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Biomedical subjects

S Gibson

Publications and source records attributed to S Gibson.

At least 127 records · Page 7Linked to original sources
PubMed1988

The pain of discography.

Lumbar discography is a commonly employed diagnostic tool, but important questions about it remain unresolved. Why is an abnormal discogram painful in one patient and not in another? This study was performed to investigate the changes in Substance P (SP) and Vasoactive-Intestinal Peptide (VIP), found in the dorsal root ganglion, following discography in normal and abnormal canine lumber intervertebral discs. The data from this study suggest that dorsal root ganglion SP and VIP are indirectly affected by manipulations of the intervertebral disc. It may be that various neurochemical changes within the intervertebral disc are expressed by sensitized (injured) annular nociceptors, and in part modulated by the dorsal root ganglion. Therefore, the concomitant pain sometimes associated with an abnormal discogram image may in part be related to the chemical environment within the intervertebral disc and the sensitized state of its annular nociceptors.

Animals↗
PubMed1988

The Nottingham Health Profile as a measure of quality of life following combined heart and lung transplantation.

This paper reports on the use of the Nottingham Health Profile as a measure of patient quality of life before and after combined heart and lung transplantation at Harefield Hospital. A total of 125 profiles from 48 patients were analysed. In both section 1 and section 2 of the profile, large and statistically significant (p less than 0.05) improvements in quality of life were associated with transplantation. The profile proved easy to use either as part of an interview during assessment for transplantation or as a postal follow-up postoperatively.

Adult↗
PubMed1988

Intestinal hypersensitivity reactions in the rat. I. Uptake of intact protein, permeability to sugars and their correlation with mucosal mast-cell activation.

We have confirmed previous observations that intestinal anaphylaxis induced in rats previously sensitized to ovalbumin (OVA) is associated with an increased uptake of an unrelated 'bystander' protein, bovine serum albumin (BSA) fed 1 hr previously. In this study, this enhanced protein uptake was associated with an increased lactulose/rhamnose excretion ratio after administration of these sugars, although there was no correlation between the two measurements. One hour after antigen challenge the serum levels of rat mast-cell protease II (RMCPII), a specific marker for mucosal mast-cell secretion, were significantly higher than both the pre-challenge levels and those of sham-challenged controls (P less than 0.002). There was a significant positive correlation between the serum levels of RMCPII and the lactulose/rhamnose excretion ratios (P less than 0.05), but no such correlation existed between RMCPII and BSA levels in the challenged rats. In other studies the urinary lactulose/rhamnose ratios of rats with cetrimide-induced gut damage were found to be significantly increased, although BSA uptake into the serum remained unaltered. We conclude that there is no simple correlation between gut permeation of low-molecular weight sugars and and the uptake of macromolecular proteins.

Animals↗
PubMed1987

Estimation by radiation inactivation of the size of functional units governing Sendai and influenza virus fusion.

The target sizes associated with fusion and hemolysis carried out by Sendai virus envelope glycoproteins were determined by radiation inactivation analysis. The target size for influenza virus mediated fusion with erythrocyte ghosts at pH 5.0 was also determined for comparison; a value of 57 +/- 15 kDa was found, indistinguishable from that reported previously for influenza-mediated fusion of cardiolipin liposomes [Gibson, S., Jung, C. Y., Takahashi, M., & Lenard, J. (1986) Biochemistry 25, 6264-6268]. Sendai-mediated fusion with erythrocyte ghosts at pH 7.0 was likewise inactivated exponentially with increasing radiation dose, yielding a target size of 60 +/- 6 kDa, a value consistent with the molecular weight of a single F-protein molecule. The inactivation curve for Sendai-mediated fusion with cardiolipin liposomes at pH 7.0, however, was more complex. Assuming a "multiple target-single hit" model, the target consisted of 2-3 units of ca. 60 kDa each. A similar target was seen if the liposomes contained 10% gangliosides or if the reaction was measured at pH 5.0, suggesting that fusion occurred by the same mechanism at high and low pH. A target size of 261 +/- 48 kDa was found for Sendai-induced hemolysis, in contrast with influenza, which had a more complex target size for this activity (Gibson et al., 1986). Sendai virus fusion thus occurs by different mechanisms depending upon the nature of the target membrane, since it is mediated by different functional units. Hemolysis is mediated by a functional unit different from that associated with erythrocyte ghost fusion or with cardiolipin liposome fusion.

Animals↗
PubMed1987

Peptide expression is altered when afferent nerves reinnervate inappropriate tissue.

Neuropeptides are found in specific subpopulations of primary afferent neurones. Peptide expression can be altered following axotomy or under the influence of nerve growth factor. Here we have examined the consequence of altering the peripheral target of afferent neurones. Many unmyelinated afferents from skin contain substance P-like immunoreactivity (SPLI) whilst those from muscle do not. We have found that fibres will innervate inappropriate tissue types. We have therefore cut and cross-anastomosed a skin and muscle hindlimb nerve in the rat and 10-12 weeks later analysed the regenerated nerves immunocytochemically for SPLI. Muscle afferents inappropriately reinnervating skin were found to contain many SPLI fibres in contrast to control nerves resutured to their own distal stumps. Conversely, skin afferents made to innervate muscle showed reduced levels of peptide staining. These results demonstrate the plasticity of peptide expression and suggest that factors in peripheral tissue or perhaps distal nerve sheaths exert a trophic influence on nervous system function.

Animals↗
PubMed1987

Resistance of nematode secretory products to cleavage by mast cell proteinases.

Mast cell proteinases are known to be released in response to helminth infection, and are, in particular, characteristic of the immune rejection of intestinal nematode parasites. In intestinal mucosal tissue the relevant enzyme is rat mast cell proteinase II (RMCP II) and that of other tissues, including the lung, is rat mast cell proteinase I (RMCP I). The function of these enzymes is unknown, and we have examined the possibility that they directly attack the parasites. This was done by examining the cleavage patterns produced by both proteinases on 125I-labelled excretory/secretory (ES) products of two intestinal nematodes (the infective larva of Ascaris suum, and adult Nippostrongylus brasiliensis) and one which has a pulmonary migration route (the third/fourth stage larva of A. suum). It was first established that all the labelled molecules were proteinaceous, by their susceptibility to broad spectrum proteinases, and that none were host components carried over into culture, by their antigenicity to infected hosts. All the nematode ES products were found to be remarkably resistant to RMCP I and II, only one major component of the infective larva of A. suum being cleaved by both enzymes. This was not found to reflect a resistance to serine proteinases in general, since selected ES components were cleaved by chymotrypsin and trypsin. This would, therefore, argue that, if the enzymes play any direct role in the immune expulsion of nematodes, it is unlikely to be successfully directed at their secretions.

Animals↗
PubMed1987

Sex differences in acetaldehyde on body temperature and open-field performance in the rat.

Using the open field, for making an overall assessment of behavioural change in response to acetaldehyde (AcH) treatment, the results from the present study demonstrate that AcH produces behavioural and physiological effects similar to those of ethanol. High doses of AcH produced a similar degree of change as high doses of ethanol. Both compounds render the animal incapable of locomotor activities, such as ambulation and rearing and both produce a decrease of 2-3 degrees C in body temperature. AcH, however, appears to be a much more potent compound as behavioural and physiological changes are exhibited even after a dose of 10 mg/kg, while the dose of ethanol necessary to produce a decrease in open-field behaviour is at least 10 times as much. Females were more sensitive to the effects of AcH at the 100 mg/kg dose.

Acetaldehyde↗
PubMed1987

The effect of muscle paralysis on the radial growth of collagen fibrils in developing tendon.

Voluntary muscle activity in chick embryos was paralysed by administration in ovo of tubocurarine hydrochloride, administered in single or multiple doses, from day 9 to day 13 after fertilization. Control eggs were given saline instead of tubocurarine or were simply incubated without operative interference. Embryos were killed at 9, 13, 14, 16 and 19 days after fertilization. Flexor digitorum tendons were removed, fixed in glutaraldehyde, embedded in plastic, sectioned, and stained with phosphotungstic acid for electron microscopy. The diameters of the tendon collagen fibrils were measured, on electron micrographs, using a Magiscan Mk II programme. Tendon collagen fibril expansion was not inhibited by tubocurarine treatment. It is concluded that the rapid increase of collagen fibril diameters, which coincides in the normal embryo with the first onset of use of the associated muscle, is not dependent on muscle activity. There remains a possibility that other ways of producing tension in the tendon could provide sufficient stimulus to fibril expansion.

Animals↗
PubMed1987

Systemic release of a mast cell proteinase following nematode infections in sheep.

An enzyme-linked immunosorbent assay (ELISA) for sheep mast cell proteinase (SMCP) has been developed. Concentrations of SMCP in homogenates of abomasal tissue from parasite-immune sheep (341 micrograms SMCP/g tissues) were raised when compared to those in normal (non-infected) abomasa (0.145 micrograms SMCP/g tissue). SMCP was not detected in sera from normal animals challenged with Haemonchus contortus but was present (less than 1.0 ng SMCP/ml) in sera from 8/11 immune sheep 2 h after intra-abomasal challenge with 1 x 10(6) exsheathed Haemonchus larvae. In two further experiments, the SMCP response in gastric lymph was monitored after homologous larval challenge in sheep immune to Ostertagia circumcincta and in normal controls. SMCP (less than 1.4 ng SMCP/ml) was detected in lymph from 2/3 and 4/5 immune animals between 1 and 4 days post-challenge with 50,000 larvae, but not from normal animals. SMCP was not detected in lymph from immune animals following challenge with 1000 Ostertagia larvae. The relatively low concentrations of SMCP in blood and lymph reflect the presence of proteinase inhibitor(s) which interfered with the ELISA.

Abomasum↗
PubMed1987

Phenotypic expression of mast cell granule proteinases. Distribution of mast cell proteinases I and II in the rat digestive system.

The distribution of mast cell granule proteinases in the rat digestive system was determined immunohistochemically. The population of toluidine blue-staining mast cells was accounted for by cells containing either rat mast cell proteinase I (RMCPI) or rat mast cell proteinase II (RMCPII). Granules in greater than 90% of RMCPI-containing cells stained red after the Alcian blue/safranin sequence, whereas all RMCPII-containing mast cells stained blue. The red/RMCPI phenotype was typical of the connective tissue mast cells (CTMC) that populate the proximal, non-mucosal, regions of the digestive system, and was also abundant in the serosa and in rectal and gastric muscularis. The blue/RMCPII phenotype, absent from non-mucosal sites except for rare cells in intestinal submucosa and muscularis, predominated in all mucosal tissues and resembled mucosal mast cells (MMC). A third, minor population of cells containing RMCPI but staining blue in the Alcian blue/safranin sequence was detected in both non-mucosal and mucosal tissues. Blue/RMCPI mast cells were rare in the small intestine but more frequent in the mucosa of the stomach and large intestine and in the connective tissues. It is suggested that granule proteinase phenotyping may provide an alternative technique in the analysis of mast cell heterogeneity.

Animals↗
PubMed1987

Characterization and mast cell origin of a chymotrypsin-like proteinase isolated from intestines of mice infected with Trichinella spiralis.

A proteinase was purified by cation exchange and affinity chromatography from the small intestines of mice infected with Trichinella spiralis. The enzyme was highly soluble and was chymotrypsin-like in its substrate specificities and susceptibility to inhibitors. It had a MW of 26,000, as determined by SDS-PAGE electrophoresis. Antibodies raised against the proteinase were affinity purified and their specificity confirmed by Western blot analysis. When used to localize the enzyme immunohistochemically, they reacted with granules of mast cells in the epithelium and lamina propria of the parasitized small intestine. The antibodies also bound to mast cell granules in a number of other sites, including tracheal epithelium, gastric mucosa, skin and tongue. Affinity-purified antibodies raised against rat mast cell proteinase II (RMCPII) cross-reacted with the mouse mast cell proteinase on Western blots.

Animals↗
PubMed1987

Roles of mucosal mast cells in intestinal cell-mediated immunity.

Mucosal mast cells in rats with GvHR have been studied by cell counts, tissue levels of the specific protease RMCPII, and, as an index of MMC activation, serum RMCPII. In semi-allogeneic GvHR without host irradiation, GvHR produced modest increases in these three indices. In contrast, irradiation profoundly depleted MMC even though enteropathy was more severe than in non-irradiated hosts. We suggest that enteropathy is not dependent on the presence of MMC. In rats given cyclosporin A, lesions of GvHR were mild and numbers of MMC were low.

Animals↗
PubMed1987

Phenotypic analysis of mast cell granule proteinases in normal rat bone marrow cultures.

Mast cells with morphological and some biochemical properties of mucosal mast cells (MMC) proliferate and mature in rat bone marrow cultures stimulated with factors from antigen- or mitogen-activated T lymphocytes. There has been much controversy over the criteria used to distinguish the different mast cell subsets, and because histochemistry of granule glycosaminoglycans does not adequately define mast cell subsets morphologically, the proteinase phenotypes of cultured mast cells were analysed. Affinity-purified cross-absorbed monospecific F(ab')2 antibodies raised against rat mast cell protease I (RMCPI) from connective tissue mast cells (CTMC) and against rat mast cell protease II (RMCPII) isolated from mucosal mast cells were used to stain granule proteinase by an immunohistochemical technique. Mast cells grown in culture from normal rat bone marrow stained exclusively with anti-RMCPII antibodies, thus providing further confirmation of their similarity to, and identity with, MMC.

Animals↗
PubMed1986

Radiation inactivation analysis of influenza virus reveals different target sizes for fusion, leakage, and neuraminidase activities.

The size of the functional units responsible for several activities carried out by the influenza virus envelope glycoproteins was determined by radiation inactivation analysis. Neuraminidase activity, which resides in the glycoprotein NA, was inactivated exponentially with an increasing radiation dose, yielding a target size of 94 +/- 5 kilodaltons (kDa), in reasonable agreement with that of the disulfide-bonded dimer (120 kDa). All the other activities studied are properties of the HA glycoprotein and were normalized to the known molecular weight of the neuraminidase dimer. Virus-induced fusion activity was measured by two phospholipid dilution assays: relief of energy transfer between N-(7-nitro-2,1,3-benzoxadiazol-4-yl)dipalmitoyl-L-alpha- phosphatidylethanolamine (N-NBD-PE) and N-(lissamine rhodamine B sulfonyl)-dioleoyl-L-alpha-phosphatidylethanolamine (N-Rh-PE) in target liposomes and relief of self-quenching of N-Rh-PE in target liposomes. Radiation inactivation of fusion activity proceeded exponentially with radiation dose, yielding normalized target sizes of 68 +/- 6 kDa by assay i and 70 +/- 4 kDa by assay ii. These values are close to the molecular weight of a single disulfide-bonded (HA1 + HA2) unit (75 kDa), the "monomer" of the HA trimer. A single monomer is thus inactivated by each radiation event, and each monomer (or some part of it) constitutes a minimal functional unit capable of mediating fusion. Virus-induced leakage of calcein from target liposomes and virus-induced leakage of hemoglobin from erythrocytes (hemolysis) both showed more complex inactivation behavior: a pronounced shoulder was present in both inactivation curves, followed by a steep drop in activity at higher radiation levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Host factors in nuclear plasmid maintenance in Saccharomyces cerevisiae.

In yeast, the initiation of DNA replication on chromosomes is believed to occur at specific sequences known as autonomously replicating sequences (ARSs). We previously isolated a number of mutants that are defective in the maintenance of minichromosomes. Analysis of these mutants suggests that although ARSs differ greatly from one another in their primary sequences, they appear to share a common enzyme complex for the initiation of DNA replication. However, this initiation enzyme complex probably binds with differential affinity to different ARSs. This idea is corroborated by our identification of an ARS-binding protein that binds to different ARSs with different efficiencies.

Cell Nucleus↗
PubMed1986

The catalytic properties of a proteinase isolated from sheep abomasal mucosal mast cells.

The catalytic properties of a sheep mast cell proteinase (SMCP), isolated from abomasal mucosal mast cells, were investigated. The enzyme was shown to have chymotrypsin-like esterase activity, with no detectable amide activity, using a range of low molecular weight substrates. Maximal activity, against Benzyloxycarbonyl-L-tyrosine-4-nitrophenol ester, was determined to be in the range pH 7.6-8.0. Inhibitor studies showed that, unlike chymotrypsin, a serine proteinase, SMCP was found to be susceptible to the action of thiol blocking agents and chelating agents, but to be unaffected by diisopropylphosphofluoridate, a serine proteinase inhibitor.

Abomasum↗
PubMed1986

The isolation and purification of a proteinase with chymotrypsin-like properties from ovine mucosal mast cells.

A mast cell granule proteinase was purified from isolated ovine mucosal mast cells by cation exchange chromatography, which defined the conditions for enzyme purification from sheep gastric mucosae. Antibodies raised against the proteinase were used in subsequent purification procedures which yielded 78 micrograms of enzyme per 5 g wet wt of abomasal tissue. Immuno-histochemistry confirmed that mucosal mast cells were the source of the enzyme. The proteinase had chymotrypsin-like esterase activity, with a molecular weight between 19,000 and 25,000.

Abomasum↗