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Biomedical subjects

D M Gibson

Publications and source records attributed to D M Gibson.

At least 73 records · Page 4Linked to original sources
PubMed1985

Rapid and automated detection of salmonella by electrical measurements.

A rapid method for determining the presence of salmonella in food is described. It consists of pre-enrichment in buffered peptone water modified by the addition of dulcitol and trimethylamine oxide, followed by selective enrichment in a selenite-cystine broth with similar modifications. Changes in the conductance of the selective enrichment broth are monitored continuously using a suitable impediometric instrument. Most of the Salmonella spp. tested gave a fast (approximately 100 microS/h) and large (greater than 600 microS) change in conductance, other enteric bacteria much less or no change. The assay is usually complete within 24 h. Samples of foodstuffs, naturally and artificially contaminated with Salmonella spp., were all correctly classified. Some strains of Citrobacter freundii produced a false positive conductance response, and they could not be selectively eliminated using antibiotics or cyanide. The conductance method is simple and easy to use, gives rapid results and involves less media and subculturing than is required for traditional methods.

Bacteriological Techniques↗
PubMed1985

Isotretinoin kinetics after 80 to 320 mg oral doses.

Twelve healthy male subjects received 80, 160, 240, and 320 mg doses of oral isotretinoin as multiples of 40 mg capsules separated by 2-week washout periods in a randomized, crossover design. Blood samples were drawn at specific times over a 72-hour period after dosing. Blood concentrations of isotretinoin as well as its major metabolite, 4-oxo-isotretinoin, were determined by a specific HPLC method. In addition to the normal laboratory battery of tests, serum triglyceride levels were determined before the first dose and again 72 hours after each of the four doses. Mean (+/- SD) maximum concentrations after 80 to 320 mg doses were 366 +/- 159, 820 +/- 474, 1056 +/- 547, and 981 +/- 381 ng/ml, whereas the respective AUC0-infinity values were 3690 +/- 1280, 7030 +/- 4140, 9780 +/- 6080, and 9040 +/- 2900 ng X hr/ml. The observed apparent elimination t1/2 remained approximately the same (14.7 hours) for each dose. The maximum concentration and AUC values for isotretinoin appear to be dose proportional from 80 to 240 mg but plateau at the 320 mg dose level. Therefore, because isotretinoin blood concentrations may not increase with higher doses in the fasting state, single, oral doses in excess of 240 mg should be used with caution. The data also suggest that elevated triglyceride levels are not a simple function of isotretinoin blood concentrations across the entire study population and dose range studied, but that in subjects with triglyceride levels in excess of the normal range triglyceride levels were positively related to isotretinoin blood concentrations.

Administration, Oral↗
PubMed1985

Pharmacokinetics of recombinant leukocyte A interferon following various routes and modes of administration to the dog.

The pharmacokinetics of recombinant leukocyte A interferon (rIFN-alpha A) were studied following intravenous (i.v.) bolus, 60 min (i.v. inf.) infusion, intramuscular (i.m.), subcutaneous (s.c.), and oral (p.o.) administrations to 15 male beagle dogs. Each animal received at least one 3 X 10(6) units/kg dose of rIFN-alpha A by one of the five routes and/or modes of administration. Blood samples were collected and the serum was separated and analyzed for rIFN-alpha A concentrations by an enzyme immunoassay, ELISA. There were no measurable rIFN-alpha A concentrations (less than 0.020 ng/ml) following oral administration. In general serum rIFN-alpha A concentrations exceeded 100 ng/ml following i.v. bolus and infusion doses then declined rapidly in a biphasic manner. The volume of distribution at steady state Vdss ranged from 0.14 to 0.21 liters/kg after i.v. infusion. Total body serum clearance (ClB) ranged from 14.6 to 23.9 ml/min, which is about 50% the estimated inulin clearance in dogs. The harmonic mean elimination half-lives ranged from 4.5 to 9.5 h. A prolonged absorption profile was seen following i.m. and s.c. administrations and the systemic bioavailability for both routes was 42% when compared with the i.v. infusion. These appear to be the first pharmacokinetic profiles of rIFN-alpha A reported in the dog.

Animals↗
PubMed1985

Polymorphism of kappa variable region (V kappa-1) genes in inbred mice: relationship to the Ig kappa-Ef2 serum light chain marker.

We describe here the cloning and complete sequence analysis of the rearranged kappa variable region gene from the V kappa-1A-producing myeloma (C.AL20-TEPC-105). A 5'-flanking region probe from the V kappa-1A gene has been used to study the V kappa-1 germ-line gene family in strains of mice differing at the Ig kappa-Ef2 locus. All Ef2a strains examined possess an identical pair of BamHI restriction fragments strongly hybridizing to the 5' probe. Surprisingly, only two of the six strains of mice previously designated Ef2b (NZB and C58) possessed clearly altered restriction fragment sizes for V kappa-1 genes. The remaining four Ef2b strains, namely BDP/J, CE/J, I/LnJ, and P/J, appear to carry V kappa-1 genes similar to those of Ef2a strains. It is suggested that these strains may carry a third form of the V kappa-1A gene, differing in the protein coding region but indistinguishable at the DNA level with the use of BamHI or EcoRI. Alternatively, these strains may fail to express V kappa-1A light chains due to a regulatory defect involving this subgroup of kappa genes.

Animals↗
PubMed1984

Phosphorylation of microsomal HMG CoA reductase increases susceptibility to proteolytic degradation in vitro.

Conversion of native, 97-100 kDa rat liver microsomal HMG CoA reductase to membrane-bound 62 kDa and soluble 52-56 kDa catalytically active forms was catalyzed in vitro by the calcium-dependent, leupeptin- and calpastatin-sensitive protease calpain-II purified from rat liver cytosol. Cleavage of the native 97-100 kDa reductase was enhanced by pretreatment (inactivation) of microsomes with ATP(Mg2+) and liver reductase kinase (compared to protein phosphatase-pretreated controls). This was reflected in a loss of the 97-100 kDa species and an increase in the soluble 52-56 kDa species (total enzyme activity and specific immunoblot recovery).

Animals↗
PubMed1984

Recombination between kappa chain genetic markers in the mouse.

In this study we report the first instance of recombination between kappa chain genetic markers in the mouse. The recombination frequency, 0.45% (95% limits, 0.12-1.61), is similar to that previously found for recombination between the kappa chain locus and the Lyt-2,3 locus (0.3%, 95% limits, 0.05-1.6), but is relatively low in comparison with that found at the heavy chain locus (0.41-5.4%). Lyt-2,3-typing of the recombinants permits a partial ordering of the kappa chain and Lyt-2,3 loci as (Lyt-2,3, Igk-Ef1) - Igk-Ef2. Light chains controlled by the two kappa markers include the Vk-(ser) subgroup (controlled by Igk-Ef1) and Vk-1 (controlled by Igk-Ef2). One of the recombinants has been recovered in a homozygous state ("NAK") and should be suitable for Vk gene mapping studies.

Animals↗
PubMed1984

Synthesis and degradation of interconvertible enzymes. Kinetic equations of a model system.

Steady-state and kinetic equations have been developed which characterize the rates of formation, interconversion, and degradation of an enzyme protein subject to reversible phosphorylation. The theoretical model system incorporates separate fractional degradative rate constants for the phosphorylated and dephosphorylated protein species. The classical models for interconvertible enzymes, and for protein turnover, are special limiting situations of the general model presented here.

Enzymes↗
PubMed1984

Guanosine triphosphate and colchicine affect the activity and the polymeric state of acetyl-CoA carboxylase.

Acetyl-CoA carboxylase was purified 300-fold from rat liver, in the absence of added citrate, by precipitation from an 18,000g supernatant in the presence of Triton X-100 at 105,000g and 20 degrees C, followed by chromatography on phosphocellulose. Acetyl-CoA carboxylase activity in this preparation was activated by preincubation with GTP (0.1-2.0 mM) and with citrate (20 mM). Colchicine (10(-6)-10(-3) M) inhibited enzyme activity and counteracted the effects of GTP and citrate. Sucrose density gradient centrifugation demonstrated that GTP and citrate preincubation promoted the formation of the polymeric, active enzyme, while colchicine engendered disassembly. Preincubation of the purified acetyl-CoA carboxylase at 4 degrees C caused inactivation and disassembly, which was countered by preincubation at 37 degrees C in the presence of GTP or citrate. These results suggest that GTP, like citrate, activates acetyl-CoA carboxylase by enhancing the conversion of the protomeric form of the enzyme to its more active, polymeric state.

Acetyl-CoA Carboxylase↗
PubMed1984

Evidence for 65 electrophoretically distinct groups of light chains in BALB/c and NZB myelomas.

Light chains of 426 randomly selected myelomas from BALB/c and NZB mice were characterized by isoelectric focusing (IF) and electrophoresis at pH 3. Approximately 50% of the light chains in each strain were found to have unique electrophoretic properties. The remaining 50% of light chains fell into groups of two or more proteins showing apparent identity. A total of 49 groups of light chains were identified in this way. On the basis of repeat frequencies, the total number of such groups has been estimated to be around 65. Analysis of light chains of known amino acid sequence suggests that many of the "IF-groups" may represent light chains belonging to single V-region subgroups. The large number of unique light chains could represent somatic mutants bearing charge differences from the germ-line-coded sequences or products of a subset of V-genes which are infrequently expressed in myeloma proteins. Comparison of the light chains of BALB/c and NZB myelomas indicated extensive overlap of IF-subgroups in the two strains. One clear difference between the two samples was the presence of three groups of light chains at an unexpectedly high frequency in the BALB/c myelomas. In contrast, no light chains belonging to these groups were found in the NZB myelomas. One of the anomalous groups corresponded to Vk-1A light chains, a subgroup previously shown to be absent in NZB serum light chain IF-profiles. A second group represented at an elevated level included MOPC467, a light chain closely related in sequence to Vk-1A. The third group included ADJPC5 and LPC1. The overrepresentation of these light chain groups in the BALB/c myelomas is not understood. Two light chain-like polypeptides were observed in approximately 3-5% of myelomas in the two mouse strains. It seems likely that in many instances this may be explained by the presence of two cell lines in early generation tumors.

Animals↗
PubMed1984

Community vs institutional care: the case of the Australian aged.

This paper analyses data, hitherto unavailable in Australia, on patterns of caring for the handicapped aged. Particular attention is given to the mechanisms enabling older handicapped persons to remain in the community. On the basis of this information, existing government policies concerning both community and institutional services are reviewed and the adequacy of present arrangements for future generations of the elderly are assessed. The findings emphasize the diverse resources and needs of the aged and the inadequacy of Australian community care programmes in terms of both variety and flexibility.

Aged↗
PubMed1984

Interferon kinetics and adverse reactions after intravenous, intramuscular, and subcutaneous injection.

Three groups of six subjects each received a single 36 X 10(6) U dose of recombinant leukocyte A interferon (rIFN-alpha A) as a 40-min infusion, an intramuscular injection, or a subcutaneous injection. Blood samples were collected at specific times after dosing for analysis of rIFN-alpha A serum concentrations by an enzyme immunoassay method, ELISA. The rIFN-alpha A was rapidly distributed and moderately eliminated (t 1/2 = 5.1 hr) after intravenous infusion. The maximum concentrations at the end of intravenous infusion were tenfold the maximum concentrations after intramuscular and subcutaneous injections. Renal tubular secretion or extrarenal elimination was suggested by clearance values of 1.8 times the glomerular filtration rate. After intramuscular and subcutaneous injection, rIFN-alpha A was absorbed slowly (time to reach maximum concentration ranged from 4 to 8 hr), which resulted in prolonged serum concentrations. Estimated bioavailability was more than 80% for both intramuscular and subcutaneous injection shares qualitatively the same adverse reactions, the reactions differ in severity and duration. The adverse effects appear to be related to route of administration of herpes labialis were also noted. There were no significant clinical laboratory abnormalities of medical concern. Although rIFN-alpha A injected by intravenous infusion or intramuscular or subcutaneous injection shares qualitatively the same adverse reactions, the reactions differ in severity and duration. The adverse effects appear to be related to route of administration.

Adult↗
PubMed1984

Subcellular Localizations of Two Dolichos biflorus Lectins.

The subcellular localizations of the Dolichos biflorus seed lectin and the structurally related lectin (cross-reactive material [CRM]) from the stems and leaves of this plant were determined by immunofluorescence, immunocytochemistry, and cell fractionation procedures. Subcellular fractionation of the cotyledons using a nonaqueous procedure to minimize disruption of the protein bodies showed that the majority of the seed lectin was associated with the protein body fraction and some lectin was also present in the starch granules. Immunofluorescence and immunocytochemistry at the light microscopic level showed that the seed lectin was mainly localized at the peripheries of these organelles. Lectin was also found in the cytoplasm of the cells, although the amount appeared to be dependent upon the degree of protein body disruption.Immunofluorescence and immunocytochemistry studies of the stem and leaf lectin (CRM) indicated that a significant portion of this lectin may be associated with the cell walls, although lectin was also seen in the cytoplasm of plasmolyzed cells. Extraction and cell fractionation studies showed that a large portion of the CRM is readily solubilized and most of the remainder is pelleted at 1000g. The CRM can be extracted from these pellets by treatment with cellulase and pectinase; other reagents such as NaCl, detergents, and EDTA could also release significant amounts of CRM. These studies suggest that the CRM is noncovalently bound to the cell walls. A comparison of the distribution of exogenously supplied [(125)I]CRM with the endogenous CRM during extraction and cell fractionation indicates that soluble CRM is not adsorbed to the 1000g pellet during fractionation.The different subcellular distributions of these two structurally related lectins suggest that different tissues of the same plant may utilize lectins for different functions.

Journal Article↗
PubMed1983

Recombination between kappa chain genetic markers and the Lyt-3 locus.

Recombination has been detected for the first time between chromosome 6 loci controlling kappa chain expression in normal mouse serum immunoglobulin and the Lyt-3 locus. The recombination event occurred at the 26th or 27th backcross generation during the derivation of the Lyt-2a, Lyt-3a-congenic line B6.PL(85NS). The line is now homozygous for the Lyt-2a, Lyt-3a allele(s) at N30F13 and homozygous animals express the Igk-Ef1b allele derived from C57BL/6. The frequency of recombination has been estimated to be 0.30% based on the present results and previous studies in which no recombination was detected. The results rule out the hypothesis that the Lyt-3 locus itself controls the light chain phenotype observed in normal serum immunoglobulin.

Alleles↗
PubMed1983

Uridine diphosphate glucose breakdown is mediated by a unique enzyme activated by fructose 2,6-bisphosphate in Solanum tuberosum.

In the presence of inorganic phosphate, uridine 5'-diphosphate glucose (UDPG) is specifically hydrolyzed to glucose 1-phosphate and UDP by a unique enzyme, UDPG phosphorylase. The activity of the enzyme was maximally stimulated by fructose 2,6-bisphosphate, a regulatory metabolite recently discovered in both plants and animals, and by 2-phosphoglyceric acid. At 1 muM, fructose 2,6-bisphosphate stimulated UDPG phosphorolysis 10-fold, whereas 2-phosphoglyceric acid was required at higher concentrations (100 muM) to produce a similar effect. Fructose 2,6-bisphosphate appears to increase the affinity of the enzyme for inorganic phosphate, with a change in K(m) from 1.6 mM to 0.3 mM. The results suggest that fructose 2,6-bisphosphate participates in the regulation of other pathways of carbohydrate metabolism in addition to playing its recognized role in glycolysis and gluconeogenesis.

Journal Article↗
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↗