Publications by authors named "Kirk C"

An accumulation of 3H-labelled inositol phosphates is observed when prelabelled rat superior cervical sympathetic ganglia are exposed to [8-arginine]vasopressin or to muscarinic cholinergic stimuli. The response to vasopressin is much greater than the response to cholinergic stimuli. The response to vasopressin is blocked by a V1-vasopressin antagonist, and oxytocin is a much less potent agonist than vasopressin.

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Stimulation of hepatocytes with vasopressin (10 nM) in the presence of 1.25 mM extracellular Ca2+ increased glycogen phosphorylase activity 4-fold within 15s and provoked a rapid efflux of cell-associated Ca2+. Vasopressin also caused a transient increase in the Ca content of a mitochondria-rich fraction separated within seconds of hormone stimulation by a rapid fractionation technique [Shears & Kirk (1984) Biochem.

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Vasopressin was among the first mammalian hormonal peptides to be identified and to have its structure determined. Its only undisputed physiological role is as a circulating neurohypophyseal antidiuretic hormone. Other notable effects of vasopressin on peripheral tissues include contraction of vascular smooth muscle, leading to elevation of blood pressure, and activation of glycogenolysis in liver.

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A rapid cellular fractionation technique [the preceding paper, Shears & Kirk (1984) Biochem. J., 219, 375-382] was employed to separate a mitochondria-rich fraction from hepatocytes within seconds.

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We have developed methods that yield estimates of the 32P content of each of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate, thus extending the information available from studies of the labelling of these lipids in intact cells or membrane preparations. The analyses are undertaken with the deacylated lipids. Assay of the 5-phosphate of phosphatidylinositol 4,5-bisphosphate is achieved by the use, under conditions of first-order kinetics, of a 5-phosphate-specific phosphomonoesterase present in isolated erythrocyte membranes [Downes, Mussat & Michell (1982) Biochem.

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The activation of Ca2+-mobilising receptors on hepatocytes and many other cells leads to a prompt reduction in the cellular content of inositol phospholipids. The primary event which underlies these changes is, most probably, a phospholipase C-catalysed attack upon phosphatidylinositol 4,5 bisphosphate. The receptor-mediated breakdown of this lipid in stimulated cells is: (i) not mediated by an increase in cytosol [Ca2+] and (ii) closely coupled to receptor occupation.

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Rat hepatocytes rapidly incorporate [32P]Pi into phosphatidylinositol 4-phosphate (PtdIns4P) and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]; their monoester phosphate groups approach isotopic equilibrium with the cellular precursor pools within 1 h. Upon stimulation of these prelabelled cells with Ca2+-mobilizing stimuli (V1-vasopressin, angiotensin, alpha 1-adrenergic, ATP) there is a rapid fall in the labelling of PtdIns4P and PtdIns(4,5)P2. Pharmacological studies suggest that each of the four stimuli acts at a different population of receptors.

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A simple and rapid assay method is described for determining the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP. Labelled ATP is incubated with cyclic AMP, cyclic AMP-dependent protein kinase and histone H2A under conditions leading to maximum phosphorylation of the histone. The specific radioactivity of the gamma-phosphate group of the 32P-labelled ATP is calculated from the total amount of [32P]Pi incorporated into a standard amount of histone.

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A double-blind crossover study was performed to compare the incidence of central nervous system (CNS) side effects with atenolol and metoprolol. Eleven women and 6 men who had previously developed such side effects on lipophilic beta-blockers were identified by means of a preliminary questionnaire. A 30-item psychiatric questionnaire was then used to detect changes in psychological status and possible CNS side effects.

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Muscarinic cholinergic agonists can provoke a wide variety of physiological responses in target tissues. This can be achieved by mobilisation of calcium from intracellular and extracellular stores. The members of the family of Ca2+-mobilizing receptors to which the muscarinic receptor belongs share the ability to provoke a calcium-independent decrease in the concentration of inositol phospholipids.

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It now appears to be generally agreed that the 'phosphatidylinositol response', discovered in 1953 by Hokin & Hokin, occurs universally when cells are stimulated by ligands that cause an elevation of the ionized calcium concentration of the cytosol. The initiating reaction is almost certainly hydrolysis of an inositol lipid by a phosphodiesterase. Phosphatidylinositol, phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate all break down rapidly under such circumstances.

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Prospective screening of 449 patients undergoing coronary or peripheral arterial revascularization detected asymptomatic cervical bruits in 44 patients (9.8%) and carotid obstruction by Doppler ultrasound in 63 individuals (14.0%).

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In isolated rat hepatocytes, vasopressin evoked a large increase in the incorporation of [32P]Pi into phosphatidylinositol, accompanied by smaller increases in the incorporation of [1-14C]oleate and [U-14C]glycerol. Incorporation of these precursors into the other major phospholipids was unchanged during vasopressin treatment. Vasopressin also promoted phosphatidylinositol breakdown in hepatocytes.

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Vasopressin stimulates the incorporation of [32P]Pi into phosphatidylinositol but not into other phospholipids in rat thoracic aorta strips. The relative abilities of three vasopressin analogues to stimulate phosphatidylinositol labelling in rat aorta are similar to their relative pressor potencies in vivo and to their relative potencies in stimulating the metabolism of rat hepatocytes, but very different from their relative antidiuretic potencies. The vasopressor antagonist [1-(beta-mercapto-beta, beta-cyclopentamethylenepropionic acid),8-arginine]vasopressin competitively inhibits [Arg8]vasopressin-stimulated phosphatidylinositol labelling in rat aorta with a pA2 of 8.

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Serum thiocyanate has shown itself to be useful in distinguishing smokers from non-smokers. Our anti-smoking attitude resulted in 12 out of 32 smokers stopping smoking during the trial. Further, our evidence suggests that a patient who says he has given up smoking entirely can probably be believed, but a claim to have reduced smoking may be less reliable.

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Rauscher leukaemia virus (RLV), extensively purified by rate and density gradient centrifugation procedures, exhibited aminopeptidase (AP) activity. The amount of virion activity was about 0.005 times the specific activity of purified hog kidney aminopeptidase M (EC 3.

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The present experiments were undertaken to define the areas of projection of pretectum and superior colliculus to the pulvinar and n. lateralis posterior, respectively, and to define other brain stem structures projecting to these thalamic nuclei in cats. For this purpose of technique of retrograde transport of horseradish peroxidase (HRP) has been used.

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342 patients referred to a neurological out-patient department in North East England between November 1975 and November 1976 were studied prospectively using two methods of psychiatric case identification, by clinical interview and by use of the General Health Questionnaire. The population of psychiatric patients identified by clinical techniques is different from that identified by the GHQ and it is concluded that GHQ cannot be used for routine screening in the neurological clinic. Clinical data were compared with a previous retrospective study.

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