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Biomedical subjects

D L Martin

Publications and source records attributed to D L Martin.

At least 55 records · Page 3Linked to original sources
PubMed1993

Primary extensor tendon reconstruction in dorsal hand defects requiring free flaps.

This study reports results in nine patients with extensive loss of soft tissue, extensor tendon, and bone, treated with an emergency free flap for skin cover, primary bone grafts, and tendon grafts passed through individual tunnels in the free flap. Four had a good result, four were fair and one poor. Six patients returned to work, two were not working and one was retired. In select patients, emergency reconstruction of severe extensor tendon injuries appears to produce better function, with fewer operations, a shorter hospital stay, minimal complications, and a shorter period of disability.

Adolescent↗
PubMed1993

Regulation of gamma-aminobutyric acid synthesis in the brain.

gamma-Aminobutyric acid (GABA) is synthesized in brain in at least two compartments, commonly called the transmitter and metabolic compartments, and because regulatory processes must serve the physiologic function of each compartment, the regulation of GABA synthesis presents a complex problem. Brain contains at least two molecular forms of glutamate decarboxylase (GAD), the principal synthetic enzyme for GABA. Two forms, termed GAD65 and GAD67, are the products of two genes and differ in sequence, molecular weight, interaction with the cofactor, pyridoxal 5'-phosphate (pyridoxal-P), and level of expression among brain regions. GAD65 appears to be localized in nerve terminals to a greater degree than GAD67, which appears to be more uniformly distributed throughout the cell. The interaction of GAD with pyridoxal-P is a major factor in the short-term regulation of GAD activity. At least 50% of GAD is present in brain as apoenzyme (GAD without bound cofactor; apoGAD), which serves as a reservoir of inactive GAD that can be drawn on when additional GABA synthesis is needed. A substantial majority of apoGAD in brain is accounted for by GAD65, but GAD67 also contributes to the pool of apoGAD. The apparent localization of GAD65 in nerve terminals and the large reserve of apoGAD65 suggest that GAD65 is specialized to respond to short-term changes in demand for transmitter GABA.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Effects of increased gamma-aminobutyric acid levels on GAD67 protein and mRNA levels in rat cerebral cortex.

Rats were injected with saline or the gamma-aminobutyric acid (GABA) transaminase inhibitor gamma-vinyl-GABA for 7 days and the effects on GABA content and glutamic acid decarboxylase (GAD) activity, and the protein and mRNA levels of the two forms of GAD (GAD67 and GAD65) in the cerebral cortex were studied. gamma-Vinyl-GABA induced a 2.3-fold increase in GABA content, whereas total GAD activity decreased by 30%. Quantitative immunoblotting showed that the decline in GAD activity was attributable to a 75-80% decrease in GAD67 levels, whereas the levels of GAD65 remained unchanged. RNA slot-blotting with a 32P-labeled GAD67 cDNA probe demonstrated that the change in GAD67 protein content was not associated with a change in GAD67 mRNA levels. Our results suggest that GABA specifically controls the level of GAD67 protein. This effect may be mediated by a decreased translation of the GAD67 mRNA and/or a change in the stability of the GAD67 protein.

Aminocaproates↗
PubMed1993

Synaptosomal glutamate uptake declines progressively in the spinal cord of a mutant mouse with motor neuron disease.

It has been suggested that the degeneration of motor neurons in amyotrophic lateral sclerosis is a consequence of excitotoxicity resulting from a loss of synaptosomal glutamate uptake. The role of synaptosomal glutamate uptake in the pathogenesis of motor neuron disease was studied in the Mnd mouse. Glutamate uptake in spinal-cord synaptosomes declined in parallel with the onset of behavioral deficits in Mnd mice but lagged considerably behind the appearance of pathology in motor neurons. Glutamate uptake did not decline significantly in corpus striatum, and GABA uptake did not change significantly in either spinal cord or striatum. The presence of pronounced histopathological changes before the loss of glutamate uptake suggests that the decline of glutamate uptake is a consequence rather than the primary cause of motor neuron disease in the Mnd mouse.

Animals↗
PubMed1992

Synthesis and release of neuroactive substances by glial cells.

Glia contain, synthesize, or release more than 20 neuroactive compounds including neuropeptides, amino acid transmitters, eicosanoids, steroids, and growth factors. The stimuli that elicit release differ among compounds but include neuropeptides, neurotransmitters, receptor agonists, and elevated external [K+]. The mechanisms of release are poorly understood in most cases. Many of the neuroactive compounds are localized in discrete subpopulations of glia. Thus, glia are equipped to send as well as receive chemical messages and appear to be present as classes of cells with differing abilities to communicate chemically. It is possible that glia are as diverse as neurons in their functional characteristics.

Animals↗
PubMed1992

Increased intracellular gamma-aminobutyric acid selectively lowers the level of the larger of two glutamate decarboxylase proteins in cultured GABAergic neurons from rat cerebral cortex.

The regulation of glutamate decarboxylase (GAD; EC 4.1.1.15) was studied by using cultures of cerebral cortical neurons from rat brain grown in serum-free medium. About 50% of the neurons in the cultures were gamma-aminobutyric acid (GABA)ergic as determined by two double-staining procedures. Immunoblotting experiments with four anti-GAD sera that recognize the two forms to varying degrees, demonstrated that the cultures contained the two forms of GAD that are present in rat brain (apparent molecular masses = 63 and 66 kDa). GAD activity was reduced by 60-70% when intracellular GABA levels were increased by incubating the cultures with the GABA-transaminase inhibitor gamma-vinyl-GABA for greater than 5-10 h or with 1 mM GABA itself. Neither baclofen nor muscimol (100 microM) affected GAD activity. Immunoblotting experiments showed that only the larger of the two forms of GAD (66 kDa) was decreased by elevated GABA levels. These results, together with previous results indicating that the smaller form of GAD is more strongly regulated by pyridoxal 5'-phosphate (the cofactor for GAD), suggest that the two forms of GAD are regulated by different mechanisms.

4-Aminobutyrate Transaminase↗
PubMed1991

Isoproterenol selectively releases endogenous and [14C]-labelled taurine from a single cytosolic compartment in astroglial cells.

Taurine is stored in and released from astroglial cells. We have investigated whether taurine is stored in multiple subcellular compartments and whether taurine is released from a distinct pool. Taurine compartmentation was examined by determining the subcellular distribution of labelled taurine and by comparing the specific radioactivities of taurine in cells with that of the taurine released into the superfusion medium during receptor stimulation. Three observations indicate that taurine is found in a single subcellular pool. First, labelled taurine was localized exclusively within a freely exchangeable pool and not sequestered in membrane-bound compartments. Second, endogenous and newly acquired [14C]taurine was released identically by the beta-adrenergic agonist isoproterenol (IPR). Third, measurements of the specific radioactivity of taurine indicate that accumulated [14C]taurine mixes homogeneously with endogenous taurine and both are simultaneously released from the same pool. Chemical analysis revealed net uptake of taurine from the medium during loading and a net loss during IPR-stimulated release. Taurine was selectively released by IPR; five other amino acids found in the superfusion medium were unaffected. One possible mechanism capable of explaining these results is that receptor-mediated taurine release from astroglia occurs by a selective transporter moving taurine from a single cytoplasmic pool to the extracellular space.

Amino Acids↗
PubMed1991

GAD and GABA in an enriched population of cultured GABAergic neurons from rat cerebral cortex.

To study various aspects of GABAergic metabolism in an easily accessible system, dissociated cells from postnatal rat cerebral cortex were cultured in a serum-based medium and characterized morphologically and biochemically. The majority (70-96%) of the neurons were GABAergic as determined by three double-labeling procedures. The specific activity of glutamine synthetase in the cultures was 4-5% of the levels in rat astrocyte cultures and intact rat brain, indicating that glia were a minor component. The developmental increase of GABA levels preceded the increase of GAD activity in both immunocytochemical and biochemical experiments. GABA turnover rates also increased with culture age and were 20-30% of GAD activity. Four anti-GAD antibodies, which recognize GAD subunits with differing molecular masses to varying degrees, were used to stain cultured neurons and make immunoblots. Immunoblots showed that the neurons contained two major subunits of GAD which differed in mass by 2 kDa. All four antibodies immunostained both neuronal perikarya and neurites but one antibody, which on the immunoblots predominantly labeled the GAD protein with the lower molecular weight, showed a somewhat more pronounced punctate staining, possibly indicating a principal localization to neurites.

Amino Acids↗
PubMed1991

GABA synthesis in brain slices is dependent on glutamine produced in astrocytes.

The rate of gamma-aminobutyric acid (GABA) synthesis in rat-brain slices was determined by inhibiting GABA transaminase with 20-microM gabaculine and measuring the increase of GABA. Added 500-microM glutamine increased the rate of GABA synthesis by 50%, indicating that glutamate decarboxylase is not saturated in brain slices. The stimulation of GABA synthesis with added glutamine in brain slices was much less than that reported for synaptosomes. The lower stimulation in slices was attributable to astrocytic glutamine production, as the rate of GABA synthesis decreased by 44% when glutamine production was inhibited with methionine sulfoximine. Added glutamine restored the rate to the maximal value observed in brain slices. The rate of GABA synthesis was decreased by 65% in slices pretreated with an inhibitor of glutaminase, and added glutamine did not reverse this effect. These results suggest that glutamine produced by astrocytes is a quantitatively important precursor of GABA synthesis in cortical slices.

4-Aminobutyrate Transaminase↗
PubMed1991

Cofactor interactions and the regulation of glutamate decarboxylase activity.

More than 50% of glutamate decarboxylase (GAD) in brain is present as apoenzyme. Recent work has opened the possibility that apoGAD can be studied in brain by labeling with radioactive cofactor. Such studies would be aided by a compound that inhibits specific binding. One possibility is 4-deoxy-pyridoxine 5'-phosphate, a close structural analog of the cofactor pyridoxal 5'-phosphate. The effects of deoxypyridoxine-P on the cyclic series of reactions that interconverts apo- and holoGAD was investigated and found to be consistent with simple competitive inhibition of the activation of apoGAD by pyridoxal-P. As expected from the cycle GAD was inactivated when incubated with glutamate and deoxypyridoxine-P even though cofactor was present, but no inactivation was observed with deoxypyridoxine-P in the absence of glutamate. Deoxypyridoxine-P also stabilized apoGAD against heat denaturation. These effects were quantitatively accounted for by a kinetic model of the apo-holoGAD cycle. Deoxypyridoxine-P inhibited the labeling by [32P]pyridoxal-P of GAD isolated from rat brain. Hippocampal extracts were labeled with [32P]pyridoxal-P and analyzed by SDS-polyacrylamide gel electrophoresis. Remarkably few bands were strongly labeled. The major labeled band (at 63 kDa) corresponded to one of the forms of GAD. Other strongly-labeled bands were observed at 65 kDa (corresponding to the higher molecular weight form of GAD) and at 69--72 kDa. Labeling of the 63- and 65-kDa bands was inhibited by deoxypyridoxine-P, but the 69-72 kDa bands were unaffected, suggesting that the latter were non-specifically labeled. The results suggest that the 63-kDa form of GAD makes up the majority of apoGAD in hippocampus.

Animals↗
PubMed1991

Regulatory properties of brain glutamate decarboxylase (GAD): the apoenzyme of GAD is present principally as the smaller of two molecular forms of GAD in brain.

The apoenzyme of glutamate decarboxylase [enzyme without bound cofactor, pyridoxal 5'-phosphate (pyridoxal-P)] serves as a reservoir of inactive glutamate decarboxylase (GAD) that can be activated when additional GABA synthesis is required. We have investigated which of two molecular forms of GAD is present as apoenzyme in synaptosomes and in cortex, caudate nucleus, hippocampus, and cerebellum of rat brain. Endogenous glutamate apodecarboxylase (apoGAD) was labeled by incubating extracts of synaptosomes or punches of each region with 32P-pyridoxal-P, followed by reduction with NaBH4, to link covalently the 32P-pyridoxal-P to GAD. Proteins were separated by SDS-PAGE. Punches from all four brain regions and forebrain synaptosomes contained two forms of GAD with apparent Mrs of 63 and 65 kDa as identified by immunoblotting with four antiGAD sera. Punches and synaptosomes contained a major 32P-pyridoxal-P-labeled band with an apparent Mr of 63 kDa that was stained on immunoblots by the antiGAD serum 1440 and the monoclonal antibody GAD-6, and a minor labeled band at 65 kDa that was stained by the 1440, 6799, and K2 antisera. Synaptosomes contained remarkably few other strongly labeled proteins, but punches contained several other labeled bands. Three additional lines of evidence indicate that the labeled 63-kDa protein is apoGAD: (1) it was purified by immunoaffinity chromatography with the GAD-1 monoclonal antibody; (2) it yielded one major labeled peptide when digested with chymotrypsin, and that peptide appeared identical in peptide-mapping experiments to the labeled active-site peptide isolated from chromatographically prepared rat brain GAD; and (3) its labeling was selectively blocked by 4-deoxypyridoxine 5'-phosphate, a competitive inhibitor of the binding of pyridoxal-P to GAD.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

The epidemiology and clinical aspects of the hemolytic uremic syndrome in Minnesota.

BACKGROUND: The frequency of the hemolytic uremic syndrome, characterized by microangiopathic hemolytic anemia, thrombocytopenia, and renal failure, is increasing. Although Escherichia coli serotype O157:H7 has been implicated as a causative agent, more information is needed about the basic epidemiology and clinical aspects of this syndrome. METHODS: We conducted a retrospective population-based study of hemolytic uremic syndrome in Minnesota residents less than 18 years of age from 1979 through 1988 to assess trends in disease occurrence, describe the clinical illness, and identify predictors of disease severity and outcome. We also conducted a case-control study of patients with onsets of illness from 1986 through 1988 to examine risk factors. RESULTS: One hundred seventeen patients were identified. The mean annual incidence increased from 0.5 case per 100,000 child-years among children less than 18 in 1979 (6 cases) to 2.0 cases per 100,000 in 1988 (26 cases) (P = 0.000004). E. coli O157:H7 was isolated from 13 of 28 patients (46 percent) who had stool specimens submitted for testing. For those who presented with typical hemolytic uremic syndrome, an elevated polymorphonuclear-leukocyte count on hospital admission, a shorter duration of prodrome, and the presence of bloody diarrhea were predictive of severe disease. In the case-control study, the patients were more likely to attend large daycare centers (more than 50 children) than were the controls (odds ratio, 10.2; P = 0.03), suggesting that day-care attendance may be a risk factor. On the basis of the population-attributable risk, however, this factor could account for no more than 16 percent of the cases. CONCLUSIONS: This study provides evidence for an increase in the incidence of hemolytic uremic syndrome, which is probably related to an increased incidence of E. coli O157:H7 infections. Hemolytic uremic syndrome has become an important pediatric and public health problem.

Adolescent↗
PubMed1990

The use of semi-permeable membranes for wound management.

This paper reports a study comparing bags made from expanded polytetrafluoroethylene semi-permeable membranes and standard plastic bags to treat burns of the hands. There was much less maceration in the PTFE bag, which allowed easier assessment of the extent and depth of burn, and only one daily dressing change was needed. The two types of bag were similar with regard to ease of use, low bacterial contamination and allowing good mobilisation of the hand. Ease of use has meant that a greater number of patients with superficial burns can be treated as outpatients and many are able to do their own daily dressing change, so fewer attendances at the clinic are needed.

Adult↗
PubMed1990

Effects of ethanol and acetaldehyde on collagen synthesis, prostaglandin release and resorption of fetal rat bone in organ culture.

We tested the effect of ethanol and its metabolite, acetaldehyde, on bone formation as measured by [3H]proline incorporation into collagenase digestible protein (CDP) and noncollagen protein (NCP), and on DNA synthesis as measured by [3H]thymidine (TdR) incorporation in fetal rat calvaria. We also determined the effects of ethanol and acetaldehyde on prostaglandin E2 (PGE2) release from calvaria and on bone resorption as measured by 45Ca release from fetal rat long bones. Bones were cultured in multiwell plastic dishes (open system) or in stoppered Erlenmeyer flasks (closed system) for 24 to 96 h. In the open system, 1% ethanol (v/v; 172 mM) resulted in a 31% decrease in TdR incorporation at 24 h with no effect on CDP and NCP. At 0.1% (17.2 mM), ethanol increased TdR by 22%, CDP by 73% and NCP by 67% at 24 h, but these effects were not sustained at 96 h. At 24 h, 1% and 0.3% ethanol decreased PGE2 release by 88% and 75% respectively. This effect was sustained for 96 h only at the higher concentration. In the closed system, 0.1% ethanol increased TdR incorporation by 38% at 24 h. However, there was no effect on the labeling of CDP or NCP. Because its boiling point is 21 degrees C, acetaldehyde could only be tested in the closed system. Acetaldehyde markedly inhibited bone metabolism. At 24 h, 0.003% (0.54 mM) to 0.01% (1.79 mM) acetaldehyde caused a dose-related inhibition of TdR incorporation from 23 to 45%. At 0.01% and 0.03% acetaldehyde inhibited proline incorporation into CDP by 48% and 94% and NCP by 40% and 74% respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetaldehyde↗