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

Hyaluronan Synthase Elevation in Metastatic Prostate Carcinoma Cells Correlates with Hyaluronan Surface Retention, a Prerequisite for Rapid Adhesion to Bone Marrow Endothelial Cells

Bone marrow is the primary site of metastasis in patients with advanced stage prostate cancer. Prostate carcinoma cells metastasizing to bone must initially adhere to endothelial cells in the bone marrow sinusoids. In this report, we have modeled that interaction in vitro using two bone marrow endothelial cell (BMEC) lines and four prostate adenocarcinoma cell lines to investigate the adhesion mechanism. Highly metastatic PC3 and PC3M-LN4 cells were found to adhere rapidly and specifically (70-90%) to BMEC-1 and trHBMEC bone marrow endothelial cells, but not to human umbilical vein endothelial cells (15-25%). Specific adhesion to BMEC-1 and trHBMEC was dependent upon the presence of a hyaluronan (HA) pericellular matrix assembled on the prostate carcinoma cells. DU145 and LNCaP cells were only weakly adherent and retained no cell surface HA. Maximal BMEC adhesion and RA encapsulation were associated with high levels of HA synthesis by the prostate carcinoma cells. Up-regulation of HA synthase isoforms Has2 and Has3 relative to levels expressed by normal prostate corresponded to elevated HA synthesis and avid BMEC adhesion. These results support a model in which tumor cells with up-regulated HA synthase expression assemble a cell surface hyaluronan matrix that promotes adhesion to bone marrow endothelial cells. This interaction could contribute to preferential bone metastasis by prostate carcinoma cells

The effect of proteoglycans on the formation of fibrils from collagen solutions

The precipitation of collagen fibrils from solutions at 37 [deg]C and approximately physiological pH and ionic strength is retarded markedly in the presence of small amounts of proteoglycan monomer (PGS), proteoglycan aggregate (PGC), reduced and alkylated PGS, or cyanogen bromide-cleavage products of PGS. The polysaccharide-peptide fragments produced from PGS with papain, trypsin, cathepsin D, or the protein core obtained by the digestion of PGS with chondroitinase ABC are ineffective in this regard. In the presence of the materials which affected the rate of precipitation of the collagen fibrils, the specific absorbance, [Delta]Asp, of the collagen gels was directly related to specific retardation, Rsp, when the ratio of proteoglycan to collagen was less than 25 [mu]g/mg, suggesting that the size and/or organization of the fibrils in the gels was dependent on the presence of proteoglycans. PGS binds to collagen if it is present in the solution before the collagen fibrils form and at a maximum of about 1 molecule of PGS for every 25-30 molecules of collagen. Although the protein core of PGS does not retard fibrillogenesis, it does bind to collagen and does so even in the presence of PGS. The data support the thesis that the organization of collagen fibrils in tissues may be related to amounts and kinds of proteoglycans in the tissues.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/22193/1/0000624.pd

Subcortical heterotopic gray matter brain malformations: Classification study of 107 individuals

OBJECTIVE: To better evaluate the imaging spectrum of subcortical heterotopic gray matter brain malformations (subcortical heterotopia [SUBH]), we systematically reviewed neuroimaging and clinical data of 107 affected individuals. METHODS: SUBH is defined as heterotopic gray matter, located within the white matter between the cortex and lateral ventricles. Four large brain malformation databases were searched for individuals with these malformations; data on imaging, clinical outcomes, and results of molecular testing were systematically reviewed and integrated with all previously published subtypes to create a single classification system. RESULTS: Review of the databases revealed 107 patients with SUBH, the large majority scanned during childhood (84%), including more than half before 4 years (59%). Although most individuals had cognitive or motor disability, 19% had normal development. Epilepsy was documented in 69%. Additional brain malformations were common and included abnormalities of the corpus callosum (65/102 [64%]), and, often, brainstem or cerebellum (47/106 [44%]). Extent of the heterotopic gray matter brain malformations (unilateral or bilateral) did not influence the presence or age at onset of seizures. Although genetic testing was not systematically performed in this group, the sporadic occurrence and frequent asymmetry suggests either postzygotic mutations or prenatal disruptive events. Several rare, bilateral forms are caused by mutations i

Integrating Technical Standards into ET Curricula to Meet ABET Standards and Industry Needs

With technical standards affecting nearly every aspect of our daily lives, from computers to the components and materials used in car engines, it is critical that undergraduate students are educated on the importance of standards and provided with opportunities to locate and apply relevant technical standards to real world situations. In addition, with ABET accreditation requiring students to have a “basic understanding and familiarity with,” and experience “using” codes and standards, faculty need to consider how such material can be naturally integrated into the curriculum. At Purdue University, education about codes and standards has been integrated into the mechanical engineering technology (MET) curriculum for decades with significant success. This paper discusses how standards are incorporated into mechanical design and quality control courses, as well as strategies for integrating standards into more courses in an MET curriculum. In addition, a discussion of standards resources that are freely available is included. Finally, a call to action for industry is presented, explaining the need and potential areas where industry can increase involvement in teaching students about technical standards

MVB-12, a Fourth Subunit of Metazoan ESCRT-I, Functions in Receptor Downregulation

After ligand binding and endocytosis, cell surface receptors can continue to signal from endosomal compartments until sequestered from the cytoplasm. An important mechanism for receptor downregulation in vivo is via the inward budding of receptors into intralumenal vesicles to form specialized endosomes called multivesicular bodies (MVBs) that subsequently fuse with lysosomes, degrading their cargo. This process requires four heterooligomeric protein complexes collectively termed the ESCRT machinery. In yeast, ESCRT-I is a heterotetrameric complex comprised of three conserved subunits and a fourth subunit for which identifiable metazoan homologs were lacking. Using C. elegans, we identify MVB-12, a fourth metazoan ESCRT-I subunit. Depletion of MVB-12 slows the kinetics of receptor downregulation in vivo, but to a lesser extent than inhibition of other ESCRT-I subunits. Consistent with these findings, targeting of MVB-12 to membranes requires the other ESCRT-I subunits, but MVB-12 is not required to target the remaining ESCRT-I components. Both endogenous and recombinant ESCRT-I are stable complexes with a 1:1:1:1 subunit stoichiometry. MVB-12 has two human homologs that co-localize and co-immunoprecipitate with the ESCRT-I component TSG101. Thus, MVB-12 is a conserved core component of metazoan ESCRT-I that regulates its activity during MVB biogenesis

Esperanto for histones : CENP-A, not CenH3, is the centromeric histone H3 variant

The first centromeric protein identified in any species was CENP-A, a divergent member of the histone H3 family that was recognised by autoantibodies from patients with scleroderma-spectrum disease. It has recently been suggested to rename this protein CenH3. Here, we argue that the original name should be maintained both because it is the basis of a long established nomenclature for centromere proteins and because it avoids confusion due to the presence of canonical histone H3 at centromeres