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Biomedical subjects

E J Thompson

Publications and source records attributed to E J Thompson.

At least 199 records · Page 11Linked to original sources
PubMed1980

Effect of sample preparation on cerebrospinal fluid protein patterns in polyacrylamide gels.

The method of preparing CSF and dilute serum samples for polyacrylamide gel electrophoresis has a marked effect on the pattern of the resolved proteins. The incorporation of sucrose or glycerol into the sample seriously impairs the quality of the resolution and affects the actual number of bands resolved and the relative mobility of certain proteins. The use of sucrose as an anti-convection medium in dense samples such as serum and CSF is not recommended.

Blood Protein Electrophoresis↗
PubMed1979

Interaction of di-iodinated 125I-labelled alpha-bungarotoxin and reversible cholinergic ligands with intact synaptic acetylcholine receptors on isolated skeletal-muscle fibres from the rat.

1. Intact synaptic acetylcholine receptors on freshly isolated rat skeletal-muscle fibres were characterized by their interaction with di-iodinated 125I-labelled alpha-bungarotoxin, acetylcholine and other cholinergic ligands at room temperature (22 deggrees C). 2. The time course and concentration dependence of 125I-labelled alpha-bungarotoxin association conformed to a bimolecular mechanism. In time-course experiments with different concentrations of 125I-labelled alpha-bungarotoxin (1.4--200 nM) the bimolecular-association rate constant, k + 1, was (2.27 +/- 0.49) x 10(4)M-1.S-1 (mean +/- S.D., N = 10). In concentration-dependence experiments, k + 1 was 2.10 x 10(4)M-1.S-1 and 1.74 x 10(4) M-1.S-1 with 10 and 135 min incubations respectively. In association experiments the first-order rate constant was proportional to the 125I-labelled alpha-bungarotoxin concentration. 125I-Labelled alpha-bungarotoxin dissociation was first order with a dissociation constant, k-1, less than or equal to 3 x 10(-6)S(-1) (half-life greater than or equal to 60 h.) The results indicated a single class of high-affinity toxin-binding sites at the end-plate with an equilibrium dissociation constant, Kd, equal to or less than 100 pM. The number of toxin-binding sites was (3.62 +/- 0.46) x 10(7) (mean +/- S.D., n = 22) per rat end-plate. 3. The apparent inhibitor dissociation constants, Ki, for reversible cholinergic ligands were determined by studying their effect at equilibrium on the rate of 125I-labelled alpha-bungarotoxin binding. There was heterogeneity of binding sites for cholinergic ligands, which were independent and non-interacting with antagonists. In contrast agonist affinity decreased with increasing receptor occupancy. Cholinergic ligands in excess inhibited over 90% of 125I-labelled alpha-bungarotoxin binding. 4. Cholinergic ligand binding was accompanied by an increase in entropy, which was greater for the agonist carbachol (delta So = +0.46 kJ.mol-1.K-1) than the antagonist tubocurarine (delta So = +0.26 kJ.mol-1.K-1). 5. The entropy and affinity changes that accompanied agonist binding suggested that agonists induced significant conformational changes in intact acetylcholine receptors. 6. The affinity and specificity of 125I-labelled alpha-bungarotoxin and tubocurarine binding to synaptic acetylcholine receptors from slow and fast muscle fibres were the same. 7. The study of binding only requires milligram amounts of tissue and may have application to other neurobiological studies and to the study of human neuromuscular disorders.

Acetylcholine↗
PubMed1979

Oligoclonal immunoglobulins and plasma cells in spinal fluid of patients with multiple sclerosis.

A new modification of polyacrylamide gel electrophoresis (PAGE) was applied to cerebrospinal fluid proteins from patients with multiple sclerosis (MS). The same spinal fluids were also examined by a cytological technique. Over 90% of patients with clinically definite or early probable or latent MS showed abnormal PAGE patterns in the form of oligoclonal gammaglobulin bands. Reactive (atypical, large) lymphocytes or typical plasma cells were found in some patients. In all such cases an oligoclonal pattern was present. The findings of oligoclonal bands provides valuable supporting evidence for the diagnosis of MS in the less definite clinical categories.

Electrophoresis, Polyacrylamide Gel↗
PubMed1979

Cystic intracranial tumours. Cyst fluid, biochemical changes and computerised tomographic findings.

The concentrations of five biochemical substances were studied in cyst fluid from 49 cystic intracranial tumours and the findings were compared with the computerised tomographic (CT) scan and histological appearances of the tumours. The attenuation of the cyst wall on unenhanced CT was found to be related to cyst fluid alkaline phosphatase levels. Enhancement of the cyst wall with contrast medium was associated with higher levels of protein, cholesterol, alkaline phosphatase, phosphohexose isomerase (PHI) and lactate dehydrogenase (LDH) in the cyst fluid. Lesions of high grade malignancy had higher levels of protein, cholesterol, alkaline phosphatase, LDH and PHI than lesion of low grade malignancy and also higher levels of alkaline phosphatase, LDH and PHI than benign lesions. Oedema around a lesion was associated with increased malignancy.

Alkaline Phosphatase↗
PubMed1979

An abnormality of cell behaviour in human dystrophic muscle cultures: a time-lapse study.

The initiation of monolayer mass cell cultures from adult human biopsies has revealed a striking abnormality in the growth and development pattern of muscle cultures from Duchenne-type dystrophy. This abnormality in cell behaviour was seen as early as 4 days in culture, well before myotube formation or confluence, and consisted of areas where cells clustered together in a multilayered mass rather than showing the typical monolayer distribution normally observed. To gain some insight into the mechanism of cell cluster development, we have examined such a culture by time-lapse cinematography and also the cell behaviour of other control cultures. The results of this study show that the clusters enlarged primarily by cell division and, to a lesser extent, by the acquisition of neighbouring cells. Furthermore, none of the single cells surrounding the clusters exhibited contact inhibition of movement. This behaviour was not observed in the other cultures examined. These findings could be indicative of an abnormality in the cell surface or cell-locomotory machinery of dystrophic cells.

Adolescent↗
PubMed1978

Rapid quantitative surface immunofixation of proteins in polyacrylamide gels.

A new method is described which allows rapid identification of sub-microgram amounts of protein in polyacrylamide gels. The technique has been applied to samples of cerebrospinal fluid and serum and is suitable for localisation of both discrete protein bands and diffuse zones. Results from cerebrospinal fluid confirm that IgG is localised in the oligoclonal bands seen in multiple sclerosis. Given antibody excess, there is a linear relationship between the amount of protein applied to the gel and the density of the stained immune complexes on the gel surface.

Blood Proteins↗
PubMed1978

beta-Bungarotoxin. Separation of two discrete proteins with different synaptic actions.

beta-Bungarotoxin, a specific presynaptic blocking agent, was prepared in two stages from the crude venom of Bungarus multicinctus by ion-exchange chromatography on the weakly acidic ion exchanger, CM-Sephadex, and on the strongly acidic ion exchanger, sulphopropyl-Sephadex. By these procedures it was purified to a single protein, which was shown by reduction to contain two polypeptide chains with mol.wts. of less than 15000. During purification of beta-bungarotoxin three other proteins were isolated. Two of these proteins have similar molecular weights, subunit structure and physiological properties to the major protein component. This latter is referred to as beta-bungarotoxin, since it has the same physiological properties as those described for unpurified beta-bungarotoxin by other workers. The first protein has very different physiological effects and biochemical properties from beta-bungarotoxin. This protein has a single class of polypeptide chains with an apparent molecular weight that is lower than the main beta-bungarotoxin protein, and appears to block synaptic transmission by a predominantly postsynaptic effect. It has been suggested [Oberg & Kelly (1976) J. Neurobiol. 7, 129-141] that the action of beta-bungarotoxin depends on its phospholipase A activity; however, in this preparation of the toxin less than 50 muunits of phospholipase A activity were detected (1 unit of activity is the amount of enzyme forming 1 mumol of L-alpha-phosphatidylcholine/min per mg of protein).

Animals↗
PubMed1978

beta-Bungarotoxin. The binding of [3H]pyridoxylated beta-bungarotoxin to a high-molecular-weight protein receptor.

beta-Bungarotoxin was labelled with pyridoxal 5'-phosphate (incorporating 3H). The kinetics of beta-bungarotoxin binding to several tissue subfragments of nervous tissue was studied. The dissociation constant of 3H-pyridoxylated beta-bungarotoxin in this reaction was 0.21-0.37 micron and that of unlabelled beta-bungarotoxin was 25 nM. Hill [(1910) J. Physiol. (London) 40, iv-vii] and Scatchard [(1949) Ann. N.Y. Acad. Sci. 51, 660-672] analyses demonstrated no co-operativity of binding and only a single class of receptor sites, consistent with a bimolecular association of beta-bungarotoxin and its receptor. The iodinated toxin was physiologically inactive. Toxin was bound in non-specific unsaturable fashion by glass and/or plastic. This low-affinity binding was corrected by addition of bovine serum albumin to a final concentration of 30 mg/ml. A soluble protein receptor of beta-bungarotoxin was isolated and the mol.wt. is approx. 200000.

Animals↗