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Biomedical subjects

R J Gay

Publications and source records attributed to R J Gay.

13 recordsLinked to original sources
PubMed1993

Effects of UV irradiation on a living skin equivalent.

The Living Skin Equivalent (LSE) is an organotypic coculture composed of human dermal fibroblasts interspersed in a collagen-containing matrix and overlaid with human keratinocytes forming a stratified epidermis. The LSE has a dry, air-exposed epidermal surface suitable for the application of oils, creams and emulsions. These features suggested its feasibility as an in vitro skin model for studying the protective effects of sunscreens. Using the thiazolyl blue (MTT) conversion assay as a measure of mitochondrial function, the extent of cytotoxicity induced by various doses of UV-R (280-400 nm) or UV-A (320-400 nm) was evaluated in the LSE. The doses of UV radiation that caused 50% reductions in MTT conversion (UV-R50 or UV-A50) in different lots of LSE were 0.053 +/- 0.021 J/cm2 (n = 29) and 11.6 +/- 4.9 J/cm2 (n = 17) for UV-R and UV-A, respectively. The protective effects of an 8% homosylate standard and of five UV-A sunscreens, topically applied to the LSE, were determined and compared with their reported protection factors in human skin. Morphological changes and the release of proinflammatory mediators (interleukin-1-alpha, tumor necrosis factor-alpha and prostaglandin E2) implicated in UV-induced erythema were also demonstrated in the LSE exposed to UV-A or UV-B. The data suggest that the LSE can be used for studying the effects of UV radiation on skin and may have utility for assessing the efficacy of certain sunscreens against UV-B and UV-A.

Cell Line↗
PubMed1984

Deprival of nicotinamide leads to enhanced glucose transport in chick embryo fibroblasts.

Chick embryo fibroblasts growing in medium free of pyridine ring precursors of NADH and NADPH replicate several times before multiplication ceases. The rate of glucose transport is progressively enhanced, finally reaching levels several times higher than those normally observed in cells severely depleted of NADH. Whereas normal cells respond to additional glucose by further reducing transport, the NADH-depleted cell is refractory to glucose even at five times the normal glucose concentration. Readdition of nicotinamide does little to restore normal transport within 24 h. On the other hand NAD+ or NADP+ provided simultaneously with glucose results in a sharp decline in measurable transport within 2-4 h. The role of the pyridine nucleotides in this reduction of transport function is for the moment unknown.

3-O-Methylglucose↗
PubMed1983

Purines as 'hyper-repressors' of glucose transport. A role for phosphoribosyl diphosphate.

Under selected conditions the rate of glucose transport and the intracellular phosphoribosyl diphosphate (PPRibP) concentrations of chick-embryo fibroblasts are inversely correlated. This relationship holds when cells are incubated with mannose, fructose, xylose or various concentrations of glucose. The metabolic inhibitors 2,4-dinitrophenol, rotenone and Methylene Blue increased glucose transport and decreased PPRibP. The addition of any pyrimidine or purine base or ribonucleoside dramatically depleted PPRibP pools, regardless of the carbon source. Addition of guanine (10 microM) or hypoxanthine (100 microM) decreased transport in glucose-grown chick cells to barely detectable values, but did not affect increases observed in cells depressed by substitution of xylose for glucose. Guanosine, inosine and the purine analogues 6-thioguanine, 6-thioguanosine, 8-azaguanine and 6-methylmercaptopurine riboside sharply decreased transport in glucose-grown cells and blocked the increase in transport resulting from the replacement of glucose by fructose or xylose in the culture medium.

3-O-Methylglucose↗
PubMed1982

Counter-transport in chick embryo fibroblasts. A significant factor in measurement of glucose entry.

Enhanced rates of carrier-mediated 3-O-methyl-D-glucose (0.1 mM) transport were observed in primary cell cultures of chicken embryo fibroblasts deprived of glucose for 1 day. The addition of 5.5 mM-glucose, glucosamine or 2-deoxy-D-glucose for 15 min (37 degrees C) to glucose-starved cultures followed by washing and immediate measurement of 3-O-methyl-D-glucose transport resulted in an apparent further stimulation of transport. Transport stimulation increased with increasing concentrations of the added preincubation sugar and was observed at test concentrations ranging from 0.1 mM- to 10 mM-3-O-methyl-D-glucose. This enhancement occurred when the preloaded sugar was rapidly effluxing from cells and was eliminated by allowing cultures to incubate in buffer without sugar for 30 min (37 degrees C) after the removal of hexose and before measuring transport. A transient overshoot in the cumulative uptake of 3-O-methyl-D-glucose was observed in glucose-starved cultures that were pre-incubated in the presence of 55 mM-glucose or -glucosamine for 15 min (37 degrees C). These data suggest that counter-transport accounts for the apparent enhancement of glucose-transport capability observed in glucose-starved cells when they are briefly re-exposed to hexose.

3-O-Methylglucose↗
PubMed1980

Influence of proliferative rates and A system substrate availability on proline transport in primary cell cultures of the R3230AC mammary tumor.

Regulation of A system amino acid transport was studied in primary cultures of the R3230AC mammary adenocarcinoma. Higher rates of carrier-mediated Na+-dependent proline transport, vc, was decreased and was attributed to a two-fold decrease in Vmax and a two-fold increase in Km. When compared to cells grown in standard media (Eagle's minimal essential medium, MEM), cells grown in media supplemented with A system substrates (alanine, serine, glycine, and proline) demonstrated adaptive decreases in proline transport; the decrease was due to two-fold reduction in Vmax, with no change in Km for proline. Even in the presence of preferred substrates for the A system, a density-dependent decrease in proline transport was manifested. Both fast- and slow-growing cultures maintained in MEM exhibited rapid increases in proline transport when switched to buffers devoid of amino acids; two-fold increases in Vmax were seen within 4 hr, but Km was unchanged. This starvation-induced adaptation was completely prevented by inclusion in the buffer of 10 mM proline, 0.1 mM alpha-(methylamino)-isobutyric acid (MetAIB) or 10 mM serine, whereas inclusion of the poorer A system substrate, phenylalanine (10 mM), had no effect. The effects of MetAIB to prevent starvation-induced increases in proline transport were dose-related, rapid, and reversible. Amino acid starvation-induced increases in proline transport were partially blocked by cycloheximide or actinomycin D. Data were obtained demonstrating a temporal relationship between increasing intracellular [proline] and decreasing vc for proline uptake. In addition, efflux of proline from preloaded cells preceded the increase in initial rates of proline entry. Taken together, we concluded that: 1) A system transport in primary cultures of this mammary adenocarcinoma is regulated by cell density as well as by availability of A system substrates, but these two types of regulation are kinetically distinct; and 2) starvation-induced enhancement of proline transport appears to be due to release from transinhibition, but may also involve a derepression-repression type of mechanism.

Amino Acids↗