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Biomedical subjects

R G Taylor

Publications and source records attributed to R G Taylor.

At least 19 recordsLinked to original sources
PubMed2005

Calpain 3/p94 is not involved in postmortem proteolysis.

Studies on the correlation between expression and/or autolysis of calpain and postmortem proteolysis in muscle have provided conflicting evidence regarding the possible role of calpain 3 in postmortem tenderization of meat. Thus, the objective of this research was to test the effect of postmortem storage on proteolysis and structural changes in muscle from normal and calpain 3 knockout mice. Knockout mice (n = 6) were sacrificed along with control mice (n = 6). Hind limbs were removed and stored at 4 degrees C; muscles were dissected at 0, 1, and 3 d postmortem and subsequently analyzed individually for degradation of desmin. Pooled samples for each storage time and mouse type were analyzed for degradation of nebulin, dystrophin, vinculin, and troponin-T. In a separate experiment, hind-limb muscles from knockout (n = 4) and control mice (n = 4) were analyzed for structural changes at 0 and 7 d postmortem using light microscopy. As an index of structural changes, fiber detachment, cracked or broken fibers, and the appearance of space between sarcomeres were quantified. Cumulatively, the results of the first experiment indicated that postmortem proteolysis of muscle occurred similarly in control and in calpain 3 knockout mice. Desmin degradation did not differ (P > 0.99), and there were no indications that degradation of nebulin, dystrophin, vinculin, and troponin-T were affected by the absence of calpain 3 in postmortem muscle. Structural changes were affected by time postmortem (P < 0.05), but not by the absence of calpain 3 from the muscles. In conclusion, these results indicate that calpain 3 plays a minor role, if any, in postmortem proteolysis in muscle.

Animals↗
PubMed2003

Retained common-duct stones after open cholecystectomy and duct exploration in children.

A retrospective study was performed of 250 patients with cholelithiasis treated at the Royal Children's Hospital, (RCH) Melbourne, over 25 years by open operation; 32 (12.8%) had proven choledocholithiasis on either preoperative imaging, operative cholangiography (OpCG), or postoperative investigation. A further 3 had underlying congenital biliary abnormalities and were excluded from further study. Thirty-one of the 32 were explored at open operation, 27 after OpCG and 4 on clinical grounds. One retained common-bile-duct (CBD) stone was undetected until the postoperative period (1/250, 0.25%). Seven ducts were not cleared, giving a duct exploration failure rate of 22.6% (7/31). All 8 retained CBD stones were identified in the early postoperative period and managed with a variety of techniques, including endoscopic retrograde cholangio-pancreatography (ERCP). The incidence of retained stones after open CBD exploration was high (22.6%), and can be attributed to difficulties in operative technique dealing with the smaller paediatric CBD. In addition, haemolytic disease seems to induce a propensity for choledocholithiasis. Given the small numbers presenting with cholelithiasis to RCH (10 per year), it is suggested that a selective approach to CBD exploration is appropriate in children. With the increasing use of laparoscopic cholecystectomy in children and the inherent technical difficulties of laparoscopic operative cholangiography, ERCP may offer an alternative solution in dealing with CBD stones rather than open or laparoscopic CBD exploration.

Adolescent↗
PubMed2002

The genome sequence of Schizosaccharomyces pombe.

We have sequenced and annotated the genome of fission yeast (Schizosaccharomyces pombe), which contains the smallest number of protein-coding genes yet recorded for a eukaryote: 4,824. The centromeres are between 35 and 110 kilobases (kb) and contain related repeats including a highly conserved 1.8-kb element. Regions upstream of genes are longer than in budding yeast (Saccharomyces cerevisiae), possibly reflecting more-extended control regions. Some 43% of the genes contain introns, of which there are 4,730. Fifty genes have significant similarity with human disease genes; half of these are cancer related. We identify highly conserved genes important for eukaryotic cell organization including those required for the cytoskeleton, compartmentation, cell-cycle control, proteolysis, protein phosphorylation and RNA splicing. These genes may have originated with the appearance of eukaryotic life. Few similarly conserved genes that are important for multicellular organization were identified, suggesting that the transition from prokaryotes to eukaryotes required more new genes than did the transition from unicellular to multicellular organization.

Base Sequence↗
PubMed2001

The physical maps for sequencing human chromosomes 1, 6, 9, 10, 13, 20 and X.

We constructed maps for eight chromosomes (1, 6, 9, 10, 13, 20, X and (previously) 22), representing one-third of the genome, by building landmark maps, isolating bacterial clones and assembling contigs. By this approach, we could establish the long-range organization of the maps early in the project, and all contig extension, gap closure and problem-solving was simplified by containment within local regions. The maps currently represent more than 94% of the euchromatic (gene-containing) regions of these chromosomes in 176 contigs, and contain 96% of the chromosome-specific markers in the human gene map. By measuring the remaining gaps, we can assess chromosome length and coverage in sequenced clones.

Chromosomes, Human, Pair 1↗
PubMed2000

High-pressure effects on lysosome integrity and lysosomal enzyme activity in bovine muscle.

This study was conducted to determine whether the application of high hydrostatic pressure could modify the enzymatic activity and membrane integrity of lysosomes in muscle. Several combinations of pressure (0-600 MPa) and time (0-300 s) were applied to two types of samples: purified enzymes (cathepsin D and acid phosphatase) in buffer solution and intact muscle (biceps femoris). The enzymes studied showed varying degrees of susceptibility depending on the level of pressure, holding time, and environment. Acid phosphatase activity was minimally affected by pressure in buffer solution, whereas cathepsin D was modulated significantly by the pressure and time applied. The activities of the enzymes extracted from meat increased with pressure. The cytochemical observations showed the presence of primary and secondary lysosomes in muscles. After pressurization, the membrane integrity of the lysosomes was modified. A correlation could be established between lysosomal enzymatic activities and the lysosome membrane breakdown.

Acid Phosphatase↗
PubMed1999

mRNA levels of dipeptidyl peptidase IV decrease during intestinal adaptation.

BACKGROUND: Glucagon-like peptide 2 (GLP-2) has recently been shown to be a potent enterotrophic factor that may mediate mucosal hyperplasia during intestinal adaptation. The intestinal brush-border protease dipeptidyl peptidase IV (DPP IV) cleaves GLP-2 to an inactive form. It has been postulated that DPP IV activity limits the enterotrophic activity of GLP-2 in rats and humans. Massive small bowel resection (MSBR) in rats is an animal model of intestinal adaptation that has been used successfully to characterize factors involved in the modulation of adaptation. METHODS: Total RNA was extracted from normal terminal ileum or terminal ileum post-MSBR from Sprague-Dawley rats which were sacrificed 2, 4, and 7 days postresection. A partial rat DPP IV clone was isolated by reverse transcription polymerase chain reaction, and Northern blot analysis of rat DPP IV mRNA levels in normal small bowel and small bowel post-MSBR was performed. RESULTS: Within normal small bowel, DPP IV mRNA levels were greatest in the terminal ileum; levels in the duodenum and jejunum were approximately 50% of those in the terminal ileum. DPP IV mRNA levels decreased in terminal ileum post-MSBR 2, 4, and 7 days after resection. CONCLUSION: The decreased DPPIV gene expression suggests a novel mechanism by which the effects on mucosal growth of GLP-2 may be further enhanced, and further that GLP-2 may be a more useful therapeutic agent in humans than currently anticipated.

Adaptation, Physiological↗
PubMed1998

Tissue distribution of rat glucagon receptor and GLP-1 receptor gene expression.

The regulation of glucose metabolism by glucagon and GLP-1 is well established, but novel functions for these and other proglucagon-derived peptides are less well defined. This paper highlights the diversity of both GLP-1 and glucagon activity by studying the tissue distribution of glucagon and GLP-1 receptor gene expression by both Southern blot analysis of RT-PCR products and nuclease protection assays. By Southern blot analysis of RT-PCR products, GLP-1 receptor mRNA was detected in lung, hypothalamus, hippocampus, cerebral cortex, kidney, pancreas, and throughout the gastrointestinal tract. Glucagon receptor expression was detected in liver, kidney, spleen, thymus, adrenal glands, pancreas, cerebral cortex, lung, and throughout the gastrointestinal tract. Nuclease protection assay revealed glucagon receptor expression to be highest in liver and kidney, whereas GLP-1 receptor expression was only detected by protection assay in lung, stomach, and large bowel. Despite previous evidence that other receptors for proglucagon-derived peptides may exist, no evidence of novel receptors or multiple isoforms of the glucagon and GLP-1 receptors was found, indicating that the two cloned receptors may mediate all the effects of proglucagon-derived peptides, or that novel receptors may share less homology with the glucagon and GLP-1 receptors than previously anticipated.

Animals↗
PubMed1998

Driver stress and performance on a driving simulator.

Effects of stress on driving performance can depend on the nature of driver's stress reactions and on the traffic environment. In an experimental study, we assessed multiple dimensions of vulnerability to driver stress by a questionnaire that was validated in previous field studies and related those dimensions to performance on a driving simulator. Results were broadly consistent with prediction. A dimension of habitual dislike of driving was associated with reduced control skills, greater caution, and disturbance of moods. A measure of aggressive driving predicted more frequent and more error-prone overtaking, which are effects attributed to the use of confrontive coping strategies in interaction with other vehicles. An alertness measure predicted speed of reaction to pedestrian hazards. This research has practical applications for system design, automated monitoring of driver performance, selection and assessment of drivers, and training.

Adolescent↗
PubMed1998

Effects of postmortem storage on the ultrastructure of the endomysium and myofibrils in normal and callipyge longissimus.

These experiments were conducted to examine ultrastructural changes in longissimus from normal and callipyge lamb during 14 d of postmortem storage at 4 degrees C. Six crossbred ewe lambs (1/2 Dorset x 1/2 Romanov) were grain-fed and slaughtered at approximately 250 d of age. Leg conformation score was the basis for classifying carcasses into normal and callipyge. The normal and callipyge longissimus had mean Warner-Bratzler shear force of 2.8 (2.7, 2.4, and 3.4) and 9.0 (12.2, 6.9, and 7.9) kg, respectively, after 14 d of postmortem storage. The results of transmission electron microscopy demonstrated ultrastructural changes, including sarcolemma detachment, loss of myofibril lateral attachments, and I-band breaks in normal longissimus. Detachment of sarcolemma from myofibrils occurred in both phenotypes, but it was delayed by several days in callipyge longissimus. Thus, the sarcolemma detachment seems not to contribute significantly to postmortem tenderization. The endomysium of both phenotypes did not change with postmortem storage. In normal longissimus, the percentage of fractured I-bands increased from 0% at d 1 to 11% at d 3 (P<.05) and did not change between 3 and 14 d (15%) postmortem (P>.05). However, postmortem storage did not affect (0 to 3%) the frequency of the I-band breaks in the callipyge longissimus (P>.05). Therefore, the break in the I-band region in postmortem muscle is a change that is associated with postmortem tenderization. We conclude that the major factor responsible for the toughness of meat from callipyge longissimus is the postmortem stability of myofibrils.

Animals↗
PubMed1997

Association of muscle-specific kinase MuSK with the acetylcholine receptor in mammalian muscle.

During synaptogenesis at the neuromuscular junction, a neurally released factor, agrin, causes the clustering of acetylcholine receptors (AChRs) in the muscle membrane beneath the nerve terminal. Agrin acts through a specific receptor which is thought to have a receptor tyrosine kinase, MuSK, as one of its components. In agrin-treated muscle cells, both MuSK and the AChR become tyrosine phosphorylated. To determine how the activation of MuSK leads to AChR clustering, we have investigated their interaction in cultured C2 myotubes. Immunoprecipitation experiments showed that MuSK is associated with the AChR and that this association is increased by agrin treatment. Agrin also caused a transient activation of the AChR-associated MuSK, as demonstrated by MuSK phosphorylation. In agrin-treated myotubes, MuSK phosphorylation increased with the same time course as phosphorylation of the beta subunit of the AChR, but declined more quickly. Although both herbimycin and staurosporine blocked agrin-induced AChR phosphorylation, only herbimycin inhibited the phosphorylation of MuSK. These results suggest that although agrin increases the amount of activated MuSK that is associated with the AChR, MuSK is not directly responsible for AChR phosphorylation but acts through other kinases.

Agrin↗
PubMed1997

Fish muscle cytoskeleton integrity is not dependent on intact thin filaments.

Striated muscle cytoskeleton was studied by ultrastructure and electrophoresis. Treatment of sea bass white muscle myofibrils and glycerinated fibres with calpain caused disruption of costameres, intermediate filaments, and Z-line, without altering sarcomeres. V8 protease also caused loss of costameres and Z-line, and disrupted sarcomeres without affecting the intermediate filaments. Recombinant lipase caused loss of Z-lines and also sarcolemma detachment, without changing sarcomeres or intermediate filaments. DNase-1 removed thin filaments and partially removed Z-lines while leaving intact the sarcolemma attachments and intermediate filaments. Calpain, V8 protease, lipase and DNase-1 treatments induced extensive loss of alpha-actinin from the Z-line, which could be related to titin cleavage (calpain, V8), phosphoinositide hydrolysis (lipase), and actin depolymerisation (DNase-1). These results show that the cytoskeletal components are independent of intact thin filaments.

Actinin↗
PubMed1996

A technique for cranial bone expansion.

There are several techniques for cranial bone remodeling available to the craniofacial surgeon. We present a method for expanding the cranial bone to reconstruct large cranial defects or contour large abnormalities using a progressive series of tongue-and-groove extensions of local cranial bone plates. This technique has been used by the senior author in more than 95 cases with a 6-month to 15-year follow-up. This technique offers several advantages over traditional cranial remodeling techniques, including three-dimensional stabilization of the remodeled calvaria, use of local tissue obviating the need for distant donor tissue or nonautologous biomaterials, greater flexibility in the modeling of the cranial bone, and a controlled decrease in the size of the remaining craniectomy defects, thus improving potential for cranial bone regeneration and closure. Although cranial bone expansion is not applicable or even necessary in all craniofacial procedures, it nevertheless is a valuable tool of the craniofacial surgeon.

Adolescent↗
PubMed1996

Age and gender differences in perceived accident likelihood and driver competences.

Road traffic accident involvement rates show clear age and gender differences which may in part be accounted for by differences in risk perception and perceptions of driving competence. The present study extends and replicates that of Matthews and Moran (1986). Young (18-30 years) and older (45-60 years) male and female drivers responded to a questionnaire on perceived accident risk and driving competence (judgment and skill) with respect to themselves and four target groups, and also rated a series of videotaped driving sequences with respect to likelihood of accident occurrence and perceived driving competence. Results showed that effects of rater characteristics were generally confined to the questionnaire. Younger males were perceived as most likely to experience an accident and were judged to be lower than other groups in driving competence. Younger groups showed little bias against older groups and vice versa, but gender-related bias was apparent. The findings of Matthews and Moran were generally confirmed. The results are discussed with reference to four main issues: (1) demographic bias effects--which are generally weak; (2) stereotyping on the basis of gender and/or age of driver; (3) group-specific bias; (4) self-appraisal bias.

Accidents, Traffic↗
PubMed1995

POU-domain gene expression in the gastrointestinal tract.

The process of self-renewal which occurs in the gastrointestinal epithelium is greatly amplified and accelerated during the intestinal adaptation which occurs in the residual ileum after massive small bowel resection (MSBR). As with growth and development, these processes must involve the coordinated regulation of many genes. Several families of nuclear proteins are known to be involved in the control of gene expression during development including the POU-domain genes; their expression has not been characterized in the gastrointestinal tract during normal cellular renewal or adaptation, and POU-domain encoding cDNAs were cloned from ileal RNA. Three known genes were cloned: Oct-1, Brn-1 and Tst-1 but no novel members of this gene family were identified. The encoded sequence for rat Oct-1 differs from that previously reported. Oct-1 is relatively ubiquitously expressed with increased expression during both development and adaptation. Minimal expression of Tst-1 was observed. Brn-1 exhibits limited expression in the adult gastrointestinal tract but may play a role in the fetal gastrointestinal tract.

Adaptation, Physiological↗
PubMed1995

Hepatocyte growth factor activates quiescent skeletal muscle satellite cells in vitro.

The effect of hepatocyte growth factor (HGF) on the activation of quiescent rat skeletal muscle satellite cells was evaluated in vitro. Satellite cells from 9-month-old adult rats are quiescent in vivo and when cultured, display a protracted lag phase prior to division that is not present in satellite cells from neonatal or regenerating muscle. Under normal growth conditions, satellite cells divide for the first time between 42 and 60 hr. Hepatocyte growth factor increased proliferation in a dose-dependent fashion prior to 48 hr with half-maximal stimulation at approximately 3 ng/ml; in addition, heparin enhanced this activity. The time course of cyclin-D1 and proliferating cell nuclear antigen (PCNA) expression was accelerated in HGF-treated satellite cells, indicating that cells entered the cell cycle earlier. No significant effects on muscle-derived fibroblast proliferation was observed. The signalling receptor for HGF is the product of the c-met protooncogene, and rtPCR analysis of satellite cells 0-72 hr in culture demonstrated the presence of this message throughout this time period. The presence of c-met in quiescent satellite cells, the ability of HGF to stimulate precocious entry into the cell cycle, and the previously described localization of HGF message in regenerating muscle (Jennische et al., 1993) indicate that HGF could act as an activator of quiescent satellite cells in vivo.

Animals↗
PubMed1995

Proteolytic activity of proteasome on myofibrillar structures.

The physiologic function of proteasome remains unclear. Evidence suggests a role in degradation of ubiquitin-protein conjugates, MHC antigen presentation, and some specificity of substrate within certain cell types. To explore further the properties of proteasome we have examined its effect on a well defined structure, the myofibril. We find that despite its large size (20S) proteasome is able to degrade myofibrils and intact, permeabilized muscle fibrils. The proteins degraded showed some specificity because actin, myosin and desmin were degraded faster than alpha-actinin, troponin T and tropomyosin. Changes in ultrastructure were slow and included a general loss of structure with Z and I bands effected before the M band and costameres.

Animals↗
PubMed1995

Effects of the protein phosphatase inhibitors, tautomycin and calyculin-A, on protein phosphorylation and cytoskeleton of human platelets.

Effects of the protein phosphatase inhibitors, tautomycin and calyculin A on protein phosphorylation and cytoskeleton of human platelets. It has been discovered recently that many cytotoxic compounds isolated from a variety of sources are potent phosphatase inhibitors. Two of these, tautomycin (TM) and calyculin-A (CL-A) were applied to human platelets to investigate the role of protein phosphorylation on cytoskeletal structure and function. Exposure to 10 microM TM or 0.1 microM CL-A induced marked morphological changes. The granules were centralized and surrounded by actin filaments, but there was no evidence of granule release. Myosin became more centralized, was occluded from the granulomere, but was not confined to the microfilament ring. These changes occurred without an increase in cytosolic Ca2+ concentrations, as determined by measurements with fura-2. TM and CL-A induced an overall increase in protein phosphorylation. Phosphorylation of the 20,000 dalton light chain of myosin increased markedly and multiple phosphorylation sites were indicated. Cytoskeletons were prepared from control, thrombin- and TM-treated platelets, the latter prepared in the absence of external calcium. The major difference in protein composition was the increased content of myosin associated with the cytoskeleton from TM-treated platelets where the dominant phosphoprotein was the 20,000 dalton light chain. These results suggest that myosin phosphorylation drives the initial shape changes, and via a contractile process results in the formation of the microfilament ring and centralization of granules.

Adult↗