Inhibition of gastric (H+ + K+)-ATPase by the substituted benzimidazole, picoprazole.

The substituted benzimidazole, picoprazole, inhibited the gastric (H+ + K+)-ATPase in a concentration-and time-dependent manner. Half-maximal inhibition of the (H+ + K+)-ATPase activity was obtained at about 2 . 10(-6)M under standard conditions. In addition to the inhibition of ATPase activity, parallel inhibition of phosphoenzyme formation and the proton transport activity were achieved. Radiolabelled picoprazole was found to bind to 100 kDa peptide; this peptide was shown by phosphorylation experiments to contain the catalytic centre of the (H+ + K+)-ATPase. Studies on the (Na+ + K+)-ATPase indicated that this enzyme was unaffected by picoprazole. From the data presented and from other pharmacological studies, it is proposed that this compound inhibits acid secretion at the level of the parietal cell by its ability to inhibit the gastric proton pump, the (H+ + K+)-ATPase.

Fatty acids suppress recovery of heart function after hypothermic perfusion.

Working rat hearts were perfused for 15 minutes at 37 degrees C before switching to a Langendorff perfusion (60 mm Hg aortic pressure) at 10 degrees C for 40 minutes of hypothermic arrest. Ventricular function was allowed to recover for 15 minutes at 37 degrees C by reestablishing the prehypothermic conditions. The perfusate was Krebs-Henseleit bicarbonate buffer containing 3% bovine serum albumin and either glucose (11 mmol/L) or glucose (11 mmol/L) plus palmitate (1.2 mmol/L) and gassed with 95% O2 and 5% CO2. In hearts receiving glucose alone as substrate, coronary flow was maintained constant during the 40 minutes of hypothermic arrest and returned to prehypothermic rates with rewarming. Ventricular function, as estimated by peak systolic pressure and heart rate, recovered to the prehypothermic level. When palmitate was added, coronary flow decreased continuously throughout the hypothermic perfusion (22% decrease by 40 minutes), and ventricular pressure development was lower throughout the rewarming perfusion. Tissue levels of adenosine triphosphate and creatine phosphate were well maintained and long-chain acyl coenzyme A and acyl carnitine decreased during hypothermia regardless of the substrate provided. With rewarming, tissue levels of adenosine triphosphate and creatine phosphate decreased in those hearts receiving palmitate. Omission of fatty acid either during hypothermia or during the first 5 minutes of rewarming improved recovery of function. Addition of oxfenicine to inhibit fatty acid oxidation, or inhibition of Ca2+ overload by verapamil and low perfusate Ca2+, prevented the effects of palmitate on ventricular function.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute hypotensive effect of combined beta1-adrenoceptor blockade and beta2-adrenoceptor stimulation in spontaneously hypertensive rats (SHR).

The effect on heart rate (HR) and blood pressure (BP) of metoprolol, propranolol and terbutaline, applied alone or in combinations was studied in spontaneously hypertensive rats. Terbutaline had little effect on BP and HR. Propranolol combined with terbutaline had no effect while metoprolol combined with terbutaline decreased BP by 48 mm Hg without affecting HR. Thus the beta2-mediated increase in peripheral vascular resistance after terbutaline was revealed by a drop in BP after beta1-blockade by metoprolol, but not when both beta1-and beta2-receptors were blocked by propranolol.

Cardiostimulatory effects of prenalterol, a beta-1 adrenoceptor partial agonist, in vivo and in vitro. Correlation between physiological effects and adenylate cyclase activity.

The cardiostimulatory effects of prenalterol, a beta-1-adrenoceptor partial agonist, were studied in vivo and in vitro and compared to those evoked by isoprenaline, a full agonist, and to those of other partial agonists. In the anaesthetized rat, prenalterol and terbutaline were found not to elevate the myocardial cyclic AMP content; this was in sharp contrast to isoprenaline. Both partial agonists did, however, produce significant effects on heart rate. In the anaesthetized cat, prenalterol exhibited chronotropic and inotropic intrinsic activities of 88 and 76% respectively in relation to isoprenaline. No statistically significant increase in myocardial cyclic AMP content could however be detected. Prenalterol did not stimulate adenylate cyclase significantly in the cat myocardial homogenate. This was also true of the beta-2-adrenoceptor selective partial agonist procaterol. In this preparation, isoprenaline, noradrenaline and adrenaline acted as full agonists. Furthermore, prenalterol produced a concentration-dependent inhibition of isoprenaline-activated adenylate cyclase. Our data indicate that maximal cardiac stimulation occurs at a low level of adenylate cyclase activation and low myocardial cyclic AMP concentration when provoked by a full beta-adrenoceptor agonist. The maximal physiological effects of a partial agonist such as prenalterol may consequently be achieved at a marginal activation of the adenylate cyclase. The present data may thus support the hypothesis of a large beta-adrenoceptor reserve for full agonists in the heart.

Hyaluronan improves the healing of experimental tympanic membrane perforations. A comparison of preparations with different rheologic properties.

Preparations of hyaluronan at various concentrations and molecular weights were topically applied to experimental tympanic membrane (TM), perforations in the rat and their ability to improve the healing pattern was elucidated. The perforation occupied the upper posterior quadrant of the TM. Alterations in healing rate and quality of the healed TM were examined by otomicroscopy and in histologic sections. Hyaluronan enhanced the healing rate and resulted in less opacity and structural alteration of the scar area. Closure time for the TM perforation was correlated to the concentration of hyaluronan but not to its molecular weight or viscosity. Improved scar quality was obtained in the presence of hyaluronan irrespective of its rheologic properties. Hyaluronan improved the restoration of the fibrous connective tissue layer. Treatment of TM perforation in man with hyaluronan at high concentrations should be considered as an alternative to myringoplasty.

Metabolic rates in normal and infarcted myocardium.

The effects of myocardial infarction in rat hearts on the utilization of fatty acids and glucose by the surviving, non-infarcted tissue were studied. Hearts were removed from the animals one-week post-infarcted and perfused in the isolated working heart preparation. Oxygen consumption and oxidation of palmitate and glucose were determined at two levels of cardiac work. Rates of substrate oxidation were estimated by 14CO2 production from [14C]-labeled substrates. This approach to measuring metabolic rates may seriously under-estimate the true oxidative rate particularly when measuring oxidation of long-chain fatty acids. Because of the large endogenous stores of fatty acids in tissue lipids, the [14C]-specific activity of the intracellular metabolites involved in the free fatty acids oxidation pathway do not equilibrate with the specific activity of the perfusate fatty acids oxidation pathway do not equilibrate with the specific activity of the perfusate fatty acid even when 14CO2 production has reached an apparent steady state. When comparing an experimental condition to the normal heart, a lower rate of 14CO2 production may not necessarily indicate a lower rate of oxidation of free fatty acids, but a difference in the rate of turnover of endogenous sources of unlabeled fatty acid such that the specific activity of intracellular metabolites equilibrate with extracellular labeled substrate to a lesser extent than in the normal heart. At physiological concentrations of fatty acids, a shift from fatty acid to carbohydrate oxidation occurred in the hypertrophied, surviving tissue following myocardial infarction in the rat.(ABSTRACT TRUNCATED AT 250 WORDS)

Possible causes of increased lung cancer incidence among butchers and slaughterhouse workers.

An excess of lung cancer among butchers and slaughterhouse workers has been reported in several record-linkage studies. In this case-referent investigation on the possibility of occupational exposures being related to the lung cancer excess, cases and referents were selected from butchers and slaughterhouse workers registered in the Swedish national census of 1960. The case group comprised all men in the study population dying from lung cancer between 1971 and 1982. Two reference groups were formed, ie, all individuals dying from other cancers and a random sample of all dead men in the study population during the same time period. The history of occupations, occupational exposures, and smoking habits was obtained from the next-of-kin by questionnaire. None of the occupational exposures that were studied (work with live animal care, in the bleeding area, on the killing floor, or with meat cutting, processing, curing, smoking, chilling and packaging) were associated with an increased lung cancer rate. Tobacco smoking habits may have contributed to the overall excess of lung cancer found previously for this occupational group.

Coxarthrosis and physical work load.

A case-referent study was performed on the possible relationship between physical work loads and an increased risk of developing coxarthrosis. The cases were 239 male recipients of a hip prosthesis as a result of severe idiopathic coxarthrosis; the referents were 302 men randomly selected from the general population. The work load was assessed through an interview and a self-administered questionnaire on the men's specific work periods. Men highly exposed to dynamic or static work loads had an increased relative risk of 2.42 (95% confidence interval 1.45-4.04) for developing coxarthrosis when compared with men with low exposure. Men with high exposure to heavy lifting between the ages of 30 and 49 years had the highest relative risk, 3.31 (95% confidence interval 1.97-5.57). Long-time exposure to physical work loads seems to be a risk factor for severe coxarthrosis among men.

Towards a valid technique of sampling fish muscle to determine redox substrates.

The redox substrates--lactate, malate, alpha-glycerophosphate, dihydroxy acetonephosphate and pyruvate--have been determined in liver and muscle tissue from young salmon. The redox quotients have also been calculated. The freeze clamping technique was used and reliable samples of fish muscle for the determination of lactate were obtained with the aid of a pair of pliers with a gear mechanism. It was established that the lactate content in the body muscle is in close agreement with that in the blood from rested salmon parr. The concentrations of redox substrates are in good agreement with those found in mammalian tissue. The determination of the content of glycogen in fish muscle is discussed in the light of the results obtained in this study.

The effect of beta-receptor blockade on adrenaline-induced changes in redox and energy state in m. vastus lateralis and m. soleus of the rat in vivo.

The metabolic changes in blood, red (m. soleus) and white (m. vastus lateralis) skeletal muscle fibres were investigated after short-term (3 min) infusion of adrenaline with or without prior treatment with propranolol or metoprolol. The adrenaline-induced increase in plasma lactate levels was totally prevented by prior treatment with metoprolol or propranolol, whilst the beta-blockers had no effect on blood glucose levels. Similar effects on lactate levels were found in the m. soleus, while metoprolol was less effective than propranolol in m. vastus lateralis. Adrenaline decreased the level of muscle creatinine phosphate and ADP, causing the equilibrium of the creatinine kinase reaction to change in the direction of ATP synthesis, although the level of ATP usually decreased. This effect was more pronounced in m. vastus lateralis compared with m. soleus. The [ATP]/[ADP] [Pi]-ratio tended to increase during infusion of adrenaline. This effect was counteracted by metroprolol but not by propranolol. The effects on the "phosphate potential" ([ATP]/[ADP] [Pi]) and the equilibrium within the creatine kinase were more pronounced in m. vastus lateralis than in m. soleus. The results demonstrate the possible role of receptors other than beta-receptors, i.e. alpha-receptors, in mediating changes in plasma glucose levels, while plasma lactate levels are regulated by the beta-adrenergic system. The role of beta-receptors in mediating changes in muscle lactate levels may differ in m. soleus and m. vastus lateralis, with a relative predominance of beta 2-receptors in m. vastus lateralis. Quantitative and qualitative differences in the adrenergic control of the energy state in the two types of muscle fibre were obvious, although it was not possible to distinguish clearly between the relative importance of alpha, beta 1 and beta 2-receptors.

Functional and metabolic effects of terbutaline and propranolol in fast- and slow-contracting skeletal muscle in vitro.

The soleus, a slow-contracting, and the extensor digitorum longus (EDL), a fast-contracting skeletal muscle from guinea-pig were prepared for isometric recording of sub-tetanic contractions in vitro. The contents of adenosine-triphosphate (ATP) and creatinephosphate (CP) together with their metabolites and the contents of lactate, pyruvate and cyclic adenosine-monophosphate (c-AMP) in the muscles were determined. It was found that the energy and redox state of the isolated soleus and EDL muscles is very stable and does not significantly differ from the normal state in vivo. Moreover, there were no consistent changes in these variables after treatment with terbutaline (a beta 2-adrenoceptor agonist) or propranolol or both. Thus, effects on energy metabolism do not seem to cause the changes in muscle contraction, characteristic for beta-adrenoceptor stimulation. On the other hand, the functional effects were accompanied by elevation of the c-AMP level of the muscles.

The metabolism of acetate in the perfused hind-quarter of the rat.

The elimination kinetics of acetate, the main end product of ethanol metabolism in the liver and the influence of acetate oxidation on the redox- and energy state of the isolated perfused hind-quarter of the rat were studied. The rate of acetate uptake increased with increasing initial concentration of acetate in the perfusion medium, suggesting that the plasma level of free acetate may be one factor in the regulation of acetate uptake in the skeletal muscle. Addition of acetate as a single dose did not affect the net production of lactate or the uptake of glucose. In continuous infusion experiments at a constant concentration of 2 mM of acetate in the medium, the lactate/pyruvate ratio was unaffected in the medium and in the muscle tissue. Addition of acetate did not affect the oxygen uptake. Experiments with 14-C-acetate showed that about 50% of added radioactivity was found in form of 14-CO2 accounting for 25 to 45% of the oxidative metabolism in the muscle tissue. It was calculated that about 25% of the acetate produced in the liver during ethanol oxidation can be consumed in the resting, perfused hind-quarter of the rat. The tissue content of high-energy phosphate compounds was not significantly affected by acetate.

Differences in metabolic responses to beta-adrenergic stimulation after propranolol or metoprolol administration.

Isoprenaline, or the beta 2-agonist terbutaline, was infused in healthy male volunteers and the plasma levels of insulin, glucose and free fatty acids (FFA) were determined. Saline, propranolol, or the selective beta 1-receptor antagonist, metoprolol, was administered i.v. prior to the infusion of the beta-stimulants. The two beta-receptor blockers inhibited isoprenaline-induced increase in chronotropy to about the same extent, while the effects on systolic and diastolic blood pressure were in accordance with a selective beta1-blocking effect of metoprolol and a non-selective beta-blocking action of propranolol. Quantitative differences were found between metoprolol and propranolol on the metabolic parameters. The effects can best be described in terms of beta 1- or beta 2-receptors, where effects on plasma FFA and glycerol levels seem to be mainly beta1-mediated. An apparent beta 2-mediated effect was found for insulin release and hepatic glucose output.

Influence of acetate on the metabolism of beta-hydroxybutyrate in the perfused hind-quarter of the rat.

The effect of acetate, at concentrations normally found during ethanol combustion in intact animals and man in vivo, on the uptake and oxidation of beta-hydroxybutyrate by the perfused rat hind-quarter was studied. The addition of acetate did not significantly affect the total uptake of beta-hydroxybutyrate, but caused a 40% decrease in the oxidation of beta-hydroxybutyrate to CO2. The oxidation of 14C-beta-hydroxybutyrate to 14CO2 accounted for about 10% of the total oxygen consumption by the perfused muscle in the absence of acetate. In the presence of acetate this figure was reduced to about 5%. The addition of beta-hydroxybutyrate did not significantly affect the metabolic fate of 14C-acetate. It is concluded that acetate is preferred as oxidative substrate to beta-hydroxybutyrate. The inhibited oxidation of beta-hydroxybutyrate by acetate did not affect the concentration ratio between beta-hydroxybutyrate and acetoacetate in the medium at the end of the perfusion, indicating that the ability of the muscle tissue to restore an increased redoxlevel, "exported" from the liver during ethanol combustion to extrahepatic tissues, was not impaired by acetate.

Influence of acetate on the metabolism of palmitate in the perfused hind-quarter of the rat.

The metabolism of 1-14C-palmitate and its metabolic interaction with U-14C-acetate were studied in the perfused hind-quarter of the rat. 9% of 1-14C-palmitate taken up was oxidized to 14CO2 accounting for 7% of total oxygen consumption by the perfused tissue. Most label from 1-14C-palmitate was found in the lipid fraction of the muscle tissue. In spite of a 40% inhibition of palmitate oxidation, acetate only caused minor changes in the overall metabolism of palmitate. U-14C-acetate was mainly oxidized to 14CO2 and the oxygen consumption due to oxidation of acetate accounted for 20-30% of the total oxygen uptake. Minor amounts of 14C-acetate were found in muscle lipids. The addition of palmitate did not alter the metabolism of acetate. It is concluded that the presence of palmitate did not affect 14C-acetate metabolism, while the presence of acetate inhibited 14C-palmitate oxidation. The possible sites of interaction are discussed. The found interaction will probably not contribute to any major extent to the disturbed lipid metabolism found in animals and man during ethanol intake. No major changes in the tissue content of high-energy phosphate compounds were found in the presence of palmitate or acetate or both.

Influence of acetate on glucose metabolism in the perfused hind-quarter of the rat.

The metabolism of U-14C-glucose and U-14C-acetate and the interaction between the two substrates in the perfused hind-quarter of the rat was studied. 5% of glucose taken up was oxidized to CO2, accounting for 15% of total oxygen consumption. Glucose was mainly incorporated into glycogen, while incorporation into lipids was negligible. Acetate did not significantly alter glucose uptake, 14C-glucose oxidation or the incorporation of 14C-glucose into glycogen and lipids. 45% of acetate taken up was oxidized to CO2, accounting for 20-25% of total oxygen consumption. Insulin did not affect acetate uptake but increased 14C-acetate oxidation. Oxygen consumption was slightly increased by simultaneous oxidation of glucose and acetate and in this situation the tissue content.of high-energy phosphate compounds was slightly elevated. It is concluded that only minor effects by acetate on glucose metabolism in the perfused skeletal muscle were found. The insignificant effects compared to previously reported studies on heart tissue (Neely and Morgan 1974) can be explained by differences in acetate metabolism between the two tissues.

The effects of secretagogues of isolated human gastric glands.

The function of isolated human gastric glands has been studied in vitro by measuring the 14C-aminopyrine accumulation (RAP) in basal, unstimulated, conditions and after stimulation with different secretagogues. A microscale technique was used which enabled determinations of RAP in tissue obtained as gastroscopic biopsies. In addition, oxyntic-gland-containing mucosa was obtained at gastric resections for gastric or prepyloric ulcer disease. Histamine and cAMP derivative both induced maximal stimulation; RAP was approximately three times larger than in basal states. Carbachol induced a smaller but still significant stimulation. Pentagastrin did not increase RAP above the unstimulated level. Combinations of histamine and carbachol or pentagastrin did not induce a larger response than carbachol alone. The peak acid response to pentagastrin or betazole in vivo did not correlate with the maximum RAP in vitro.

Inhibition of acid secretion in isolated gastric glands by substituted benzimidazoles.

A new class of gastric acid inhibitors, substituted benzimidazoles (H 83/69 and H 149/94), have been tested in an isolated rabbit gastric gland preparation. Acid formation in the glands was stimulated by histamine, dibutyryl cAMP (DBcAMP), and high extracellular K+ concentrations, and the glandular secretory response was measured by changes in oxygen consumption and in accumulation of the weak base [14C]aminopyrine (AP). The substituted benzimidazoles inhibited AP accumulation induced by all stimulants in a dose-dependent noncompetitive manner. In contrast, cimetidine only inhibited histamine-induced AP accumulation. Basal AP accumulation, not affected by cimetidine, was also inhibited by the substituted benzimidazoles, as was the increase in glandular oxygen consumption produced by the addition of histamine and DBcAMP. Basal oxygen consumption was inhibited by about 15%. The substituted benzimidazoles, like AP, are weak bases and were also found to accumulate in the glands. Semiquantitative morphological studies of glands stimulated by histamine plus theophylline did not show any change in the enlarged secretory surface area after stimulation in the presence of inhibitory concentrations of H 149/94 (10(-4) M). The results suggest that substituted benzimidazoles have a mechanism of action different from that of H2-receptor antagonists and indicate a very distal site of action in the events leading to acid formation.