Psychological effects of vulvectomy.
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Biomedical subjects
Publications and source records attributed to J Robinson.
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A rat hepatoma cell line was shown to synthesize heparan sulfate and chondroitin sulfate proteoglycans. Unlike cultured hepatocytes, the hepatoma cells did not deposit these proteoglycans into an extracellular matrix, and most of the newly synthesized heparan sulfate proteoglycans were secreted into the culture medium. Heparan sulfate proteoglycans were also found associated with the cell surface. These proteoglycans could be solubilized by mild trypsin or detergent treatment of the cells but could not be displaced from the cells by incubation with heparin. The detergent-solubilized heparan sulfate proteoglycan had a hydrophobic segment that enabled it to bind to octyl-Sepharose. This segment could conceivably anchor the molecule in the lipid interior of the plasma membrane. The size of the hepatoma heparan sulfate proteoglycans was similar to that of proteoglycans isolated from rat liver microsomes or from primary cultures of rat hepatocytes. Ion-exchange chromatography on DEAE-Sephacel indicated that the hepatoma heparan sulfate proteoglycans had a lower average charge density than the rat liver heparan sulfate proteoglycans. The lower charge density of the hepatoma heparan sulfate can be largely attributed to a reduced number of N-sulfated glucosamine units in the polysaccharide chain compared with that of rat liver heparan sulfate. Hepatoma heparan sulfate proteoglycans purified from the culture medium had a considerably lower affinity for fibronectin-Sepharose compared with that of rat liver heparan sulfate proteoglycans. Furthermore, the hepatoma proteoglycan did not bind to the neoplastic cells, whereas heparan sulfate from normal rat liver bound to the hepatoma cells in a time-dependent reaction. The possible consequences of the reduced sulfation of the heparan sulfate proteoglycan produced by the hepatoma cells are discussed in terms of the postulated roles of heparan sulfate in the regulation of cell growth and extracellular matrix formation.
We obtained spontaneous formation of Epstein-Barr virus-transformed B lymphocyte colonies from the blood of four patients with mononucleosis with the use of soft-agar medium. The colonies were propagated into separate cell lines and analyzed for their immunoglobulin secretion. Of 52 such lines, 44 produced immunoglobulin composed of a single class of heavy chain and single type of light chain. Among these clonal transformants, 27 (62%) of 44 produced mu-chain, 13 (29%) of 44 produced gamma-chain, and 4 (9%) of 44 produced alpha-chain. Analysis of light-chain secretion revealed that of the 44 cell clones secreting complete, monoclonal immunoglobulin products, 31 produced kappa-chain and 13 produced gamma-chain. One clone secreted mu-heavy chain and no light chain, an observation suggesting the clone is a pre-B cell phenotype. Seven lines derived from clones had aberrant patterns of immunoglobulin secretion that produced either two heavy chains or two light chains. B lymphocytes in different states of immunoglobulin gene expression are, therefore, transformed in vivo by Epstein-Barr virus.
Five solid media were evaluated for isolation of Aeromonas spp. from faeces: desoxycholate citrate agar (DCA), MacConkey's agar (MAC), xylose-desoxycholate-citrate agar (XDCA), Rogol's medium (ROG), which contained ampicillin 20 mg/L and p-nitrophenyl-glycerine 25 mg/L as inhibitors, and blood agar (BA) with ampicillin 10 mg/L. False negative oxidase tests limited the usefulness of DCA and MAC and, although the use of XDCA avoided the problem of fermentation of lactose, some Aeromonas spp. failed to grow on XDCA or produced minute colonies unsuitable for oxidase tests. BA yielded the highest rate of isolation for Aeromonas spp. from 323 faecal samples--15.2% for all Aeromonas spp. and 9.3% for enterotoxigenic (ENT+) strains. This compares with 10.8% for all strains and 6.5% for ENT+ strains isolated on DCA, 7.1% for all strains and 4% for ENT+ strains on MAC and 4% for all strains and 1.5% for ENT+ strains on ROG. Blood agar with ampicillin is recommended for isolation of Aeromonas spp. from faeces.
Biochemical characteristics and virulence factors were compared in 147 Aeromonas spp. isolated from patients with diarrhea and in 94 strains isolated from metropolitan water supplies in the same area during the same period. Fermentation of arabinose occurred with 58.5% of the environmental strains and 15% of the clinical isolates; 39.4% of the strains from water and 6.8% of the fecal isolates fermented salicin. The frequency of esculin hydrolysis was the same in both groups. Ninety-one percent of clinical isolates and 70.2% of environmental strains were enterotoxigenic and, except for four clinical isolates, all of these strains also produced hemolysins. Hemagglutination that was inhibited by fucose and mannose but not by galactose was found in 67% of the water isolates and 10.2% of the clinical strains. Although the distribution of several characteristics differs in clinical and environmental strains, many of the strains found in water have properties identical with those of the clinical isolates. We suggest that such strains may be potential enteric pathogens.
The occurrence of Aeromonas spp. in the metropolitan water supply of Perth, Western Australia, Australia, was monitored at several sampling points during a period of 1 year. Water within the distribution system conformed to international standards for drinking water but contained Aeromonas spp. in numbers comparable to those in raw surface water, although this water was free of Escherichia coli. Coliforms and E. coli were found in raw surface waters, and Aeromonas spp. were found in raw water from surface and underground sources. Chemical treatment, followed by chlorination at service reservoirs, resulted in water free of E. coli and a decrease in the number of Aeromonas spp. Aeromonas spp. were found in the greatest numbers in summer. Multiple regression analysis showed that growth of Aeromonas spp. in chlorinated water was related to water temperature, residual chlorine, and interaction between these variables. The incidence of Aeromonas-associated gastroenteritis, determined from isolates referred to us for enterotoxin testing, paralleled the pattern of isolation of Aeromonas spp. in water within the distribution systems. We suggest that the presence of Aeromonas spp. in drinking water needs public health appraisal and that further work should be undertaken to permit reevaluation of standards for the quality of drinking water.
The recovery of Aeromonas spp. from the unchlorinated water supply for a Western Australian city of 21,000 people was monitored at several sampling points during a period of 1 year. Membrane filtration techniques were used to count colonies of Aeromonas spp., coliforms, and Escherichia coli in water sampled before entry to service reservoirs, during storage in service reservoirs, and in distribution systems. Aeromonas spp. were identified by subculture on blood agar with ampicillin, oxidase tests, and the use of Kaper medium and then were tested for production of enterotoxins and hemolysins. During the same period, two-thirds of all fecal specimens sent for microbiological examination were cultured on ampicillin-blood agar for Aeromonas spp. Recovery of Aeromonas spp. from water supplies at distribution points correlated with fecal isolations and continued during autumn and winter. Coliforms and E. coli were found most commonly in late summer to autumn. This pattern differs from the summer peak of Aeromonas isolations both from water and from patients with Aeromonas spp.-associated gastroenteritis in Perth, Western Australia, a city with a chlorinated domestic water supply. Of the Aeromonas strains from water, 61% were enterotoxigenic, and 64% produced hemolysins.
Aeromonas spp. show patterns of hemagglutination with human group O cells in the presence of fucose, galactose, and mannose. These patterns are related to biotype as well as to the source of isolates. There was good correlation between hemagglutination pattern and the presence of diarrhea among strains isolated in Western Australia, which was the only source with adequate data for classification of children with an without diarrhea. Most of the environmental and other nonfecal isolates produced patterns different from those in strains associated with diarrhea. These results suggest that hemagglutinins should be considered with enterotoxins as virulence factors in Aeromonas spp.
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The interaction of coat proteins from coated vesicles with model lipid membranes was examined using small unilamellar vesicles of dipalmitoylglycerophosphocholine as model membranes. Changes in membrane permeability were measured by the leakage of entrapped fluorescent dye, carboxyfluorescein. Both clathrin and the 55000-Da protein were found to be active. Density gradient centrifugation showed the formation of an irreversible protein-lipid complex. Dynamic light-scattering measurements showed that this complex is significantly larger than the original vesicles, suggesting that fusion is induced. The effects of pH, urea, Tris and ionic strength were studied and the possible biological relevance of the results is discussed.
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Aeromonas hydrophila was isolated from the faeces of 32 patients during a nine month period in three hospitals in Western Australia. All 32 isolates produced enterotoxin which was detected by the suckling-mouse test, and all patients except one had diarrhoea. Treatment should be considered in patients with chronic diarrhoea and in those with malignant disease or with hepatobiliary disease who are at risk of developing aeromonas septicaemia.
Bovine corneal endothelial (BCE) cells seeded and grown on plastic dishes were labeled with 35S-sulfate or 3H-glucosamine for 48 h at various phases of growth of the cultures. Newly synthesized proteoglycans were isolated from the culture medium and from the extracellular matrix (ECM) produced by the BCE cells, and the glycosaminoglycan (GAG) component of the proteoglycans was analyzed. Cells actively proliferating on plastic surfaces secreted an ECM that contained heparan sulfate as the major 35S-labeled GAG (86%) and dermatan sulfate as a minor component (13%). Upon reaching confluence, the BCE cells incorporated 35S-labeled chondroitin sulfate (20%), as well as heparan sulfate (66%) and dermatan sulfate (14%), into the EC. Seven-day postconfluent cells incorporated newly synthesized heparan sulfate and dermatan sulfate into the matrix in approximately equal proportions. Dermatan sulfate was the main 35S-labeled GAG (60-65%) in the medium of both confluent and postconfluent cultures. 35S-Labeled chondroitin sulfate (20-25%) and heparan sulfate (15%) were also secreted into the culture medium. The type of GAG incorporated into newly synthesized ECM was affected when BCE cells were seeded onto ECM-coated dishes instead of plastic. BCE cells actively proliferating on ECM-coated dishes incorporated newly synthesized heparan sulfate and dermatan sulfate into the ECM in a ratio that was very similar to the ratio of these GAGs in the underlying ECM. Addition of mitogens such as fibroblast growth factor (FGF) to the culture medium altered the type of GAG synthesized and incorporated into the ECM by BCE cells seeded onto ECM-coated dishes if the cells were actively growing, but had no effect on postconfluent cultures.
A year-long, prospective study of Aboriginal children with diarrhoea showed a high rate of isolation of bacterial pathogens, viruses and parasites in faecal specimens. Seventeen per cent of patients had enterotoxigenic bacteria in their stools; none of the 132 Aboriginal children without diarrhoea studied at the same time had enterotoxigenic bacteria isolated. Salmonellae and shigellae were also much more common in the diarrhoea group. Bacterial pathogens including enterotoxigenic E. coli and Aeromonas, salmonella, shigella, and campylobacter occurred in 39% of the diarrhoea group but were found in only 4.5% of the controls. Rotaviruses were detected in 12% of children with diarrhoea and in 8% of controls. Intestinal parasites, particularly Giardia lamblia, were present in over 25% of children in each group.
A prospective, 12-month study of 975 non-Aboriginal children with diarrhea and age- and sex-matched children without diarrhea, in Perth, Western Australia, was designed to investigate the significance of enterotoxigenic Aeromonas species as a cause of diarrhea. Enterotoxigenic Aeromonas species were found in the fecal specimens of 10.8% of the patients with diarrhea but in only 0.7% of those without diarrhea. Most Aeromonas species were isolated during the summer. Other important bacterial pathogens included Campylobacter, Salmonella, Shigella, and enterotoxigenic Escherichia coli; rotavirus infections appeared to be much less important in the Western Australian environment. Most of the patients were younger than two years of age and about one-quarter had mixed bacterial and/or viral intestinal infections. Enterotoxigenic Aeromonas species can be identified with 97% accuracy using a simple hemolysin assay which should be considered for use by routine diagnostic laboratories, particularly in children's hospitals.
Enterotoxin production correlated with biotype in a study of 686 strains of Aeromonas spp. from Indonesia, Thailand, the United States, and Western Australia. Most strains were isolated from feces but nonfecal human isolates and environmental strains were also included. More than 80% of Voges-Proskauer (VP)-positive strains, classified as A. hydrophila, were enterotoxigenic in the suckling mouse assay as were 90% of VP-positive, arabinose-negative strains. An association between positive VP, arabinose fermentation, and failure to produce enterotoxins was found only with environmental strains. VP-negative strains which did not oxidize gluconate or produce gas from glucose were classified as A. punctata subsp. caviae. Only 2 of the 286 strains produced enterotoxins, and both were from Indonesian fecal samples. There were few remaining VP-negative strains, classified as A. punctata subsp. punctata and, of these, about half were enterotoxigenic. Regardless of source and species, 97% of Aeromonas spp. were correctly classified in relation to enterotoxin production with a hemolysin assay. A combination of biochemical testing and hemolysin assay should be suitable for diagnostic laboratories to identify enterotoxigenic Aeromonas spp. which, in children, are associated with diarrhea, unlike non-enterotoxigenic strains.