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Biomedical subjects

M J Duggan

Publications and source records attributed to M J Duggan.

At least 37 records · Page 2Linked to original sources
PubMed1990

Biochemical evidence for the existence of gamma-aminobutyrateA receptor iso-oligomers.

Polyclonal antibodies were raised against synthetic peptides whose sequences were from unique regions of the bovine gamma-aminobutyrateA receptor alpha 1, alpha 2, and alpha 3 subunits. The anti-alpha 1 324-341, anti-Cys alpha 2 414-424, and anti-Cys alpha 3 454-467 antibodies all specifically immunoprecipitated [3H]flunitrazepam and [3H]muscimol binding activities in parallel from Na+ deoxycholate extracts of bovine cerebral cortex. The maximum number of benzodiazepine binding sites immunoprecipitated by each antibody in three brain regions, cerebral cortex, cerebellum, and hippocampus, was investigated. Differences were found for both the maximum number of sites immunoprecipitated by each antibody in one brain region and for the percentage of benzodiazepine binding sites immunoprecipitated by one specificity antibody between the different brain regions. Furthermore, it was found that co-immunoprecipitation with either anti-alpha 1 324-341, anti-Cys alpha 2 414-424, and anti-Cys alpha 3 454-467 or anti-alpha 1 324-341 and anti-Cys alpha 3 454-467 antibodies resulted in an increase in the percentage of benzodiazepine binding sites immunoprecipitated, the sum of which was equal to the percentages pelleted by the individual antibodies. These results demonstrate for the first time the existence in mammalian brain of gamma-aminobutyrateA receptor alpha subunit iso-oligomers.

Amino Acid Sequence↗
PubMed1989

Mapping the benzodiazepine photoaffinity-labelling site with sequence-specific gamma-aminobutyric acidA-receptor antibodies.

The gamma-aminobutyric acidA (GABAA) receptor purified from adult bovine cerebral cortex was photoaffinity-labelled with the agonist benzodiazepine [3H]flunitrazepam and the radioactivity shown to be coincident with a band with Mr 53,000 that was recognized by three anti-(GABAA receptor alpha 1 subunit sequence)-specific antibodies. Complete and limited CNBr cleavage of the purified photoaffinity-labelled receptor was carried out. The products of this reaction were analysed for radioactivity, for immunoreactivity with anti-[alpha 1-(1-15)-peptide], anti-[alpha 1-(324-341)-peptide] and anti-[alpha 1-(413-429)-peptide] polyclonal antibodies and for carbohydrate by biotinylated concanavalin A lectin overlay. Complete CNBr cleavage gave a radioactive peptide with Mr 10,000-12,000 that was not recognized by the above-mentioned specific antisera. By using the deduced amino acid sequence of the alpha 1 subunit [Schofield, Darlison, Fujita, Burt, Stephenson, Rodriguez, Rhee, Ramachandran, Reale, Glencorse, Seeburg & Barnard (1987) Nature (London) 328, 221-227], it is proposed that the site of the benzodiazepine-agonist photoaffinity-labelling reaction does not lie within the amino acid sequences alpha 1 1-58 and alpha 1 149-429.

Affinity Labels↗
PubMed1989

Identification of the alpha 3-subunit in the GABAA receptor purified from bovine brain.

Polyclonal antibodies have been raised to synthetic amino acid sequences of the bovine GABAA receptor alpha 1 and alpha 3 subunits. Anti-alpha 1 subunit antibodies recognise a polypeptide of 53 kDa whereas anti-alpha 3 subunit antibodies recognise a polypeptide of 59-60 kDa, in Western blots of GABAA receptor purified from adult bovine cerebral cortex, cerebellum and 12-day calf cerebral cortex.

Animals↗
PubMed1989

Bovine gamma-aminobutyric acidA receptor sequence-specific antibodies: identification of two epitopes which are recognised in both native and denatured gamma-aminobutyric acidA receptors.

Polyclonal antibodies have been raised against synthetic peptides whose sequences correspond to the N-terminal 15 amino acids and the C-terminal 17 amino acids of the bovine gamma-aminobutyric acidA (GABAA) receptor alpha 1 subunit. These antibodies were shown to react with the denatured GABAA receptor alpha subunit, Mr 53,000, in Western blots with both purified receptor and brain membranes as antigens. Also, both antibodies recognised both the purified and detergent-solubilised GABAA receptor as demonstrated by dose-dependent specific immunoprecipitation of the GABA and benzodiazepine binding sites from solution. Evidence is also presented to show brain-regional distribution of the expression of the alpha 1 subunit.

Amino Acid Sequence↗
PubMed1988

Benzodiazepine binding site heterogeneity in the purified GABAA receptor.

The displacement of [3H]flunitrazepam binding activity by ethyl-beta-carboline-3-carboxylate (beta CCE) was studied in both membrane-bound and purified GABAA receptors from adult bovine cerebral cortex, hippocampus and cerebellum. It was found that the best fit for the displacement of benzodiazepine binding in the cerebellar membranes was a single site with IC50 = 0.55 +/- 0.21 nM, whereas the best fit for cortical and hippocampal membranes was a two-site model with respective values of IC50 = 0.2 +/- 0.09 nM (high affinity), IC50 = 21 +/- 6 nM (low affinity) (cortex) and IC50 = 0.25 +/- 0.05 nM, IC50 = 20 +/- 2 nM (hippocampus). These same properties were retained in the purified GABAA receptor from the three brain regions. Thus, we have demonstrated that binding site heterogeneity as defined by the displacement of beta CCE is preserved in purified GABAA receptors and we suggest that this provides evidence for the existence of GABAA receptor isoforms.

Animals↗
PubMed1988

Bone marrow findings in immunodeficiency syndromes.

We have summarized the findings in selected congenital and acquired immunodeficiency disorders that frequently display blood and bone marrow abnormalities. Many other immunodeficiency syndromes were not discussed, as bone marrow abnormalities are infrequently seen in them. Examination of the bone marrow is not usually critical for diagnosing most immunodeficiency disorders. However, it may be diagnostic in the evaluation of fever of unknown origin, to rule out other causes of peripheral cytopenias, and to screen for virus-associated malignancies frequently seen in immune-deficient patients (for example, B cell lymphoma, Kaposi's sarcoma). Examination of the bone marrow is critical in the diagnosis of the virus-associated hemophagocytic syndrome and is often important in evaluating patients with the acquired immunodeficiency syndromes who display hematologic disturbances, such as those being treated with AZT, or those who might have opportunistic infections or malignant neoplasms.

Bone Marrow↗
PubMed1987

Collagen-binding of von Willebrand's factor antigen in the classification of von Willebrand's disease.

Analysis of the multimeric structure of von Willebrand's factor (vWf) was compared with vWf-binding to collagen in subjects' plasma. The percentage binding of vWf to collagen was determined by both an enzyme-linked immunosorbent assay (ELISA) method and quantitative immunoelectrophoresis. Normal subjects, type I von Willebrand's disease (vWd), and type II vWd were studied. The authors have found by correlating collagen-binding results with multimeric analysis that normal controls exhibit 85.4 +/- 5.1% collagen absorption of their vWf, patients with type I vWd 80.8 +/- 5.3%, whereas patients exhibiting a deficiency of high molecular weight vWf have 32.3 +/- 16.6% collagen absorption of vWf. The results suggest that this functional assay of vWf can be used in the classification of vWd and in the detection of new dysfunctional forms of vWf.

Antigens↗
PubMed1987

Bone marrow patterns in aplastic anemia: observations with 1.5-T MR imaging.

Six patients diagnosed as having aplastic anemia underwent magnetic resonance (MR) imaging. The bone marrow in the lumbar spine, pelvis, and proximal femora was examined in each case. One patient had a normal marrow signal intensity (SI) and pattern and was considered cured of disease. The other patients had varying numbers of focal low-SI areas interspersed with high-SI areas in the marrow of the spine. The pelves and proximal femora of all patients had diffuse high-SI marrow without focal abnormalities. Biopsy of a lumbar vertebral body in one patient demonstrated that the focal low-SI areas may have been islands of active hematopoietic cells scattered in otherwise fatty marrow. MR may prove to be a valuable, noninvasive method of following the response of patients with aplastic anemia to therapy; more investigation is necessary before this can be definitely concluded.

Adolescent↗
PubMed1987

Familial aortic dissection in absence of ascending aortic aneurysms: a lethal syndrome associated with precocious systemic hypertension.

The interaction of elevated blood pressure and aortic metabolism in the genesis of aortic dissection is uncharacterized. A kindred with fatal familial aortic dissection in association with precocious systemic hypertension and in absence of a definable connective tissue syndrome has undergone genealogical, clinical, pathological, and biochemical evaluation. Six family members spanning three generations have died of acute dissection. Five men died at a mean age of 28 years (range 22-34), while the proband's paternal grandmother died at 62 years of age. All were hypertensive. A constellation of subtle clinical features points toward deficient integrity of connective tissues; however, major hallmarks of known connective tissue syndromes including aortic root ectasia or aneurysms are absent. Studies of cultured dermal and aortae fibroblasts of two of the proband's brothers mitigate against Ehlers-Danlos IV syndrome. This family's susceptibility to aortic dissection reflects the synergistic liability of coexistent elevated blood pressure and metabolic abnormalities in the genesis of aortic degeneration.

Adult↗
PubMed1986

Lead pollution from the external redecoration of old buildings.

Lead concentrations before, during and after external redecoration have been measured in seven school playgrounds. All playgrounds showed increases in lead concentration during redecoration but the extent of contamination and duration varied enormously. The geometric mean lead value per school playground, taken on the day of the highest concentration during redecoration, ranged from 370 to 53000 micrograms g-1. The two post-war schools had significantly lower lead in dust concentrations than the other older schools.

Air Pollutants↗
PubMed1985

Lead in playground dust and on the hands of schoolchildren.

Measurements of the quantity of lead on the hands of schoolchildren and of the concentration of lead in dust from school playgrounds have been carried out; the hand-lead was collected by wiping with a moist tissue. When hand-lead and dust-lead values were averaged for each school, a statistically significant relationship between the two sets of means was obtained. Some limited measurements of size distribution of the dust particles on the children's wipes were also made; these indicated that most of the particles were less than 10 micron in diameter.

Child↗
PubMed1983

Contribution of lead in dust to children's blood lead.

The importance of urban dust as a source of lead for young children is still disputed. Although blood-lead data from various population surveys usually show a peak concentration in early childhood, there is evidence that such a peak is small or absent altogether in children without much access to the general environment. An examination of those studies where groups of people in regions of low and high lead contamination have been compared shows that the child/adult blood-lead ratio is almost always enhanced in the more exposed groups. This implies a route of lead uptake which is important for children but less so for adults, and it is likely that this route is the dust-hand-mouth one. There are sufficient data to suggest a quantitative relationship between raised levels of blood lead and lead in dust. There is a strong case for a lead-in-dust standard but some will probably remain unpersuaded unless or until there are reliable data for blood lead and environmental lead involving matched groups of young people from urban and rural areas.

Adolescent↗
PubMed1982

Synthesis and biological properties of 2'-deoxy-5-vinyluridine and 2'deoxy-5-vinylcytidine.

Rapid and efficient syntheses for the preparation of 2'-deoxy-5-vinyluridine and 2'-deoxy-5-vinylcytidine are described starting from nucleoside precursors. Contrary to some previous reports, 2-deoxy-5-vinyluridine has be found to be quite stable under normal laboratory conditions but when tested in animals shows neither toxicity nor anti-leukemic (L1210 cells) or anti-parasitic (Plasmodium berghei) activity. 2'-Deoxy-5-vinylcytidine appears to polymerise readily. It is much less toxic to cell cultures than 2'-deoxy-5'vinyluridine but is almost as active against herpes virus replication (ID50:0.2 microgram/ml) for both type 1 and type 2 herpes virus (selectivity index:225).

Animals↗