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Biomedical subjects

W C Russell

Publications and source records attributed to W C Russell.

At least 55 records · Page 3Linked to original sources

Adherence of Ureaplasma urealyticum to human epithelial cells.

Adherence of Ureaplasma urealyticum cells to eukaryotic cell monolayers was quantified using the Bertholet assay to monitor ammonia produced from urea by ureaplasma urease. Adherence was abolished by pre-treatment of ureaplasmas with HeLa cell extracts and inhibited to varying degrees by pretreatment of the ureaplasmas with N-acetylneuraminic acid, specific antisera and monoclonal antibodies. The data suggest the presence of several ureaplasma adhesins, some of which are species- or serotype-specific and some of which are proteinaceous and antigenic. The serotype-8-specific 96 kDa surface-expressed antigen may be one adhesin. Pre-treatment of HeLa cell monolayers with neuraminidase significantly reduced ureaplasma adherence and, using a novel 'immunoblot adherence assay', ureaplasmas were shown to bind to a number of HeLa cell components, three of which appear to terminate in sialic acid.

Adhesins, Bacterial↗

Recognition of non-B DNA structures by cellular proteins.

By utilising a gel retardation assay, cytoplasmic extracts of Hela cells have been shown to contain a number of polypeptides which bind preferentially to non-B forms of DNA. These polypeptides vary in apparent molecular weights and are present as minor components of the total extract. Similar properties were demonstrated by purified preparations of eukaryotic topoisomerase I and by a DNA binding subunit of the transcription factor NFkB. These results suggest that non-B DNA binding proteins may represent families of proteins which recognise more open forms of DNA as a cognate parameter to their cellular functions.

Chromatography, Affinity↗

The effect of incisional infiltration of bupivacaine upon pain and respiratory function following open cholecystectomy.

A controlled, prospective, double-blind trial of wound infiltration with bupivacaine in elective open cholecystectomy was performed to determine if this was an effective method of pain relief and reduced respiratory complications. Additionally, dextran was added to the bupivacaine in an attempt to prolong the effect. The solutions used were, bupivacaine alone 0.25% (n = 14), bupivacaine 0.25% with dextran 70 (n = 16) and saline (n = 16) as a control. To determine the effect of each solution, the subjects were assessed for pain perception and respiratory function before and after surgery. Pain was assessed using a visual analogue scale and narcotic usage, and respiratory function was assessed by spirometry, chest X-rays and arterial blood gases. The study did not demonstrate any objective improvement in either pain relief or respiratory function. This may reflect inadequate infiltration by the surgeons in the study or that infiltration should have been performed prior to incision.

Adult↗

Hydrolysis of urea by Ureaplasma urealyticum generates a transmembrane potential with resultant ATP synthesis.

When urea is added to Ureaplasma urealyticum, it is hydrolysed internally by a cytosolic urease. Under our measuring conditions, and at an external pH of 6.0, urea hydrolysis caused an ammonia chemical potential equivalent to almost 80 mV and, simultaneously, an increase in proton electrochemical potential (delta p) of about 24 mV with resultant de novo ATP synthesis. Inhibition of the urease with the potent inhibitor flurofamide abolished both the chemical potential and the increase of delta p such that ATP synthesis was reduced to approximately 5% of normally obtained levels. Uncouplers of electrochemical gradients had little or no effect on these systems. The electrochemical parameters and ATP synthesis were measured similarly at three other external pH values. Any change in delta p was primarily via membrane potential (delta psi), and the level of de novo ATP synthesis was related to the increase in delta p generated upon addition of urea and more closely to the ammonia chemical potential. Although the organisms lack an effective mechanism for internal pH homeostasis, they maintained a constant delta pH. The data reported are consistent with, and give evidence for, the direct involvement of a chemiosmotic mechanism in the generation of around 95% of the ATP by this organism. Furthermore, the data suggest that the ATP-generating system is coupled to urea hydrolysis by the cytosolic urease via an ammonia chemical potential.

Adenosine Triphosphate↗

Urea-hydrolysis-dependent citrulline synthesis by Ureaplasma urealyticum.

Some of the ammonia produced by hydrolysis of urea by Ureaplasma urealyticum is channelled into an anabolic pathway with resultant 'de novo' synthesis of citrulline. The organism appears to possess ornithine carbamoyltransferase and carbamoyl phosphate synthetase or some modified form of these enzymes.

Ammonia↗

Western blotting analysis in patients with MS using human brain vessels as antigen.

Serum samples from patients suffering from multiple sclerosis, other neurological diseases and normal controls were screened by "western blotting" for antibody directed against proteins of human brain vessels purified from a post mortem brain. A small number of sera contained autoantibodies against some of the proteins of the brain vessels, particularly in patients suffering from MS, epilepsy and migraine. The significance of these results is discussed.

Antibody Specificity↗

Characterization of the immunoglobulin A protease of Ureaplasma urealyticum.

Ureaplasma urealyticum strains of all serotypes express a specific human immunoglobulin A1 protease that cleaves immunoglobulin A1 to produce intact Fab and Fc fragments. The use of a variety of inhibitors suggests that the enzyme is a serine protease. N-terminal sequencing of the Fc digestion product showed that the enzyme cleaves between the proline and threonine residues 235 and 236 in the hinge region of the heavy chain of immunoglobulin A1.

Amino Acid Sequence↗

Estimation of genetic parameters for wool fiber diameter measures.

Genetic and phenotypic correlations and heritability estimates of side, britch, and core diameters; side and britch CV; side and britch diameter difference; and clean fleece weight were investigated using 385 western white-faced ewes produced by 50 sires and maintained at two locations on a selection study. Data were analyzed using analysis of variance procedures, and effects in the final model included breed of sire-selection line combination, sire within breed-selection line, and location. Heritabilities were estimated by paternal half-sib analysis. Sires within breed-selection line represented a significant source of variation for all traits studied. Location had a significant effect on side diameter, side and britch diameter difference, and clean fleece weight. Age of ewe only affected clean fleece weight. Phenotypic and genetic correlations among side, britch, and core diameter measures were high and positive. Phenotypic correlations ranged from .68 to .75 and genetic correlations ranged from .74 to .89. The genetic correlations between side and britch diameter difference and side diameter or core diameter were small (-.16 and .28, respectively). However, there was a stronger genetic correlation between side and britch diameter difference and britch diameter (.55). Heritability of the difference between side and britch diameter was high (.46 +/- .16) and similar to heritability estimates reported for other wool traits. Results of this study indicate that relatively rapid genetic progress through selection for fiber diameter should be possible. In addition, increased uniformity in fiber diameter should be possible through selection for either side and britch diameter difference or side or britch CV.

Age Factors↗

Proteolysis is a key process in virus replication.

Proteases were amongst the first enzymes to be isolated and crystallized, and the discovery of their existence in viruses and the realization of the vital role they play has given a new lease of life to one of the oldest topics in biochemistry. Already we have seen the discovery of new variations on well-studied reaction mechanisms and there is the promise of others to come that may be totally novel. When this is allied to the prospect of developing the knowledge which is beginning to accrue into a much needed antiviral therapy, it is clear that viral proteases and their key role in viral replication will be an increasing focus of attention for some time to come.

Animals↗

Cross-reacting antigens between Mycoplasma ovipneumoniae and other species of mycoplasma of animal origin, shown by ELISA and immunoblotting with reference antisera.

Using enzyme-linked immunosorbent assays with Mycoplasma ovipneumoniae as antigen, the cross-reactivity of antigens between this species and 22 other mycoplasma species was examined using reference polyclonal antisera. Significant cross-reactivity with M. ovipneumoniae was demonstrated by five species, only, viz. M. bovoculi, M. dispar, M. flocculare, M. hyopneumoniae and M. hyorhinis. Using one-dimensional SDS-PAGE and immunoblotting techniques with homologous and heterologous antisera, cross-reacting antigens of M. dispar, M. flocculare, M. hyopneumoniae and M. ovipneumoniae were further investigated. Cross-reacting antigens with apparent molecular weights of 64, 44 and 32 kDa were common to all and a 184 kDa cross-reacting antigen occurred in all except M. ovipneumoniae. Further cross-reacting antigens (one-way and two-way) between two of the four species are reported. Four monoclonal antibodies against different antigens of M. ovipneumoniae did not recognise any antigen in the other three species examined.

Animals↗

Palmitoylated proteins in Ureaplasma urealyticum.

After incubation of Ureaplasma urealyticum serotype 8 in the presence of 3H-labeled palmitic acid, about 25 acylated proteins were detected by electrophoresis and fluorography. Of these, at least six were shown to be antigenic by immunoprecipitation of solubilized palmitate-labeled cells with a homologous polyclonal serum. These six included the serotype 8-specific surface-expressed 96-kDa antigen. After phase partition of palmitate-labeled cells with Triton X-114, all but six acylated proteins partitioned entirely into the detergent phase. The others, including the 96-kDa antigen, partitioned preferentially into the detergent phase and were apparently amphipathic. These results are consistent with the acylated proteins being mainly membrane associated.

Acylation↗

Isolation and detection of urease genes in Ureaplasma urealyticum.

Urease from ureaplasmas was purified by immunoaffinity chromatography, and the N-terminal amino acid sequence was determined for two of the three subunits. These sequences were used to design primers for a polymerase chain reaction (PCR) that amplified most of the gene coding for one of the subunits. By using a novel "PCR walking" technique, we synthesized almost the complete locus on two overlapping PCR products. We present here a partial nucleotide sequence of the urease locus from Ureaplasma urealyticum (serotype 8), which agrees with our N-terminal amino acid data but differs slightly from the sequence previously reported (A. Blanchard, Mol. Microbiol. 4:669-676, 1990). Also described are PCR primers, intended for diagnostic use, that amplify a sequence from all Ureaplasma strains tested but not from any other mycoplasmas or urease-positive bacteria.

Amino Acid Sequence↗

Effect of spray-chilling on quality of beef from lean and fatter carcasses.

Carcasses from five trim cows and five choice steers were used to study the effects of spray-chilling on cooler shrink, chill rate, purge loss from vacuum-packaged cuts, cook loss, shear values and bacterial growth. Spray-chilling reduced cooler shrink but had no effect on chill rate, purge loss from vacuum-packaged cuts, cook loss or shear values. Aerobes, facultative anaerobes, aerobic psychrotrophs, facultative anaerobic psychrotrophs and lactic acid bacteria all tended to be higher on rounds from spray-chilled sides. Leaner (and lighter) cow carcasses chilled faster and had lost a higher percentage of their weight at 24 h than fatter and heavier steer carcasses. The leaner carcasses had higher bacterial counts initially and throughout storage. This difference may have been due to differences in the level of initial contamination during dressing and not due to the carcasses' leanness. Purge-weight loss for each carcass increased and cooking weight loss decreased with increased storage times, making the total weight loss from meat aged 5 vs 10 wk similar.

Animals↗

Polypeptide and antigenic variability among strains of Mycoplasma ovipneumoniae demonstrated by SDS-PAGE and immunoblotting.

Comparison of the polypeptide patterns of 22 isolates of M. ovipneumoniae by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a marked degree of heterogeneity with only limited groupings identifiable. Of the 50 major polypeptides identified in one strain (956/2), 35 were shown to be antigenic using immunoblotting with a homologous polyclonal serum. Radioimmune precipitation of 125I-surface-labelled proteins and phase partition using Triton X-114 detergent indicated that these were membrane associated. Cross-reactivity between the isolates was examined by immunoblotting using one polyclonal serum and four monoclonal antibodies (MAbs), all raised against strain 956/2. The polyclonal serum revealed considerable antigenic heterogeneity, but at least nine major antigens were conserved across all isolates. Two MAbs cross-reacted with all 22 strains, but the other two MAbs allowed some differentiation of the strains. One (MO/3) divided the isolates into groups of 16 and 6 based on the presence of absence of a 26-kDa antigen. All strains isolated from sheep with pulmonary adenomatosis fell into the smaller group and did not possess the 26-kDa antigen.

Animals↗

The humoral immune response of lambs experimentally infected with Mycoplasma ovipneumoniae.

Using sera from lambs experimentally infected with Mycoplasma ovipneumoniae and Pasteurella haemolytica, the development of a good humoral immune response to M. ovipneumoniae was detected by ELISA. The antibody titres peaked 41 days post-infection and good antibody titres were maintained over the 16-week experimental period. Immunoblotting revealed that antibodies to specific antigens appeared in the sera in a sequential manner, some being seen shortly after infection and others developing only after a substantial time lag. Antibodies were raised against almost all the major antigens detected in one laboratory strain (956/2) and against all antigens previously shown to be conserved in 22 Scottish field isolates of M. ovipneumoniae.

Animals↗

Effect of plane of nutrition and age on carcass maturity of sheep.

One hundred twenty ewe and wether lambs were fed high (76% TDN, 12.3% crude protein) or low (70.6% TDN, 16.1% crude protein) energy diets and slaughtered at average ages of 261, 356 or 469 d. High planes of nutrition for lambs at 356 or 469 d of age resulted in heavier carcasses, thicker fat depth and higher flank streaking scores compared to lambs fed low planes of nutrition. Rib bone, break joint and overall maturity scores were not influenced (P greater than .05) by diet. Metacarpal and metatarsal bones from lambs fed high planes of nutrition were slightly longer and heavier and required more force to break (P less than .05) compared with bones from lambs fed low planes of nutrition. The majority of the ewes fed high or low planes of nutrition had fused growth plates and possessed spool joints by 469 d; none of the wethers had spool joints. An additional 27 wethers were slaughtered at an average of 480, 578 or 662 d. Rib bone, break joint and overall maturity scores were not different (P greater than .05) for animals fed the high or low energy diets, but bone weight and shaft break force values were higher for those fed high energy diets. At 662 d of age, only 20% of the wethers on each dietary regimen possessed fused growth plates. Overall, plane of nutrition in market-weight lambs and yearlings had very little influence on the presence or absence of a break joint or on other bone maturity scores.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗