Vascular smooth muscle cell detachment from elastin and migration through elastic laminae is promoted by chondroitin sulfate-induced “shedding” of the 67-kDa cell surface elastin binding protein

Impaired elastin fiber assembly is observed in the fetal ductus arteriosus (DA), associated with a reduced concentration of elastin binding protein (EBP), a 67-kDa galactolectin. It is also seen in cultured aortic (Ao) smooth muscle cells (SMC) following the release of the EBP by glycosaminoglycans rich in AN-acetylgalactosamine, such as chondroitin sulfate (CS). In the DA, impaired elastin fiber assembly is observed in conjunction with intimal thickening associated with increased migration of SMC into the subendothelium, a feature we previously related to increased production of fibronectin. In this report, we determined whether SMC use the EBP to attach to an elastin substrate, whether shedding of the EBP promotes SMC migration through a threedimensional network of pure elastic laminae prepared from sheep aorta, and whether the latter is associated with increased production of fibronectin. We observed reduced attachment to elastin-coated surfaces of DA SMC deficient in EBP compared to Ao SMC. Addition of CS but not heparan sulfate (a glycosaminoglycan which does not induce EBP shedding) decreased Ao SMC attachment to elastin, as did preincubation with VGVAPG elastin-derived peptides which saturate the EBP. The immunolocalization of cell surface EBP suggested that cells can quickly replace EBP released from their surfaces by CS treatment. The magnitude of CS-induced impaired attachment of SMC to elastin was dose dependent and could be further increased by the administration of cyclohexamide and sodium azide. Also, the reversibility of CS-induced detachment was prevented by monensin. This suggests that a process of new synthesis and intracellular transport of the EBP was necessary to replace the EBP molecules released from the cell surface by CS treatment. In the migration assay, both DA and Ao SMC attached to the top of an elastin membrane, but only DA SMC deficient in EBP migrated through the laminae. Addition of CS, which induced shedding of EBP, resulted in Ao SMC migration associated with increased synthesis of fibronectin. We postulate that CS-induced release of EBP from SMC surfaces causes cell detachment from elastin and an increase in fibronectin synthesis, processes which may be critical in promoting SMC migration associated with intimal thickening developmentally in the DA and perhaps also in vascular disease.     

A P Element Chimera Containing Captured Genomic Sequences Was Recovered at the Vestigial Locus in Drosophila following Targeted Transposition

A P element carrying the Dopa decarboxylase gene, P[Ddc], was targeted into vg21, a cryptic P element induced mutant allele of the vestigial (vg) locus. The resulting allele, vg28w, contained the expected P[Ddc] plus an additional 9.5 kb of DNA, captured from elsewhere on chromosome II. Reversion of the vg28w mutant allele demonstrated that the entire insert can excise but cannot reinsert at an appreciable frequency. We explain the targeted transposition as the repair of a double stranded gap, created by the excision of the P element at vg21, and suggest that the formation of chimeric elements may be an important component of P element dependent genomic instability.     

A review of the bioavailability of petroleum constituents

Exposure to petroleum constituents at contaminated sites may occur through a variety of pathways, including inhalation of vapors and particulates, ingestion of water and soils, and dermal contact with water and soils. Accurately assessing the human health risks from such exposures requires information on the medium‐ and route‐specific bioavailability of petroleum constituents (e.g., how well these chemicals enter the body via the gastrointestinal tract and skin). For example, when the medium or exposure route in an animal toxicity assay (e.g., ingestion of water) differs from the actual route of human exposure at the petroleum contaminated site (e.g., dermal contact with soil), adjustments should be made that reflect the relative bioavailability of the chemical in the different media. The focus of this article is on (1) the availability of oral and dermal absorption data for one PAH (benzo[a]pyrene, (B[a]P) and three VOCs in soil (benzene, toluene, and xylene); (2) factors affecting the uptake of these PAHs and VOCs from soil; and (3) ways to incorporate bioavailability data into human health risk assessments. Based on our review, we recommend the following default values for the oral and dermal absorption of B[a]P, benzene, toluene, and xylene from soil:

A review of the bioavailability of petroleum constituents

All authors

Jennifer Bramard & Barbara D. Beck

https://doi.org/10.1080/15320389209383417

Published online:

02 December 2008

Display Table

Site‐specific information such as chemical concentrations in soil, soil characteristics, soil loadings on the skin, contact site, and contact time could result in modifications of these numbers. As shown, our default absorption values are generally less than those recommended by the U.S. EPA (1991a,b,c). The implications of these estimates of bioavailability for risk assessment and for the selection of soil cleanup levels at petroleum‐contaminated sites are discussed.     

Evolutionary significance of the renal excretion of transferrin half-molecule fragments

It is generally thought that the duplicated structure of serum transferrin in vertebrates arose by gene duplication and fusion from a small ancestral protein. We have found that the isolated domains of transferrin are rapidly lost from the bloodstream via the kidneys. Therefore we suggest that the ancestral transferrin was not a serum protein or, alternatively, that it was not as small as the half-molecule.     

Two approaches to obtaining low,extracellular deoxyribonuclease activity in cultures of heterocyst-forming cyanobacteria

Six out of 158 axenic strains of heterocyst-forming cyanobacteria consistently failed to produce circles of clearing in agar medium containing DNA-methyl green. When tested with [³H]DNA and coliphage DNA, supernatant fluids from cultures of two of these strains [University of Texas Culture Collection (UTEX) strain 2014 and 19-6C-C] showed no detectable deoxyribonuclease activity, and such fluids from another two of the six, and four others, showed low but detectable deoxyribonuclease activity. Covalently closed circular (plasmid) DNA was not detectably degraded by supernatant fluids from UTEX 2014 and 19-6C-C and from four of the other strains. When DNA was incubated with whole cells of certain strains, a sereis of fragments of discrete size was produced, perhaps by cell-bound, periplasmic, restriction endonucleases. Inclusion of one-tenth strength saline sodium citrate (SSC) in an eight-fold dilution of the medium of Allen and Arnon had little effect on growth of Anabaena variabilis American Type Culture Collection (ATCC) strain 29413 yet prevented all but slight degradation of plasmid pBR322 or of DNA.     

An autosomal gene assignment in a marsupial: The gene for LDH-A is on chromosome 5 of the red kangaroo,Macropus rufus

A number of somatic cell hybrids between red kangaroo (Macropus rufus) and mouse cells, which lose marsupial chromosomes, were found to express the kangaroo form of LDH-A. Concordance between the expression of marsupial LDH-A and the presence of chromosome 5 in the hybrid cells and selected subclones enabled the gene for LDH-A to be assigned to this chromosome. This is the first autosomal gene assignment in a marsupial and should prove important for chromosome mapping in the red kangaroo and in many other species of marsupials.     

Analysis of development and growth in a mutant of Physarum polycephalum with defective cytokinesis

Cultures of amoebae of the mutant strain ATS23 isolated from strain CLd of Physarum polycephalum contain multinucleate cells and cells with increased nuclear DNA content. Plasmodia derived from ATS23 clones show abnormal morphology and defective sporulation. All abnormalities are enhanced by high incubation temperature (31 °C). Genetic analysis suggested that all the abnormalities were caused by a single mutation, denoted hts-23. The kinetics of plasmodium formation were followed in cultures of apogamic amoebae carrying hts-23 and hts⁺ (wild type) respectively. Results indicated that, relative to wild type, hts-23 did not increase the rate of plasmodium formation. There was evidence that, in both mutant and wild-type strains, commitment to plasmodium development occurred in uninucleate cells. Analysis of cell pedigrees by time-lapse cinematography indicated that the primary abnormal event in cultures of hts-23 amoebae was failure of cytokinesis; an apparently complete cleavage furrow was formed but cell separation failed, resulting in a binucleate cell. This event occurred randomly in pedigrees in which the majority of divisions were completed normally; its frequency increased during incubation at 31 °C. All other abnormalities in hts-23 amoebal cultures could be attributed to this primary event, assuming that DNA synthesis continued in the absence of cytokinesis and that the binucleate cells underwent the amoebal type of “open” mitosis, allowing the possibility of spindle fusion. This implies that the acquisition of “closed” mitosis is an essential early step in plasmodium development.     

Diel variations of selected physico-chemical parameters in Lake Kissimmee,Florida

Every two months, diel depth profiles were made of dissolved oxygen, pH, temperature and conductivity at two stations in Lake Kissimmee, Florida, from July 1974 through June 1975. Results suggest that stratification does not occur in the large (137 km²), shallow (mean depth 2.5 m) lake, though steep vertical gradients in these parameters may develop. The data also suggest that when these gradients do occur, they rapidly degenerate at night or under slight wind stress. Since many Florida lakes are relatively shallow (mean depth < 5 m) and topography offers little obstruction to wind action, it is believed that the holomictic condition observed in Lake Kissimmee may be typical of the state's lakes, particularly those in the central portion of the peninsula.     

A yellow polysaccharide-producing bacterium with unusual characteristics

Summary A polysaccharide-producing Gram-negative bacterium was isolated from a sample of hay. It grew best on nitrate-containing media with sucrose as carbon source; the colony form was highly unusual. No polysaccharide was formed on glucose. A spontaneous mutant producing the same polysaccharide on both sucrose- and glucose-containing media was isolated. The polysaccharide has been characterized as an acid heteropolymer containing D-galacturonic acid, D-glucose, D-galactose and D-mannose in the approximate molar ratio 1:1:3:1, together with about 2 mol of acetate. The polysaccharide in aqueous solution was highly viscous with pseudoplastic characteristics.