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Biomedical subjects

C S Stewart

Publications and source records attributed to C S Stewart.

At least 37 records · Page 2Linked to original sources

Interactions between rumen bacterial strains during the degradation and utilization of the monosaccharides of barley straw cell-walls.

Pure cultures and pair-combinations of strains representative of the rumen cellulolytic species Ruminococcus flavefaciens, Fibrobacter succinogenes and Butyrivibrio fibrisolvens were grown on cell-wall materials from barley straw. Of the pure cultures, R. flavefaciens solubilized straw most rapidly. The presence of B. fibrisolvens, which was unable to degrade straw extensively in pure culture, increased the solubilization of dry matter by R. flavefaciens and the solubilization of cell-wall carbohydrates by both R. flavefaciens and F. succinogenes. During fermentation, both R. flavefaciens and F. succinogenes released bound glucose and free and bound arabinose and xylose into solution. The accumulation of these sugars, especially arabinose and xylose, was greatly reduced in co-cultures containing B. fibrisolvens, suggesting that significant interspecies cross feeding of the products of hemicellulose hydrolysis (particularly soluble bound xylose released by F. succinogenes) occurs during straw degradation by mixed cultures containing this species.

Animal Feed↗

Inhibition of enterotoxigenic Escherichia coli by the microflora of the porcine ileum, in an in vitro semicontinuous culture system.

An. in vitro fermentation system capable of maintaining at least part of the microbial population of the contents of the porcine ileum has been developed. The system was tested over the pH range 6.0-8.5, anaerobically and at dissolved oxygen concentrations within the ranges detected in the ileum of piglets at weaning (50 and 100 mumol l-1). The results demonstrated that changes in pH and dissolved oxygen within these ranges had relatively little effect on the total numbers of aerobic and anaerobic bacteria. Lactic acid bacteria, enumerated anaerobically, showed changes in viable counts in response to pH changes but were apparently unaffected by changes in dissolved oxygen, although the proportion of aerotolerant species within this group was increased at high concentrations of dissolved oxygen. The relative proportions of lactobacilli and coliforms were similar to those reported in vivo when dissolved oxygen was present at a concentration of 50 mumol l-1. Under these conditions the simulated population was consistently found to resist colonization by an enterotoxigenic Escherichia coli, of a serotype known to cause weaning scours in the piglet.

Animals↗

Analysis of bacterial phospholipid markers and plant monosaccharides during forage degradation by Ruminococcus flavefaciens and Fibrobacter succinogenes in co-culture.

Marker components of the phospholipids of Ruminococcus flavefaciens and Fibrobacter succinogenes were identified for studies on the degradation of forage by these bacteria growing in mixed culture. The principal fatty acid methyl esters and dimethyl acetals detected varied between strains and were influenced by the addition of a mixture of higher volatile fatty acids and vitamins to the medium, but these effects were small compared to the differences between the species. When two strains of R. flavefaciens were grown on a mixture of clover and ryegrass, and on barley straw in the presence or absence of two strains of F. succinogenes, the solubilization of plant material tended to be lowered by the presence of F. succinogenes. R. flavefaciens was the predominant bacterium among colonies recovered from roll tubes, and the phospholipids were primarily those of R. flavefaciens. Analysis of the culture supernatant liquids showed that F. succinogenes produced greater amounts of free and bound xylose from both clover and straw than did R. flavefaciens. With both forages, cultures containing the two species produced more soluble free arabinose, and less soluble-bound arabinose, than either species grown alone.

Animal Feed↗

Binding and degradation of lectins by components of rumen liquor.

The binding of 15 different plant lectins to feed particles and microbes in rumen liquor, and their degradation were studied in vitro. The rate of degradation assessed from the label released when radioactive iodine-labelled lectins were incubated with rumen liquor conflicted with the rates calculated from measurements of the survival of the antigenic structure (immuno-rocket electrophoresis) or the biological function (haemagglutination) of the lectins. Thus solubilization of the radioactive label indicated that Concanavalin A (Con A), but not the soyabean agglutinin, SBA, or kidney bean phytohaemagglutinin, PHA-E3L, was stable to rumen proteolysis. In contrast, both SBA and PHA-E3L were shown by immuno-rocket electrophoresis or haemagglutination tests to be highly resistant to breakdown, while the degradation of Con A proceeded at a constant slow rate under the same conditions. This was in accord with the previously established general stability of lectins in the gut of single-stomach animals. Of the 15 lectins, SBA, favin (Vicia faba lectin) and Con A were bound by hay and the particle fraction of rumen liquor. This was, in part, specific and reversible in the presence of appropriate sugars. Most pure bacterial strains preferentially bound lectins with specificity for glucose/mannose (favin and Con A), while rumen fungi reacted with SBA. The level of binding was low with other lectins. However, inter-strain differences of lectin-binding were found in Selenomonas ruminantium and Ruminococcus flavefaciens. Clearly, as some lectins were not fully degraded in the rumen, they could be expected to depress the utilization of the diet not only in single-stomach animals but, possibly, also in ruminants.

Animals↗

Evidence for heterogeneity in facioscapulohumeral muscular dystrophy (FSHD).

Facioscapulohumeral muscular dystrophy (FSHD) is a slowly progressive primary disease of muscle which is usually inherited as an autosomal dominant disorder. FSHD has been localized to the long arm of chromosome 4, specifically to the 4q3.5-qter region. Initially published linkage studies showed no evidence for heterogeneity in FSHD. In the present study we have examined individuals in seven FSHD families. Two-point lod scores show significant evidence for linkage for D4S163 (lod score 3.04 at recombination fraction .21) and D4S139 (lod score 3.84 at recombination fraction .20). D4S171 also gave a positive score (lod score 2.56 at recombination fraction .24). Significant evidence for heterogeneity was found for each of the three markers. Multipoint linkage analysis in this region resulted in a peak multipoint lod score of 6.47. The multipoint analysis supported the two-point studies with odds of 20:1 showing linkage and heterogeneity over linkage and homogeneity. Five of the seven families gave a posterior probability of > 95% of being of the linked type, while two families appeared unlinked to this region of 4q (P < .01%). Individuals in the two unlinked families met the clinical criteria for the diagnosis of FSHD, including facial weakness, clavicular flattening, scapula winging, proximal muscle weakness, and myopathic changes on muscle biopsies without inflammatory or mitochondrial pathology. This study demonstrates genetic heterogeneity in FSHD and has important implications for both genetic counseling and the elucidation of the etiology of FSHD.

Chi-Square Distribution↗

The inhibition of fungal cellulolysis by cell-free preparations from ruminococci.

The degradation of filter paper by the anaerobic fungus Neocallimastix frontalis strain RE1 was reduced by the addition of cell-free supernates from cultures of Ruminococcus albus strain J6 and R. flavefaciens strains 17 and 007. Fungal uptake of, and growth on, glucose was not affected. After gel permeation and anion exchange chromatography, inhibitory activity towards fungal cellulolysis was recovered in a fraction from strain 17 that contained at least five negatively charged polypeptide components, molecular mass 45-68 kDa, on SDS-PAGE.

Animals↗

Linkage studies in facioscapulohumeral muscular dystrophy (FSHD).

Facioscapulohumeral muscular dystrophy (FSHD) has been localized to the 4q35-qter region of chromosome 4. Linkage analyses of two polymorphic markers from the region, D4S139 and D4S163, have been carried out using four large multigenerational FSHD families. The results indicate that both markers are closely linked to FSHD, with D4S139 being the closest proximal marker to FSHD.

Chromosome Mapping↗

Parathyroid hormone-related protein of malignancy: immunohistochemical and biochemical studies in normocalcaemic and hypercalcaemic patients with cancer.

Immunohistochemical staining for parathyroid hormone-related protein was performed in 27 tumours from 19 normocalcaemic and eight hypercalcaemic patients with cancer. All the tumours from hypercalcaemic patients stained positively for the protein, as did 17 tumours from normocalcaemic patients. Only hypercalcaemic patients had biochemical evidence of increased bone resorption and abnormalities of renal tubular reabsorption of calcium and phosphate, consistent with the presence of parathyroid hormone-related protein. While tumour mass was higher in hypercalcaemic patients, only one of the initially normocalcaemic patients with positively staining tumours subsequently went on to develop hypercalcaemia and more advanced disease. These data confirm the importance of parathyroid hormone-related protein as a mediator of humoral hypercalcaemia in patients with solid tumours and suggest that low tumour mass may be one reason why serum calcium values are not increased in all patients with tumours containing parathyroid hormone-related protein. None the less normocalcaemia, despite tumour progression in patients whose tumours stained positively for parathyroid hormone-related protein, suggests that other factors may also be important, such as differences in the rate of secretion of the protein by different tumours, or the production of different forms of parathyroid hormone-related protein with varying biological effects.

Humans↗

Prevention of fungal colonization and digestion of cellulose by the addition of methylcellulose.

When the attachment of cellulolytic rumen fungi to cellulose is blocked by the addition of methylcellulose, cellulose digestion is entirely inhibited. Even after these fungi have colonized and penetrated the cellulosic fibers of filter paper, the addition of methylcellulose effectively halts cellulose digestion. This effect of methylcellulose is accompanied by the complete inhibition of fungal attachment to cellulose fibers; the addition of methylcellulose does not affect the growth of these organisms on soluble substrates. We conclude that fungal cellulose digestion, like bacterial cellulose digestion, requires the spatial juxtaposition of the cellulolytic organism and its insoluble substrate. The simultaneous inhibition of both attachment and digestion by the same inhibitor suggests that these two processes are functionally linked in the fungi.

Animals↗

The properties of forms of Ruminococcus flavefaciens which differ in their ability to degrade cotton cellulose.

Two mutant strains of Ruminococcus flavefaciens strain 007 that differ in their ability to hydrolyse cotton fibres have been shown also to differ in their cell-surface topology, in that the cotton degrading form possessed larger and more protuberant cell surface structures. The strains had similar CMCase, cellobiosidase and beta-glucosidase activities. The results indicate the importance of cell-surface properties in cotton degradation by R. flavefaciens.

Animals↗

Effect of coumarin on glucose uptake by anaerobic rumen fungi in the presence and absence of Methanobrevibacter smithii.

The effect of coumarin (1,2 benzopyrone) on glucose utilisation by the anaerobic rumen fungi Neocallimastix frontalis and N. patriciarum has been compared with the effect of p-coumaric acid. Both compounds largely inhibited glucose utilisation by N. patriciarum strain Cx when present in the medium at a concentration of 2.5 mM, and had a similar effect on N. frontalis strain RE1 at 5 mM. Although in earlier studies co-culturing rumen fungi with Methanobrevibacter smithii enhanced resistance to ionophores, no comparable protective effect of M. smithii was found in the present study.

Animals↗

Influence of hydrogen-consuming bacteria on cellulose degradation by anaerobic fungi.

The presence of methanogens Methanobacterium arboriphilus, Methanobacterium bryantii, or Methanobrevibacter smithii increased the level of cellulose fermentation by 5 to 10% in cultures of several genera of anaerobic fungi. When Neocallimastix sp. strain L2 was grown in coculture with methanogens the rate of cellulose fermentation also increased relative to that for pure cultures of the fungus. Methanogens caused a shift in the fermentation products to more acetate and less lactate, succinate, and ethanol. Formate transfer in cocultures of anaerobic fungi and M. smithii did not result in further stimulation of cellulolysis above the level caused by H2 transfer. When Selenomonas ruminatium was used as a H2-consuming organism in coculture with Neocallimastix sp. strain L2, both the rate and level of cellulolysis increased. The observed influence of the presence of methanogens is interpreted to indicate a shift of electrons from the formation of electron sink carbon products to H2 via reduced pyridine nucleotides, favoring the production of additional acetate and probably ATP. It is not known how S. ruminantium exerts its influence. It might result from a lowered production of electron sink products by the fungus, from consumption of electron sink products or H2 by S. ruminantium, or from competition for free sugars which in pure culture could exert an inhibiting effect on cellulolysis.

Anaerobiosis↗

Linkage of Charcot-Marie-Tooth neuropathy type 1a to chromosome 17.

Charcot-Marie-Tooth disease Type 1 (CMT) is an inherited neuropathy with known genetic heterogeneity, with at least one autosomal dominant form (CMT Type 1b) linked to the Duffy region of chromosome 1. Autosomal dominant families not demonstrating linkage to the Duffy blood group marker have been designated CMT Type 1a. We report linkage of six CMT Type 1a families to the chromosome 17 markers EW301 (D17S58) and pA10-41 (D17S71) with maximum LOD scores of zeta = 10.49 at theta (maximum recombination fraction) = 0.05 and zeta = 7.36 at theta = 0.06, respectively.

Charcot-Marie-Tooth Disease↗

Enhanced resistance of anaerobic rumen fungi to the ionophores monensin and lasalocid in the presence of methanogenic bacteria.

The presence of Methanobrevibacter smithii altered the susceptibility of the anaerobic fungi Neocallimastix frontalis and Piromonas communis to the carboxylic ionophores monensin and lasalocid. The ionophores depressed growth (measured by chitin accretion), the uptake of glucose and the production of H2, formate and acetate by the fungi growing axenically in semi-solid medium. In the presence of M. smithii, the sensitivity of the fungi to monensin and lasalocid was decreased. For example, the uptake of glucose by N. frontalis strain RE1 in the culture was reduced to 50% of the control value by monensin at 0.5 microgram/ml. In the presence of M. smithii strain PS, approximately three times as much monensin was needed to bring about the same effect. In similar tests, the sensitivity of strain RE1 to lasalocid was decreased about nine-fold in the presence of M. smithii. The effect was not observed if the methanogens were killed by autoclaving before inoculation. It is suggested that the enhanced resistance to ionophores in the presence of M. smithii is a consequence of changes in the energy metabolism of the fungi growing in co-culture.

Acetates↗

Degradation of barley straw, ryegrass, and alfalfa cell walls by Clostridium longisporum and Ruminococcus albus.

The recently isolated ruminal sporeforming cellulolytic anaerobe Clostridium longisporum B6405 was examined for its ability to degrade barley straw, nonlignified cell walls (mesophyll and epidermis) and lignified cell walls (fiber) of ryegrass, and alfalfa cell walls in comparison with strains of Ruminococcus albus. R. albus strains degraded 20 to 28% of the dry matter in barley straw in 10 days, while the clostridium degraded less than 2%. A combined inoculum of R. albus SY3 and strain B6405 was no more efficient than SY3 alone, and the presence of Methanobacterium smithii PS did not increase the amount of dry matter degraded. In contrast, with alfalfa cell walls as the substrate, the clostridium was twice as active (28% weight loss) as R. albus SY3 (15%). The percentages of dry matter degraded from ryegrass cell walls of mesophyll, epidermis, and fiber for the clostridium were 50, 47, and 32%, respectively, and for R. albus SY3 they were 77, 73, and 63%, respectively. Analyses of the predominant neutral sugars (arabinose, xylose, and glucose) in the plant residues after bacterial attack were consistent with the values for dry matter weight loss. Measurements of the amount of carbon appearing in the fermentation products indicated that R. albus SY3 degraded ryegrass mesophyll cell walls most rapidly, with epidermis and fiber cell walls being degraded at similar rates. Strain B6405 attacked alfalfa cell walls at a rate greater than that of any of the ryegrass substrates. These results indicate an unexpected degree of substrate specificity in the ability of C. longisporum to degrade plant cell wall material.

Anaerobiosis↗

Automatic analysis of the electromyographic interference pattern using the turns: amplitude ratio.

This study was performed to compare different techniques of analyzing the electromyographic interference pattern (IP). Recordings were made from the biceps muscle with a concentric needle electrode at different sites and at different constant levels of voluntary contraction. The number of turns per second (NT), the mean amplitude change between successive turns (MA) and NT:MA ratio were determined for epochs of 1 sec duration. Normal limits of individual epoch NT:MA ratios and the mean value of NT:MA ratio obtained from all epochs in each muscle were determined. The mean NT:MA ratio was less in normal males than in females. IP recordings were made in the biceps muscle of 69 patients with neuropathy and 54 patients with myopathy, though this muscle was not necessarily affected by the disease in all patients. The IP was abnormal by visual inspection in 82% of patients compared to 61% based on NT:MA ratio and 74% using a technique that automatically quantitates some features of the IP that are assessed subjectively by an electromyographer. All techniques demonstrated IP abnormalities in more than 80% of the muscles that were moderately to severely weak. Though measuring the NT:MA ratio without monitoring the force of contraction is not as sensitive as other IP analysis techniques, it may be useful in quantitating abnormalities when other techniques are not available.

Adult↗