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Biomedical subjects

S M Johnson

Publications and source records attributed to S M Johnson.

At least 181 records · Page 10Linked to original sources

Slow synaptic potentials in neurones of the myenteric plexus.

1. Intracellular recordings were made in vitro from neurones in the myenteric plexus of the guinea-pig ileum. Potential changes were recorded in response to focal stimulation of the surface of the myenteric ganglion at a distance of 30-100 microM from the impaled cell. 2. Stimuli comprising single pulses evoked cholinergic excitatory post-synaptic potentials ('fast' e.p.s.p.s) in S cells. In a small proportion (20%) of both S and AH cells such stimuli also evoked depolarizing potentials with a time course about 1000 times slower than that of the fast e.p.s.p. 3. Stimuli comprising repeated pulses (up to 20 Hz for 2 sec) evoked the slow depolarizing potentials in a higher proporton of neurones (42%). These stimuli caused a hyperpolarizing potential change in 9% of cells, and in a very few cells biphasic changes i membrane potential were observed. 4. Both the slow depolarizing and the slow hyperpolarizing responses persisted in atropine (up to 5 microM) and hexamethonium (up to 200 microM), but were reversibly abolished by changing to calcium-free solutions. 5. Evidence is presented which suggests that the slow depolarizing response is caused by inactivation of the membrane potassium conductance, and the slow hyperpolarizing response is due to activaton of the potassium conductance.

Animals↗

Multiple actions of 5-hydroxytryptamine on myenteric neurones of the guinea-pig ileum.

1. Intracellular recordings were made from neurones lying within ganglia isolated from the myenteric plexus of the guinea-pig ileum. 2. 5-hydroxytryptamine (5-HT) (200 nM-10 microM) was applied to 160 neurones by adding it to the perfusing solution. Depolarizations were observed in 27% of neurones, and were more likely to be observed in S-cells. Hyperpolarizations were observed in 30% of neurones, and were more likely to be seen in AH cells. 52% of neurones were unaffected. 2. 5-HT was applied to 104 neurones by micro-electrophoresis from an electrode with its tip placed close to the soma membrane. Depolarizations were observed in 35% of cells, and hyperpolarizations 13%; 10% of neurones showed biphasic responses in membrane potential and the remaining cells were unaffected. 4. The depolarizing responses to 5-HT were associated with an increase in neuronal input resistance and were probably due to inactivation of the membrane potassium conductance. The responses declined in amplitude during prolonged or repeated applications of 5-HT. 5. The hyperpolarizing responses to 5-HT were associated with a fall in neuronal input resistance and were probably due to activation of the membrane potassium conductance. These responses persisted throughout the duration of application of 5-HT and did not show marked tachyphylaxis. The hyperpolarizing responses were reversibly abolished by calcium-free solutions. 6. The depolarizing responses to 5-HT may underlie the release of acetylcholine caused by 5-HT in the guinea-pig ileum. The hyperpolarizing responses may contribute to the inhibition of the peristaltic reflex which has been observed when 5-HT is applied to the serosal aspect of the intestine.

Animals↗

Uptake and O-methylation of isoprenaline in the rabbit ear artery.

Factors which influence uptake on O-methylation of 3H-isoprenaline (3HISO) by isolated segments of the rabbit ear artery were investigated. The tendency of the artery to accumulate unchanged amine was enhanced either when the O-methylation of ISO approached saturation at a high substrate concentration (6 mumol 1(-1) ISO), or when O-methylation was inhibited. Under these conditions, deoxycorticosterone acetate (DOCA) partially inhibited the accumulation of ISO. DOCA exerted a stronger inhibitory effect on O-methylation, but only at low ISO concentrations (0.2 and 0.8 mumol 1(-1)). The results are interpreted as evidence that (a)O-methylation in the intact artery is a saturable process which serves to limit the accumulation of 3HISO when the bathing concentration of the amine is low, and (b) DOCA exerts its effects by inhibiting extraneuronal uptake into the compartment possessing O-methylating activity. Analysis of the kinetics of efflux of 3HISO suggested that O-methylation was restricted to only one compartment, whereas unchanged ISO was accumulated within at least two compartments in addition to the extracellular compartment. Attention is drawn to a small but significant inhibitory effect of chronic sympathetic denervation on the accumulation, although not the O-methylation of ISO, raising the possibility that there may also be distribution of 3HISO within the sympathetic nerves.

Animals↗

Lymphocyte 5'-nucleotidase in primary hypogammaglobulinaemia and cord blood.

The 5'-nucleotidase (5'-N) deficiency on circulating mononuclear cells of patients with 'common variable' hypogammaglobulinaemia (CV-H) was shown to be the result of one or more of three factors: reduced T cell 5'-N activity, a reduced percentage of circulating B cells, and a low B cell 5'-N activity. A lack of circulating B lymphocytes, together with a low T cell 5'-N activity in some cases, was found to be responsible for the deficiency of lymphocyte 5'-N in patients with X-linked hypogammaglobulinaemia (X-H). The low levels of 5'-N activity in CV-H and X-H patients were not due to abnormal compartmentalization of the enzyme, altered enzyme kinetics, or the presence of a regulatory factor. Cord Blood B and T cells have a lower 5'-N activity than adult lymphocytes. The patient and cord blood data are discussed in relation to the stage of cellular maturity.

Adolescent↗

The composition and fluidity of normal and leukaemic or lymphomatous lymphocyte plasma membranes in mouse and man.

The lymphocyte surface membranes from normal and leukaemic or lymphomatous cells from man and mouse were isolated, characterized, and analyzed both biochemically and by diphenyl hexatriene fluorescence polarization. The cholesterol/phospholipid molar ratio for all the pure lymphocyte plasma membranes was 0.45--0.50, and the fluorescence polarization results showed that values much higher than this were not credible. The lipid composition of all the plasma membranes was remarkably similar, except for the concentration of free fatty acids and glycerides. The latter two were particularily high in the mouse lymphoma membrane and these, rather than a low cholesterol concentration, were responsible for the increased fluidity of the cells. The most prominent protein in most of the plasma membrane preparations was actin. This is found only by some authors, and its presence probably depends on the method of lymphocyte disruption.

Adult↗

Hemodynamic effects of oxprenolol and propranolol in hypertension.

Oxprenolol is a beta-adrenergic blocker with intrinsic sympathomimetic activity. Such drugs are not currently available in the United States, although they have the advantage of less negative inotropic effect than the available propranolol. In 18 patients with mild essential hypertension, oxprenolol (9 patients) or propranolol (9 patients) was added to thiazide in random double-blind fashion and continued for 7 wk during which supine heart rate, blood pressure, and noninvasively measured cardiac output (by CO2 rebreathing) were determined weekly. With thiazide dosage constant throughout, maximal dose titration to 386. +/- 52.1 (SEM) mg/day of oxprenolol and 360.0 +/- 45.4 mg/day of propranolol was achieved over the first 5 wk. Blood pressure fell with both (141.8 +/- 4.8/96.0 +/- 2.3 to 128.0 +/- 5.1/87.2 +/- 1 mm Hg on oxprenolol, p less than 0.01; 150.8 +/- 5.5/98.0 +/- 1.7 to 129.9 +/- 5.5/86.8 +/- 3.4 mm Hg on propranolol, p less than 0.01). Cardiac output fell from 6.85 +/- 0.63 to 5.77 +/- 0.45 1/min (p less than 0.01) on oxprenolol, and from 6.79 +/- 0.61 to 5.37 +/- 0.37 1/min (p less than 0.02) on propranolol. Oxpranolol. Oxprenolol reduced heart rate from 76.4 +/- 2.0 to 65.6 +/- 2.1 beats/min (p less than 0.001) and it fell from 82.0 +/- 3.8 to 65.3 +/- 3.7 beats/min (p less than 0.001) with propranolol; the fall in heart rate was less but not significantly so for oxprenolol (-14.2 +/- 1.8% and -19.8 +/- 2.8%, p less than 0.1). Thus oxprenolol is equivalent to propranolol in antihypertensive action; minor hemodynamic differences between the two drugs might reflect intrinsic sympathomimetic activity of oxprenolol. Oxprenolol should be considered as an alternative to propranolol.

Adult↗

5'-nucleotidase of B and T lymphocytes isolated from human peripheral blood.

The ecto-5'-nucleotidase activities of highly purified T and B lymphocytes from human peripheral blood have been investigated using biochemical and histochemical techniques. The enzyme activity of the purified B cells was about 3.5 times that of the T cells. Using a histochemical assay, 21--55% of the B cells stained positively for 5'-nucleotidase, but only 2--22% of the T cells were positive. These results are discussed in relation to the low 5'-nucleotidase activities found on peripheral blood lymphocytes from patients with chronic lymphatic leukaemia and some patients with primary hypogammaglobulinaemia.

Agammaglobulinemia↗

Slow reacting substance (SRS) from ionophore A23187-stimulated peritoneal mast cells of the normal rat. II. Evidence for a precursor role of arachidonic acid and further purification.

The generation of slow reacting substance (SRS) from ionophore A23187-stimulated rat peritoneal mast cells was enhanced by arachidonic acid (AA). This SRS generation was inhibited by 5,8,11,14-eicosatetraynoic acid (ETYA), an acetylenic analogue of AA and an inhibitor of both fatty acid cyclooxygenase and lipoxygenase. Indomethacin, a fatty acid cyclooxgenase inhibitor, had an enhancing effect upon SRS generation. This suggests SRS generation occurred through an ETYA sensitive step--perhaps a lipoxygenase. Radiolabel from [14C]-AA was incorporated into SRS with comigration of radioactivity and bioreactivity in silicic acid and thin layer chromatographies. Upon silicic acid chromatography, the active principle was eluted in the methanol fraction. Two-dimensional thin layer chromatography revealed chromatographic separation from other known spasmogenic substances and phospholipids. Mast cell SRS was found to display physiochemical properties similar to those of rat basophilic leukemia cell SRS, namely: that mast cell SRS generation was 1) enhanced by arachidonic acid; 2) inhibited by ETYA but not by indomethacin; 3) incorporation of [14C]-AA into the active principle; and 4) similar behavior during purification in silicic acid and thin layer chromatographies.

5,8,11,14-Eicosatetraynoic Acid↗

Slow reacting substance (SRS) from ionophore A23187-stimulated peritoneal mast cells of the normal rat. I. Conditions of generation and initial characterization.

When rat peritoneal mast cells were exposed to the ionophore A23187, a principle was released that possessed the biologic properties of slow reacting substance (SRS) from various sources. The response was dose, time, and temperature dependent with no activity being demonstrated in unstimulated cells. Supporting evidence that the mast cell product was similar or identical to SRS obtained from other sources include: 1) appropriate differential bioassay profile, 2) resistance to lipolysis and proteolysis, 3) acid lability and base stability, 4) inactivation by limpet arylsulfatase, and 5) inhibition by low concentrations FPL 55712. These data demonstrate that the isolated rat peritoneal mast cell contains the biosynthetic capacity to produce a bioreactive substance with the properties of SRS.

Animals↗

Ecto 5'-nucleotidase deficiency in primary hypogammaglobulinaemia.

The activity of the lymphocyte ectoenzyme 5'-nucleotidase is very low in the majority of patients with primary 'common variable' hypogammaglobulinaemia. In order to test whether this can be explained by lymphocyte subpopulation deficiencies we measured 5'-nucleotidase activity, using both biochemical and histochemical techniques, in purified T and B cells from patients and healthy subjects. Purified B cells from normal subjects have about four times the activity of T cells. This explains why the levels of lymphocyte 5'-nucleotidase activity are at the lower limit of the normal range in patients with X-linked hypogammaglobulinaemia who lack B cells. The low levels in the 'common variable' group can be explained by low activity in their T lymphocytes associated with either low activity in their B cells or depletion of B cells. The finding that inhibition of the enzyme does not interfere with in vitro lymphocyte transformation or immunoglobulin production in normal subjects indicates that the enzyme deficiency is not directly responsible for the hypogammaglobulinaemia. These and other studies suggest that this enzyme appears on lymphocytes at a certain stage of development and that both T and B lymphocytes in some patients with 'common variable' hypogammaglobulinaemia are developmentally immature.

Agammaglobulinemia↗

The effect of feeding on the bile salt-independent canalicular secretion in dogs.

Mongrel dogs were prepared by cholecystectomy, ligation of the lesser pancreatic duct, and insertion of cannulas into the duodenum and stomach. During experiments on the unanesthetized dogs, bile was collected by cannulating the common bile duct through the duodenal cannula. Canalicular secretion was estimated by the biliary clearance of erythritol. Feeding caused increased bile flow, increased erythritol clearance, decreased bile salt concentration, and unchanged bile salt output. These findings indicate that feeding stimulated the bile salt-independent canalicular secretion.

Animals↗