Search PubMed⌕ Search

Biomedical subjects

D W Barnes

Publications and source records attributed to D W Barnes.

At least 73 records · Page 4Linked to original sources
PubMed1984

Human serum spreading factor: relationship to somatomedin B.

Human serum spreading factor (SF) is a blood glycoprotein that promotes the attachment and spreading of a variety of cell types in serum-free culture, as well as affecting migration, proliferation and differentiation of some cell types under appropriate conditions. The amino acids occupying 17 of the first 23 positions from the NH2-terminus of SF were determined, and this sequence was found to be identical to that reported for somatomedin B (SmB) by Fryklund and Sievertsson (FEBS Letters 87:55). Immunoassay of SF in plasma from subjects with conditions related to altered GH levels indicated that serum SF levels were not GH-responsive to any marked degree. No effect of purified SF was observed in a cell growth assay used previously to detect mitogenic activity in SmB preparations. These results support the conclusion that SF acts as a substratum molecule to elicit its biological effects in cell culture, and does not act in a manner similar to peptide growth factors. These data also are consistent with the conclusion of Heldin et al. (Science 213:1122) that the mitogenic activity of SmB preparations is derived from a contaminating factor, and is not due to SmB.

Amino Acid Sequence↗
PubMed1984

Suppression of humoral antibody production by exposure to 1,2,3,6,7,8-hexachlorodibenzo-p-dioxin.

The day 4 IgM antibody (AB) response to sheep red blood cells was suppressed in adult female B6C3F1 mice subchronically (14 day) exposed to 1,2,3,6,7,8-hexachlorodibenzo-p-dioxin (1,2,3,6,7,8-HCDD) at concentrations reflecting the levels of this dioxin isomer as a contaminant in technical-grade pentachlorophenol. Hepatic microsomal parameters were also measured in these mice and indicated the characteristic induction of mixed-function oxidase enzyme activities, particularly aryl hydrocarbon hydroxylase. The response to sheep red blood cells was also suppressed when measured in vitro by culturing the antigen with spleen cells from a second group of mice subchronically exposed to 1,2,3,6,7,8-HCDD. Enumeration studies with fluorescent-labeled antisera in these animals indicated a dose-related suppression in the number of T-lymphocytes with no effect on B-lymphocytes. 1,2,3,6,7,8-HCDD directly suppressed several in vitro AB responses when added to cultures of spleen cell suspensions from untreated B6C3F1 mice. The rank order of sensitivity was determined to be: polyclonal AB response to LPS greater than or equal to T-independent AB response to DNP-Ficoll greater than T-dependent AB response to sheep red blood cells. Subsequent studies indicated that the suppression by 1,2,3,6,7,8-HCDD could be produced after only a 30-min preincubation. The suppression of all in vitro AB responses was abolished when the dioxin was preincubated with a metabolic activation system: crude liver homogenate (B6C3F1 mice pretreated with the mixed-function oxidase inducer; Aroclor 1254) plus NADP and isocitrate. The suppression of the T-dependent AB response was still evident when the metabolic activation system was heat-inactivated (as verified by an inability to activate cyclophosphamide) prior to the preincubation.

Animals↗
PubMed1984

Quantitation of spreading factor in human biologic fluids.

Spreading factor (SF) is a glycoprotein isolated from human plasma or serum that promotes attachment and spreading of a wide variety of mammalian cells in culture. Under appropriate conditions, this factor also affects cell proliferation and differentiation in vitro. An immunoassay employing a previously characterized monoclonal antibody to SF, with purified SF as a reference standard, was utilized to quantitate levels of SF in human plasma, serum, amniotic fluid, and urinary protein. Specific anti-SF monoclonal antibody-binding activities in individual samples of human serum were identical to specific anti-SF monoclonal antibody-binding activities in the plasma samples from which the sera were derived. Specific anti-SF monoclonal antibody-binding activity in plasma from adults, when compared with purified SF reference standards, represented an average of 4 micrograms of SF per mg plasma protein (range, 2.5 to 7 micrograms/mg). Specific anti-SF monoclonal antibody-binding activity in sera from fetal cord blood, infants, children, or adolescents was not significantly different from that of adults. Specific anti-SF monoclonal antibody-binding activities in amniotic fluid samples from pregnancies of 12 to 23 weeks' gestation were within the range observed in sera of adults. Anti-SF monoclonal antibody-binding activity in concentrated urinary protein, when compared with purified SF reference standards, represented a specific activity of 10.9 micrograms of SF per mg protein.

Adolescent↗
PubMed1983

Isolation of human serum spreading factor.

Serum spreading factor (SF) was isolated from human serum by a four-step procedure employing affinity chromatography on glass beads, concanavalin A-Sepharose, DEAE-agarose, and heparin-agarose. The final product was purified approximately 260-fold from the starting material and was maximally active in assays of cell spreading-promoting activity at 300 ng/ml. The isolated human SF preparation consisted of two proteins of apparent molecular weights approximately 65,000 (SF65) and 75,000 (SF75). Both SF65 and SF75 have been shown previously to exhibit cell spreading-promoting activity and to bind monoclonal antibody to human serum SF.

Amino Acids↗
PubMed1983

Characterization of human serum spreading factor with monoclonal antibody.

Serum spreading factor is a glycoprotein isolated from human serum that promotes spreading of a variety of cell types on culture dishes. We developed mouse hybridoma lines secreting monoclonal antibody to serum spreading factor that markedly inhibited the rate of serum spreading factor-promoted spreading of both fibroblastic and epithelial cells in culture. Fibronectin-promoted cell spreading was unaffected by monoclonal antibody to serum spreading factor, and the factor appeared to be distinct by several criteria from fibronectin or laminin. Human serum-promoted cell spreading was partially inhibited by monoclonal antibody to serum spreading factor. The antibody recognized primarily two forms of serum spreading factor that migrated in NaDodSO4/polyacrylamide gel electrophoresis in a manner consistent with molecular weights of 65,000-70,000 and 75,000-78,000. In addition to being found in plasma, serum spreading factor was also found associated with washed human platelets.

Animals↗
PubMed1983

Subchronic toxicology of diethystilbestrol in the mouse.

This study evaluated the subchronic (14-day) toxicity of selected (0.2, 1.0, and 4.0 mg/kg) daily subcutaneous injections of diethylstilbestrol (DES) in female (C57B1/6 X C3H)F1 mice. Parameters observed included body and organ weights, gross organ morphology, histopathology, clinical chemistry, and hepatic microsomal enzyme activities. The liver, bone marrow, and thymus are major target organs for DES. Liver enlargement, with associated histopathological changes consistent with mild hepatitis, centrolobular necrosis, and sinusoidal changes were observed. Supporting the histological changes were alterations in serum enzyme levels and microsomal enzyme activity. Bone marrow changes included decreases in the number of cells as well as the number of colony forming units per gram stem cells. Toxicity to the thymus was evidenced by decreased thymic weights and lymphocyte depletion. The hepatic and thymic effects were observed at the lowest (0.2 mg/kg) dose. Although all parameters were not assessed for recovery, those that were evaluated returned to control levels by thirty days after treatment.

Animals↗
PubMed1982

The role of lysosomes in exercise-induced hepatic protein loss.

Previous reports have shown that exercise causes a loss of liver protein. The purpose of the present study was to elucidate the mechanism of this exercise-induced protein loss. Exercise caused: (1) an increase in mechanical and osmotic lysosomal fragility; (2) a significant loss of hepatic water, glycogen, protein, phospholipid and RNA; (3) loss of protein from the soluble, mitochondrial and microsomal fractions: (4) loss of mitochondrial, microsomal and cytosolic, but not lysosomal, enzyme activity; (5) an increase in the number of autophagic vacuoles; (6) an increase in the lysosomal size. Taken together, these results suggest that the autophagolysosomal system is responsible for the exercise-induced hepatic protein loss.

Animals↗
PubMed1982

The relative biological effectiveness of fast neutrons (42MeVd leads to Be) for early and late normal tissue injury in the pig.

Early and late radiation damage has been investigated in a number of normal tissues in the pig after irradiation with single doses of neutrons produced by 42MeV deuterons on beryllium. The results have been compared with data obtained after irradiation with single doses of 250kV X rays. In the skin a low RBE value of approximately 1.2 was obtained for the early (3-9 week) epithelial reaction. For the subsequent dermal vascular response, higher RBE values in the range of 1.35-1.6 were obtained: the RBE decreasing with an increase in the neutron dose. For late skin damage, assessed by the relative reduction in the linear dimensions of an irradiated field, a RBE value of approximately 1.5 was obtained. In the kidney the RBE value, for a neutron dose level (550 cGy) at which renal function was just preserved, was 2.0. A lower value of 1.7 was found for doses resulting in a loss of renal function. The results of 133Xenon clearance studies showed two waves of impaired ventilation function in the irradiated lung. In the acute reaction (3-9 months), at a dose level consistent with just preserving normal ventilation function, the RBE value was less than 1.2. For late lung damage (15-24 months) the RBE value was higher, 1.4. For the rectum, methods are presently only available for assessing acute damage. A RBE of 2.0 was found for neutron doses in the range 350-575 cGy. The RBE values for early endpoints in the skin, lung and gut of the pig are comparable with those published previously for other species, including man. The values for late effects in pig skin and lung were higher than for early damage in those tissues.

Animals↗
PubMed1982

Epidermal growth factor inhibits growth of A431 human epidermoid carcinoma in serum-free cell culture.

A medium consisting of a rich basal nutrient mixture supplemented with bovine insulin (10 micrograms/ml), human transferrin (10 micrograms/ml), human cold-insoluble globulin (5 micrograms/ml), and ethanolamine (0.5 mM) supported the growth of the A431 human epidermoid cell line in the absence of serum with a generation time equal to that of cells in serum-containing medium. Addition of epidermal growth factor (EGF) to this culture medium at concentration mitogenic for other cell types resulted in a marked inhibition of A431 cell growth. Inhibitory effects of EGF were observed at 1 ng/ml and near-maximal effects were observed at 10 ng/ml. The inhibitory effect of EGF could be reversed by the omission of EGF in subsequent medium changes and could be prevented by the addition of anti-EGF antibody to the culture medium. Inhibition of A431 cell growth by EGF also could be demonstrated in serum-containing medium.

Carcinoma, Squamous Cell↗
PubMed1982

Toxicology of chloral hydrate in the mouse.

Chloral hydrate has been found in our drinking water supplies at levels up to 5 micrograms/1. The purpose of this study was to evalute the acute and subchronic toxicology of chloral hydrate in the random-bred CD-1 mouse, to provide data for risk assessment. The acute oral LD50 of this compound was 1442 and 1265 mg/kg in male and female mice, respectively. Acute toxicity appeared to be related to depression of the central nervous system. Fourteen-day exposure by gavage in male mice at doses 1/10 and 1/100 the LD50 caused an increase in liver weight and a decrease in spleen weight at the highest dose level. Based on the data derived from 14 days of exposure, a 90-day study was performed. The compound was delivered via the drinking water; levels of the compound delivered per day were equivalent to those dosed in the 14-day study. The target organ in both sexes appeared to be the liver, with the males most affected. Male mice demonstrated a dose-related hepatomegaly accompanied by significant changes in serum chemistries and hepatic microsomal parameters. The females did not demonstrate the hepatomegaly observed in males, but did show alterations in hepatic microsomal parameters. No other significant toxicological changes were observed in either sex following 90 days of exposure.

Administration, Oral↗
PubMed1982

Toxicology of organic drinking water contaminants: trichloromethane, bromodichloromethane, dibromochloromethane and tribromomethane.

This study evaluated the subchronic toxicity of selected halomethanes which are drinking water contaminants. The compounds studied were trichloromethane, bromodichloromethane, dibromochloromethane and tribromomethane. Subchronic 14-day gavage studies were performed with the use of doses encompassing one-tenth the LD50 for the compounds. A 90-day gavage study of one of the compounds, trichloromethane, was also done. Parameters observed included body and organ weights, histopathology, hematology, clinical chemistries, and hepatic microsomal enzyme activities. Toxicity to the humoral immune system was assessed by measuring the number of splenic IgM antibody-forming cells and the serum antibody level to sheep erythrocytes. Cell-mediated immunity was evaluated by measuring the delayed type hypersensitivity response and popliteal lymph node proliferation response to sheep red blood cells. The functional activity of the reticuloendothelial system, as measured by the vascular clearance rate and tissue uptake of 51Cr sheep red blood cells was also determined. The major effects of the halomethanes were increased liver weights, elevations of SGPT and SGOT, decreased spleen weights and a decrease in the number of splenic IgM antibody-forming cells. The humoral immune system appeared to be an indicator of halomethane toxicity. There is evidence that subchronic 14-day exposure may be of greater value than long-term studies in determining the toxicity of these compounds.

Administration, Oral↗
PubMed1978

Inhibition of sugar uptake in adenosine-treated 3T3 cells.

Addition of 5 to 250 micromolar adenosine to the culture medium resulted in a 30-80% inhibition of the rate of uptake of 2-deoxyglucose or 3-0-methylglucose by sparse or confluent 3T3 cells within three hours. The inhibition of deoxyglucose uptake could be reversed partially by changing the cells to medium without adenosine for two hours and could be prevented completely by the addition of persantin, an inhibitor of nucleoside uptake. The adenosine effect is not due to inhibition of pyrimidine synthesis, since it is not prevented by uridine. It is not seen in 3T6 cells lacking adenosine kinase. The inhibition could be observed on confluent cells whose deoxyglucose uptake was stimulated by insulin, epidermal growth factor (EGF), calf serum or calcium phosphate. Although the percentage stimulation over control by these factors varied, the percentage inhibition by addition of adenosine of the stimulated rates, as well as the unstimulated rate, was relatively constant. EGF, insulin and calcium phosphate caused little or no stimulation of deoxyglucose uptake by sparse cells, whether adenosine treated or untreated. The results suggest that adenosine acts intracellularly after phosphorylation to regulate sugar uptake through a mechanism which is independent of the regulation by hormones and cell density.

Adenosine↗
PubMed1978

Relationship of molecular weight to antiviral and antitumor activities and toxic effects of maleic anhydride-divinyl ether (MVE) polyanions.

The molecular weight (MW) and dose dependency of several of the toxic effects and antitumor and antiviral activities of a new series of five maleic anhydride-divinyl ether copolymers (MVE) were established. Each polyanion preparation was relatively homogeneous and exhibited a narrow MW range, from 12,500 (MVE-1) to greater than 52,000 (MVE-5). All of the polyanions were effective as adjuvants to surgery against the metastatic Lewis lung carcinoma, and also exhibited marked antitumor activity against the P815 mastocytoma. MVE-1 retained antitumor activity while losing considerable antiviral activity. This polyanion also exhibited the least toxicity with regard to criteria such as sensitization to the lethal effects of endotoxin, inhibition of reticuloendothelial function, and depression of the microsomal mixed functional oxidase system. The MVE-4 (MW, 32,000) and MVE-5 (MW, 52,600) polyanions exhibited potent antitumor and antiviral activity, but also demonstrated dose-dependent toxic effects.

Animals↗
PubMed1977

Stimulation of sugar uptake and thymidine incorporation in mouse 3T3 cells by calcium phosphate and other extracellular particles.

Evidence is presented that the marked stimulation of sugar uptake and thymidine incorporation by addition of extra Ca2+ to stationary phase mouse 3T3 cells in culture is phosphate dependent and due to the action of the calcium phosphate precipitate formed in the medium. The cells are similarly stimulated by a variety of particulate materials, including calcium pyrophosphate, barium sulfate, kaolin, and polystrene beads. The precipitate effects on sugar uptake are of the same magnitude as those seen with certain hormones (insulin, epidermal growth factor) or with fresh 10% calf serum. The effect of barium sulfate on thymidine incorporation is also of the same magnitude as seen with these hormones, but much less than half that found with fresh calf serum. The stimulation by barium sulfate or hormones of thymidine incorporation is not phosphate dependent.

Barium Sulfate↗