Identification of novel chondroitin proteoglycans in Caenorhabditis elegans: embryonic cell division depends on CPG-1 and CPG-2.

Vertebrates produce multiple chondroitin sulfate proteoglycans that play important roles in development and tissue mechanics. In the nematode Caenorhabditis elegans, the chondroitin chains lack sulfate but nevertheless play essential roles in embryonic development and vulval morphogenesis. However, assignment of these functions to specific proteoglycans has been limited by the lack of identified core proteins. We used a combination of biochemical purification, Western blotting, and mass spectrometry to identify nine C. elegans chondroitin proteoglycan core proteins, none of which have homologues in vertebrates or other invertebrates such as Drosophila melanogaster or Hydra vulgaris. CPG-1/CEJ-1 and CPG-2 are expressed during embryonic development and bind chitin, suggesting a structural role in the egg. RNA interference (RNAi) depletion of individual CPGs had no effect on embryonic viability, but simultaneous depletion of CPG-1/CEJ-1 and CPG-2 resulted in multinucleated single-cell embryos. This embryonic lethality phenocopies RNAi depletion of the SQV-5 chondroitin synthase, suggesting that chondroitin chains on these two proteoglycans are required for cytokinesis

Loss of the Heparan Sulfate Sulfotransferase, Ndst1, in Mammary Epithelial Cells Selectively Blocks Lobuloalveolar Development in Mice

Considerable evidence indicates that heparan sulfate is essential for the development of tissues consisting of branching ducts and tubules. However, there are few examples where specific sulfate residues regulate a specific stage in the formation of such tissues.We examined the role of heparan sulfation in mammary gland branching morphogenesis, lactation and lobuloalveolar development by inactivation of heparan sulfate GlcNAc N-deacetylase/N-sulfotransferase genes (Ndst) in mammary epithelial cells using the Cre-loxP system. Ndst1 deficiency resulted in an overall reduction in glucosamine N-sulfation and decreased binding of FGF to mammary epithelial cells in vitro and in vivo. Mammary epithelia lacking Ndst1 underwent branching morphogenesis, filling the gland with ductal tissue by sexual maturity to the same extent as wildtype epithelia. However, lobuloalveolar expansion did not occur in Ndst1-deficient animals, resulting in insufficient milk production to nurture newly born pups. Lactational differentiation of isolated mammary epithelial cells occurred appropriately via stat5 activation, further supporting the notion that the lack of milk production was due to lack of expansion of the lobuloalveoli.These findings demonstrate a selective, highly penetrant, cell autonomous effect of Ndst1-mediated sulfation on lobuloalveolar development

Genetic Vaccination-Induced Immune Responses to the Human Immunodeficiency Virus Protein Rev: Emergence of the Interleukin 2-Producing Helper T Lymphocyte

Overview summary The immune system poses a major obstacle to the long-term success of in vivo gene therapies. Immune responses to foreign transgene products and/or the vectors that facilitate gene transfer may neutralize the transgene product, eliminate transfected cells, and culminate in inflammation within transfected tissues. The majority of studies that address these issues have focused on cytotoxic T lymphocyte (CTL) and antibody responses induced by gene transfer. However, the IL-2-producing helper T lymphocyte (HTL) represents a critical regulatory cell that likely influences the inductive phase of the immune response following gene transfer. The current study employed limiting dilution analysis (LDA) techniques to characterize the development of IL-2-producing HTLs induced by genetic vaccination with a plasmid encoding the mutated HIV protein Rev M10. Further, we assessed the ability to inhibit the transgene-induced HTL response by cotransfer of a plasmid encoding the immunosuppressive cytokine TGFβ1.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63282/1/hum.1998.9.15-2187.pd

Deletion of the Basement Membrane Heparan Sulfate Proteoglycan Type XVIII Collagen Causes Hypertriglyceridemia in Mice and Humans

Background: Lipoprotein lipase (Lpl) acts on triglyceride-rich lipoproteins in the peripheral circulation, liberating free fatty acids for energy metabolism or storage. This essential enzyme is synthesized in parenchymal cells of adipose tissue, heart, and skeletal muscle and migrates to the luminal side of the vascular endothelium where it acts upon circulating lipoproteins. Prior studies suggested that Lpl is immobilized by way of heparan sulfate proteoglycans on the endothelium, but genetically altering endothelial cell heparan sulfate had no effect on Lpl localization or lipolysis. The objective of this study was to determine if extracellular matrix proteoglycans affect Lpl distribution and triglyceride metabolism. Methods and Findings: We examined mutant mice defective in collagen XVIII (Col18), a heparan sulfate proteoglycan present in vascular basement membranes. Loss of Col18 reduces plasma levels of Lpl enzyme and activity, which results in mild fasting hypertriglyceridemia and diet-induced hyperchylomicronemia. Humans with Knobloch Syndrome caused by a null mutation in the vascular form of Col18 also present lower than normal plasma Lpl mass and activity and exhibit fasting hypertriglyceridemia. Conclusions: This is the first report demonstrating that Lpl presentation on the lumenal side of the endothelium depends on a basement membrane proteoglycan and demonstrates a previously unrecognized phenotype in patients lacking Col18.National Institute of Health (NIH)[HL087228]National Institute of Health (NIH)[GM33063]National Institute of Health (NIH)[HL67255]CEPID/FAPESPCNPqUniversity of Colorado Denver Department of MedicineLeducq FoundationAmerican Heart Association[0735038N

Prisoners Teaching ESL: A Learning Community among “Language Partners”

A program in which prisoners teach ESL classes, supported by volunteer teacher-trainers, is a learning community with immense and sometimes unforeseen value

Translating statistical species-habitat models to interactive decision support tools

Understanding species-habitat relationships is vital to successful conservation, but the tools used to communicate species-habitat relationships are often poorly suited to the information needs of conservation practitioners. Here we present a novel method for translating a statistical species-habitat model, a regression analysis relating ring-necked pheasant abundance to landcover, into an interactive online tool. The Pheasant Habitat Simulator combines the analytical power of the R programming environment with the user-friendly Shiny web interface to create an online platform in which wildlife professionals can explore the effects of variation in local landcover on relative pheasant habitat suitability within spatial scales relevant to individual wildlife managers. Our tool allows users to virtually manipulate the landcover composition of a simulated space to explore how changes in landcover may affect pheasant relative habitat suitability, and guides users through the economic tradeoffs of landscape changes. We offer suggestions for development of similar interactive applications and demonstrate their potential as innovative science delivery tools for diverse professional and public audience

Boynton-Delray Coastal Water Quality Monitoring Program

This report discusses a sequence of six cruises in the vicinity of the Boynton-Delray (South Central) treated-wastewater plant outfall plume (26°27\u2743 N, 80°2\u2732 W), the Boynton Inlet (26°32\u2743 N, 80°2\u2730 W), and the Lake Worth Lagoon, Palm Beach County, Florida. The sampling cruises took place on June 5-6, 2007; August 28-29, 2007; October 18-19, 2007; February 14 and 18, 2008; May 19-20, 2008; and July 11-13, 2008. Water was sampled at 18 locations at the surface, middle, and near the seafloor (where there was sufficient depth) for a total of 45 samples; these samples were analyzed for a variety of nutrients and related parameters. The water sampling unit contained a conductivity-temperature-depth (CTD) instrument from which data were obtained at each sampling site. Synchronal ocean current data were measured by a nearby acoustic Doppler current profiler (ADCP) instrument. The inlet measurements were consistently lower in salinity and more acidic (lower in pH) than the coastal ocean and were warmer during the May and, especially, during the February cruises. For most analytes, viz., nitrite+nitrate (N+N), total suspended solids (TSS), chlorophyll-a, silica (Si), and total dissolved nitrogen (TDN), the lagoon concentrations were significantly higher than the coastal ocean; the inlet concentrations appeared to be consistent with lagoon water with partial mixing with the coastal ocean, as expected. Estimates of the nutrient flux to the coastal ocean were computed: approximately 1,500 kg of dissolved nitrogen (N), 2,350 kg of silicate (Si), 33 kg of orthophosphate (P), and 59 kg of ammonium (NH4) per day were delivered to the coastal ocean through the inlet. The outfall boil at South Central outfall (the smallest in volume of the six outfalls in southeast Florida) is only visible under ideal conditions. In the six cruises described in this document, the outfall boil could be found in only one cruise (August 28-29, 2007). Elevated concentrations of nutrients (N+N, P, Si, and P) at the outfall vicinity were measured, and these concentrations decreased rapidly away from the outfall for most analytes, to become undistinguished from the background within 3 km or less. Not finding the boil, however, in five of six cruises meant that the waters with the highest concentrations were probably missed. When the boil was sampled in August 2007, N+N, P, and total dissolved phosphorus (TDP) concentrations at the boil were roughly the same as from the inlet. For other analytes (chlorophyll-a, TSS, Si, and dissolved organic carbon [DOC]), the concentrations at or near the outfall were significantly less than those from the lagoon and inlet on most of the cruises. The coastal ocean appeared to be significantly impacted by the Boynton Inlet and less so from the inlet. A suggestion of a source to the south was seen in some analytes. Measurements from the Gulf Stream Reef area were the lowest in the study, and may provide “background” concentrations for this region. As expected, the coastal ocean was warmer and more stratified in the summer compared to the winter, e.g., whereas no thermocline was noted in the CTD data from February 2007, a strong thermocline was observed in most casts during July 2008. In certain cases (e.g., N+N in June 2007, pH in July 2008), an increase in the concentration (decrease for pH) from north to south implied a source from the south, e.g., the Boca Raton Inlet or Boca Raton outfall

Earliest Archaeological Evidence of Persistent Hominin Carnivory

The emergence of lithic technology by ∼2.6 million years ago (Ma) is often interpreted as a correlate of increasingly recurrent hominin acquisition and consumption of animal remains. Associated faunal evidence, however, is poorly preserved prior to ∼1.8 Ma, limiting our understanding of early archaeological (Oldowan) hominin carnivory. Here, we detail three large well-preserved zooarchaeological assemblages from Kanjera South, Kenya. The assemblages date to ∼2.0 Ma, pre-dating all previously published archaeofaunas of appreciable size. At Kanjera, there is clear evidence that Oldowan hominins acquired and processed numerous, relatively complete, small ungulate carcasses. Moreover, they had at least occasional access to the fleshed remains of larger, wildebeest-sized animals. The overall record of hominin activities is consistent through the stratified sequence – spanning hundreds to thousands of years – and provides the earliest archaeological evidence of sustained hominin involvement with fleshed animal remains (i.e., persistent carnivory), a foraging adaptation central to many models of hominin evolution