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Biomedical subjects

R G Lewis

Publications and source records attributed to R G Lewis.

54 records · Page 3Linked to original sources
PubMed1973

Biosynthesis of glycosaminoglycans by cultured mastocytoma cells.

Biosynthesis of glycosaminoglycans by several lines of cultured neoplastic mouse mast cells was studied by incorporation of [(35)S]sulphate (and in some cases [6-(3)H]glucosamine) into macromolecular materials found in both the cells and their growth media. Such intracellular and extracellular radioactively labelled materials (shown to be glycosaminoglycans by susceptibility to digestion with heparinase) were further characterized by ion-exchange chromatography and by digestion with testicular hyaluronidase and chondroitinase. All but one cell line produced chondroitin sulphate as the major sulphated glycosaminoglycan; the remainder of the glycosaminoglycan was heparin-like material. No [(3)H]hyaluronic acid was synthesized. Cells of a newly derived line, termed P815S, synthesized more glycosaminoglycan than the other lines. This glycosaminoglycan, found in both cells and growth medium, was almost entirely chondroitin 4-sulphate. No chondroitin 6-sulphate was found. The chondroitin 4-sulphate from the cells was shown by gel filtration to be smaller than the chondroitin 4-sulphate in the media of these cultures. This discovery of relatively high proportions of chondroitin 4-sulphate in these mastocytoma-derived cells is noteworthy, since mast cells have generally been considered to produce heparin as their major glycosaminoglycan.

Journal Article↗
PubMed1973

Biosynthesis of glycosoaminoglycans by microsomal preparations from cultured mastocytoma cells.

Neoplastic mast cells of mice (including long-established and newly derived lines) were grown in large-volume suspension cultures to provide enough cells for preparation of microsomal fractions. Microsomal preparations from P815Y and P815S cells synthesized (14)C-labelled glycosaminoglycan when incubated with UDP-[(14)C]glucuronic acid and UDP-N-acetylgalactosamine. No significant amount of (14)C-labelled glycosaminoglycan was formed when UDP-N-acetylglucosamine was substituted for the UDP-N-acetylgalactosamine. Microsomal preparations from X163 cells synthesized (14)C-labelled glycosaminoglycan when incubated with UDP-[(14)C]glucuronic acid and either UDP-N-acetylgalactosamine or UDP-N-acetylglucosamine. The (14)C-labelled glycosaminoglycan formed in the presence of UDP-N-acetylgalactosamine was degradable by testicular hyaluronidase, indicating that it was chondroitin-like. The (14)C-labelled glycosaminoglycan formed in the presence of UDP-N-acetylglucosamine was not degradable by testicular hyaluronidase. Microsomal preparations from P815S cells were tested for sulphating activity by incubation with adenosine 3'-phosphate 5'-sulphatophosphate, as well as UDP-[(14)C]glucuronic acid, and UDP-N-acetylgalactosamine. The resulting newly synthesized polysaccharide was shown by chondroitinase ABC digestion to be 70% chondroitin 4-sulphate and 30% chondroitin. The molecular size of this newly synthesized glycosaminoglycan was determined by gel filtration to be larger than 40000 mol.wt. In general, the glycosaminoglycan-synthesizing ability of the microsomal preparations appeared to reflect glycosaminoglycan synthesis by the intact cells.

Journal Article↗
PubMedDate not supplied

Experimental methodologies and preliminary transfer factor data for estimation of dermal exposures to particles.

Developmental efforts and experimental data that focused on quantifying the transfer of particles on a mass basis from indoor surfaces to human skin are described. Methods that utilized a common fluorescein-tagged Arizona Test Dust (ATD) as a possible surrogate for housedust and a uniform surface dust deposition chamber to permit estimation of particle mass transfer for selected dust size fractions were developed. Particle transfers to both wet and dry skin were quantified for contact events with stainless steel, vinyl, and carpeted surfaces that had been pre-loaded with the tagged test dust. To better understand the representativeness of the test dust, a large housedust sample was collected and analyzed for particle size distribution by mass and several metals (Pb, Mn, Cd, Cr, and Ni). The real housedust sample was found to have multimodal size distributions (mg/g) for particle-phase metals. The fluorescein tagging provided surface coatings of 0.11-0.36 ng fluorescein per gram of dust. The predominant surface location of the fluorescein tag would best represent simulated mass transfers for contaminant species coating the surfaces of the particles. The computer-controlled surface deposition chamber provided acceptably uniform surface coatings with known particle loadings on the contact test panels. Significant findings for the dermal transfer factor data were: (a) only about 1/3 of the projected hand surface typically came in contact with the smooth test surfaces during a press; (b) the fraction of particles transferred to the skin decreased as the surface roughness increased, with carpeting transfer coefficients averaging only 1/10 those of stainless steel; (c) hand dampness significantly increased the particle mass transfer; (d) consecutive presses decreased the particle transfer by a factor of 3 as the skin surface became loaded, requiring approximately 100 presses to reach an equilibrium transfer rate; and (e) an increase in metals concentration with decreasing particle size, with levels at 25 microns typically two or more times higher than those at 100 microns--consistent with the earlier finding of Lewis et al. for the same sample for pesticides and polycyclic aromatic hydrocarbons (PAHs).

Administration, Cutaneous↗
PubMedDate not supplied

Evaluation of gel permeation chromatography for clean up of human adipose tissue samples for GC/MS analysis of pesticides and other chemicals.

Gel permeation chromatography (GPC) has been evaluated for clean up of human adipose tissue sample extracts in preparation for confirmatory analysis by gas chromatography-mass spectrometry (GC/MS). Studies were conducted with standard solutions, fortified chicken fat, and actual human adipose tissue samples. Nearly quantitative removal of lipid material was achieved with minimal losses of 16 organochlorine pesticides and metabolites: two polychlorinated biphenyl (PCB) mixtures, two chlorinated naphthalene (PCN) mixtures, and a polybrominated biphenyl (PBB) mixture. Mass spectra free from interferences were obtained over the mass range from 46 m/z to 560 m/z.

Adipose Tissue↗
PubMedDate not supplied

Identification of polybrominated biphenyls in the adipose tissues of the general population of the United States.

Hexabromobiphenyl has been identified by gas chromatography/mass spectrometry (GC/MS) in pooled extracts of adipose tissue samples collected from the general population of the conterminous United States. Mass spectra derived from tissue extracts subjected to gel permeation chromatography were compared with those obtained from an authentic PBB mixture containing 2,4,5,2',4',5'-hexabromobiphenyl as the principal component. GC retention times and unique characteristics of the mass spectrum permitted confirmation of identity of the hexabromo isomer. Levels in one tissue sample were determined to be in the 1 to 2 ppb range.

Adipose Tissue↗
PubMedDate not supplied

Chemical analysis of human blood for assessment of environmental exposure to semivolatile organochlorine chemical contaminants.

A chemical method for the quantitative analysis of organochlorine pesticide residues present in human blood was scaled-up to provide increased sensitivity and extended to include organochlorine industrial chemicals. Whole blood samples were extracted with hexane, concentrated, and analyzed without further cleanup by gas chromatography with electron capture detection. The methodology used was validated by conducting recovery studies at 1 and 10 ng/g (ppb) levels. Screening and confirmational analyses were performed by gas chromatography/mass spectrometry on samples collected from potentially exposed residents of the Love Canal area of Niagara Falls, New York and from volunteers in the Research Triangle Park area of North Carolina for 25 specific semivolatile organochlorine contaminants including chlorobenzene and chlorotoluene congeners, hexachloro-1,3-butadiene, pesticides, and polychlorinated biphenyls as Aroclor 1260. Dichlorobenzene, hexachlorobenzene, and beta-hexachlorocyclohexane residues fell in the range of 0.1 to 26 ppb in a high percentage of both the field and volunteer blood samples analyzed. Levels of other organochlorine compounds were either non-detectable or present in sub-ppb ranges.

Chromatography, Gas↗
PubMedDate not supplied

Environmental equity and pesticide exposure.

Although people of color and low-income groups bear a disproportionate share of the health risks from exposure to pesticides, research attention has been meager, and data on acute and chronic health effects related to their toxic exposures are generally lacking. Increased resources are needed both to study this issue and to mitigate problems already identified. People of color should be a major research focus, with priority on long-term effects, particularly cancer, neurodevelopmental and neurobehavioral effects, long-term neurological dysfunction, and reproductive outcome. Suitable populations at high risk that have not been studied include noncertified pesticide applicators and seasonal and migrant farm workers, including children.

Body Burden↗