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Biomedical subjects

C S Stewart

Publications and source records attributed to C S Stewart.

At least 55 records · Page 3Linked to original sources
PubMed1985

The effect of ammonia treatment on the solubilization of straw and the growth of cellulolytic rumen bacteria.

Pre-treatment of straw with anhydrous ammonia increased its susceptibility to solubilization by the predominant cellulolytic bacteria from the rumen, Bacteroides succinogenes, Ruminococcus albus and R. flavefaciens. Ammonia treatment also increased the production of microbial protein and fermentation products by all three species. Scanning electron microscope observations of straw during digestion suggested that the attack of straw by these bacteria was accompanied by the formation of substantial numbers of adherent microcolonies.

Ammonia↗
PubMed1984

Polyacrylamide gel electrophoresis of Bacteroides succinogens.

Polyacrylamide gel electrophoresis (PAGE) of sodium dodecyl sulphate (SDS) extracts of bacterial cells was used to assess the similarity of polypeptide patterns in 15 isolates of the rumen cellulolytic bacterium Bacteroides succinogens. Analysis of densitometer traces of the polypeptide band patterns suggested that the majority of the isolates tested were closely related to the type strain S85. Representatives of other species of Gram-negative rumen bacteria had distinct SDS-PAGE patterns, and it is concluded that SDS-PAGE is a useful aid to the identification of B. succinogenes.

Animals↗
PubMed1982

Preparation of the cellulase from the cellulolytic anaerobic rumen bacterium Ruminococcus albus and its release from the bacterial cell wall.

1. Most of the cellulase (CM-cellulase) elaborated by the rumen bacterium Ruminococcus albus strain SY3, which was isolated from a sheep, was cell-wall-bound. 2. The enzyme could be released readily by washing either with phosphate buffer or with water. 3. The amount of enzyme released was affected by the pH and ionic strength of the phosphate buffer. 4. The cell-wall-bound enzyme was of very high molecular weight (>>1.5x10(6)) as judged by its chromatographic behaviour on Sephacryl S-300. 5. The molecular weight of the extracellular enzyme was variable and depended on the culture conditions. 6. When cellobiose was used as the energy source and the medium contained rumen fluid (30%), the extracellular enzyme was, in the main, of high molecular weight. 7. When cellulose replaced the cellobiose, the cell-free culture filtrate contained only low-molecular-weight enzyme (M(r) approx. 30000) in late-stationary-phase cultures (7 days). 8. Cultures that did not contain rumen fluid contained mainly low-molecular-weight enzyme. 9. Under some conditions the high-molecular-weight enzyme could be broken down to some extent into low-molecular-weight enzyme by treatment with dissociating agents. 10. Cell-free and cell-wall-bound enzymes showed the same relationship when the change in fluidity effected by them on a solution of CM-cellulose was plotted against the corresponding increase in reducing sugars, suggesting that the enzymes were the same. 11. It is possible that R. albus cellulase exists as an aggregate of low-molecular-weight cellulase components on the bacterial cell wall and in solution under certain conditions.

Carboxymethylcellulose Sodium↗
PubMed1982

Short-term regulation of hydroxymethylglutaryl coenzyme A reductase by reversible phosphorylation: modulation of reductase phosphatase in rat hepatocytes.

Hydroxymethylglutaryl CoA reductase catalyzes the limiting step in cholesterol synthesis in liver and other tissues. Beginning in 1973 studies with subcellular systems established that microsomal reductase is inactivated with ATP(Mg) and reductase kinase, and restored to full activity with phospho-protein phosphatase. By contrast reductase kinase is inactivated with phosphatase and reactivated with a second protein kinase (reductase kinase kinase). This bicyclic system has now been confirmed in terms of homogeneous enzyme components and by direct reversible phosphorylation with [gamma 32P]ATP in several laboratories. Short-term endocrine control of reductase and reductase kinase has been demonstrated in intact rat hepatocytes. Preincubation of cells with glucagon brought about a fall in the expressed activity of reductase and a rise in reductase kinase consistent with net phosphorylation of both enzymes. Total reductase levels were also severely depressed after glucagon. Addition of insulin to suspensions of hepatocytes had the reverse effect on expressed activity of reductase (elevated) and reductase kinase (depressed). Insulin also prevented the decay in total reductase activity. Since both protein kinases identified in this system are cAMP-insensitive, it was possible that hormonal signaling is mediated through the protein phosphatase that acts on both reductase kinase and reductase. In recent studies we have shown that the rate of activation of endogenous reductase in hepatocyte extracts (microsomes plus cytosol) is responsive to hormonal modulation. Pretreatment of hepatocytes with insulin increases apparent reductase phosphatase activity in extracts while glucagon diminishes the rate of reductase activation. HMG CoA is converted to mevalonate by the reductase enzyme. In hepatocytes mevalonate is rapidly converted to cholesterol and to a variety of isoprene derivatives. Expressed reductase activity falls precipitously when hepatocytes are incubated with mevalonate (added in the form of mevalono-lactone). As in the case with glucagon pretreatment reductase phosphatase is rapidly diminished. (Mevalonate itself is not inhibitory to reductase or reductase phosphatase activity in subcellular systems.) It is probable that a product of mevalonate metabolism generated in intact cells may act as a reductase phosphatase inhibitor. Among these added inorganic pyrophosphate inhibited reductase phosphatase at low concentrations.

Animals↗
PubMed1982

Influence of plant phenolic acids on growth and cellulolytic activity of rumen bacteria.

Isolated rumen bacteria were examined for growth and, where appropriate, for their ability to degrade cellulose in the presence of the hydroxycinnamic acids trans-p-coumaric acid and trans-ferulic acid and the hydroxybenzoic acids vanillic acid and 4-hydroxybenzoic acid. Ferulic and p-coumaric acids proved to be the most toxic of the acids examined and suppressed the growth of the cellulolytic strains Ruminococcus albus, Ruminococcus flavefaciens, and Bacteroides succinogenes when included in a simple sugars medium at concentrations of >5 mM. The extent of cellulose digestion by R. flavefaciens and B. succinogenes but not R. albus was also substantially reduced. Examination of rumen fluid from sheep maintained on dried grass containing 0.51% phenolic acids showed the presence of phloretic acid (0.1 mM) and 3-methoxyphloretic acid (trace) produced by hydrogenation of the 2-propenoic side chain of p-coumaric and ferulic acids, respectively. The parent acids were found in trace amounts only, although they represented the major phenolic acids ingested. Phloretic and 3-methoxyphloretic acids proved to be considerably less toxic than their parent acids. All of the cellulolytic strains (and Streptococcus bovis) showed at least a limited ability to hydrogenate hydroxycinnamic acids, with Ruminococcus spp. proving the most effective. No further modification of hydroxycinnamic acids was produced by the single strains of bacteria examined. However, a considerable shortfall in the recovery of added phenolic acids was noted in media inoculated with rumen fluid. It is suggested that hydrogenation may serve to protect cellulolytic strains from hydroxycinnamic acids.

Journal Article↗
PubMed1981

Growth and rumen function of gnotobiotic lambs fed on starchy diets.

Four gnotobiotic lambs were fed, after weaning, on a pelleted concentrate diet containing 90% barley. They were inoculated with a defined flora of 11 species of bacteria designed to reproduce the feed digestion of the conventional lamb. Cultural assessments of the growth of the bacteria and analysis of rumen fermentation products showed that rumen function was near normal, and the lambs grew steadily. However, at an age of about 115 to 130 d, rumen function seemed to fail in three of the lambs and they rapidly lost weight and died. The fourth lamb was removed from isolation at about the same time, when there was some slight indication of loss of rumen function. Kept among conventional lambs it then continued to gain weight as did the conventional animals, but its rumen flora changed. The main species in the defined flora remained as major components of the rumen population after the lamb had been exposed to other animals, but in addition a mixed population of other bacteria, like that in conventional animals, quickly developed. Other experiments showed that gnotobiotic lambs could be routinely weaned on to a concentrate feed containing 45% barley and 50% grass and that, when the lambs were inoculated with a defined flora similar to that given to the lambs in the first experiment, an adequate rumen fermentation could be established and the animals would grow. On changing to an all-grass feed, however, rumen function declined and the lambs ceased to grow. A change back to the barley/grass diet restored rumen fermentation and lamb growth, but whereas amylolytic bacteria were present in normal numbers, cellulolytic bacteria were absent and there appeared to be little, if any, fibre digestion. However, the rumen function again appeared to fail at about the same age of lamb as in the first experiment.

Animal Feed↗
PubMed1981

Selective isolation and characteristics of Bacteriodes succinogenes from the rumen of a cow.

Eleven isolates of Bacteriodes succinogenes were obtained from the rumen of a cow by an enrichment method with dewaxed cotton fibers as the selective substrate. All of the isolates degraded cotton fibers, but none formed clear zones in cellulose agar, having only a limited ability to degrade the type of cellulose powder used. One isolate, BL2, was studied in greater detail and was found to accumulate a glycogen-like polysaccharide when excess (0.5 to 1.0%) soluble carbohydrate was supplied in the nutrient medium. Although the pattern of growth and polysaccharide accumulation by strain BL2 changed during maintenance of the organism in the laboratory, the maximum amount of carbohydrate found in the cells was constant, at around 74% of the cell dry weight. The findings are discussed in relation to the methods of assessing the role of B. succinogenes in the rumen fermentation.

Animals↗
PubMed1979

Specific and nonspecific cytotoxicity of leukocytes from human renal allograft recipients against donor fibroblasts.

A microcytotoxicity assay was used to search for cell-mediated cytotoxicity, serum-blocking factors, and antibody-dependent cell-mediated cytotoxicity (ADCC) against fibroblasts of donor origin in 19 human renal allograft recipients. Peripheral mononuclear cells (PMC) were frequently cytotoxic when they were obtained within 6 days prior to rejection, following sustained rejections, and within 1 month after removal of rejected grafts, but the toxicity was usually nonspecific. Recipient PMC were noncytotoxic when they were obtained within 6 days after the onset of an acute reversed rejection and during quiescent intervals when there was no evidence of rejection within 6 days before or after the assay. These data suggest that, regardless of its lack of specificity, there is some relationship between cytotoxicity of recipient PMC and allograft rejection. In none of 69 post-transplant serum samples was ADCC against donor target cells detected with the microcytotoxicity assay.

Cytotoxicity Tests, Immunologic↗
PubMed1979

Maceration of Clover and Grass Leaves by Lachnospira multiparus.

A strain of Lachnospira multiparus, a pectin-hydrolyzing bacterium from the rumen, was incubated in nutrient media in the presence of surface-disinfected clover leaflets. When the culture flasks containing the leaflets together with Lachnospira were shaken after overnight incubation, extensive maceration of the leaflets was seen, although uninoculated control leaflets remained intact during a similar treatment. Examination of inoculated leaflets by transmission electron microscopy showed extensive invasion of intercellular areas of the mesophyll tissue but only minor invasion of vascular tissue. Cutting the leaves before incubation greatly increased the ability of L. multiparus to colonize and macerate the leaflets. Similar experiments with grass leaves are also described, and the possible role of maceration in the digestion of plant material in the rumen is discussed. Although Lachnospira stains gram variable and often gram negative, the ultrastructure of the cell wall was that of a gram-positive bacterium.

Journal Article↗
PubMed1978

Patterns of aspirin use among American youth.

The use of aspirin was ascertained in several population cohorts. In the college age samples, aspirin had been used in the previous 48-hour period by one of every 4 students. Females reported a higher rate (33.2%) than males (16.9%). In the 4th to 12th grade samples, aspirin use was higher than other drugs with 39.6% of the 3,306 students sampled indicating that they had ingested aspirin during the previous week. In a 9--11 year-old category, 25% of these 4th to 6th graders used aspirin without parental knowledge. The relationship of aspirin use and drug education is discussed.

Adolescent↗
PubMed1977

Effect of fat supplementation on voluntary food intake and rumen metabolism in sheep.

1. In an experiment in which a high-fat supplement was given in the dry form to lambs offered dried grass ad lib., both the voluntary intake and digestibility of the dried grass were reduced. When the high-fat supplement was given in a liquid suspension so that the rumen was by-passed, the voluntary intake and digestibility of the dried grass were not significantly altered. 2. The effect of injecting an emulsion of tallow into the rumen of sheep on rumen metabolism was studied in another experiment. Increasing the fat supplementation lowered the rate of digestion of both dried grass and cotton thread, lowered markedly the concentration of rumen ammonia, and raised the proportion of propionic acid in the rumen.

Animal Nutritional Physiological Phenomena↗
PubMed1977

Factors affecting the cellulolytic activity of rumen contents.

The cellulolytic activity of rumen contents was assayed by measuring losses in weight and tensile strength of cotton yarn incubated in rumen contents in the presence of dietary additives (barley, tallow) and at different pH values. The addition of barley depressed cellulolysis and the titer of filter paper-degrading bacteria only if the pH was allowed to fall. Lowering the pH from 7.0 to 6.0 by addition of HCl almost completely inhibited attack of cotton and greatly reduced the titer of filter paper-degrading bacteria. The layering of tallow on cotton inhibited attack of cotton but did not decrease the titer of filter paper-degrading bacteria. The results are discussed with special reference to the importance of the study of cellulosic substrates, which require a known enzyme or mixture of enzymes for attack.

Journal Article↗
PubMed1976

Defined bacterial populations in the rumens of gnotobiotic lambs.

Five gnotobiotic lambs were fed on sterile diets until they were killed at 13 to 21 weeks of age. They were dosed orally with different combinations of 11 species of rumen bacteria. The biochemical reactions of each of the bacteria inoculated had been determined in pure culture in vitro, and they were chosen to perform the main reactions known to be associated with digestion in the normal mature rumen. Two of the bacteria could not be reisolated, but the remainder had established readily in the rumen, forming stable, mixed, defined populations. The total numbers of bacteria in the rumen, and the viable counts of most of the individual species were comparable to those of normal sheep. The concentration of volatile fatty acids was lower, however, and in four of the lambs there was a higher proportion of butyric acid and a lower proportion of propionic acid than in normal sheep. Cellulolytic, ureolytic, and methanogenic activities appeared to be taking place and lactate-utilizing bacteria appeared to reverse the accumulation of lactate which resulted from the activity of lactate-producing bacteria. Some of the bacteria also established at high levels in the caecum.

Acetates↗