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Biomedical subjects

A Hunt

Publications and source records attributed to A Hunt.

At least 73 records · Page 4Linked to original sources
PubMed1991

Complete cDNA sequence of human complement pro-C5. Evidence of truncated transcripts derived from a single copy gene.

Two truncated human C5 clones, pHC5A and pHC5B, were isolated from an adult human liver cDNA library, and contained inserts of 2930 and 2181 bp, respectively. Both clones were polyadenylated and encoded the 5'-end of the C5 pro-molecule, thereby completing the human pro-C5 cDNA sequence. However, near the 3'-ends, at exon/intron boundaries, the nucleotide sequences of pHC5A and pHC5B diverged from each other and from the full-length 6.0-kb C5 cDNA sequence. Clone pHC5A, which overlapped the first human C5 clone described (J-16), encoded most of the C5 signal peptide, the complete beta-chain, the linker peptide, 177 amino acids of the alpha-chain, and contained 144 bp of Alu family consensus sequence encoding 48 amino acids of divergent protein sequence in an open reading frame. Clone pHC5B encoded the entire C5 signal peptide, the beta-chain, the linker peptide, nine amino acids of the alpha-chain, and six amino acids of divergent protein sequence in an open reading frame. Northern blot experiments demonstrated the presence of a 3.0-kb truncated C5 mRNA in adult human liver and a 4.8-kb truncated C5 mRNA in HepG2 cells in addition to the 6.0-kb full-length transcript. Truncated C5 mRNA were not detected in Raji, MOLT-4, human fibroblast or U937 cells, although the full-length 6.0-kb transcript was seen in MOLT-4 cells. Southern blot analyses indicated that the human C5 structural gene is large, complex, and is present in the human genome in a single copy, thereby demonstrating that the truncated C5 clones and mRNA are derived from a single C5 gene by alternative processing events.

Amino Acid Sequence↗
PubMed1991

Development and use of a pneumoconiosis database of human pulmonary inorganic particulate burden in over 400 lungs.

Over 400 cases with data from in situ electron microprobe quantitation of non-fibrous inorganic particles (e.g., silica, alumino-silicates, talc, metals) in pulmonary tissue sections, and data from quantitative digestion analyses for fiber content (e.g., asbestos, silica, alumino-silicates, man-made fibers, talc) comprise an extensive microcomputer data set of lung particle burden. When allied with demographic and histopathologic information the result is a comprehensive database of occupational pulmonary pathology. Examples of the kinds of information which can be extracted from the database include: 1) summary information on the types sizes and associations of particles in lungs with a variety of exposures, 2) concentrations of etiologic particle type in cases with recognized pneumoconioses, and 3) correlations between particle type, pathology, occupation and social history. The database provides a powerful tool for assessing such information on statistically meaningful sample sets.

Asbestos↗
PubMed1990

Development and regulation of excitatory amino acid receptors involved in the release of arachidonic acid in cultured hippocampal neural cells.

Release of [3H]arachidonic acid mediated by excitatory amino acid (EAA) receptors was investigated from prelabelled primary cultures of hippocampal neurons and astroglial cells. Treatment with N-methyl-D-aspartate (NMDA), quisqualate (QA) and kainate resulted in age- and dose-dependent stimulation of [3H]arachidonic acid release. During development, the maximum response for NMDA was observed relatively earlier (at 7 days) than those for QA and kainate (at 14 days) in the hippocampal neuronal cultures. The half maximal effects were obtained at about 15 microM NMDA at all ages studied and about 0.5 microM QA at 14 and 20 days. At optimum concentrations NMDA- and QA-induced releases were additive. Unlike with neurons, treatment with all the 3 EAA receptor agonists, NMDA, QA and kainate, had no significant effect on [3H]arachidonate release in hippocampal astroglial cells. In cultured 14-day-old neurons, the increases in NMDA- and QA-mediated [3H]arachidonic acid release were completely blocked by the NMDA receptor antagonist, 2-amino-5-phosphonovaleric acid, and the ionotropic QA receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, respectively. But the iontropic QA receptor agonist alpha-amino-3-hydroxy-5-methyl-isoxazole-4- propionic acid (AMPA) had no significant effect on [3H]arachidonate release, indicating that interaction between ionotropic QA and metabolotropic QA receptors may be essential for optimal QA-mediated arachidonic acid release. At physiological concentrations of Mg2+ (1.2 mM), AMPA was found to potentiate NMDA-induced release of [3H]arachidonic acid; the effect appeared to be related to a removal of Mg2+ blockade mediated by mild depolarisation.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1990

Exposure to N-methyl-D-aspartate increases release of arachidonic acid in primary cultures of rat hippocampal neurons and not in astrocytes.

The release of [3H]arachidonic acid (ARA) was investigated from prelabelled primary cultures of hippocampal neurons and astroglial cells. The activation of N-methyl-D-aspartate (NMDA) subtype of glutamate receptors resulted in a dose-dependent stimulation of [3H]ARA release. The half maximal effect was obtained at about 15 microM NMDA, whereas the maximum concentration (50 microM NMDA) produced about a 2-fold increase in 7-day-old cultures. This elevation in [3H]ARA release was blocked in a dose-related manner by the NMDA receptor antagonist, 2-amino-5-phosphonovaleric acid (APV), and by Mg2+ which blocks NMDA receptor-linked Ca2+ ion channels. The removal of external Ca2+ inhibited NMDA-induced release, whereas treatment with calcimycin (A 23187, a Ca2+ ionophore) greatly increased the [3H]ARA release. The inhibitors of phospholipase A2, nordihydroguaiaretic acid and mepacrine, decreased the NMDA-dependent [3H]ARA release in a dose-related manner, maximum inhibition reaching to about 90% at high doses. Entry of Ca2+ brought about by opening the voltage-sensitive channels by high K+ had no effect on the release of [3H]ARA, indicating that NMDA gated channels are situated in a part of the neuron where Ca2+ entry through this route is more efficiently coupled to the activation of phospholipase A2. Treatment with NMDA had no significant effect on [3H]ARA release in hippocampal astroglial cells as opposed to neurons. This was not due to inability of astrocytes to release ARA, for ATP still evoked [3H]ARA release, and this was markedly inhibited by mepacrine. It is suggested that ARA act as both intracellular and intercellular messengers in the functioning of NMDA receptors in synaptic transmission and plasticity in the hippocampus.

Adenosine Triphosphate↗
PubMed1990

Quinolinic acid promotes the biochemical differentiation of cerebellar granule neurons.

The possible involvement of quinolinic acid in the biochemical differentiation of cultured glutamatergic cerebellar granule neurons was studied in terms of the activity of phosphate-activated glutaminase (GLNase) and aspartate aminotransferase (ASP-AT). Treatment with quinolinate elevated the specific activity of GLNase and amount of protein per culture dish in a dose-dependent manner. The half maximal effect was obtained at about 0.5 mM quinolinate, whereas the maximum concentration, which produced about a 2.3-fold increase in GLNase activity, was about 2 mM. Quinolinate, like N-methyl-D-aspartate (NMDA), had no significant effects on the activities of ASP-AT and lactate dehydrogenase enzymes. The increases in the activity of GLNase and amount of protein were completely blocked by the NMDA receptor antagonist, 2-amino-5-phosphonovaleric acid. The result would indicate that, (a) contrary to an earlier proposal, ASP-AT does not appear to be a good marker for studying dynamic responses of glutamatergic neurons, and (b) the trophic effect of quinolinic acid on the development of cerebellar granule neurons is mediated by selective activation of NMDA subtype excitatory amino acid receptors.

Animals↗
PubMed1990

Histamine increases ornithine decarboxylase activity in primary cultures of cerebellar granule cells.

The effect of histamine on the activity of ornithine decarboxylase (ODC) of cerebellar granule neurons was studied using primary cultures grown both in serum-containing medium and in chemically defined medium. In comparison with granule neurons grown in chemically defined medium, the activity of ODC was about twice as great in the neurons grown in serum-containing medium. Treatment of cultured cerebellar neurons with histamine caused a dose-dependent increase in ODC activity. The maximum elevation was observed at 500 nM of histamine, when the increase in ODC activity was about 50% and 120% over controls in granule cells grown in serum-containing medium and in chemically defined medium, respectively. Histamine had no significant effect on the activity of lactate dehydrogenase in these cultures. The present findings provided direct evidence for the involvement of histamine in the regulation of ODC-related non-mitotic growth of granule neurons in the cerebellum.

Animals↗
PubMed1990

Death of subcortical cholinergic neurons in certain neurodegenerative disorders may not be due to an overstimulation of N-methyl-D-aspartate receptors.

Stimulation of N-methyl-D-aspartate (NMDA) receptors increases the activity of enzymes involved in the synthesis of nerve cell specific neurotransmitters. In the present study this phenomenon has been used to identify the neurons in the septal-diagonal band region having NMDA receptors. Exposure of cultures relatively enriched in subcortical cholinergic neurons to a depolarizing concentration of K+ (40 mM) significantly enhanced the expression of choline acetyltransferase (ChAT) activity. In contrast, when these septal cells were treated with as much as 100 microM NMDA no significant increase was observed in the activity of choline acetyltransferase, although there was a marked enhancement in glutamate decarboxylase activity. The results would indicate that subcortical cholinergic neurons do not possess excitatory amino acid receptors of the NMDA subtype, and that therefore neurotoxicity mediated through NMDA receptors may not be involved in the death of cholinergic neurons in degenerative disorders of the brain, such as Down's syndrome and Alzheimer's disease, which entail major losses of these neurons.

Animals↗
PubMed1990

Cell-type specific effects of N-methyl-D-aspartate on biochemical differentiation of subcortical neurons in culture.

The possible involvement of N-methyl-D-aspartate (NMDA) receptors in the biochemical differentiation of cultured neurons derived from the medial frontal part of the forebrain containing the septum-diagonal band region was studied in terms of the activities of enzymes important in the synthesis of neurotransmitter compounds. The activity of choline acetyltransferase (ChAT) was used as a marker for cholinergic neurons, glutamate decarboxylase (GAD) for GABAergic neurons and phosphate-activated glutaminase (GLNase) and aspartate aminotransferase (ASP-AT) for glutamatergic neurons, while lactate dehydrogenase (LDH) was included as an ubiquitous enzyme. The exposure of cultures to a depolarizing concentration of K+ (40 mM) for the last 3 days (i.e. between 2 and 5 days in vitro) significantly enhanced the expression of ChAT, GAD and GLNase activities, but high K+ caused little alteration in the activities of ASP-AT and LDH. On the other hand, treatment with NMDA markedly elevated the specific activities of GAD and GLNase only, and the compound had no significant effects on the activities of ChAT, ASP-AT and LDH enzymes. The enhancements of the specific activities of GAD and GLNase were completely blocked by the NMDA receptor antagonist, 2-amino-5-phosphonovaleric acid, and by the NMDA receptor-linked Ca2+ ion channel blocker, MK-801. On the basis of the present findings it is concluded that, (a) contrary to an earlier proposal, ASP-AT does not appear to be a good marker for the glutamatergic neurons, (b) the failure of the subcortical cholinergic neurons to respond by an increase in ChAT activity to NMDA may indicate that these nerve cells lack NMDA subtype excitatory amino acid receptors, and (c) as the septal GABAergic input in the hippocampus is involved in the modulation of long-term potentiation, the presence of NMDA receptors on these neurons would now suggest that NMDA receptors are linked to both the initiation and the modulation of hippocampal plasticity in the mammalian brain.

Animals↗
PubMed1990

A simple protocol for the screening and preliminary identification of antibodies to human IgA using unpurified proteins in the passive haemagglutination test.

Experiments have shown that suitably diluted, unpurified IgA paraprotein sera can be substituted for chemically purified IgA proteins in the passive haemagglutination test for the detection of anti-IgA antibodies. The substitution of unpurified IgA paraprotein was made possible by the enhanced serological activity obtained by pre-heating the unpurified immunoglobulin preparations at 56 degrees C for 60 min. Using a small screening panel of suitably coated tanned red cells, various population groups were investigated for the presence of anti-IgA antibodies. Results were similar to those obtained by other workers using purified IgA preparations in the passive haemagglutination test.

Antibodies, Anti-Idiotypic↗
PubMed1990

Sustained improvement in left ventricular function after successful coronary angioplasty.

The short and long term effects of successful percutaneous transluminal coronary angioplasty on left ventricular function, at rest and on exercise were investigated in 49 patients. Thirty-four had had no previous infarction (group 1) and 15 had (group 2). Technetium-99m gated blood pool images were obtained at rest and during exercise before, six weeks after, and a mean of fifteen months after successful angioplasty. Before angioplasty the mean (SD) ejection fraction fell significantly on exercise in both groups from 58 (10)% to 53 (13)% in group 1 and from 48 (10)% to 40 (16)% in group 2. This change was paralleled by a worsening wall motion score (from 0.6 (0.4) to 1.6 (1.2) in group 1 and from 2.3 (1.9) to 3.3 (2.4) in group 2). Six weeks after the procedure there was little change in resting ejection fraction but it increased significantly on exercise (to 62 (11)% in group 1 and to 53 (13)% in group 2). There was a concomitant significant improvement in the exercise wall motion score (to 0.4 (0.6) in group 1 and to 1.8 (1.1) in group 2). This improvement in exercise ejection fraction and wall motion was maintained at later follow up with no significant deterioration in either variable and a clearly sustained improvement in ejection fraction (60 (10)% in group 1 and 51 (10)% in group 2) and wall motion score (0.2 (0.2) in group 1 and 1.3 (0.8) in group 2) compared with values before angioplasty. The initial improvement in left ventricular function on exercise after successful angioplasty was maintained for at least 9-24 months both in patients with previous myocardial infarction and in those without.

Angioplasty, Balloon, Coronary↗
PubMed1990

Hemiparkinsonism-hemiatrophy syndrome: clinical and neuroradiologic features.

We evaluated 11 patients with hemiparkinson-hemiatrophy syndrome, 6 with body and contralateral cerebral hemispheric hemiatrophy, 4 with only body hemiatrophy, and 1 with just brain hemiatrophy. The mean age of symptom onset was 38.1 years (range, 18 to 54) with 5.2 +/- 3.1 (mean +/- SD) years of illness until the last follow-up visit. The presenting symptom was unilateral tremor in 6 patients, hand dystonia in 2, bradykinesia in 2, and abnormal gait in 1 patient. Three patients had a good response to levodopa, 4 had moderate response, and 2 patients had a poor response. During a mean follow-up period of 1.7 years (range, 4 months to 5 years), the Hoehn and Yahr score changed in only 3 patients: 2 gained 1.5 points and 1 gained 3 points over 2.5 years. We discuss the association between hemiparkinsonism-body hemiatrophy and contralateral hemispheric hemiatrophy, and raise the possibility of early childhood brain insult with delayed-onset parkinsonism.

Adolescent↗
PubMed1989

Regulation of production by primary cultures of rat forebrain astrocytes of a trophic factor important for the development of cholinergic neurons.

The possibility was examined for the production of a trophic factor by astrocytes important for the biochemical development of cholinergic neurons. The activity of choline acetyltransferase (ChAT) was used as a marker to study the maturation of cholinergic neurons, while the metabolic state of astrocytes was monitored in terms of glutamine synthetase activity. When the dissociated cells, derived from the septal-diagonal band region of embryonic rat brain, were cultured either on a preformed layer of highly enriched astrocytes or in the presence of astrocyte-conditioned medium, a marked increase was observed in the expression of ChAT activity. Similar effect was not noticed when cerebellar granule cells replaced the astrocytes. The production of cholinergic factor was decreased when the astroglial cells were induced to proliferate by epidermal growth factor, whereas its production was increased when quiescent astrocytes were treated with a low dose of cytosine arabinoside. The results would indicate that a trophic factor important for the development of cholinergic cells is produced by astrocytes, and that exogenously added agents influencing astroglial metabolism have a marked regulatory effect on the production of this trophic factor.

Animals↗
PubMed1989

Balloon occlusion during coronary angioplasty as a model of myocardial ischaemia: reproducibility of sequential inflations.

In order to evaluate the potential of balloon occlusion during coronary angioplasty as a model of myocardial ischaemia in man we have measured coronary sinus blood flow (CSBF), myocardial oxygen consumption (MVO2), lactate extraction (LER) and electrocardiographic changes in 11 patients undergoing left anterior descending artery (LAD) angioplasty. Baseline measurements were made before balloon crossing and between inflations. Four consecutive inflations each of 60 s duration were made; 5 min return to baseline was allowed between inflations. There was a significant reduction in CSBF and MVO2 (ml min-1) during inflations 2, 3 and 4 (CSBF: 121 +/- 66----94 +/- 53, 113 +/- 49----99 +/- 42, 124 +/- 66----102 +/- 41, P less than 0.02; MVO2: 11.3 +/- 6.6-9.1 +/- 3.9, 10.4 +/- 3.7-8.7 +/- 2.4, 12.2 +/- 4.4----9.4 +/- 2.8, P less than 0.05). However during the first period of balloon occlusion there were inconsistent changes in coronary flow with an overall rise in mean flow (97 +/- 35----128 +/- 80 ml min-1, P = NS) and an overall rise in mean myocardial oxygen consumption (9.6 +/- 3.8----12.5 +/- 7.5 ml min-1, P = NS). There was lactate production during all four inflations but the changes during the first one did not achieve statistical significance. These inconsistent changes during the first inflation were thought to be due to partial obstruction of the stenosis by the deflated balloon before primary dilatation. The changes due to crossing and during the first two inflations were further investigated in another group of 12 patients undergoing LAD angioplasty. Great cardiac vein flow (GCVF), CSBF, MVO2 and LER were recorded at baseline, during crossing and during the first two inflations. With the deflated balloon across the stenosis there were no changes in CSBF or MVO2 but there was a fall in GCVF (103 +/- 28----77 +/- 50, P = NS) and a significant fall in LER (77 +/- 57----16 +/- 37, P less than 0.01). Although there was a fall during the first inflation in CSBF, GCVF, MVO2 and lactate extraction none of these changes were significant. During the second inflation these changes were of greater magnitude and achieved statistical significance. While balloon occlusion during coronary angioplasty has the potential of providing a model of ischaemia in man we have found the first inflation period unreliable, due to the variable degree of occlusion by the deflated balloon. We suggest that only subsequent inflations after the primary dilatation are used for observations.(ABSTRACT TRUNCATED AT 400 WORDS)

Angioplasty, Balloon, Coronary↗
PubMed1989

Isolation and characterization of a mutant of Haemophilus influenzae type b deficient in outer membrane protein P1.

The gene for outer membrane protein P1 of Haemophilus influenzae type b has been previously cloned and expressed in Escherichia coli. To investigate the physiologic role of the P1 protein, the cloned P1 gene was insertionally inactivated with the Tn5 derivative Tn5tac1, and an isogenic P1-deficient Haemophilus mutant was then generated by transformation with linearized plasmid DNA containing the insertionally inactivated gene. The P1-deficient strain grew normally in vitro and induced bacteremia in the infant rat model.

Bacterial Outer Membrane Proteins↗
PubMed1988

A novel way of removing quiescent astrocytes in a culture of subcortical neurons grown in a chemically defined medium.

A new method has been described for removing a very small number of contaminating astrocytes in neuronal cultures (derived from the septal-diagonal band region of 17-day-old embryonic rat brain) grown in a chemically defined medium. The proportion of these glial fibrillary acidic protein (GFAP)-positive cells was usually less than 1.5% up to 10 days, but thereafter their number increased rapidly reaching 10-15% by 22 days in vitro. A prolonged exposure to normally used concentration of cytosine arabinoside (Ara-C; 10 microM) was toxic to both astroglial and neuronal cells, while a brief treatment (48 h) with a low level (4 microM) of Ara-C failed to eliminate these astrocytes, as judged by glutamine synthetase activity and GFAP-positive cell count. However, these quiescent astroglial cells could be easily eliminated if they were induced to proliferate by epidermal growth factor before exposure to Ara-C. The combined treatment with these agents had no effect on the number of acetylcholinesterase-positive cells, and on the development of cholinergic and GABA-ergic neurons, as measured in terms of choline acetyltransferase and glutamate decarboxylase activity, respectively.

Animals↗
PubMed1988

The mechanism of cytosine arabinoside toxicity on quiescent astrocytes in vitro appears to be analogous to in vivo brain injury.

Neuronal cultures derived from the septal diagonal band region of the embryonic rat brain and grown in a chemically defined medium contained a very small number of contaminating astroglial cells. During the first week in culture, these cells were well dispersed in the form of a single isolated cell with fine fibrous branched processes. Treatment with 4 microM cytosine arabinoside for 24 h failed to kill these astrocytes (most probably present in quiescent form), as judged by glutamine synthetase activity and glial fibrillary acidic protein-positive cell count. On the other hand, the exposure of cultures to cytosine arabinoside resulted in a marked increase in choline acetyltransferase enzyme activity. The overall results, together with our previous findings, are consistent with the proposal that a brief exposure to a relatively low concentration of cytosine arabinoside induces quiescent astrocytes to produce a large quantity of a neurotrophic factor that is involved in the regulation of cholinergic cells.

Animals↗
PubMed1988

Topographical distribution of neurochemical changes in Alzheimer's disease.

Biochemical indices of cortical nerve cells affected in Alzheimer's disease have been proposed (excitatory dicarboxylic amino acid, EDAA, sodium-dependent carrier; phosphate-activated glutaminase activity; serotonin type 2 recognition site; somatostatin-like immunoreactivity). These and the content of EDAAs and two related amino acids, and choline acetyltransferase (ChAT) activity have been measured in up to 13 areas of cerebral cortex and the cerebellar cortex from 16 patients with Alzheimer's disease and 17 controls. Reduction of the index of the serotonin recognition site, somatostatin content and another biochemical index of interneurones coincide and indicate a rather unexpected focal loss of such neurones from the parietal lobe. No unequivocal measure of the integrity of pyramidal neurones could be established as the content of no amino acid was reduced, the index of the EDAA carrier showed evidence of change in few brain regions and glutaminase activity was subject to unexplained variability. ChAT activity alone closely paralleled a previous report of the distribution of morphological degeneration. The results are discussed in relation to therapy and positron emission tomography.

Aged↗