my mom's Eastern European roots. To do this, I deconstructed corned beef and cabbage and reassembled it into something similar to my mom and grandma's Hungarian specialty: stuffed cabbage rolls. Read the story behind this recipe for Stuffed Cabbage Rolls with Mustard Sauce and Carrots on Small Kitchen College. As my little green-clad buddies might say, "They're magically delicious!"
Archive for March, 2012
Eat Under $10: Budget Corned Beef & CabbageTuesday, March 13th, 2012
Posted in Holidays, Irish, Small Kitchen College |
You're Invited: Potluck Fondue PartyFriday, March 2nd, 2012
Is it totally weird that I get more excited about planning the food I'll be making at my parties more than anything else? I couldn't care less about the drinks! I recently threw a Potluck Fondue Party as a fun way to relax and reunite with friends after midterm season. I have to say, this is the perfect party theme for winter! There's nothing like gathering your friends around the ol' fondue pot, whether it be cheesy or chocolatey goodness. This if my second article for Small Kitchen College, and I especially had a lot of fun with the photo shoot (and, of course, eating the fondue afterwards). Read on for tips on how to throw the perfect potluck fondue party. You never know, I may have a few fondue tricks up my sleeve in the article. Chile con queso fondue in a bread bowl, anyone?
Posted in Comfort Food, Desserts, Small Kitchen College |
Microwaved Eggs: Gross or Perfect for a Breakfast Tostada?Thursday, March 1st, 2012
Microwaved eggs. If you just cringed at the thought of that, then you're normal. If you're like me, then you'd think microwaved eggs would be the perfect challenge to take on for Student Health 101's March issue: How to cook a fast but nutritious breakfast…using only a microwave.
To be honest, getting this assignment was a total bummer at first. How can I come up with a tasty, microwave-only recipe when I love experimenting with so many different kinds of tools in my kitchen? When I discovered it was possible to microwave eggs, I knew I wasn't going to just settle on boring old scrambled eggs. Oh no, I was going to go big. I was going to attempt to make those oh-so delectable poached eggs with a runny yolk that bursts open when you puncture the egg white (If you love poached eggs as much as I do, then you'll understand the satisfaction I take in doing this). (more…)


text: abs welding rod 5/32 black coil abs welding rod 5/32 black coil Share This Story, Choose Your Platform! FacebookTwitterLinkedInRedditGoogle+TumblrPinterestVkEmail About the Author: Bruce Leave A Comment Cancel reply Comment Save my name, email, and website in this browser for the next time I comment. Notify me of new posts by email.


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автор: DM•
Jul 10, 2017
Really a good course, learning lot of new things , thanks to the instructors.. both the instructors make me understand through videos by their simple and easy way of teaching. Thank you
автор: DP•
May 10, 2016
This was a wonderful course and I feel that I am more confident now while drafting emails, preparing minutes and memos. Thank you so much Coursera.com
Рецензии: 43
автор: Angelina Wong•
Oct 16, 2018
Very informative and useful.
автор: Al Mohaiminul Alam Mehedie•
Jun 26, 2018
People who are looking for a course to write professional emails and know the types of all emails and memos must take the course.
автор: Mel•
May 18, 2018
Very helpful materials provided by thoughtful teachers. Information was given in the most suitable and convenient format(s). The course enabled me to sharpen my business writing skills and learn more about office etiquette.
автор: Daniela Cárcamo Sáez•
Apr 09, 2018
It is a shame that I have to pay to resolve the quizzes. I am going to do the course but not the quizzes, I do not earn enough money to pay
автор: Pedro Cabrera•
Mar 21, 2018
Very difficult to navigate, this is the first course that I have taken where there is no interaction with the instructors. I imagine for the sake of saving work for you the grading is done by other students that are also in the process of learning. I guess you are making a lot of money with little effort.
автор: Erin Foy•
Mar 06, 2018
Found it incredibly basic and almost patronising in places - if you dont know what CC means on an email this course is perfect... To be fair i didnt finish the course - as i found the first three weeks to be almost useless.
автор: Mostafa Salama Shakir Khalifa•
Feb 23, 2018
very useful
автор: Bassam ALmahywi•
Feb 09, 2018
its nice
автор: Arefin•
Feb 07, 2018
It is a very well designed course with relevant real life implications. I took it as a knowledge refresher course so that my written content can still display assertiveness and gravitas, or carry empathetic tone where required . It also taught me some basics of online communication that I never realized I had done wrong until it was pointed out to me by this course. I owe my improvements to the dedicated staff, tutors, and instructors who made it possible for me to learn from a far-away place. Thank you!
автор: Xiangfeng Niu•
Dec 19, 2017
The slides can not draw my attention.


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Transport Assessment
Stage 2: Influencing Travel to the Development: Incentives to individuals
Incentives to individuals can complement physical and organisational measures as part of a package of measures to influence travel patterns.
Appropriate incentives will vary and are likely to be most effective with a package of incentives to walk, cycle and use public transport combined with financial disincentives to car use and constraints on car parking.
Various types of financial incentives can be offered to those who walk and cycle, or agree not to take a car to work.. Examples are, company supported cycle purchase schemes or relocation packages which can favour employees who choose housing, which has easy non-car access to the site. Alternatively, local public transport operators may offer special benefits for new season tickets on local services.
Many organisations offer direct incentives to use public transport by providing loans or subsidising season tickets. Such tickets normally provide benefits to holders beyond the journey to work, with some bus operators offering transferable tickets for use on any service within the operating areas. Alternatively, companies might find it more effective to organise their own works buses directly, which can then be tailored to the needs of the individual organisation. Incentives to encourage more sustainable travel benefit from increasingly favourable taxation arrangements.
Company car policy has often encouraged individuals to drive to work or on work-based trips. The taxation arrangements for company cars has changed significantly so that it can be better for individuals to give up company cars and take other travel incentives instead. Company car policy can also be changed by:
- identifying those staff with most need of company car benefits;
- changing the nature of the car-use benefits, for example the travel mileage rates;
- replacing the benefits with other measures relating to other modes, such as cycle allowances; and
- encouraging the use of taxis as an alternative to car use.
For housing developments, discounted public transport tickets may be offered, and special introductory offers to new residents can be beneficial to operators. Negotiations with operators may also redefine ticket zone boundaries to benefit residents of the development.


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Not long ago, the unthinkable — a Scott Brown victory over Martha Coakley — became the distinctly possible and, following the latest gaffe by the Coakley campaign, it may now have ascended to the realm of the "about as likely as not."
The gaffe occurred when the Coakley campaign and the Democratic Senatorial Campaign Committee decided to attack Brown as the agent of "Wall Street greed." Not terribly original, but not a bad idea either.
Unfortunately for Coakley, the picture selected by the DSCC to symbolize Wall Street greed shows the World Trade Center. Talk about a negative campaign ad.
Meanwhile, President Obama has decided to head for Massachusetts on Sunday to campaign for Coakley. He apparently believes that the unthinkable has indeed become the "about as likely as not."
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"Arise and take our stand for freedom as in the olden time." Winston Churchill
"Proclaim Liberty throughout All the land unto All the Inhabitants Thereof." Inscription on the Liberty Bell


text:
If you have received a notice from the Department of Revenue that it has intercepted all or part of your tax return to satisfy a court debt, you are entitled to dispute the submission of the intercept itself. You must notify the Tax Refund Intercept Program of your intention to contest the intercept within thirty (30) days of the date of the letter from the Department of Revenue.
Under the law, the underlying judgment of the court may not be disputed. The TRIP program is governed by Sections 48-7-160 through 48-7-170 of the Georgia Code. Legitimate reasons to dispute or contest an intercept are:
The debt to the court was paid in full.
The amount of the debt to the court may be incorrect.
I am not the person named in the judgment.
The judgment is against my spouse, a joint filer.
I have emerged from bankruptcy proceedings since the judgment was entered.
The judgment is more than six years old.
A dispute form has been created to help disputants submit the necessary information to refer the dispute to the court that requested the intercept. You may fill in the boxes and print a copy of the form, and the Program will accept submissions that are emailed, faxed or sent by regular mail. Please do be mindful of the thirty (30) day deadline.
Click here to download the Dispute Form
The court will contact you regarding your dispute and to schedule a hearing with an independent hearing officer if you desire. Once a hearing has been held, the hearing result may be challenged by filing an appeal in superior court. If the requesting court directs TRIP to do so, it will send you all or part of the intercepted amount.


text:
Part Number: 4724
1½"-2" Positive Seal, 2 Port NeverLube Valve
For over 20 years this has been Jandy's best selling valve. It never needs lubrication.
Your customers will love that! In fact, it's totally maintenance free. It is extremely reliable; designed to never leak, crack or break. The NeverLube valve is built from the toughest CPVC material and can withstand extreme chemical and temperature fluctuations. For optimum performance, use in conjunction with the Jandy Valve Actuator for automatic operation. Available in 2" by 2½" or 2" by 1½" sizes.
Available as 3-port diverter and 2-port isolation valve.
Exceptional quality and versatility make Jandy valves the preferred choice of pool professionals. Jandy sets the benchmark in quality and innovation for pool and spa valves. For over 30 years Jandy valves have been the undisputed leader in the pool and spa industry. Our commitment to quality and innovation makes for easier, problem-free installations.
Jandy valves share one thing in common - dependability! Dependability begins with top-of-the-line materials and rigorous quality control standards. We offer the widest variety of valves for a multitude of pool and spa applications. Superior quality and plumbing versatility make Jandy valves the #1 choice of pool and spa professionals everywhere!
• Best selling valve
• Extremely reliable
• Never requires lubrication
• Lifetime Guarantee
Technical Details
• Designed for easy automation
• Made from durable CPVC
• Proprietary NeverLube technology with Lifetime Guarantee


text:
Holy Man of Steel! Here are the photos from the Man of Steel press junket. I want to point out that Henry Cavill (Clark Kent/Superman) and Zach Snyder (Director of Man of Steel) were gracious, delightful men who seem very grateful for their work and success. The sound stage was decorated beautifully. There were many costumes, still photos and props from the film on display and staged in visually striking ways. Click on photos for a larger view. Enjoy
|Man of Steel--View from the catwalk
|Man of Steel--View from the catwalk
|Man of Steel--Superman logo
|Man of Steel--Clark Kent (Henry Cavill) costume
|Man of Steel--Stills from the film
|Man of Steel--Clark Kent costume
|Man of Steel--General Zod (Michael Shannon) and Kryptonian costumes
|Man of Steel--General Zod and Kryptonian costumes close up
|Man of Steel--Kryptonian Carbine Pistols
|Man of Steel--Smallville Bank and Trust
|Man of Steel-Daily Planet globe, Clark Kent and Lois Lane (Amy Adams) costumes and stills from the film


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Subscribe To Dakota Fanning Playing Princess Margaret In Girls' Night Out Updates
I've already subscribed
Princesses Margaret and Elizabeth were icons for a certain generation of girls growing up before and after World War II, the two daughters of King George VI living the kind of palace life everyone envied. And though it's been a while since anyone referred to the two princesses as a unit-- Elizabeth went on to be Queen, of course, while Margaret died in 2002-- they're making something of a comeback these days. The two daughters play small but significant roles in The King's Speech, which focuses on their father, and now there's a new film in the works about what happened to them when they were freed from the confines of Buckingham Palace for one night as teenagers, on VE Day at the end of World War II.
According to Variety Dakota Fanning has been set to star as Margaret, who was nearly 15 at the end of the war; casting for Elizabeth, who was 19 at the time, will begin shortly. The film will be directed by Michael Hoffman, who last made The Last Station and will be working with King's Speech star Colin Firth and Cameron Diaz on Gambit, the remake of the 1960s caper film that was scripted, oddly enough, by the Coen Brothers.
Sales for both films will take place at the European Film Market next week, so Hoffman is going to be a busy guy. With Fanning on board, and what seems like a renewed international interest in the royal family, the Princess Margaret film-- titled, kind of goofily, Girls' Night Out-- should be a fairly easy sell. I mean, is there anything Dakota Fanning can't make more appealing with her mere presence?


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I should have stuff up this week. As feared life and Enterprise caught over took my will and desire to be a cool person and make posts. Looking to rectify this nicely this week.
Archive for the 'Upkeep' Category
Slight case of laziness
Posted in General, Upkeep, tagged Lazy, update on 03/20/2012|
Break time!
Posted in Music Videos, Upkeep, tagged Indy, Music, Music Videos, Rant, rock, video on 02/08/2012|
Taking a break from all this since it's been mostly status quo and that's no good. I shan't be long away, perhaps a week or two as to just settle some projects and my life a bit, nothing to worry about.
But before I kick off, my good buddies Mushy Callahan recently compiled a nice new video for their song 'Loose Lips'. It's a mighty fine video and best of all I was able to contribute some footage to towards it. Also follow their Facebooks and Twitters.
Getting back to Art
Posted in Rant, Upkeep, tagged Art, Art Projects, artist, Rant, Upkeep on 01/10/2012|
I think the biggest and boldest problem affecting me is that in recent months is I've not been getting to do the things that really challenge me. I think I've become too comfortable doing virtually nothing hard, which is terrible. Overall I think there's a lack of conflict which I can turn into something creative.
The source of this? I hate my job. It does take a lot of time and energy from me, but I don't think that's a good enough excuse. Like 110% of everyone hates their job at some point. I think there's also a lack of anyone in my life pushing me, someone special. I probably need a muse, or a girlfriend.
Still, can't be looking at excuses. The other day I did stumble on a website that houses a collective of artist work which will make perfect fodder for my 'Artist Talks' since the website doesn't go into much detail or much of anything. So I guess I can shine there.
Probably what I really need to do is finish some old projects before I start some new ones. Also axe the ones which are no good. A shame, but I can't toil on things which will go no where. Speaking of which, no where is where that one final pigeon is.
Lets start with that.
And I'm away.
More posts coming on their way. Got a little book called '5 good reasons to punch a dolphin in the nose', but more on that tomorrow in a post.
Also since I've had a lack of taking pictures (thanks having a thing called a job) I'm going to start posting / tweaking old pictures that have been sitting stale and unused on my compies hard drive. I'm going to work hard to post something up for ever Thursday (which really isn't hard given that I can schedule like the dickens when I have free time).
And that aside, see the movie Drive and I'll make a decent quip about 50/50 which I go to see tomorrow.
Ta for now.
P.S. sorry for the near endless 'Art of Music Video' postings.
Mostly Operational
Posted in Upkeep on 09/13/2011|
Main computer is operating again, however in the process of data recovery / compiling witty things to say and post.
It is however a little ironic that some of my busier time on this blarg has been when I'm not posting things. But that just makes things stale. Will do my darnedest to get an Art of Music Videos done up for Friday. Yes, there's a lot of irony with my last AMV posting…
Lack of posts stems from being drained from working too many long hours. I'm tossing this up just to combo breaker my Art of Music Videos streak.
Hope to be able to write about some nice neat things that have been happening, but time will tell with that. For now, another Art of Music Videos will be posted up.
Making some changes around here. I've decided I need to be fancier and more attractive and flashy to get the attention of the internet generation. So to do so I figured I'd roll out some sort of unifying banner for my Art of Music Video postings rather than keep on using random images (which I still will, but this won't make as mandatory).
Also if you have not noticed WordPress or Youtube finally worked some magic and now all videos will play on my site without you having to view them on Youtube. As well, there's a new menu page with a listing of all the videos I have done. Use it with the Search box to find the post. Anyways here are the new post headers I worked oh so hard on making.
YES! A new 'as often as I can' weekly posting on 'Mondays' about 'hating'. This is to get back on to my whole 'you are fail' category because it's not getting much loving or attention. Haven't been venting frustrations and it would be best if I did it here in this environment than taking an office stapler to someone's head.
I'm in the works on refining the Artist Talk format as well with its own custom header, but I'm spent for this week and have a bit of work to do doing some things retro actively and in another blog space. Expect things to be a bit slower than usual until then.
Special Shout-out to Music Video Stills who did all the heavy lifting way back in 2006 to get me all the images I used to make the new Art of Music Videos banner. That was some epic mysterious work done so I could get this done faster. Also got some ideas for upcoming Art of Music Video entries.


text:
Next Episode of Scarecrow and Mrs. King is
not planed. TV Show was canceled.
A single mother in suburban Washington D.C. discovers she has a talent for espionage work when she meets a dashing undercover agent--but balancing her duties as a spy with her responsibility as a parent proves constantly entertaining in this romantic-adventure series.
Bruce Boxleitner(Lee Stetson)
Kate Jackson(Mrs. Amanda King)
Beverly Garland(Dorothy 'Dotty' West)
Martha Smith(Francine Desmond)
Mel Stewart(Billy Melrose)
Paul Stout(Philip King)
Greg Morton(Jamie King)
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