Enhanced hypothalamic noradrenaline biosynthesis in Goldblatt I renovascular hypertension. 1. Hypertension was induced in rats by renal artery clip with the contralateral kidney removed (Goldblatt I) or left intact (Goldblatt II). 2. Plasma noradrenaline was increased 62% in the Goldblatt I animals after 3 weeks. 3. Hypothalamic tyrosine hydroxylase and dopamine beta-hydroxylase activities, and the concentration of noradrenaline were increased in the Goldblatt I animals after 3 weeks. 4. Enhanced hypothalamic noradrenaline synthesis may be a pathogenic factor in Goldblatt I renovascular hypertension.


Definitive evidence for renin in rat brain by affinity chromatographic separation from protease. 1. Angiotensin I-generating activity of rat brain extract was separated into two components by affinity chromatography on a casein-Sepharose gel column. 2. The component without affinity to the gel was identified as true renin on the basis of its sensitivity to anti-renin antibody and the lack of protease activity. 3. The second renin-like component with affinity to the gel was a protease insensitive to the anti-renin antibody. Its renin-like activity examined with sheep substrate was pronounced compared with the rate of angiotensin I generation from the rat substrate. 4. It was concluded that rat brain contains true renin, which can be detected by the use of rat substrate but can be masked when examined with sheep substrate.


Renal release of active and inactive renin in essential and renovascular hypertension. 1. Active and acid-activable inactive renin were measured in renal venous and arterial plasma of 18 patients with essential hypertension (EHT) and 19 patients with renovascular hypertension (RVHT). In seven patients with EHT and in 11 patients with RVHT measurements were made before and 25-35 min after an intravenous injection of 300 mg of diazoxide. 2. Under basal conditions the renal vein to artery ratios for active and inactive renin in EHT ranged from 0.71 to 1.96 and from 0.68 to 1.44 respectively. In 14 patients with RVHT the renal vein to artery ratio for active renin on the affected side was above the range found in EHT and in six of them the renal vein to artery ratio for inactive renin was also elevated. 3. The diazoxide-induced release of active renin from kidneys, which had a stenotic artery but were not seriously contracted, was associated with a fall of the renal vein to artery ratio for inactive renin to a value below 1.00. 4. The results indicate that changes in the release of active and inactive renin do not always run in parallel. The findings are compatible with the hypothesis that circulating inactive renin can be activated in the kidney.


An endogenous protease activating plasma inactive renin. 1. The protease inhibitors Trasylol and soyabean trypsin inhibitor prevented the activation of plasma inactive renin by acid. 2. N-Ethylmaleimide inhibited acid-activation to some extent but o-phenathroline had no effect. 3. Acid-activation of the inactive renin in human plasma is mediated by a serine protease.


Activation of inactive plasma renin: evidence that both cryoactivation and acid-activation work by liberating a neutral serine protease from endogenous inhibitors. 1. We have found that 'acid'-activation of inactive human plasma renin is a two-phase process. About 30% of activation occurs during dialysis to pH 3.3; the remaining 70% occurs at alkaline pH. 2. The 'alkaline phase' of activation has a pH optimum between 7.5 and 8.4. It is inhibited by unacidified plasma and by soya-bean or lima-bean trypsin inhibitors. 3. 'Cryoactivation' of inactive plasma renin, which occurs at -4 degrees C and alkaline pH, is also inhibited by soya-bean or lima-bean trypsin inhibitors and by the serine protease inhibitors diisopropylphosphorofluoridate and benzamidine. 4. Thus endogenous neutral serine proteases participate in the activation of inactive plasma renin in vitro. Their action is prevented in the circulation by inhibitors which are inactivated by acid or cold.


The interaction between prazosin and clonidine. 1. The interaction between prazosin and clonidine was studied in anaesthetized rats, pithed rats and in anaesthetized cats. 2. Prazosin diminished the clonidine-induced hypotensive effect in anaesthetized rats, probably via an antagonism at the level of central alpha-adrenoreceptors. 3. In pithed rats, stimulation of the Nervi accelerantes caused tachycardia, which was diminished considerably by clonidine. The antagonism by clonidine was partly reversed by prazosin, suggesting that prazosin possesses a certain degree of presynaptic activity apart from its predominant effect at the postsynaptic alpha-receptor. Piperoxan was more active than prazosin. 4. The central hypotensive effect of clonidine, injected into the left vertebral artery of cats was significantly reduced by prazosin, administered before clonidine via the same route. Intravenously injected prazosin did not diminish the central hypotensive effect of clonidine. The antagonism is, therefore, caused by a central mechanism. 5. The combined application of clonidine and prazosin in antihypertensive treatment is probably not only irrational but ought to be discouraged in view of the interaction between the drugs, which leads to a reduced antihypertensive potency of clonidine.


Labetalol in hypertensive patients with angina pectoris: beneficial effect of combined alpha- and beta-adrenoreceptor blockade. 1. Eight hypertensive patients with angina pectoris had placebo added to their existing medications for 8 weeks, then incremental doses of active labetalol with simultaneous stepwise reduction in other medicines until blood pressure was satisfactorily controlled; after that only labetalol and thiazide (8 weeks) and finally labetalol-placebo together with previous beta-adrenoreceptor antagonists and thiazide for 4 weeks were administered. 2. During the labetalol plus thiazide period resting blood pressures and measurements obtained during isotonic exercise, isometric exercise and the cold pressor test were significantly lower than during the initial placebo addition period. Angina scores were significantly reduced during this period. 3. During the final treatment with placebo, beta-adrenoreceptor antagonist and thiazide, blood pressures remained reduced, but angina was significantly worse. 4. Labetalol which antagonizes both alpha- and beta-adrenoreceptors produced better relief of angina pectoris than beta-adrenoreceptor antagonists during improvement in blood pressure in hypertensive patients.


beta-Adrenoreceptor-blocking agents and lipid metabolism. 1. Chlorothiazide twice a day plus atenolol, metoprolol, pindolol and propranolol in single daily doses administered to patients with essential hypertension achieved effective control of blood pressure. 2. Each beta-adrenoreceptor-blocking drug was associated with small, but significant, increases in plasma triglyceride concentrations and suppression of fasting immuno-reactive glucagon concentrations.


Sympathetic and parasympathetic components of reflex cardiostimulation during vasodilator treatment of hypertension. 1. Haemodynamic responses to diazoxide (300 mg intravenously) were studied in 15 hypertensive patients before and after chronic beta-adrenoreceptor blockade by 320 mg of propranolol daily. After diazoxide alone, mean arterial pressure and total peripheral resistance were lowered by 24 +/- 3 and 35 +/- 5% (mean +/- SEM) respectively. Cardiac output and heart rate rose by 25 +/- 9 and 21 +/- 3%. During beta-adrenoreceptor blockade, the percentage changes of mean arterial pressure, heart rate, cardiac output and total peripheral resistance after vasodilatation were not significantly different from those after diazoxide alone. 2. Atropine, 0.04 mg/kg body weight, was given to 12 hypertensive patients chronically treated with beta-adrenoreceptor blockade, before acute vasodilatation by diazoxide. Diazoxide caused no increase in heart rate after combined beta-adrenoreceptor and parasympathetic blockade. However, cardiac output rose by 14 +/- 5%. 3. We conclude that withdrawal of parasympathetic tone is an important determinant of circulatory homeostasis after acute vasodilatation during beta-adrenoreceptor blockade.


Relation between prognosis and the blood pressure before and during treatment of hypertensive patients. 1. A study was conducted amongst 1247 treated hypertensive patients to determine the predictive power of untreated baseline and achieved treated blood pressures in the development of the complications of hypertension. In addition the relative importance of systolic and diastolic pressures was calculated. 2. Statistical analysis was done by calculating univariate differences in blood pressure between cases with and without complications. The higher the univariate distance, the greater the predictive power. 3. Blood pressures achieved during treatment were more important than baseline pressures for predicting stroke in both men and women, confirming the benefits of antihypertensive therapy in preventing strokes. 4. There was some evidence of prevention of myocardial infarction in men and of angina in women as a result of therapy. 5. There was no evidence to suggest that any one group of drugs, including beta-adrenoreceptor-blocking drugs and thiazides, conferred any extra benefit in preventing coronary heart disease. 6. The systolic blood pressures achieved during treatment predicted stroke better than diastolic pressure, but no consistent trends were found for coronary heart disease.


Plasma adrenaline and noradrenaline in essential hypertension and after long-term treatment with beta-adrenoreceptor-blocking agents. 1. Plasma adrenaline and noradrenaline were measured in supine and upright positions in 27 essential hypertensive patients and in 12 age-matched control subjects. 2. Plasma adrenaline was markedly increased in a large proportion of patients, compared with control subjects, either in supine or in upright positions, whereas no significant differences were found in plasma noradrenaline. 3. Six hypertensive patients were chronically treated with beta-adrenoreceptor-blocking agents (five with propranolol and one with pindolol). Plasma noradrenaline was not significantly changed in the supine position but was markedly increased on standing in four out of six patients. Plasma adrenaline was decreased in five out of six patients either in suprine or upright positions. 4. These findings support the view that the adrenergic system participates in the mechanism of essential hypertension.


[Studies on staphylococcal enterotoxin B. II. Production and regulation (author's transl)]. Enterotoxigenic strain of Staphylococcus aureus (ATCC 14458) was grown under various conditions with constant shaking to determine the requirements for maximum toxin production. It was evident that 3% tryptic soy broth, 3% NZ-Amine NAK + 3% casein hydrolysate, 3% NZ-Amine NAK + 1% yeast extract, and 3% NZ-Amine NAK + 1% yeast extract + 0.2% glucose are most available toxin production media. But concentration of glucose could strictly triggered the enterotoxin producing efficiency. When glucose concentration was less than 0.5%, although with higher yield, the toxin production was delayed for certain period of time. However, if glucose concentration was up to more than 0.5%, the enterotoxin production was almost inhibited. Some metabolites of glucose to elucidate the inhibitory effect have also investigated. Our results indicated that glycerol and citric acid inhibited the toxin production directly, while the inhibitory effect of lactic acid and acetic acid were due to those acidic metabolites, decreased the pH value of media, and adversely suppressed the bacterial growth.


The role of the septo-hippocampal system and its noradrenergic afferents in behavioural responses to none-reward. Our experiments were designed with two purposes: (i) to examine the effects on one behaviour of differing interventions in the septo-hippocampal system; (ii) to compare these effects with those of minor tranquillizers. The behaviour studied (in rats) is extinction in the alley after continuous (CRF) or partial (PRF) reinforcement. Minor tranquillizers and large septal lesions produce three effects: (1) resistance to extinction is increased after CRF; (2) resistance to extinction is decreased after PRF; (3) the partial reinforcement extinction effect (PREE) is abolished. Small septal lesions fractionate this syndrome: either effect (1) or an actual increase in the size of the PREE is produced by medial septal lesions abolishing hippocampal theta; effects (2) and (3), but not (1), are produced by lateral septal lesions sparing theta. Dorso-medial fornix section, abolishing theta, reproduces the effects of medial septal lesions. Fimbrial section, sparing theta, reproduces some of the effects of lateral septal lesions. Minor tranquillizers produce a rise in the threshold for septal driving of hippocampal theta specifically at 7.7 Hz. This effect is reproduced by blockade of noradrenergic transmission or destruction of the dorsal noradrenergic bundle with 6-hydroxydopamine. This lesion reproduces all three behavioural changes listed above. These results suggest a model for the role of the septo-hippocampal system and its noradrenergic inputs in the PREE. This model is compared with other approaches to the septo-hippocampal system.


Localization of putative transmitters in the hippocampal formation: with a note on the connections to septum and hypothalamus. Biochemical assays on microdissected samples, denervation studies, subcellular fractionation, and light and electron microscopic autoradiography of high affinity uptake have been performed to study the cellular localization of transmitter candidates in the rat hippocampal formation. High affinity uptake of glutamate and aspartate is localized in the terminals of several excitatory systems, such as the entorhino-dentate fibres (perforant path), mossy fibres (from granular cells) and pyramidal cell axons. Thus, in stratum radiatum and oriens of CA1, 85% of glutamate and asparate uptake and 40% of glutamate and aspartate content are lost after lesions of ipsilateral plus commissural fibres from CA3/CA4. Hippocampal efferents also take up aspartate and glutamate, since these activities are heavily reduced in the lateral septum and mamillary bodies after transection of fimbria and the dorsal fornix. The synthesis (by glutamic acid decarboxylase), content and high affinity uptake of gamma-aminobutyrate (GABA) are not reduced after lesions of these or other projection fibre systems. A localization in intrinsic neurons is confirmed by a selective loss of glutamic acid decarboxylase after local injections of kainic acid. Peak concentrations of the enzyme occur near the pyramidal and granular cell bodies, corresponding to the site of the inhibitory basket cell terminals, and in the outer parts of the molecular layers. Some 85% of glutamic acid decarboxylase is situated in 'nerve ending particles'. Acetylcholine synthesis (by choline acetyltransferase) disappears after lesions of septo-hippocampal fibres. Since 80% of the hippocampal choline acetyltransferase is in 'nerve ending particles', the characteristic topographical distribution of this enzyme should reflect the distribution of cholinergic septo-hippocampal afferents. Serotonin, noradrenaline, dopamine and histamine are located/synthesized in afferent fibre systems. Some monoamine-containing afferents to the hippocampal formation pass via the septal area, others via the amygdala. The hippocampal formation also contains nerve elements reacting with antibodies against neuroactive peptides, such as enkephalin, substance P, somatostatin and gastrin/cholecystokinin.


Origins of specificity in the binding of small molecules to dihydrofolate reductase. Dihydrofolate reductase is the target for the therapeutically important 'anti-folate' drugs such as methotrexate and trimethoprim. Methotrexate is a powerful inhibitor of the enzyme, binding up to 10,000 times more tightly than the structurally similar substrate, folate. Two contributions to this striking difference in affinity have been identified: the two ligands bind in different charge states, and there are conformational differences between the two complexes. The origins of the tight binding of methotrexate have been explored further by studying the binding of 2,4-diaminopyrimidine and p-aminobenzoyl-L-glutamate, which may be considered as 'fragments' of methotrexate. These two compounds bind simultaneously but also cooperatively, the binding of one 'fragment' leading to a 50-fold increase in the affinity for the other. Studies of structural analogues of these fragments show that the specificity as well as the strength of binding can be altered by the presence of the other 'fragment'; both positive and negative cooperativity are observed. The relation of these observations to methotrexate binding, and the notion of intramolecular cooperativity in ligand binding are discussed.


Gamma glutamyl transferase: application of a new radiochemical assay to the analysis of its subcellular distribution in the rat liver. gamma-Glutamyl transferase (gamma-GT) is a key catalyst in the metabolism of glutathione. Its activity in the rat liver is usually very low but it increases significantly during the process of chemical hepatocarcinogenesis. A new radiochemical assay is reported which measures the amount of 3H-aniline liberated from gamma-glutamyl-3H-anilide. This assay is highly specific and no biological substances interferes. By applying this new assay to the analysis of the subcellular distribution of the rat liver gamma-GT, it is shown that it distributes between two pools, one which corresponds to a membrane-bound enzyme, the other to a soluble enzyme. The subcellular distribution of gamma-GT is discussed by comparison with the patterns of distribution of classical marker enzymes.


Influence of detergents and organic solvent extraction on human gamma-glutamyltransferase activity. Human serum and urinary gamma-glutamyltransferase (gamma-GT) have been found to react differently towards the detergents Triton X-100 and sodium lauryl sulphate (SDS). Serum gamma-GT was virtually unaffected by Triton X-100 at a concentration of 5% whereas urinary gamma-GT was 10-15% activated under similar conditions. There was a 100-fold difference in the response of serum and urinary gamma-GT to SDS. The enzyme activity in urine was completely destroyed by 0.02% SDS, whereas it required 2.0% to destroy the serum enzyme. These latter differences, however, were found to result from the environments of the serum and urinary enzyme rather than to intrinsic factors within the molecule. Extraction of serum gamma-GT from normal individuals with n-butanol resulted in little or no loss of activity from the aqueous phase, whereas more than 30% activity was lost from normal urines. Extraction using various mixtures of butanol and di-isopropyl ether (DIPE) produced largely similar results, except that 25% DIPE: 75% butanol mixture produced a marked loss of activity from serum. It is suggested that gamma-GT activity in human body fluids may be dependent on the presence of a lipid fraction.


Human skeletal muscle enolase and factors influencing its activity. Enolase activity was determined in human skeletal muscle extracts using two procedures to measure the phosphoenolpyruvate formed. The results closely agreed, as did double determinations on bipartite muscle pieces. Activities in m. vastus lateralis quadricipitis did not differ significantly in adult males, females and 11- to 14-year-old girls. M. vastus medialis and deltoideus exhibited significantly higher activities than vastus lateralis. Endurance-trained athletes had lower enolase activities when compared to untrained subjects. The effects of changes in temperature, pH and K+ concentration in the medium were investigated and the possible physiological implications discussed.


Autoxidation of native oxymyoglobin. Kinetic analysis of the pH profile. The rate of autoxidation of native oxymyoglobin to metmyoglobin has been examined over the pH range of 4.8--12.6 in 0.1 M buffer at 25 degrees C, and some 40 values of the observed first-order rate constant, kobs, are plotted against pH of the solution. In order to understand the kobs--pH profile thus obtained, some mechanistic models are proposed for the autoxidation reaction. The fitting of their rate equations as a function of pH has been examined to the experimental kobs-pH plot by a least-squares method with the use of a digital computer. The complicated pH-profile can be best explained by the 'acid-base catalyzed three states model', which reveals not only the catalytic role of hydrogen ions and hydroxyl ions, but also the involvement of two dissociation groups of myoglobin molecule in the autoxidation reaction.


Agarose-bound horse-liver alcohol dehydrogenase. Dependence of molecular properties and activity on coupling conditions. 1. Spectroscopic methods for protein and active-site determination with the same sample of immobilised horse liver alcohol dehydrogenase have been developed. 2. The influence of pH, active-site protection of the soluble enzyme and protein concentration on coupling of alcohol dehydrogenase with cyanogen-bromide-activated Sepharose has been investigated. In phosphate buffer (pH 8.0) products with over 90% active-site retention have been synthesized. The binary complex alcohol-dehydrogenase . NADH gives a preparation with the same active-site content but a lower apparent specific activity compared to the unprotected enzyme. Increase in protein concentration yields products with the same active-site content relative to bound protein but the apparent specific activity is decreased. 3. The great similarity in spectroscopic properties of soluble and immobilised enzyme, as well as of their ternary complexes, shows that no significant conformational change has taken place during immobilisation. 4. Exchange of the non-catalytic Zn2+ against Co2+ yields a hybrid Sepharose--Co2Zn2-alcohol-dehydrogenase with over 90% active-site retention during metal exchange. The absorption spectra of the soluble and immobilised hybrid are identical.


Isolation and characterization of the acidic phosphoproteins of 60-S ribosomes from Artemia salina and rat liver. Eucaryotic L7/L12-type proteins are present in ethanol/salt extracts (P1 protein) of ribosomes from Artemia salina and rat liver. These proteins are partially phosphorylated and occur in two forms of closely related structure: a major form eL12 having methionine at the N-terminal position and a minor form of eL12 (eL12') which seems slightly elongated and contains a blocked N terminus. Purification of the four different forms of this protein, eL12, eL12-P, eL12' and eL12'-P, was performed by ion-exchange chromatography on carboxymethyl-cellulose and DEAE-cellulose. Using a radioimmuno assay, 1.8 copies of eL12 and 0.9 of eL12' were found on the 80-S A. salina ribosome. In ribosomes of both rat liver and A. salina, eL12 is present in a larger quantity than eL12'. 40-S and 60-S ribosomal subunits extracted with ethanol/salt were essentially free of eL12 proteins. A large pool of eL12 was found in the cytosol after removal of the ribosomes by centrifugation or molecular sieving. The proteins of rat liver and A. salina are similar with regard to their isoelectric points and molecular weights. Sedimentation equilibrium studies indicated that the isolated protein eL12 occurs as a dimer.


The cycling of calcium, sodium, and protons across the inner membrane of cardiac mitochondria. A method is described that permits simultaneous determination of the net charge transfer associated with Ca2+ transport by the ruthenium-red-sensitive carrier and the ionized internal [Ca2+] in heart mitochondria. The data indicate that this carrier catalyses a charge-uncompensated flux of Ca2+. Full charge compensation for Ca2+ influx is provided by the respiration-dependent efflux of H+. The net efflux of Ca2+ induced by Na+ is analysed in terms of two other carriers, a Na+-Ca2+ antiporter and a Na+-H+ antiporter. Evidence is presented that these two carriers are separate and that the Na+-H+ exchange is the more rapid. The fluxes of Ca2+, Na+ and H+ during the Na+-induced efflux of Ca2+ support a series of events in which the Na+-H+ exchange enables unidirectional Ca2+ fluxes via the uniport and antiport systems to be integrated into a cycle.


Purification and physicochemical properties of fatty acid synthetase and acetyl-CoA carboxylase from lactating rabbit mammary gland. Fatty acid synthetase was purified 13-fold from lactating rabbit mammary glands by a procedure which involved chromatography on DEAE-cellulose, ammonium sulphate precipitation and gel filtration on Sepharose 4B. The preparation was completed within two days and over 100 mg of enzyme was isolated from 100--150 g of mammary tissue, which represented a yield of over 40%. The preparation was homogeneous by the criteria of polyacrylamide gel electrophoresis and ultracentrifugal analysis. The sedimentation constant, S20,w was 13.3 S, the absorption coefficient, A280nm1%, measured refractometrically was 10.0 +/- 0.1, and the amino acid composition was determined. The subunit molecular weight determined by gel electrophoresis in the presence of sodium dodecyl sulphate was 252,000 +/- 6,000, and the molecular weight of the native enzyme measured by sedimentation equilibrium was 515,000. These experiments indicate that at the concentrations which exist in mammary tissue (2--4 mg/ml) fatty acid synthetase is a dimer. The purified enzyme did however show a tendency to dissociate to a monomeric 9-9S species on storage for several days or following exposure to a low ionic strength buffer at pH 8.3. There was only a small quantity of alkali labile phosphate (0.2 molecules per subunit) bound covalently to the purified enzyme. Acetyl-CoA carboxylase was purified 300-fold in a 50% yield within 24 h by ammonium sulphate and polyethylene glycol precipitations [Hardie, D.G. and Cohen, P. (1978) FEBS Lett. 91, 1--7]. The preparation was in a state approaching homogeneity as judged by polyacrylamide gel electrophoresis, gel filtration on Sepharose 4B and ultracentrifugal analysis. The sedimentation constant, S20,w, was 50.5 S, the absorption index, A280nm1%, was  14.5 +/- 0.7, and the amino acid composition was determined. The subunit molecular weight of acetyl-CoA carboxylase determined by gel electrophoresis in the presence of sodium dodecyl sulphate was identical to that of fatty acid synthetase (252,000) as shown by electrophoresis of a mixture of the two proteins. The preparations also contained two minor components of molecular weight 235,000 and 225,000, which appear to be derived from the major species of mol. wt 252,000. A large emount of phosphate (3.2 molecules per subunit) was found to be bound covalently to the purified enzyme. The properties of fatty acid synthetase and acetyl-CoA carboxylase are compared to those obtained by other workers.


Properties of human hemoglobin immobilized on Sepharose 4B. This paper reports the properties of human hemoglobin covalently bound to Sepharose 4B both in 'high-affinity' and 'low-affinity' conformations. The results suggest that the coupling reaction is strongly affected by the conformational changes linked to oxygenation of the protein. The rate and the extent of the reaction are different for the oxy and deoxyderivatives, probably due to the change in reactivity of the amino groups in the liganded and unliganded tetramer. The data on the equilibrium which is established between matrix-bound and soluble subunits, measured by the 'subunit-exchange chromatography', indicate that the system displays a minimal heterogeneity when hemoglobin is coupled to the gel in the deoxy state at intermediate protein concentration and pH 8. Maxtrix-bound hemoglobin is characterized by a higher oxygen affinity and by decreased homotropic and heterotropic interactions with respect to hemoglobin in solution, but the changes depend strongly on the conditions used in the coupling procedure.


Study of the tryptophan residues of lysozyme using 1H nuclear magnetic resonance. The identification and complete assignment of the C-2 and N-1 proton nuclear magnetic resonances (NMR) of the six tryptophan residues of hen lysozyme are reported. Identification of the resonances required a detailed examination of the spectra of the protein in H2O and in 2H2O, and involved the application of spin-echo and Carr-Purcell-Meiboom-Gill pulse sequences. Assignment was achieved by observing the effects on the NMR spectra of performing specific chemical modifications, of binding paramagnetic species (lanthanide ions and spin labels), of binding inhibitors and protons and of carrying out solvent exchange experiments. The problems involved in completion of assignment are fully discussed. In the course of performing experiments to make assignments, several interesting aspects of the behaviour of the tryptophan residues in the protein structure were observed and are discussed.


Acute influence of different beta-blocking agents upon left heart hemodynamics at rest and during exercise in patients with coronary artery disease. The study investigated the acute hemodynamic changes induced in patients with angiographically confirmed coronary artery disease by 3 beta-blockers: metoprolol, cardioselective without intrinsic sympathomimetic activity (ISA), group I, 11 patients; bunitrolol, noncardioselective with ISA, group II, 11 patients; oxprenolol, noncardioselective with ISA, group III, 11 patients. Hemodynamic variables were obtained at rest and during exercise, before and 45 min after 10 mg i.v. of the drug. Changes in LVEDP and cardiac indexes were such as LV function was improved in 1 patient of group I, 7 patients of group II and 5 patients of group III; impaired in 4 patients of group I and in 1 patient of group III; unchanged in the others. Contractility indexes were less influenced by bunitrolol and oxprenolol. During exercise there was a significant difference between groups for LVEDP which was lower in group II (P less than 0.01). The data seem to indicate that the choice of the beta-blocker may be of importance when it is desirable that an already compromised cardiac function be not further impaired by pharmacological intervention.


Comparison of disposition and effect of timolol and propranolol on exercise tachycardia. The kinetic disposition and beta-adrenergic blocking action in relation to plasma level of a single oral dose of either timolol or propranolol has been compared in healthy male volunteers. The disposition profiles clearly disclosed different properties of the two drugs, although their half-lives were similar. The available fraction of timolol in the systemic circulation was estimated to be approximately 60% of the dose, and 17.4% was exereted unchanged in urine. The logarithm of plasma concentration showed a significant correlation with the beta-blocking activity assessed by an exercise test. The mean potency ratios of timolol to propranolol as an antagonist of chronotropic effects on exercise tachycardia were 11 to 17 and 3.6 to 5.5 in dose- and concentration-effect relationships, respectively. The absolute reduction of exercise heart rate gave the best coefficient of all measures of beta-blockade. When drug action was measured as beta-blockade assessed by a given response to exercise tachycardia, the effect declined linearly with time, even though plasma levels fell exponentially. This results suggest that the pharmacokinetic t1/2 is much shorter than the pharmacological t1/2.


Influence of food on the absorption of acetylsalicylic acid from enteric-coated dosage forms. The absorption of acetylsalicylic acid (ASA) from two different enteric-coated dosage forms, tablets (Premaspin) and granules (Reumyl), was studied in healthy volunteers under fasting and non-fasting conditions by following the plasma concentration and urine recovery of salicylates after single doses of ASA 1 g. Conventional tablets (Aspirin) were used as the reference. Under fasting conditions the absorption of ASA from the two different enteric-coated preparations was complete. Taken with food the enteric-coated tablets gave much lower plasma concentrations than under fasting conditions, and absorption was not complete in all subjects. In contrast, absorption from the enteric-coated granules was not influenced by the intake of food. It was concluded that enteric-coated granules of ASA permit more reproducible absorption than enteric-coated tablets.


A relatively specific and quantitative assay for histamine H2-receptor blocking activity by determination of inhibition of histamine-induced gastric acid secretion in the rat. A relatively specific method for the quantitative assay of histamine H2-receptor antagonists has been developed. The method is based on the antagonism of the histamine-induced, and the spontaneous, gastric acid secretion in the stomach-perfused, urethane-anesthetized acute rat. For the induced gastric acid secretion, the animal received two consecutive equal injections of histamine (2 mg/kg, intrajugularly), the second 1.7 h after the termination of the first histamine-induced acid hypersecretion. Atropine (5 mg/kg), chlorisondamine (2 mg/kg), imipramine (10 mg/kg) and tripelennamine (5 mg/kg), administered i.p., failed to inhibit the histamine-induced, or the spontaneous, gastric acid secretion. Metiamide inhibited the histamine-induced acid secretion (ED50 1.85 mg/kg, i.p.) and the spontaneous acid secretion (10 mg/kg, i.p.). These results suggest that in the rat model employed in this study, histamine and H2-receptors play a very important role in the regulation of the gastric acid secretion. Other mechanisms involving cholinergic tone, biogenic amine uptake and histamine H1-receptors do not seem to intervene.


Comparative effects of clozapine and alpha-adrenoceptor blocking drugs on regional noradrenaline metabolism in rat brain. Clozapine increased brain noradrenaline (NA) metabolism, as indicated by changes in 3-methoxy-4-hydroxyphenylglycol sulfate content, in brain regions corresponding to the predominance of alpha- over beta-receptors, i.e., hypothalamus, medulla, midbrain and cortex, but not corpus striatum or cerebellum. Phenoxybenzamine had a stronger effect in the hypothalamus than did clozapine, but did not change cortical NA metabolism within a 60 min treatment time; however, cortical NA metabolism was increased 150 min after phenoxybenzamine. The delayed effect of phenoxybenzamine may be due to either a poor affinity for some central receptors or a slow rate of entry into certain brain regions. Thioridazine and the benzodioxane, dibozane, had regional effects similar to clozapine. The similarity between clozapine and dibozane in ther effects on regional brain NA metabolism may reflect a preference for presynaptic alpha-receptors. It is unlikely that the antipsychotic activity of clozapine is related to a specific adrenolytic effect, but may reflect the combined activity of this drug on several transmitter systems.


Irreversible beta-adrenoceptor blockade of atrial rate and tension responses. The competitive reversible beta-adrenoceptor antagonist activity of Ro 03-5255 [1-(5-acetylaminobenzfuran-2-yl)-2-isopropylaminoethanol] upon isoprenaline-induced increases of the rate and tension of guinea-pig isolated atria is described. The chlorinated derivative [Ro 03-7894; 1-[5-chloracetylaminobenzfuran-2-yl)-2-isopropyl-aminoethanol] in contrast exhibited concentration-dependent non-competitive irreversible blocking activity as measured by depression of the maximum responses which were not restored by a washout period that successfully reversed Ro 03-5255. When orciprenaline was used as a weak agonist of low efficacy, the maximum responses were depressed to a greater extent. The blockade by Ro 03-7894 was relatively specific for beta-adrenoceptors since it did not antagonize histamine or calcium chloride. The depression of the maximum responses to orciprenaline was reduced by the presence of sodium thiosulphate. Sodium thiosulphate was ineffective in reversing an established blockade. The blockade by Ro 03-7894 was therefore assumed to involve irreversible binding to the beta-adrenoceptor after conversion to an appropriate electrophilic ligand. The significance of this is discussed.


Blockade by WB 4101 of alpha-adrenoceptors in the rat vas deferens and guinea-pig taenia caeci. The effectiveness of WB 4101, a recently described alpha-adrenoceptor antagonist, in blocking an excitatory and two inhibitory responses to alpha-receptor activation was studied. One of the inhibitory responses was the reduction by the selective alpha-agonist amidephrine of carbachol contractures of isolated guinea-pig taenia caeci. WB 4101 antagonised this inhibition with a Schild plot slope of 0.99 and a pA2 of 8.9. The same pA2 value was obtained for blockade of the contractile effect of amidephrine and noradrenaline on the rat vas deferens. WB 4101 was, however, several hundred times less active in antagonising the inhibitory effect of clonidine on the twitch response of the vas deferens to field stimulation. Incidental observations were that the twitch was increased by low concentrations of amidephrine, and by relatively high concentrations of WB 4101. Because of its potency and postsynaptic selectivity, WB 4101 should be useful for adrenoceptor classification.


pH and temperature dependence of adenosine uptake in human erythrocytes. Kinetic analysis of the saturable adenosine uptake in human erythrocytes suggests the existence of two saturable components, distinguished by different Km values (1.4 and 260 micron, respectively, at pH 7.4 and 25 degrees C). Both components were abolished by p-nitrobenzylthioguanosine or dipyridamole. Total uptake was significantly higher at pH 8 than at pH 7 at adenosine concentrations above 2 micron. The increase in uptake at the higher pH was brought about mainly by an increase in the maximum rate of transport of the low-affinity uptake system. With rising temperature the Km and the V of both uptake components increased. No transition temperature was observed between 12 and 37 degrees C.


A difference in effects of physiological Ca2+ concentrations on activity of guanylate cyclase preparations obtained from the taenia caecum of guinea pig and from the longitudinal muscle of rat duodenum. A cholinergic stimulant, butyltrimethylammonium bromide and serotonin increased the tissue levels of cyclic GMP in the taenia caecum of guinea pig but not those in the longitudinal muscle of rat duodenum. On the other hand, physiological Ca2+ concentrations enhanced the activity of a guanylate cyclase preparation obtained from the taenia caecum of guinea pig, while guanylate cyclase in the longitudinal muscle of rat duodenum was not influenced by Ca2+. The difference in the effects of the smooth muscle stimulants on the tissue levels of cyclic GMP in two different smooth muscles in attributed to differences in the properties of guanylate cyclase of smooth muscles.


Measurement of beta-adrenoceptor antagonists in biological fluids using a radioreceptor assay. A new assay for plasma levels of beta-adrenoceptor antagonists is described. The assay depends upon the ability of the drug to compete with a labelled beta-adrenoceptor antagonist (-)-[3H]dihydroalprenolol for beta-adrenoceptor binding sites on lung membranes. The assay is simple to perform, very sensitive and does not require prior extraction of plasma. The assay can also detect bioactive metabolites of these agents and is clearly not limited to a single beta-adrenoceptor antagonist.


Antagonism of central histamine H1 receptors by antipsychotic drugs. The activity of 8 widely antipsychotic drugs as antagonists of central histamine H1 receptors was determined from the inhibition of the binding of 3H-mepyramine to a membrane fraction from guinea-pig brain. The phenothiazines examined, clorpromazine, fluphenazine, thioridazine and trifluoperazine, were all potent H1 antagonists. Both alpha- and beta-flupenthixol were potent inhibitors, but butaclamol, although less potent, showed stereospecificity. Haloperidol and spiperone were markedly weaker antihistamines than the phenothiazines.


Antagonism of morphine-induced catalepsy by L-prolyl-L-leucyl-glycinamide. In view of the recently demonstrated extra-endocrine central actions of hypothalamic releasing hormones, we have investigated the effects of prolyl-leucyl-glycinamide (PLG) and thyrotropin releasing hormone (TRH) on morphine-induced catalepsy. Although acute administration of PLG (10 mg kg-1 s.c.) slightly attenuated the cataleptic response, chronic PLG treatment (10 mg kg-1 s.c. for 10 days) virtually abolished morphine-induced catalepsy. TRH, administered subcutaneously, exhibited little or no anti-cataleptic activity. These results are discussed in relation to the possible central site of narcotic-induced catalepsy and the therapeutic potential of PLG in Parkinson's disease.


Serotonin antagonists and central hyperthermia produced by biogenic amines in conscious rabbits. Previous studies have shown that the hyperthermia produced by intracerebroventricular injection of 5-hydroxytryptamine (5-HT) to conscious rabbits was antagonized by cyproheptadine and increased by LSD. Other putative antagonists, i.e. cinanserin, methiothepin, 2-bromo LSD, methysergide and dimetiotazine are investigated in the present report. Cinanserin and methiothepin resembeld cyproheptadine, 2-bromo LSD had almost the same effects as LSD and methysergide exhibited a mixed pattern of action, being depressant or potentiating as a function of dose. Dimetiotazine had no specific action. Cinanserin, however, differed from cyproheptadine in selectively antagonizing and early component of the 5-HT rise, unmasking an important fall and leaving a late hyperthermic component unaffected, thus disclosing three distinct effects of 5-HT action. The cinanserin, methiothepin and methysergide antagonism of the 5-HT-induced temperature rise was greater than the antagonism of the noradrenaline (NA)-induced rise. Methiothepin and methysergide inhibited both the 5-HT and DA hyperthermia; cinanserin--like cyproheptadine--was more effective on the 5-HT rise. The potentiation of the 5-HT temperature rise by 2-bromo LSD and methysergide was more developed than was the potentiation of the NA and DA rises. The effects of the drugs studied on 5-HT action argue in favour of the existence of several types of central 5-HT receptors. The dissociation observed between the antagonism to 5-HT and that to DA does not favour a mediation of DA hyperthermia by 5-HT; antiserotonin drug antagonism of DA hyperthermia is more simply accounted for by interactions at the level fo specific DA receptors. The potentiation of the 5-HT-induced temperature rise by 2-bromo LSD and methysergide might result from an antagonism of the hypothermic component. As with LSD, 2-bromo LSD and methysergide alone also produced hyperthermia, the origin of which is briefly discussed.


Effects of the unilateral nigral application of dopaminergic drugs on the in vivo release of dopamine in the two caudate nuclei of the cat. The effects of the unilateral application of d-amphetamine, benztropine, haloperidol and thioproperazine to one substantia nigra on the release of 3H-dopamine (3H-DA) in the two caudate nuclei were examined in halothane-anesthetized cats. For this purpose animals were implanted with push-pull cannulae and 3H-DA was estimated in superfusates during the continuous delivery of L-3,5-3H-tyrosine. The nigral application of d-amphetamine (10-6 M) or benztropine (10-6 M) reduced the release of 3-H-DA in in the ipsilateral caudate nucleus and induced an opposite effect in the contralateral side. In contrast, the nigral application of haloperidol (10-6 M) or thioproperazine (10-6 M) slightly increased the release of 3H-DA in the ipsilateral caudate nucleus and induced a reduction of 3H-transmitter release in the contralateral side. These results emphasize the role of the dendritic release of DA in the control of the activity of dopaminergic neurons and confirm our previous findings concerning the existence of a reciprocal control in the activity of the two dopaminergic pathways.


Analysis of the action of 4-aminopyridine during repetitive stimulation at the neuromuscular junction. 4-Aminopyridine (4-AP) increased the quantal content (m) of end-plate potentials (e.p.p.s) evoked by continual stimulation (0.2--25 Hz) in frog end-plates depressed by Mg2+. The increase in m was due to an increase in the binomial parameter n. This was interpreted to mean that 4-AP increased the number of activated release sites. In junctions blocked by d-tubocurarine, 4-AP first increased and then decreased the amplitude of e.p.p.s. elicited during a train of stimuli of increasing frequency, indicating that 4-AP increased transmitter release more than mobilization.


Responses of olfactory bulb neurones to the dipeptide carnosine. Carnosine has been applied by microiontophoresis ot identified neurones in the olfactory bulb of the rat from solutions of different pH. Although mainly without effect when compared with conventional excitatory and inhibitory amino acids, the dipeptide tended to be depressive when ejected as a cation and excitant when ejected as an anion. The results obtained are not in favour of this substance being an excitatory transmitter in the primary olfactory pathway.


Glutathione peroxidase in dried blood spots. A new procedure utilizing dried blood spots was developed for detecting glutathione peroxidase deficiency. Samples from a known patient with a partial defect and from rats with an induced deficiency were distinguished from respective control groups by their longer defluorescence endpoints. Samples from 100 patients with anemia and 2 phenyl-ketonuric infants on low-protein diets contained glutathione peroxidase activity similar to that in 82 controls, when screened for the enzyme defect by the new procedure.


Factors influencing the distribution of 99m-technetium methylene diphosphonate in bone and soft tissues. The influences of dilution, storage and pH on the distribution of 99m-technetium methylene diphosphonate (99m-Tc-MDP) in bone and various soft tissues have been investigated in the rat. The results show a marked departure from normality when large dilution factors and very acidic or basic preparations are used.


Cholinergic and adrenergic innervation of the penis artery of the bull: transmitter concentrations and synaptic vesicles. The penile artery of the bull contained significant amounts of acetylcholine, noradrenaline and dopamine, and its axon profiles contained either numerous small granular or agranular vesicles, as well as some large granular vesicles. In the dorsal metatarsal artery, only noradrenaline and dopamine were detectable, and the axon profiles observed contained numerous small granular vesicles. In the penile artery, the axons with small agranular vesicles, probably cholinergic axons, were in close contact with axons containing small granular vesicles. It is suggested that, in the penile artery of the bull, one of the functions of the cholinergic nerves is suppression of excitatory adrenergic neurotransmission.


Transplantation of allogeneic bone marrow from thymus-deprived monkeys and certain characteristics of "graft-versus-host" cells. Transplantation of allogeneic bone marrow both from normal and thymus-deprived donors (thymectomized lethally irradiated and restored with autologous bone marrow) was followed 6--7 days later by the appearance of large numbers of GVH cells in bone marrow and peripheral blood which were one of the characteristic features of acute GVHR. GVH cells isolated from peripheral blood 7 days after transplantation showed a high labelling index (about 30%) after 40 min incubation with 3H-thymidine. The prolongation of incubation failed to increase the proportion of labelled cells due to the low viability of GVH cells. The results obtained with T and B cell tests suggested that GVH cells had the properties of T lymphocytes: about 60% of the cells could form rosettes with non-sensitized sheep red blood cells; there were virtually neither Ig-bearing cells nor cells with receptors for C3 and Fc portion of Ig.


[Hormone and metabolic profile in diabetic hyperosomolar coma. Plasma insulin response to intravenous tolbutamide (author's transl)]. Fifteen patients with non-ketotic hyperosmolar diabetic coma were investigated and compared with ketoacidotic patients. Basal plasma insulin levels were low in all patients (14.8 +/- 1.0 micronU/ml in hyperosmolar coma, 11.0 +/- 1.3 in keto-acidosis), but insulin level increased after intravenous tolbutamide (between 30 and 105 micronU/ml) in eight hyperosmolar comas. Insulin showed no increase in seven hyperosmolar comas and in none of the ketoacidotic patients. In hyperosmolar coma plasma free fatty acids (1710 +/- 197 micronEq/1), triglycerides (3,4 +/- 0,4 g/1) and cortisol levels (49,7 +/- 9,0 microgram/100 ml) were increased, must as in keto-acidosis. Growth hormone (1,7 +/- 0,1 ng/ml) was normal, unlike the case in keto-acidosis. Plasma lactate concentrations were elevated and account for the frequent mild acidosis found in hyperosmolar coma. In spite of the low peripheral "insulin/glycemia ratio", the positive response to tolbutamide in half of the hyperosmolar cases suggests a less complete pancreatic deficiency than in keto-acidosis. The plasma high free fatty acid and triglyceride levels suggest that the lack of ketosis is not due to inhibition of lipolysis but could be a consequence of inhibition of hepatic ketogenesis.


Rectovaginal fistula: management by intravenous feeding and surgical repair. Two patients, each with a large rectovaginal fistula, were managed successfully by intravenous feeding and surgical repair of their fistulae. Intravenous feeding eliminated the need for food from the gastrointestinal tract, thus minimizing the fecal flow. Both fistulae healed following surgical repair. Neither patient required a diverting colostomy.


Microinvasive carcinoma of the uterine cervix. There has been remarkable improvement in the early diagnosis of cervical carcinoma in recent years. There is, however, disagreement regarding the definition and treatment of microinvasive carcinoma of the uterine cervix. This review analyzes a large number of reports representing current opinion about the definition, diagnosis and appropriate management of early cervical carcinoma.


Serum immunoglobulin levels in normal, premature and postmature newborns and their mothers. Immunoglobulin (Ig) levels (IgG, IgA, IgM and IgD) were measured in 92 paired maternal and cord sera by the modified single radial immunodiffusion method. There was a progressive increase in mean IgG levels as the gestational age increased which reached a maximum of 1771 mg/100 ml in the postmature babies. The mean IgG level of infants of 32--36 weeks' gestation was lowest (1395 mg/100 ml). The maternal IgG levels were generally higher than those of the cord sera. The mean maternal IgG level (1773 mg/100 ml) was considerably higher than the corresponding level in the cord sera of the lowest gestational age group. Such marked difference was not present in newborns who were of normal gestational ages. The mean IgA and IgM levels of the cord sera were low. There were no significant differences between the mean IgA and IgM levels of newborns in different gestational age groups, even when their mothers showed variations in mean IgA and IgM levels. There was no detectable level of IgD in the cord sera.


Galactorrhea and the defective luteal phase of the menstrual cycle. Twelve patients with primary infertility due to a defective luteal phase associated with hyperprolactinemia and galactorrhea are reported. All were treated with 2-bromoergocryptine. During the first cycle of therapy, the serum prolactin level decreased in all cases, and galactorrhea disappeared during the second cycle of therapy. Seven women responded to 2-bromoergocryptine, as was reflected in their endometrial biopsies. The remaining five were given clomiphene citrate on the second and sixth days of the next 2-bromoergocryptine cycle. Three of these patients responded favorably to this combined therapy. Six pregnancies were reported during therapy. The results of follow-up after labor or abortion suggested that these cases represent a latent stage of the amenorrhea-galactorrhea syndrome.


Pregnancy and labor in Tanzanian primigravidae aged 15 years and under. The outcome of pregnancy and labor in 259 Tanzanian primigravidae 15 years and under is compared with that of 377 primigravidae between the ages of 21 and 25. The risk of antenatal complications, such as anemia, and premature labor and low birth weight babies was much higher in the younger age group. There was no difference in the incidence of preeclampsia or complications during labor between the two groups. The younger women had a lower perinatal mortality rate than the controls.


A comparison of socio-demographic and fertility characteristics of women sterilized in hospitals and camps. Socio-demographic characteristics of 5846 women undergoing sterilization in hospitals and 1752 women undergoing sterilization in camps in India are analyzed. The average age of women accepting sterilization was 29.8 years; the mean number of living children was 4.2. Women sterilized in hospitals were of significantly higher age and parity than those sterilized in camps. The mean parity for all women was 4.6. A steep increase in median parity was observed with increasing age in both hospitals and camps. The study indicates that, while the level of education may affect acceptance of sterilization, there is no minimum level of education necessary for its acceptance. Most of the women in the study were not gainfully employed. As expected most of the women were Hindus. Muslims were underrepresented in the hospital series; in camps, however, Muslims showed a much higher rate of acceptance of sterilization. The rate of pregnancy wastage was significantly higher in the hospital and the child loss rate was significantly higher for the camp cases. While the rate of previous abortions was higher for low parity women, the child loss rate was higher for high parity women. The child loss parity ratio was significantly higher for the camps than for the hospital cases. The vast majority of women in both hospitals and camps reported no previous contraceptive practice. Sterilization appears to have been the method of choice for most of these women.


Tubal sterilization and ovarian perfusion: selective arteriography in vivo and in vitro. To determine whether there are changes in the utero-tubo-ovarian circulation after tubal sterilization, selective arteriograms were taken in vivo every 0.3--7.0 seconds with a Puck-type seriograph and also in vitro. A control arteriographic study was made. The operations performed were fimbriectomy and modified Pomeroy. The 30 patients studied were divided into two groups. One was formed by 20 nonpregnant women. The other ten patients were in the early puerperium. Arteriographic images with and without tubal sterilization were prepared, and the different techniques were compared. With the Pomeroy operations, the various tubal sites where surgery was performed were also compared. It was concluded that neither the modified Pomeroy sterilization nor fimbriectomy alters ovarian perfusion in either nonpregnant women or those in the early puerperium.


First trimester abortion by vacuum aspiration: interphysician variability. This study compares the performances of four physicians using two types of plastic cannulae (flexible and rigid) for 1100 vacuum aspiration procedures. The criteria for assessing physician performance by cannula type were: (a) frequency of cannula obstructions, (b) amount of retained tissue obtained by sharp curettage after vacuum aspiration, (c) estimated blood loss during the procedure and (d) time required to perform the procedure. Data for each physician were compared and yielded significantly different results. The length of time required to use the cannula differed significantly among physicians; two of them had significantly shorter cannula times with the flexible instrument. The others had the lowest distributions of blood loss and the most difference between cannula usage. Consistent grouping over several variables suggests that differences in these criteria are more likely to be attributable to subtle distinctions in operator technique than to variations in equipment.


A comparative study of the tubal ring applied via minilaparotomy and laparoscopy in postabortion cases. Sterilization with tubal rings applied via minilaparotomy or laparoscopy was performed on 300 randomly selected postabortion subjects to evaluate the safety and effectiveness of the two surgical methods. One hundred and forty-nine procedures were performed by minilaparotomy and 151 by laparoscopy. Gas leakage due to equipment problems was the most common technical difficulty during laparoscopy, and difficulty in exteriorizing the tubes was the most common surgical difficulty. Women undergoing laparoscopy experienced significantly less pain during surgery and had significantly lower rates of immediate and early postoperative complications than those women undergoing minilaparotomy. The rates of gynecologic abnormalities at six and 12 months were similar for both procedures. At this writing, no pregnancies have been reported among the study subjects. The results of our study indicate that laparoscopy is superior to minilaparotomy when it is performed in a controlled hospital situation.


Radiographic abnormalities in patients with Stein-Leventhal syndrome. Radiographic abnormalities in 254 patients with Stein-Leventhal syndrome are presented in a retrospective study of the six-year period from 1972--1977. Twenty-two (8.7%) of the patients were found to have abnormal sella turcica x-rays, eight (3.2%) patients had abnormal hysterosalpingograms and five (2.0%) patients had abnormal intravenous pyelograms. In addition, three patients with an abnormal sella turcica had serum prolactin levels greater than 45 ng/ml, which should be regarded as evidence of a pituitary adenoma. These findings suggest that serum prolactin assays and skull x-rays with polytomography should be routinely performed on patients with Stein-Leventhal syndrome.


A comparison of the clip and ring techniques for laparoscopic sterilization of postabortion and postpartum patients. The spring-loaded clip and the tubal ring techniques of laparoscopic female sterilization are compared for ease of performance, safety and effectiveness at the Kandang Kerbau Hospital in Singapore. The two techniques were randomly assigned to 299 postpartum patients and 300 postabortion patients. The assigned procedure could not be performed for two of the ring cases and three of the clip cases. Bleeding and equipment problems made laparotomy necessary in two clip cases and equipment problems caused a change in technique in the other clip case. Hydrosalpinx and adhesions were responsible for a change to laparotomy in the two ring cases. Surgical difficulties, chiefly the result of equipment problems, were significantly higher for both postpartum and postabortion clip procedures. Complications and complaints reported immediately after surgery and at the 7- to 21-day follow-up visit were low for all four groups. For both postabortion and postpartum patients, six- and 12-month data show that pregnancy rates are significantly higher for patients sterilized with the clip than with tubal rings.


Systemic reactions with total dose infusion of iron dextran complex in obstetric patients. Iron dextran complex (Imferon) total dose infusion (TDI) was used to treat iron deficiency anemia in 310 obstetric patients. Systemic reactions were reported for 13.5% of the group. Most (90.5%) of the severe reactions occurred during the test dose, while 66.6% recorded during or after TDI were mild. Antihistamine premedication included administration of promethazine hydrochloride to 104 patients and intramuscular and intravenous drip chlorpheniramine hydrogen maleate to 206 patients. Total systemic reactions were statistically greater  with intramuscular promethazine hydrochloride than with intramuscular and intravenous chlorpheniramine maleate (p less than 0.001). On the other hand, the rates of severe reactions and reactions during the test dose were similar for both antihistamine schedules (p greater than 0.05). The incidence of all systemic reactions was the same for pregnant and postnatal women (p greater than 0.05), but severe reactions were more frequent among pregnant than postnatal patients (p less than 0.001). There were no fatalities.


Comparative IUD study: Lippes Loop D, Dalkon Shield and TCu-200. A study was undertaken to look at the clinical behavior of the Dalkon Shield, the Lippes Loop D and the TCu-200 inserted by physicians and midwives. Differences among the three devices were not statistically significant, although lower expulsion rates and higher pregnancy rates were reported for the Dalkon Shield. No differences were observed between insertions performed by physicians and those performed by midwives; this emphasizes the efficiency of using paramedical personnel in the field of intrauterine contraception.


Plasma concentrations of methylergometrine after intravenous and intramuscular administration. The concentrations of methylergometrine (M) in the plasma were determined by a new radioimmunoassay after a single intravenous and intramuscular administration of two brands. Methergin (Sandoz Pharmaceuticals, Inc, East Hanover, NJ, USA) and Myomergin (Leiras Pharmaceuticals, Turku, Finland). No significant differences between the two brands were found. After the intravenous injection, M distributed quickly from the plasma to the tissues, and according to the elimination phase half-life M has no cumulative properties. After the intramuscular injection, M was absorbed quickly with peak plasma concentrations at one-half hour.


The incretin concept today.

The incretin concept today. 1. The insulinogenic factor of the gastrointestinal mucosa named "incretin" is only one part of the complex enteroinsular axis. --2. Of the chemically defined gastrointestinal hormones GIP is the strongest incretin candidate. --3. Because of the dual function of GIP as gastrone and insulinotropic substance several safeguards against GIP-mediated insulin hypoglycaemia exist. --4. No pathological condition has yet been found which is causally related to hyper- or hyposecretion of GIP. However, an exaggerated GIP response (usually secondary to the disease) may participate in the pathogenesis of hyperinsulinaemia of patients with obesity and duodenal ulcer. --5. The injection of GIP antibodies only partially abolishes the incretin effect. Therefore, GIP, although important, is not the only incretin.


The prevalence and persistence of group B streptococcal colonization among hospital personnel. A prospective study was performed to determine the prevalence and persistence of group B streptococcal colonization among obstetric (high-risk) and nonobstetric (low-risk) personnel. Seventy-four individuals participated in the study and the following sites were sampled: throat, rectum, vagina (females) and anterior urethra (males). The overall colonization rate was 32.4% and no statistical difference was found between high- and low-risk groups. The most frequently recovered serotypes were type III (37.5%) and type II/Ic (33.3%). Individuals older than 30 years were more likely to carry type II/Ic, whereas personnel in their twenties were most frequently colonized with type III. The rectum was the most frequently colonized site (83.3%). The vagina/urethra was colonized in 62.5% and the throat in 8.4% of carriers. Twenty-three culture-positive individuals were recultured from all sites three to six months later and persistent colonization was found in 56.5%. There was no statistical difference in persistence between the high- and low-risk groups. Type III carriers tended to become culture-negative, whereas type II/Ic carriers were significantly more likely to remain colonized with group B streptococci.


Histamine receptors in the gastric microcirculation. The types and functions of histamine receptors in the submucosal arterioles of the corpus and antrum of the cat and rat stomach were studied using an in vivo microscopy technique. Change in arteriolar diameter in response to superfusion of histamine with and without antagonists was measured by an image-splitting technique. H1 and H2 histamine receptors subserving vasodilatation were demonstrated in both the antral and corpus submucosal arterioles of the cat and rat. However, the H1 effect was predominant in the antrum (the H2 antagonist inhibited histamine dilatation only in the presence of the H1 antagonist), while H1 and H2 effects were approximately equal and independent in the corpus.


[Presynaptic receptor systems in central noradrenergic neurons]. Slices of rat occipital cortex, hypothalamus or cerebellar cortex were stimulated either electrically or by high potassium (in few experiments by tyramine). 1. Electrical or potassium stimulation elicit physiologically noradrenaline release in contrast to tyramine. 2. Stimulation-induced overflow of tritium, reflecting noradrenaline release, was diminished by a) alpha-adrenoceptor agonists, b) morphine and enkephaline, c) prostaglandin E1. The effect of agonists was abolished only by specific antagonists. Metabolism of 3H-noradrenaline was unchanged. 3. It is concluded that transmitter release from the noradrenergic neurones of several brain areas is modulated by drugs acting on alpha-adrenoceptors, opiate receptors and prostaglandin receptors. In contrast to other tissues there was no evidence obtained for presynaptic dopamine, beta-, muscarine, nicotine and angiotensin receptors. It seems likely that the receptors, involved in the modulation of noradrenaline release, are located at the noradrenergic nerve ending themselves, i.e. that they are presynaptic receptors.


[Clinical and experimental histochemical studies on the activities of liver lysosomal enzymes and gamma-glutamyl transpeptidase (gamma-GTP) (author's transl)]. The effects of acute and chronic administration of D-Galactosamine (GalN), Ethanol and Phenobarbital were investigated on the activities of lysosomal enzymes, i.e.; acid phosphatase, beta-glucuronidase and n-acetyl-beta-glucosaminidase, and others such as gamma-GTP and adenosine triphosphatase. The histochemical distribution of gamma-GTP in the liver was also studied on biopsy specimens from patients with chronic hepatitis, and gamma-GTP levels in the serum of patients receiving drugs inductable of hepatic microsomal enzymes. 1) After a single intraperitoneal injection of GalN, the lysosomal enzyme activities were lowered in the necrotic areas, but raised in the perinecrotic areas, the proliferative Kupffer cells and intra- and/or extra-cellular eosine bodies. 2) gamma-GTP activities in rat liver after chronic administration of GalN were markedly increased in bile canalicular membrane of periportal parenchymal cells, the epithelium of bile duct and ductules, and som inflammatory cells of portal fields. Levels of serum gamma-GTP were also elevated. On histochemical studies with biopsy specimens from patients with chronic active hepatitis showing elevated gamma-GTP activity, the activity was revealed a similar localization to GalN-treated rats. These data suggested that the increased activities might be reflected on the active stage in chronic hepatitis. 3) Chronic ethanol treatment in rats induced clearly-stained lysosomes varied in size, especially large-sized. The activities of hepatic gamma-GTP were slightly increased in the bile canalicular membrane of periportal parenchymal cells and the epithelium of proliferative bile ductules. 4) It has been shown by histochemical and biochemical techniques that hepatic gamma-GTP activity was increased after phenobarbital administration in rats. A significant rise in serum gamma-GTP was observed in patients on long-term treatment with anti-epileptic drugs. These data indicated that the increased activities of serum gamma-GTP might be accompanied with induction of hepatic microsomal drug-metabolizing enzymes.


The hospital and optimal chemotherapy in schizophrenia. With some schizophrenics, response to antipsychotic drugs is best monitored in the hospital. They include dysphoric responders, those whose illness has been only minimally responsive to antipsychotic drugs, and habitual noncompliers with drug regimen. Dysphoric responses need to be acknowledged and regarded as a danger signal. Patients whose illness is only modestly sensitive to drugs are particularly in danger of psychotoxicity due to excessive dosage. Habitual noncompliers have little investment in staying well, and cannot be expected to bear even mild drug side-effects.


Pharmacists expand their roles through clinical services. This article highlights various educational forums that were held at the 13th annual Midyear Clinical Meeting of the American Society of Hospital Pharmacists. Among the areas covered by the article are justification for clinical pharmacy services, administrative considerations in a drug utilization review program, and the expanding role of the pharmacist in the emergency department.


Immunogenicity of gonococcal Gc2 polysaccharide: comparative studies with pneumococcal type III polysaccharide and Salmonella typhosa Vi antigen. A plaque assay technique was used to assess the immunogenicity of a gonococcal cell wall polysaccharide (Gc2 antigen) in BALB/c mice. The Gc2 antigen was shown to be immunogenic, and the kinetics of the response differed from that of a pneumococcal polysaccharide (SSS-III) and a polysaccharide antigen of Salmonella typhosa (Vi antigen). In addition, using antithymocyte sera, the T-lymphocyte dependency of these antigens was investigated. The immune response to the Gc2 antigen was demonstrated to be dependent on a population of helper T cells, whereas the response to SSS-III appears to be regulated by suppressor T cells. There appears to be marked differences in the immune response of mice to different bacterial polysaccharides.


Degradation of levan by Actinomyces viscosus. Actinomyces viscosus ATCC 15987 was examined for its ability to hydrolyze its own levan. Washed whole cells and an ammonium sulfate fraction from cell-free culture fluids were shown to possess levan hydrolase activity. Analyses of reaction mixtures by gel filtration and thin-layer chromatography demonstrated that the product of levan hydrolysis was free fructose. The cell-associated and extracellular enzyme preparations also hydrolyzed inulin and the levans synthesized by Aerobacter levanicum and Bacillus subtilis. Growth of A. viscosus in media supplemented with 0.1% A. viscosus levan resulted in a 33-fold increase and a 7-fold increase in the specific activities of the respective extracellular and cell-associated enzymes when compared with those from 55 mM glucose cultures. Growth in the presence of 29.2 mM sucrose resulted in a 28-fold increase and a 5-fold increase in the specific activities of the respective enzymes when compared with those from the glucose cultures. The extracellular enzyme exhibited high activity over a wide pH range, with 87 and 89% of its pH 6.0 optimum activity at pH 5.0 and 7.0, respectively. The cell-associated enzyme also exhibited optimum activity at pH 6.0, but this was decreased to 10 and 20% at pH 5.0 and 7.0, respectively. Analysis for the presence of extracellular levan during growth of A. viscosus in sucrose broths demonstrated that peak levan concentrations occurred during the mid-exponential to late-exponential phase of growth followed by a rapid decline in extracellular levan as a result of levan hydrolase activity.


Bordetella pertussis does not induce beta-adrenergic blockade. Bordetella pertussis organisms induce histamine sensitivity and diminish the normal hyperglycemic response to epinephrine in experimental animals. These effects have been attributed to beta-adrenergic blockade. However, under conditions in which the decrease in epinephrine-induced hyperglycemia after B. pertussis administration was demonstrable, there was no change in rat reticulocyte beta-adrenergic receptor number or affinity measured by iodohydroxybenzylpindolol binding or in isoproterenol-stimulated adenylate cyclase activity. Therefore, there was no generalized beta-adrenergic blockade induced by B. pertussis. The observed effects can be explained by the hypersecretion of insulin resulting from B. pertussis administration.


Binding of cholesterol by Neisseria gonorrhoeae. The binding of [1,2-3H]cholesterol to Neisseria gonorrhoeae CS-7, Pseudomonas aeruginosa, and Salmonella typhimurium (smooth and rough strains) was investigated. The kinetics of cholesterol binding to N. gonorrhoeae CS-7 demonstrated that binding occurred slowly with maximum binding by 10 h. Under optimum conditions, a large percentage (65%) of the added cholesterol was associated with the cells. Chemical fractionation revealed that ca. 98% of the labeled cholesterol was associated with the cell membrane(s). The bound cholesterol was not esterified and was associated primarily with the cytoplasmic membrane. Intact gonococci bound 4 to 30 times more cholesterol than the deep rough mutant S. typhimurium TA1535, the wild-type S. typhimurium DB-21, and P. aeruginosa. In contrast, isolated cell membranes from all organisms rapidly bound cholesterol to the same extent. Therefore, the outer membrane can function as a permeability barrier to cholesterol. Cholesterol binding to both whole cells and isolated cell membranes was influenced by the incubation temperature. The rate of cholesterol binding by whole cells of N. gonorrhoeae decreased markedly at lower temperatures, with almost complete cessation of binding at 0 degrees C. A similar temperature effect on the binding of cholesterol to isolated membranes was not observed. Thus, the effect of temperature on the binding of cholesterol to whole cells was an effect not on the actual binding process but rather on the ability of the cholesterol molecule to penetrate the lipid domain of the gonococcal outer membrane.


Extractability of cell wall polysaccharide from lactobacilli and streptococci by autoclaving and by dilue acid. Autoclaving cell wall of Streptococcus mutans Ingbritt for 15 min under the Rantz and Randall conditions released one-tenth of the total cell wall carbohydrate, whereas two-thirds was extracted after autoclaving for 180 min. The extract contained the serotype c-specific antigen but lacked the lipoteichoic acid component extracted when whole cells were autoclaved. Autoclaving cell wall preparations from other strains of S. mutans and also Streptococcus salivarius and Streptococcus mitis in 0.85% NaCl for 180 min released the major proportion of the wall polysaccharide fraction. Approximately 50 to 90% of wall carbohydrate of Lactobacillus fermentum and Lactobacillus casei was released when cell wall preparations were autoclaved in 0.85% NaCl for 180 min. For wall preparations from several strains of S. mutans, autoclaving for 60 min at pH 3.75 released only 39 to 62% of wall carbohydrate, whereas almost total release could be achieved with the lactobacilli. Heating S. mutans Ingbritt cell wall for 24 h at 60 degrees C in 0.1 N H(2)SO(4) released only two-thirds of the wall carbohydrate; by comparison nearly all of the wall carbohydrate was released in 3 h from L. casei and L. fermentum. Autoclaving L. casei cell wall and purified soluble wall fractions hydrolyzed the phosphodiester bond between the polysaccharide and peptidoglycan. This was shown by the release of reactive N-acetylhexosamine in both cases and the presence of a phosphomonoester in the autoclaved soluble wall fractions. The results indicate that autoclaving can hydrolyze covalent linkages, and this must be considered when the Rantz and Randall procedure is used to obtain antigen preparations.


Proteolytic activity of Candida albicans: action on human salivary proteins. The susceptibility of human salivary proteins to degradation by Candida albicans was studied. The organism was cultivated in either whole-salivary supernatant or parotid fluid, both of which were supplemented with glucose (0.1%). The culture pH's were at, or above, neutrality. After growth, the culture supernatant solutions were examined by polyacrylamide gel electrophoresis for alterations in their profiles of salivary proteins. No evidence of proteolysis of whole-saliva or parotid fluid proteins was found. Salivary proteins, however, are susceptible to degradation by preparations of C. albicans protease. Candida protease was incubated with parotid fluid adjusted to several pH values. After incubation the reaction mixtures were subjected to polyacrylamide gel electrophoresis. Extensive degradation of parotid proteins was found at pH 4, very slight proteolysis at pH 5, and no degradation at pH 6 or 7. No selectivity in proteolysis of the several parotid proteins was noted. These results indicate that C. albicans protease is strictly dependent upon low (ca. 4) pH for activity on salivary proteins. Furthermore, it is suggested that due to the pH requirements of the enzyme, it is unlikely to be of major significance to the pathogenesis of Candida-induced oral inflammatory lesions.


Phytohemagglutinin stimulation of enhanced immunoglobulin G production in mice inoculated with type III pneumococcal polysaccharide. In BALB/c mice previously inoculated intraperitoneally with an immunogenic dose of the T-independent antigen type III pneumococcal polysaccharide, the intravenous administration of the T-cell activating agent phytohemagglutinin P causes a pronounced increase in the number and relative proportion of immunoglobulin G-producing cells. These results, detected by a modified hemolytic plaque assay, were supported by finding increased levels of serum immunoglobulin G anti-type III pneumococcal polysaccharide in the treated mice. A comparable stimulation of immunoglobulin G antibody-producing cells was not induced in phytohemagglutinin P-treated nude mice, indicating that the change in class of the predominant antibody is attributable to the activation by the phytohemagglutinin P of a T-cell population. Under the conditions of these experiments, phytohemagglutinin P also promotes a progressive suppression of the antibody-forming cells during the response to type III pneumococcal polysaccharide.


Dissociation between clinical and exercise responsiveness to beta-blockade in angina. Twenty outpatients with mild angina were prescribed placebo tablets b.i.d. for 7 weeks followed by acebutolol, a cardioselective beta-blocker, 200 mg b.i.d. for 21 weeks under single-blind conditions. One graded multistage treadmill test was carried out after each treatment period and an angina diary was filled daily for the 6 months of the trial. Attack frequency declined by 71% from 2.59 per week on placebo to 0.76 per week on acebutolol (p less than 0.05). Exercise duration on the treadmill increased by 56%, from 5.95 minutes on placebo to 9.32 minutes on acebutolol (p less than 0.001). A satisfactory clinical response (50% or greater decline in attack frequency per week) occurred in 15 out of 19 patients (79%; a 100% or greater increase in exercise duration on the treadmill was observed in 10 out of 19 cases (53%). Exercise responsiveness was well predicted by exercise duration on placebo (r = 0.91, p less than 0.0005), patients with the least initial tolerance being the most improved. Clinical responsiveness was not well predicted by initial exercise tolerance (r = 0.38, N;S.) or by the improvement in exercise tolerance (r = 0.33, N.S.). It is concluded that acebutolol substantially reduces anginal attack frequency even in patients in whom exercise tolerance is not significantly improved, at the dose of 400 mg/day.


Extramitochondrial protein synthesis in calf brain synaptosomes. Isolated synaptosomes of calf brain cortex incorporated labelled amino acids into their mitochondrial, membranous and soluble proteins in an approximate ratio of 1:1:0.5 Synaptosomal protein synthesis was sensitive to ATP, noradrenaline, cycloheximide and puromycin, and together with mitochondrial protein synthesis, also to chloramphenicol, 2.4-dinitrophenol, KCN and hyperosmotic conditions. The absence of Na+ and K+ ions slightly inhibited both synaptosomal and mitochondrial protein synthesis. Using incorporated radioactivity as an indicator of synthesized proteins, the synaptosomal soluble proteins could be obtained in one large peak in gel and indicator of synthesized protein, the synaptosomal soluble proteins could be obtained in one large peak in gel filtration on Sephadex G-100 and G-25, and in two components in disc electrophoresis on a 7% polyacrylamide gel. An approximate molecular weight was calculated for the synthesized proteins using known proteins as standards, giving 15000-35000 in the gel filtration eluant, and 27000 and 36000 in the disc electrophoresis bands.


Scintigraphy of the testes: a new method in the diagnosis of undescended testes. A new method of scintigraphy of the testes using 99mTc--Na-pertechnetate is described. Eleven men without clinical features of testicular disease and 38 patients with cryptorchidism were examined. The results of the radionuclide investigations were confirmed surgically. Scintigraphy of the testes provides an opportunity to work out the indications for the particular type of the operation and management of the patients with cryptorchidism.


In vitro cultivation of leprosy bacilli in hyaluronic acid-based medium. 2. Progress and developing concept of the role of hyaluronic acid suggested by culture and armadillo infection studies. Progress is summarized relating to the verification, identification of M. leprae and understanding of the process of adaptation the pathogen passes through before in vitro growth takes place. It is recognized that hyaluronic acid apparently does not serve as a source of energy but the possibility is presented that it plays a role in the reconstruction of M. leprae cell walls made "leaky" by constant intracellular life. This apparently occurs, in culture, initially by the development of coccoid forms which after a period of weeks finally give rise to acid-fast bacilli. If these understandings are correct and the bacillary cell walls are vitated by enzyme and other action occasioned by intra-macrophage existence, then cell wall antigenicity may also be vitiated or altered by intracellular parasitism and restored by in vitro cultivation. The possible importance of this hypothesis in the understanding of immunologic responses in leprosy, and in the possibilities for therapeutic use and vaccine development are discussed.


Adverse effects in maintenance therapy.

Adverse effects in maintenance therapy. Adverse effects in maintenance therapy with neuroleptic drugs and lithium salts were reviewed with special emphasis on tardive dyskinesia. The possible teratogenicity of neuroleptics and lithium salts was discussed and the chromosomal abnormalities seen in patients treated with lithium salts described.


Therapy of extrapyramidal side effects, with particular reference to persistent dyskinesia and lithium tremor. In 13 of 27 cases of persistent dyskinesia, treatment with amantadine, in an average dose of 300 mg, brought good to moderate improvement. In 14 patients who showed no response whatever, further treatment, with bromocriptine in doses raised gradually to a final daily dosage of 15 mg, was effective in four cases, though the improvement was generally only moderate. In more than 50 percent of the cases of initial tremor induced by lithium therapy, oxprenolol in daily doses of 160--240 mg produced good effects and moderate improvement was noted in a few further cases. In a series of 20 patients with initial tremor due to neuroleptic therapy, on the other hand, the same treatment proved unsuccessful in the majority of cases. This is the converse of the experience gained with the classical antiparkinson agents, which have proved more effective against tremor induced by neuroleptics than against lithium-induced tremor.


The ocular hypertensive response following experimental acid burns in the rabbit eye. Following application of 50 microliter of 2N hydrochloric acid to the rabbit cornea, the intraocular pressure rapidly increases and remains markedly elevated for up to 3 hr. The initial rapid increase in intraocular pressure appears to be the result of acid-induced shrinkage of the outer collagenous coats of the eye. The sustained rise in intraocular pressure is mediated in part by prostaglandin release. Increased prostaglandin-like activity, determined in the aqueous after an acid burn, was greatly inhibited by pretreatment of rabbits with indomethacin and to a much lesser extent by pretreatment with imidazole. Both indomethacin and imidazole essentially abolished the sustained elevation of intraocular pressure after an acid burn. Analysis of changes in pH and protein level in the aqueous implies that the stimulus for prostaglanding release within the eye is the penetration of hydrogen ions into the aqueous humor, with resultant intraocular trauma.


The cytochemical localization of nucleotide pyrophosphatase activity in plant tissues using naphthyl esters of thymidine-5'-phosphate. A cytochemical method for the localization of nucleotide pyrophosphatase activity in plants employing naphthol AS-BI thymidine 5'-monophosphate and alpha-naphthyl thymidine 5'-monophosphate as specific substrates is reported. Biochemical evidence for the validity of this method is presented and the synthesis of the naphthol AS-BI ester is described. The application of this cytochemical technique to shoots of Triticum sp. and roots of Vicia faba has shown nucleotide pyrophosphatase to be ubiquitous in its distribution in these organs and to occur in a structurally-bound form in the cytoplasm. The highest activity was detected in developing fibres adjacent to the leaf vascular bundles, in the coleoptile epidermal and hypodermal cells and in the coleoptile and leaf xylem.


A combined histochemical and biochemical study of myofibrillar ATPase in pectoral, leg and cardiac muscle of several species of bird. Frozen sections of the pectoral, gastrocnemius and cardiac muscles from seven different species of birds were stained for myofibrillar ATPase and for succinic dehydrogenase. Several methods of myofibrillar ATPase were used including different pre-incubation treatments. Myofibrillar ATPases were also measured biochemically and the pH profile of the activity was compared with the histochemical staining following pre-incubation at different pH. Myofibrils from the different muscles were also subjected to sodium dodecyl sulphate acrylamide gel electrophoresis in order to separate the low molecular weight components of myosin. The results demonstrated that histochemical methods can be applied, with a reasonable degree of confidence, to classifying fibres in avian muscles although the classification used for mammalian muscles needs to be modified. They also showed that avian muscles, particularly the pectoralis, varies considerably between species and their mode of locomotion.


Morphometric analyses of the electric organ of Torpedo: the influence of different fixative modes on the vesicle diameter. The influences of various fixatives on the vesicle size of the electric organ of Torpedo marmorate were investigated. Thin section and freeze etched preparations were examined under the electron microscope. In thin section increased vesicle diameters were observed compared with the freeze etched preparations. The same experiments in different torpedo fish led to significantly different vesicle sizes observed. Variations of the molarity, the pH and osmolarities result in particularly high differences in vesicle diameters. Using Karnovsky's method (1965) and a fixative consisting of 2.5% glutaraldehyde in 0.2 M cacodylate buffer, pH 7.2, results in vesicle sizes comparable to those reported by other authors. Results obtained from freeze etched preparations are not comparable in general with results from thin section experiments with the same fixative.


Analysis of PCO2 differences during rebreathing due to slow pH equilibration in blood. A quantitative analysis of the reaction and transport processes that occur in blood during and after gas exchange has been used to investigate mechanisms that might account for positive alveolar-mixed venous (A-V) and alveolar-arterial (Aa) PCO2 differences during rebreathing. The analysis was used to determine PCO2 changes that take place in blood as it travels from veins to arteries under conditions in which no CO2 is exchanged in the lung. The predicted A-V and Aa PCO2 differences are all positive and lie within the range of reported measured values. The differences are due to disequilibrium of [H+] between plasma and red blood cells, and to disequilibrium of the reactions CO2 in equilibrium HCO3- + H+ in plasma, as blood leaves the tissue and/or lung capillaries. The differences are increased with exercise and with continued O2 uptake in the lung, the latter due to the Haldane shift. We conclude that the two disequilibria and the Haldane shift contribute to the reported PCO2 differences in rebreathing animals but may not fully account for them. These mechanisms cannot explain any PCO2 differences that might exist during net CO2 elimination from blood in the lung.


Slow postcapillary pH changes in blood in anesthetized animals. To investigate the hypothesis that blood pH and PCO2 continue to change after the blood leaves an exchange capillary, we used a rapidly responding, pressure-insensitive, stopped-flow pH electrode apparatus. Arterial blood from an anesthetized dog or cat is drawn through the apparatus into a syringe. Syringe movement is then suddenly stopped. Temperature and pH of the blood in the electrode assembly are continuously monitored, both before and after blood withdrawal ceases. Hemolysis was reduced by coating all blood contact surfaces with silicone and fasting the animal overnight, anesthetizing it with crystalline pentobarbital sodium, and allowing it to ventilate spontaneously. After stopping withdrawal, pH of blood in the electrode chamber continued to change, rising 0.01 unit with t1/2 of 4.4 s. After lysed blood was returned to the animal to provide carbonic anhydrase to the plasma, no pH change was seen after stopping the flow. The small pH rise occurring in arterial blood in vivo is probably due in large part to disequilibrium of [H+] between red blood cells and plasma at the end of the pulmonary capillary, the equilibration process being rate-limited by the extracellular CO2 hydration-dehydration reaction.


Sensitivity of the carotid body to within-breath changes in arterial PCO2. Respiration, sinus nerve chemoreceptor discharge, and carotid arterial pH were monitored in cats. Chemoreceptor discharge frequency showed oscillations that had a respiratory period when averaged over many respiratory cycles. These oscillations disappeared when pH oscillations of respiratory period were eliminated from the carotid arterial blood. The maximum sinus nerve discharge was associated with the most acid point of the recorded pH oscillation. Briefly increasing PCO2 by giving CO2-rich saline into the aortic root resulted in brief reduction in carotid arterial pH, and when this reduction occurred during inspiration tidal volume increased, even with a pH change no larger than the pH oscillations. However, increased chemoreceptor discharge could only be demonstrated when each pH change had twice the amplitude of the pH oscillations. Injections of fixed acid mixed with free carbonic anhydrase transiently increased chemoreceptor frequency, whereas injections of fixed acid alone had no effect. The carotid body is therefore sensitive to small rapid changes in arterial PCO2, and the pH electrode record indicates the size of the stimulus except when fixed acid changes are produced too closely upstream.


Mixing technique for study of oxygen-hemoglobin equilibrium: a critical evaluation. In the mixing technique for study of oxygen-hemoglobin equilibrium, the O2 saturation (SO2) of a blood mixture is calculated from the volume ratio at which an oxygenated sample is mixed with a deoxygenated sample, and the PO2 in the mixture is measured polarographically. Any predetermined level of SO2 may be obtained by proper choice of the volume ratio. It is shown that the volume and oxygen saturation of the mixed samples are by far the most critical parameters in calculating SO2, and a method is suggested by which the volume ratio is accurately measured by weighing the blood samples before mixing. Other parameters that influence determination of SO2, e.g., the O2 capacity of the blood, are much less important. The method has been applied to establish the O2 dissociation curve in human blood, and good reproducibility and agreement with standard curves were obtained. Measurements in rabbit blood yielded similarly satisfactory results. The technique is particularly applicable to problems that require exact adjustment of SO2 to a predetermined value, such as determination of the half-saturation pressure or of the Bohr effect at various levels of O2 saturation.


Temperature-induced changes in blood acid-base status in the alligator, Alligator mississipiensis. Gas exchange and arterial blood acid-base status were measured in 13 conscious alligators, Alligator mississipiensis, at 15, 25, and 35 degrees C. Arterial pH decreased by 0.250 units (from 7.635 to 7.385) and arterial carbon dioxide partial pressure increased by 11.4 Torr (from 11.8 to 23.2) as body temperature increased from 15 to 35 degrees C. No statistically significant changes occurred in arterial bicarbonate concentration. When OH-/H+ and alpha-imidazole were compared at each temperature, more variability was observed in OH-/H+, which increased from 8.7 to 12.0 as temperature increased from 15 to 35 degrees C. alpha-Imidazole remained essentially constant (0.76 at 15 degrees C and 0.80 at 35 degrees C). Body temperature increase caused marked increases in minute ventilation (VE), oxygen consumption (VO2), and carbon dioxide production (VCO2). The relative changes in these parameters resulted in a decrease in both VE/VO2 and VE/VCO2. The data of the present study are consistent with the concept that poikilotherms regulate their alveolar ventilation with changes in body temperature in order to keep OH-/H+ or alpha-imidazole constant.


Affinity chromatography of trypsin and related enzymes. V. Basic studies of quantitative affinity chromatography. A detailed study of the quantitative affinity chromatography of trypsin [EC 3.4.21.4] is reported here. Frontal chromatography using an enzyme solution of very low concentration on an affinity adsorbent gave the dissociation constant of the enzyme-immobilized ligand complex (Kd). Kd values determined under various conditions enabled us to discuss in detail the interaction of trypsin and affinity adsorbents (mainly Gly-Gly-Arg Sepharose). The pH dependence of Kd was consistent with that of the interaction of trypsin and product-type compounds. The effects of changes in temperature, ionic strength, dielectric constant, etc., were also studied. The Ki values of soluble competitive inhibitors can be determined by analysis of their effects on the elution volume of the enzyme. The values obtained were in good agreement with those obtained by kinetic analysis. The present method proved to be useful as a general procedure to investigate the interaction of a protein and a specific ligand.


Crystalline reduced nicotinamide adenine dinucleotide phosphate-adrenodoxin reductase from pig adrenocortical mitochondria. Essential histidyl and cysteinyl residues of the NADPH-binding site and environment of the adrenodoxin-binding site. Pig NADPH-adrenodoxin reductase was crystallized from pig adrenocortical mitochondria and its physicochemical properties were investigated. Pig NADPH-adrenodoxin reductase is a typical flavoprotein. Its optical absorption spectrum showed peaks at 272, 377, and 450 nm in the oxidized form. The adrenodoxin reductase contained one FAD per mol. The molecular weight was 49,000. The isoelectric points of the adrenodoxin reductase and its complex with adrenodoxin were 5.3 and 4.6, respectively. Pig NADPH-adrenodoxin reductase, unlike bovine NADPH-adrenodoxin reductase, was found to be free of carbohydrate. The fluorescences of tryptophanyl residues and FAD of the adrenodoxin reductase were quenched by holo- and apo-adrenodoxins. The NADPH-binding site of the adrenodoxin reductase was examined by photooxidation and selective chemical modifications with diethyl pyrocarbonate and sulfhydryl reagents. The results indicate that a histidyl and a cysteinyl residue of the adrenodoxin reductase are essential for the NADPH-binding site. The circular dichroism spectrum of the adrenodoxin reductase showed negative ellipticity in the visible region. Spur formation was observed between pig and bovine NADPH-adrenodoxin reductases against the antibody to bovine NADPH-adrenodoxin reductase in Ouchterlony double-diffusion agar plates. The antibody did not interact with spinach ferredoxin-NADP+ reductase.


A D-serine dehydratase acting also on L-serine from Klebsiella pneumoniae. D-Serine dehydratase [EC 4.2.1.14] was purified from a strain of Klebsiella pneumoniae 140-fold from crude extract with a yield of 5%. This enzyme catalyzed formation of pyruvate and ammonia not only from D-serine but also from L-serine, and also catalyzed the formation of alpha-ketobutyrate and ammonia from D-threonine. Km values for D-serine, L-serine, and D-threonine were 2.8 mM, 20 mM, and 3.6 mM, respectively. Km for pyridoxal 5'-phosphate was 2.5 micron. The molecular weight was estimated to be 46,000 by Sephadex G-150 gel filtration and 40,000 by SDS-polyacrylamide gel electrophoresis. This enzyme was inducible by D-serine. Induction by casamino acids appeared to depend on the presence of D-serine.


Partial purification and characterization of an enzyme involved in the formation of beta-aspartyl dipeptides in rat kidney. The formation of beta-aspartyl-glycine from asparagine and glycine was demonstrated in the supernatant of rat kidney. The enzyme involved in this process was partially purified. Based on the properties of the enzyme reaction and the coincidence of purification rates of this activity and asparaginase, it can be speculated that the enzyme is a kind of asparaginase. Examination of the preference for beta-aspartyl donors and acceptors showed that asparagine and glycine were the preferred donor and acceptor, respectively. beta-Aspartyl dipeptides also transferred their aspartyl residues to amino acids. Amino acids other than glycine also accepted the aspartyl moiety from the donors.