Mechanism of action of beta-blocking drugs in angina pectoris: a review. The effect of beta-blockers in improving exercise tolerance in angina pectoris can be accounted for by the reduction in heart rate and arterial pressure than they produce. The failure of a few patients to respond despite reduction in heart rate and the failure of those who do respond to obtain the full expected benefit suggests that beta-blockade has some action that offsets in while or in part the advantage gained from reduction in heart rate and pressure. This action might include an increase in left ventricular size with consequent increase in work, or an increase in left ventricular diastolic pressure with resultant impairment of blood flow to the inner layers of the myocardium.


Effect of propranolol on mortality in patients with angina. Two hundred and seventeen patients from general practice suffering from angina pectoris were studied over a three year period; 91 patients had beta-blocker treatment, the other 126 did not. The two groups were comparable for age, sex, blood pressure and length of history of angina. The frequency of infarction and mortality from infarction in the two groups was compared. The annual infarct rate in the 126 patients not treated with beta-blockers is shown to be more than three times the rate in the 91 patients treated by beta-blockers. The annual death rate from myocardial infarction is almost four times greater in the group not treated by beta-blockers compared with the beta-blocked group. It is concluded that this provides further evidence that treatment which includes beta-blockade in patients suffering from ischamic heart disease reduces the risk of myocardial infarction and death.


Adverse reactions to beta-blocking drugs: a review. Adverse reactions may occur as a predictable or unpredictable drug-induced illness can be reduced if the awareness of the prescribing doctor of clinically relevant drug and patient factors, which affect the response to the drug, can be increased. However, unexpected and unpredictable adverse reactions, sometimes reported as a single case in an anecdotal case report, are more difficult to evaluate and explain. Beta-blocking agents such as propranolol, oxprenolol and practolol have been asssociated with numerous adverse reactions. The influence of drug factors, such as range of action, absorption, metabolism, excretion, interactions, and patient factors such as disease states, idiosyncrasy, and personality are discussed.


Discovery of the hypotensive effect of propranolol. Following the observation that pronethalol lowered the blood pressure a similar effect was demonstrated with propranolol. Over a period of years propranolol was demonstrated to produce a fall in blood pressure comparable with other potent drugs with the advantage of good control of the supine blood pressure and with the absence of postural and exercise hypotension. These observations were slow to be accepted, as besides a novel mode of action, most attempts to repeat the work failed to pay enough attention to adequate dosage.


The use of propranolol in hypertension: a review. A review if given of the use of propranolol in hypertension emphasizing the importance of the studies by Prichard and Gillam in 1964 and 1969. The current use of propranolol in hypertension as regards dosage, dose intervals and starting doses is also outlined. Moreover, the combined treatment with propranolol and vasodilators is briefly described, as are attempts to make predictions of the therapeutic response by studying various haemodynamic and laboratory parameters. Finally, an attempt is made to assess the future role of propranolol in the treatment of hypertension in comparison with other antihypertensive drugs.


Indications for beta-adrenoceptor blocking drugs in hypertension. The past 10 years have firmly established the role of beta-adrenoceptor blocking agents in the treatment of hypertension. They have been shown to lower systolic and diastolic blood pressure in the lying and standing position in mild, moderate and severe hypertension. Precise indications for beta-blockade have not yet been completely defined. Some authorities regard them as the drug of first choice in the management of most grades of idiopathic hypertension. There are in addition certain situations where beta-blockade seems especially suitable. These include the presence of associated coronary heart disease manifest either as angina pectoris or dysrhythmia. These agents can be introduced when side effects from other drugs are severe or intolerable and are valuable in the management of hypertensive young males since beta-blocking drugs do not interfere with sexual function. Compared with normotensive subjects 'stress' has been shown to produce excessive rise of blood pressure in those with labile or sustained idiopathic hypertension. After therapy with beta-blocking agents the rise in blood pressure after 'stress' is reduced. If labile and/or mild hypertension are the precursors of subsequently more severe sustained hypertension, then long term beta-blockade may help to control this response.


Flurazepam ('Dalmane') in the treatment of insomnia in patients with respiratory disorders. The effects on subjective aspects of sleep of flurazepam 15 mg and amylobarbitone sodium 100 mg were compared in insomniac patients with chest disease. A double-blind technique was used. The results indicate a significant superiority of flurazepam over the other drug.


Implications of cross inhibitory interactions of potential mediators of hormone and neurotransmitter action. Mediators of hormone and neurotransmitter action may exert negative control on the accumulation of one another (adenosine 3':5'-cyclic monophosphate, or conversely). A model in which one agonist stimulates the formation of two mediators which inhibit each other's accumulation has been simulated. Three types of agonist-receptor interaction, five mechanisms of inhibition, and three types of basal activity have been considered which lead to a set of 45 descriptions of the general cross inhibition model. For these submodels, pattern of the relation of agonist to mediator concentrations at steady state conditions have been defined. Some patterns are complex with mediator curves exhibiting extrema, one a maximum and the other a minimum. The complexity of the pattern depends on the submodel and on the degree of asymmetry between the parameters of each of the mediator pathways (e.g.,affinities for the agonist, strength of the inhibitions, etc.). This simple model can thus account for complex experimental results without requiring the postulation of elaborate molecular models of agonist-receptor interaction. The simulations presented emphasize the necessity of investigating a wide range of agonist concentrations.


Supersaturation in sickle cell hemoglobin solutions. The kinetic inhibition of the gelation of hemoglobin S is compared to the change in hemoglobin S soulbility, when the solubility is altered by carbon monoxide, pH, or urea. By means of a new technique, the delay time and the extent of gelation are measured on the same sample. They delay time, td, is found to be proportional to a high power (30-40) of the hemoglobin S solubility. Togehter with the previously reported concentration dependence, this result demonstrates that the rate is proportional to a high power of the supersaturation, S, defined as the ratio of the total hemoglobin S concentration to the equilibrium solubility. The results obey the supersaturation equation td-1 = gammaSn, where gamma is an empirical constant (about 10(-7) sec-1) and n is about 35. The supersaturation equation can successfully account for observations on the kinetics of cell sickling and is therefore used to estimate the increase in the delay time for sickling necessary to produce significant clinical benefit to patients with sickle cell disease.


Mechanism of uncoupling in mitochondria: uncouplers as ionophores for cycling cations and protons. Classical uncouplers such as 2,4-dinitrophenol have been shown to be ionophores with the capability for transporting monovalent or divalent cations with equal efficiency. The conditions appropriate for the maximal expression of this ionophoric capability have been explored. Two critical factors are the polarity of the organic phase and the pH of the aqueous phase that is equilibrated with the organic phase. The demonstrated cationic ionophoric capability of uncouplers, taken in conjunction with the known ability of uncouplers to cycle protons across a membrane phase, provides the experimental basis for the thesis that uncoupling of electron flow from ATP synthesis via classical uncouplers involves the substitution of one coupled process by another. Uncoupling thus reduces to the replacement of one driven reaction (ATP synthesis) by the driven reaction (cyclical transport) mediated by the uncoupler.


Suppression of lytic effect of beta lactams on Escherichia coli and other bacteria. Growth of E. coli at pH 5 protected the bacteria against the lytic effect of beta lactam antibiotics typically observed when the cells are grown at pH 7 or 7.5, i.e., the pH values routinely used in laboratory experiments. In contrast, the typical effects of beta lactam antibiotics on cellular shape and elongation and cell division appeared to be similar in cultures grown under neutral and acid pH conditions. The pH-dependent antibiotic tolerance can also be demonstrated with pneumococci, staphylococci, streptococci, and Bacillus subtilis. We suggest that the mechanism of the pH-dependent antibiotic tolerance may involve either the production of a more stable plasma membrane or the suppression of the activity of a murein hydrolase(s) that catalyzes the antibiotic-induced lysis; at least a fraction of these enzyme molecules may be localized at the cell surface and be accessible to experimental manipulation.


Bandage-backfall reaction: occurs in infancy, hypothalamic damage, and catalepsy. In cataleptic clinging, produced either by catecholamine-blocking drugs or lateral hypothalamic damage in adult cats, rats, or monkeys, bandaging the face and neck causes the head to fall backwards. Early in development, a similar reaction can be seen in normal undrugged infants.


Physiological effects of the plasminolytic derivatives of fibrinogen. The macromolecular breakdown products of fibrinogen are known mainly for their inhibitory effect on the clotting of fibrinogen by thrombin. This inhibitory effect is due to interference with both the proteolytic action of thrombin and the polymerization of the fibrin monomers. However, the action of these products is not limited to these effects. They can on the one hand inhibit the consumption of Factors II and XIII and promote the inactivation of Factor VIII by thrombin. On the other hand, they can potentiate the activation of prothrombin in purified systems via the intrinsic pathway. The incidental observation was also made that Factor IXa and or Factor Xa inactivate Factor VIII. As substrates of both thrombin and plasmin the large fragments protect these two enzymes from spontaneous inactivation, while at the same time they inhibit their respective proteolytic activities. Contradictory results were obtained regarding their effect on platelets. The micromolecular (dialyzable) breakdown products prolong the thrombin, prothrombin, and partial thromboplastin times of plasma and retard the generation of the intrinsic prothrombin activator. They can also potentiate the effects of biologically active peptides and amines on the smooth muscles and on vascular permeability.


Bromazepam versus diazepam in psychoneurotic inpatients. A double blind between-groups trial was carried out to compare the effectiveness of the new benzodiazepine compound, bromazepam, (Lexotan, Ro 5-3350) as an anti-anxiety compound with that of diazepan. The fifty-eight psychoneurotic inpatients admitted to the trial were divided into two groups, "obsessives" and "neurotics", in order to assess any specific action of the drug. Assessment was made by means of B.P.R.S. of Overall and Gorham, R.S.A.S. of Hamilton, P.R.S. of Wittenborn and I.M.P.S. of Lorr. The results showed that bromazepam has a general therapeutic effectiveness (though not always as great as that of diazepam) as an anti-anxiety agent, while no difference in the effects of the two drugs was found between the two psychopathologic groups; with both treatments the "obsessives" responded less than the "neurotics". Detailed evaluation of the individual items of the I.M.P.S. showed that bromazepam is likely to exert a disinhibiting or "activating" action, unlike most other benzodiazepines in clinical use.


[Practice and further development of the AMP-system (a report about a symposion and a seminar) (author's transl)]. During two days in November 1974 TV-tapes of psychiatric interviews were demonstrated and the psychopathology was documented on AMP-record sheet 3. It was a purpose of the discussion to get stimulation for the improvement of the AMP-manual and to decrease the interrater-variance. It was only the first tape which was not followed by a discussion. This film was presented a second time at the end of the seminar in order to examine the training effects. Both record-sheets were compared with each other. Generally the variance decreased. The discussion as to an improvement of the AMP-manual lead to the following suggestions: In defining a psychopathological item it is important to differentiate more precisely between the patient's subjective experience and his behaviour, to find better criteria concerning the rating of different intensities and to improve discrimination between singular items. Furthermore the logic of documentation procedure should be unequivocal and more practical. Following the seminar there was a symposion discussion more general and some specific methodological problems of the further development of the AMP-system. Finally study groups were formed to elaborate on the topics "AMP-manual", "rater-training", "social data" and "electronic data processing".


[Problems of prevention, registration and evaluation of post-vaccination neurogenic lesions in children from clinical and legal viewpoints. II. Problems in the evaluation of neurogenic post-vaccination lesions in childhood]. On the view of neurologist the authors describe the suppositions and the special problems of diagnostic and the importance of clinical and paraclinical findings for medical vaccination reports. They examined 13 patients with neurological cerebral defects after triple vaccination (DPT).


Effects of humoral modulators and naloxone on morphine-induced changes in the spontaneous locomotor activity of the rat. The s.c. administration of 20 mg/kg of morphine-HCl produced a decrease in the spontaneous locomotor activity (SLMA) of rats. The decrease in SLMA was significantly antagonized by p-chlorophenylalanine (p-CPA). When rats pretreated with p-CPA were given 5-hydroxytryptophan before morphine injection, the marked sedative response to morphine was restored, suggesting that the morphine-induced decrease in SLMA of rats may depend on the release of 5-hydroxytryptamine by morphine. By contrast, the s.c. administration of 5 mg/kg of morphine-HCl produced a significant increase in SLMA of rats. The magnitude of the increase was reduced by atropine, scopolamine or alpha-methyl-p-tyrosine. It appears that both adrenergic and cholinergic mechanisms participate in the increase in SLMA of rats induced by morphine. Both the increase in SLMA produced by 5 mg/kg of morphine and the decrease in SLMA induced by 20 mg/kg of morphine were completely antagonized by the s.c. administration of naloxone-HCl, 0.0625 and 0.25 mg/kg, respectively. Thus, it appears that the receptor with which morphine interacts to produce stimulation is chemically identical with or very similar to the receptor with which morphine combines to induce depression. The former receptors, however, are likely to be located on different neurons from the latter.


Treatment of phenothiazine-induced dyskinesia. Previous experience indicated the possibility that a serotonin antagonist substance, was effective in relieving dyskinesia as a reaction to phenothiazine antipsychotic drugs. Since the substances used in the original observations did not become available on the United States market, a compound known to be a serotonin antagonist, cyproheptadine, which was readily available and safe, was used to treat three patients who had phenothiazine-induced dyskinesia. In these patients, cyproheptadine was definitely effective in relieving the dyskinesia over an extended period of time. Further controlled studies are required to confirm this report.


Narcotic blockade, length of addiction and persistence of etonitazene consumption in rats. Rats were given daily trials to determine relative preference for an opiate (etonitazene, ETZ) or for water. Animals with a greater history of previous drug exposure developed ETZ preferences more rapidly than did relatively drug-naive animals. Pretreatment with adequate blocking doses of naloxone reduced drug intake to near zero in most subjects. However, animals with the greatest history of prior addiction continued to drink large quantities of ETZ, despite pretreatment with relatively large doses of naloxone. These results can be explained by assuming that stimuli associated with the reinforcing properties of the opioid solution become strong conditioned reinforcers, capable of maintaining responding for long periods of time despite blockade of the reinforcement properties of the drug.


On the supersensitivity of dopamine receptors, induced by neuroleptics. Different neuroleptics caused dopamine receptor blockade (antagonism against methylphenidate-induced compulsive gnawing) for varying lengths of time. When the receptor blockade had expired, supersensitivity to dopamine agonists (occurrence of apomorphine-induced compulsive gnawing and enhancement of methylphenidate-induced gnawing) developed and persisted for varying periods of time. The degree and duration of supersensitivity was related to the degree and duration of the preceding receptor blockade. Inhibition of catecholamine or 5-HT synthesis had no influence on development of supersensitivity. Stimulation with a dopamine agonist, apomorphine, during the period of the development of supersensitivity did not modify the enhanced receptor supersensitivity. A cholinergic-dopaminergic balance was shown to be involved in the manifestation of compulsive behavior during the supersensitivity phase. Tolerance to the dopamine antagonistic effect of a neuroleptic also developed after a single neuroleptic treatment, most likely due to increased sensitivity of the receptors for the dopamine agonist. It is concluded, that the dopamine receptor blockade induced by a single dose of a neuroleptic agent is a dynamic phenomenon which in the course of time is replaced by an increased sensitivity of the receptors to dopamine agonists. Noradrenergic or 5-HT neuron systems do not seem to be involved in the neuroleptic-induced supersensitivity, whereas a dopaminergic-cholinergic balance is operative in the supersensitivity situation.


The demonstration of a change in adrenergic receptor sensitivity in the central nervous system of mice after withdrawal from long-term treatment with haloperidol. Mice, administered haloperidol (3 mg/kg/d) in their drinking water for 21 days, displayed, 4 days after cessation of the haloperidol-treatment, marked locomotor stimulation to clonidine (100 or 500 mug/kg) which lasted for about 6 h. 25 mug clonidine/kg was inactive. Premedication with FLA-63 (25 mg/kg) blocked the difference in stimulation after clonidine between the haloperidol- and vehicle-treated animals, but locomotor activity was still present in both groups. Haloperidol-treated animals displayed a supersensitive response to dexamphetamine. The difference in stimulation produced by dexamphetamine in the two groups was completely blocked by phenoxybenzamine (2.5 mg/kg), phentolamine (10 mg/kg), which drugs did not, however, block the locomotor stimulation produced by dexamphetamine in vehicle-treated animals. Pimozide (3 mg/kg) blocked all locomotor stimulation produced by dexamphetamine in both vehicle- and haloperidol-treated groups, while 1 mg/kg completely blocked the dexamphetamine response in vehicle-treated animals but not in haloperidol-treated animals. FLA-63 (25 mg/kg) blocked the difference in response between the haloperidol- and vehicle-treated groups to dexamphetamine, but did not antagonise the stimulation in the vehicle-treated animals. The data suggest that long-term haloperidol treatment leads to the development of "supersensitive" adrenergic receptors in the central nervous system, which, appropriately stimulated, effect an increase in locomotor activity. Moreover, the results indicate that a large component of the supersensitive response to dexamphetamine observed after long-term haloperidol-treatment is due to adrenergic receptor supersensitivity. However, the dopamine receptor (which was shown to be supersensitive to apomorphine) is of fundamental importance because phenoxybenzamine and phentolamine, while blocking the supersensitive response to dexamphetamine, failed to block the response to dexamphetamine in vehicle-treated animals, which was, however, blocked by pimozide.


Changes in brain norepinephrine associated with sensitization to d-amphetamine. Pretreatment of B6AF1/J mice with d-amphetamine HCl 10 mg/kg, twice daily for 5 days, produced a 4-fold increase in the running response to a test dose of 5 mg/kg amphetamine. Amphetamine pretreatment decreased whole-brain norepinephrine levels to 50% of control values and whole-brain dopamine to 85%. The test dose of 5 mg/kg amphetamine lowered whole brain norepinephrine levels of control mice from 0.50 mug/g to 0.28 mug/g in 2 h. In amphetamine-pretreated mice, this injection caused an increase in whole-brain norepinephrine levels from 0.22 mug/g to 0.55 mug/g at 30 min, followed by a decrease to 0.22 mug/g at 60 min. No change in whole brain dopamine levels was observed in either group. Amphetamine sensitization and norepinephrine depletion were still evident 25 days after pretreatment. No cross sensitization to morphine or cocaine was observed. Reserpine pretreatment resulted in a 3-fold increase in locomotor activity following injection of d-amphetamine, 5 mg/kg. No sensitization or changes in catecholamine levels were observed in amphetamine-treated A/J mice. These results suggest that the sensitization produced by amphetamine pretreatment may be related to the depletion of brain norepinephrine.


Effects of a novel anti-aggressive agent upon two types of brain stimulated emotional behavior. The effects of anti-aggressive agent Sch 12679 were evaluated upon stable baselines of rage and predation elicited by electrical stimulation of the hypothalamus in cats. Sch 12679 depressed approach and terminal aspects of both forms of attack. This is consistent with previous reports, and suggests the drug is effective in reducing many forms of aggression including brain stimulated emotional behavior.


Effects of combined treatment with nortriptiline and lorazepam on conflict behavior and motility of rats. Lorazepam attenuated the suppressant effects of punishment on the response rate of rats in the multiple schedule of reinforcement devised by Geller and Seifter (1960). Nortriptiline alone was ineffective on punished responses. Both drugs, at certain dose levels, inhibited the nonpunished response. Combined treatment with the two drugs in a dose ratio of 1:20 attenuated the effects of punishment at all dose levels tested. The effects of the combination upon both punished and nonpunished responding was greater than might be accounted for by a simple additive effect of the individual treatments. Lorazepam and nortriptiline both induced a dose-related decrease in the locomotor activity of rats; when given together the 2 drugs antagonized each other. The results give an experimental support to the clinical observations about the usefulness of the benzodiazepine-antidepressant combination in certain depressive illnesses.


Mepiprazole, a new psychotropic drug: effects on uptake and retention of monoamines in rat brain synaptosomes. The influence of mepiprazole (EMD 16,923), a new pyrazol-ylalkyl-piperazine derivative, on the uptake of 3H-norepinephrine (NE), 3H-dopamine (DA), and 3H-serotonin (5-HT) into rat brain synaptosomes from cerebral cortex, corpus striatum, and hypothalamus was investigated in comparison with several psychotropic drugs, including oxypertine, d-amphetamine, imipramine, desipramine, chlorimipramine, amitriptyline, and chlorpromazine in vitro. Mepiprazole was a relatively weak inhibitor of monoamine uptake and exhibited its strongest action on the hypothalamic 5-HT uptake, being almost equipotent with desipramine (IC50 = 0.9 MUM). Furthermore, the influence of the drugs on the retention of 3H-amines previously taken up by whole rat brain synaptosomes was studied. Unlike the tricyclic antidepressants, mepiprazole as well as oxypertine and d-amphetamine markedly increased the efflux of radioactivity during a 20-min incubation at 37 degrees C at low concentrations (10(-6) to 10(-5) M), whereas at 10(-4) M all drugs greatly enhanced the efflux. The ability of mepiprazole to increase 5-HT concentration at the receptor level by a combination of neuronal uptake inhibition and release is discussed in relationship to the central actions of the drug.


Time course of free and N4-acetylated sulfapyridine concentrations in the plasma and saliva of man after sulfasalazine (salicylazosulfapyridine) administration: preliminary findings. The time course of free and N4-acetylated sulfapyridine (SP) concentrations in the saliva and whole plasma was determined in a healthy male volunteer after a single 2.0 g oral dose of salicylazosulfapyridine (SASP) as four-500 mg commercial, uncoated tablets. The mean (+/- S.D.) plasma: saliva concentration ratios for free and acetylated SP was 2.04 (+/- 0.24) and 3.12 (+/-0.43), respectively, and were independent of plasma concentration and saliva pH. The elimination half-lives of SP and N4-acetyl SP could be determined from either salivary or plasma concentration-time data. These preliminary results suggest that measurement of saliva concentrations of free and acetylated SP may be a convenient, noninvasive method for monitoring indirectly the plasma concentrations of SP and acetyl SP in patients treated with SASP and for determining acetylator phenotype.


Model to study long-term effects of propoxyphene napsylate (darvon-N) on narcotic-dependent mice. Five different strains of mice were investigated for their suitability to serve as models for the study of long-term effects of propoxyphene napsylate (PN) on narcotic-dependent mice. The NIH/Swiss outbred strain proved to be most suitable because of its high liability to manifest physical dependence on morphine. Following removal of subcutaneously-implanted morphine pellets from the mice, the incidence of spontaneous withdrawal jumping was used as a quantifiable criterion of physical dependence. An orally administered suspension of PN suppressed the withdrawal jumping in a dose-dependent manner. However, chronic (daily) administration of PN at doses high enough to prevent withdrawal jumping was characterized by a high degree of toxicity.


In vitro studies on the antimicrobial effects of colostrum and milk from vaccinated and unvaccinated pigs on Escherichia coli. Unsupplemented porcine colostrum and milk exhibited a powerful bactericidal effect for porcine strains of E coli incubated in vitro at 37 degrees C. This activity was independent of complement but was susceptible to acid pH, to the presence of soluble iron and to the selective immunoprecipitation of IgG, IgA and IgM. Manifestation of bactericidal activity required bacteria in an active state of metabolism and the length of incubation was an important factor in demonstrating the quality of the anticoli activity, ie, proliferation-inhibitory, bacteriostatic or bactericidal. Whey obtained by acid precipitation or by the application of rennin was devoid of bactericidal activity but was capable of slowing down proliferation of E coli. There was no correlation between lysozyme and anticoli activity although the complete removal of lysozyme by adsorption on to bentonite reduced bactericidal titres. With very few exceptions the highest bactericidal titres were recorded for colostrum, but even 28 days post partum about one half of 22 undiluted milk samples exhibired bactericidal activity.


Aquatic life at high altitude: respiratory adaptations in the Lake Titicaca frog, Telmatobius culeus. Telmatobius culeus has a combination of behavioral, morphological and physiological adaptations which allows an aquatic life in cool (10 degrees C) O2-saturated (at 100 mm Hg) waters at high altitude (3812 m). The skin surgace area is increased by pronounced folds and the cutaneous capillaries penetrate to the outer layers of the skin. The erythrocyte volume (394 mu3) is the smallest reported for amphibians. The P50 (15.6 at ph 7.65 and 10 degrees C) is the lowest, and the erythrocyte count (729 - 103/mm3) the highest for an anuran. The O2 capacity (11.7 vol%), hemoglobin (8.1 g/100 ml), hemoglobin concentration (0.281 pg/mu3) and hematocrit (27.9%) measured at 18 degrees C and 3800 m are all elevated in comparison with most amphibians. The O2 dissociation curve is sigmoid (n = 2), the Bohr factor is small (deltalog P50/deltapH = -0.30) and the buffering capacity (-8.9 m M HCO3 - 1-1) is typical for an aquatic amphibian. The metabolic rate (14.1 mul -g-1-h-u) is the lowest reported for a frog and among amphibians only the giant salamanders (Amphiuma, Necturus and Siren) have lower values. If prevent from surfacing in hypoxic waters, the frogs ventilate the skin by "bobbing" behavior; if allowed to surface, they will ventilate the small lungs and the metabolic rate increases to 23 mul-g-1-h-1.


The oxygen affinity of chicken haemoglobin in whole blood and erythrocyte suspensions. The oxygen equilibrium of whole chicken blood has a P50 of 52.3 mm Hg and a Hill coefficient of 2.6 at pH 7.4 and 37 degrees C when determined with new microtechniques which are not vitiated by cellular respiration. The apparent failure of the haemoglobin to reach full saturation at arterial PO2 IS DISCUSSED IN RELATION TO THE HAEMoglobin-oxygen equilibrium concept. The low affinity observed is due to intraerythrocytic inositol pentaphosphate (IPP), and the affinity of a haemolysate "stripped" of IPP is greatly increased (deltalog P50 =1.25). Unlike the mamalian analogue 2, 3-diphosphoglycerate, IPP concentration of whole blood does not decrease after incubation for 10 hr at 37 degreesC.


[The role of arthropods as vector agents in viral hepatitis]. The increased incidence during the last years of viral hepatitis demands a new approach to the various eventual possibilities of contamination. The author discusses the transmission of serum hepatitis by mosquitoes, especially in warm climates, and the recent results obtained in investigations on the mechanical or biological transmission by haematophage arthropods.


The value of epicillin and ampicillin in the treatment of meningococcal and pneumococcal meningitis. 96 patients with meningitis due to Neisseria meningitidis and Diplococcus pneumoniae were treated with epicillin or ampicillin according to a predesigned randomization chart. Results indicate that epicillin and ampicillin are effective single drugs in the treatment of meningococcal and pneumococcal meningitis. No adverse reactions were noted with either drug and they were comparable in their efficacy.


Disorders induced by pheochromocytoma in the cardiovascular system. A therapeutic approach. The diagnosis of pheochromocytoms is still made only post mortem in a high percentage of the cases. It is important to know both the diversity of clinical forms (of the 16 cases investigated 13 (80%) presented permanent AH, of which 12 cases with paroxysmal attacks, followed by collapse only in one case, and 3 (20%) with paroxysmal AH on a normotensive background) and the severity of cardiovascular accidents, which increases twofold during crises (arrhythmias, left ventricular failure, coronary insufficiency, cerebrovascular strokes, secondary adrenalinic shocks). These complications are preceded by the "alarm syndrome". The preoperatory treatment is associated: blocking of adrenergic alpha-receptors and beta-receptors; correction of hypovolemia, also applied during the surgical phase I (until the venous ligature is made and the tumor excised) under continuous monitoring (ECG, ABP and central venous pressure). In phase II vasoplegin should be promptly corrected by nor. E+E, and corticoids.


Cholinergic stimulation of the rat hypothalamus: effects of liver glycogen synthesis. Cholinergic stimulation of the lateral hypothalamic neurons with intrahypothalamic microinjections of acetylcholine or carbachol caused a marked increase in the content of the active form of glycogen (starch) synthase in the liver. Total activity of the enzyme (active plus inactive forms) was not increased significantly. The lowest effective dose of acetylcholine was 5 X 10(-10) mole, and the optimum dose was 5 X 10(-9) mole. Similar applications of other neurotransmitters, such as norepinephrine, dopamine, serotonin, and gamma-aminobutyric acid, did not affect the enzyme's activity.


Beta-blockers and the central nervous system. Some of the interesting points which emerged from an international symposium on beta-blockers and the central nervous system, held in St Moritz in January 1976, are presented. Fifty-eight participants from 12 countries attended the symposium.


Somatostatin, 1976.

Somatostatin, 1976. Somatostatin, a growth hormone release-inhibiting factor, isolated originally from the hypothalamus, has been shown to have widespread effects on brain and endocrine and exocrine pancreatic and gut function. Furthermore, it has a widespread distribution in the CNS, gut and C cells of the thyroid -- cells which probably migrated originally from the neural crest during development. While the pharmacological effects of somatostatin are diffuse, its physiological role is at present unknown, but in view of its concentration in synaptosomal fractions of neural tissue, it may have a neurotransmitter or a synaptic modulator function.


Severe ischemia of the hand following radial artery catheterization. Percutaneous radial artery catheterization for blood gas monitoring and continuous arterial pressure recording has become a common place procedure in the management of critically ill patients. Five patients with severe ischemia after cannulation have been encountered in the past 20 months, four of whom lost segments of one or more digits. Review of the events preceding and during radial artery cannulation allowed elucidation of the following principles: radial artery catheterization should be preceded always by a negative Allen test; the catheter should be removed after 12 to 18 hours, especially if the patient is critically ill, is hypercoagulable, or has impaired tissue perfusion; the superficial temporal artery is safer to use and permits long-term cannulation (5 to 7 days) without ill effect; and agressive approach to assessing flow and arterial reconstruction is essential if severe ischemic symptoms occur during or after radial artery catheterization. Ancillary measures, including cervicodorsal sympathetic block, intravenous low molecular weight dextran and heparin, and intra-arterial reserpine and fibrinolysin, may improve palmar circulation but should not be substituted for both noninvasive and angiographic study of arterial flow, followed by surgical restoration of flow, when indicated.


Bacterial precipitins and their immunoglobulin class in atopic asthma, non-atopic asthma, and chronic bronchitis. In a study of groups of patients with atopic (extrinsic) asthma, non-atopic (intrinsic) asthma, and chronic bronchitis, no difference could be detected in the numbers having precipitating antibodies against species specific antigens from Staphylococcus aureus or Streptococcus pneumoniae compared to suitably matched control subjects. Precipitating antibodies against species specific antigens from Haemophilus influenzae, demonstrated in this investigation by double diffusion in agar gel, were found much more frequently in patients with chronic mucopurulent or obstructive bronchitis (50%) than in either asthmatic subjects (6%) or normal controls (6%) (P = less than 0.0005). While the precipitating antibody demonstrated in these patients against the extracts of Str. pneumoniae and Staph. aureus was in the IgG class alone, IgM and IgA antibody were detected against the species specific but not the non-species specific antigens of H. influenzae. These results underline the importance of H. influenzae as an infecting agent in chronic bronchitis and suggest that the finding of precipitins against the species specific H1 and H2 antigens of this bacterium denotes infection either concurrently or in the recent past. There is no evidence to suggest from this study that infection with Staph. aureus, Str. pneumoniae or H. influenzae is any more common in asthmatics as a group compared to controls or between patients with the non-atopic (intrinsic) and atopic (extrinsic) form of the disease.


The effect of beta adrenergic blockade upon exercise-induced changes in blood coagulation and fibrinolysis. Exercise stress is known to stimulate blood coagulation and fibrinolysis possibly as a result of sympatho-adrenal stimulation of the beta adrenergic receptor. In order to test this hypothesis five men exercised on four separate occasions with and without prior beta adrenergic blockade with oxprenolol, propranolol and pindolol. The increase in plasma adrenaline and noradrenaline concentration was much greater during exercise under beta blockade but activation of fibrinolysis was enhanced as well. Factor VIII levels did not change significantly during any of the experiments. These results suggest that the activation of fibrinolysis by exercise is not mediated by the beta adrenergic receptor. The influence of beta adrenergic blockade on the change in factor VIII with exercise was inconclusive.


Studies on the action mechanism of the antihemostatic effect of lodopeptides. A synthetic iodopeptide having a glutamic acid-diiodotyrosine molar ratio of 1:1 has been shown to be an effective anticoagulant both in vivo and in vitro. Contrasted with heparin the following general conclusions may be made regarding its action. The iodopeptide does not act through the inactivation of thrombin in plasma. Iodopeptide does interact with fibrinogen to form a complex which, in vitro, is not soluble in buffered saline at physiological pH. At pH 8, iodopeptide interacts with fibrinogen to form a soluble complex in the presence of 0.9% NaCl that is not coaguable either by thrombin or Crotalus venom enzymes. All the available evidence indicates that the fibrinogen to fibrin conversion is not inhibited under these conditions, but that fibrin, once formed, is not able to polymerize due to interference by iodopeptide. Similar results were obtained with heparin in vitro with thrombin-fibrinogen mixtures in the absence of NaCl. Studies with Russell's viper venom in native PRP strongly suggest that the iodopeptide also interferes with processes in the early coagulation pathway associated with prothrombin activation.


Canine allogeneic bone marrow transplantation. Technique and variables influencing engraftment. We have studied the toxicity and immune suppression of supralethal total body irradiation (800-2000 rads, 60Co) at three dose intensities (10 rads/min, 49 rads/min, and 100 rads/min). In 79 intensively supported radiation control animals, the LD50(5) (tn 5 days) for these dose intensities is estimated to be 1556, 941, and 921 rads, respectively. A biomodal pattern of early (median 4 days) and late (median 9 days) deaths was observed corresponding to histopathological evidence of the intestinal and hematopoietic radiation syndromes. Random donor bone marrow transplants were performed in 83 animals to test immune suppression afforded by 800 rads and 1000 rads at dose intensities of either 10 rads/min or 49 rads/min. Bone marrow cell dose was varied to analyze its effect on engraftment. A greater degree of immunosuppression with less toxicity was achieved at the lower dose intensity. A minimum dose of 3-5 X 10(8) nucleated allogeneic bone marrow cells/kg (readily obtainable from living donors) resulted in a high percentage of engraftment with lethal graft-versus-host disease following conditioning with 1,000 rads midplane at 10 rads/min, the optimum regimen employed.


Graft-versus-host reactivity and renal allograft survival in rats given allogeneic spleen cells or spleen allografts. Selective recruitment of antigen-sensitive cells (ASC) into the spleen as a method of inducing specific suppression was attempted by intravenous injection of either DA or Lewis spleen cells 24 hr before a (DA X Lewis)F1 renal allograft into a Lewis or DA recipient, either with or without a splenectomy. This led to suppression of rejection in the DA recipient and delayed rejection in the Lewis recipient. Splenectomy produced a minimal augmentation effect. Assay of graft-versus-host (GVH) reactions in (DA X Lewis)F1 rats by a popliteal node assay showed that injection of allogeneic DA or Lewis spleen cells 48 hr before the assay significantly reduced the reaction produced by node lymphocytes but not spleen lymphocytes, suggesting a loss of ASC from the lymph nodes. Lewis spleen allografts did not produce such a significant reduction in the GVH reactivity of DA node lymphocytes as intravenous Lewis cells, whereas DA spleen allografts led to an increased GVH reactivity of Lewis node lymphocytes. From these studies, it is not possible to attribute the suppression produced by the intravenous injection of allogeneic cells to selective recruitment of antigen-sensitive cells to the spleen.


Disseminated systemic expression of the "local" popliteal lymph node assay in rats. In contrast to the traditional belief that the popliteal lymph node (PLN) assay of graft-versus-host (GVH) reactivity is a local phenomenon, this study describes significant systemic components of both donor and host lymphoid cell activity. Not only are host radiosensitive lymphocytes of systemic origin necessary for the manifestation of normal PLN hypertrophy, but as few as 2.5 X 10(6) parental lymph node cells (LNCs) injected into the hind footpad of adult F1 hybrid rats disseminate widely, provoking significant systemic GVH reactions, as measured by splenomegaly, and distant lymphadenopathy. Futhermore, locally injected donor LNCs ultimately engender three forms of attenuated GVH reactivity: (1) the dissipation of potentially unlimited GVH reactivity, (2) refractoriness of the host to subsequent rechallenge by GVH-inducing cells, and (3) progressive loss of GVH reactivity in donor LNCs when serially transferred to secondary F1 recipients. Whether this modulation of the cell-mediated immune response is the expression of an anti-recognition structure response by the host or the activation of some other immunoregulatory protein, it is absent or reduced in splenectomized recipients. The data suggest that the spleen provides an immunoregulatory microenvironment in which cell-mediated immune responses, such as the GVH reaction, are modulated.


Infection rate of arboviruses in Dutch recruits returning from Surinam. Paired serum samples from approximately 650 Dutch recruits residing temporarily in Surinam, the first sample being collected on arrival, the second when leaving the country, were examined for haemagglutination-inhibiting antibodies to two types of arbovirus (Mucambo and Restan) isolated from local mosquitoes, and to two types of arbovirus (Oriboca and Caraparu) isolated from human blood. In a group of inidviduals in whom mild febrile illness had been reported, antibodies had developed in 3.9 to 6.7%, and in a group without reported history of febrile illness in 3.0 to 47% for each of the viruses.


Sipple's syndrome. A urologist's viewpoint. A case of Sipple's syndrome, a triad of medullary carcinoma of the thyroid, pheochromocytoma, and parathyroid hyperplasia is presented. The cause, laboratory diagnosis, and operative management of the syndrome are discussed. The need for thorough endocrine screening of patients with pheochromocytoma and family screening of patients with the full syndrome is stressed.


Disorders of micturition: neuropharmacologic basis and results of drug therapy. The clinical use of various pharmacologic agents in problems of micturition is based on the new concepts of intrinsic urethrovesical innervation, presence and regional predominance of autonomic neuroreceptors, and experimental evidence of the effects of various drugs on the bladder and the urethra. A new concept, relating to the processes that control bladder filling and emptying, is coming into being and replacing the traditional concept based on anatomic grounds alone. On the basis of the published data, and from personal experience both experimental and clinical, pharmacologic agents singly or in combination can be effectively and safely used in various problems of micturition such as incontinence (enuresis, stress incontinence in women, postprostatectomy, urgency incontinence), and functional outflow obstruction caused by neurologic or non-neurologic disease processes.


Host factors in urinary infections.

Host factors in urinary infections. To prevent the urologist from becoming simply a urinary tract mechanic we must develop more specific tests, perfect present tests and use these tests to elicit faulty mechanisms of host resistance of patients with urinary tract infections. We must then provide, from our research information, therapeutic measures to protect the faulty host from the inconvenience of recurrent significant bacteriuria and its sequelae.


Physical and chemical changes of ACD-preserved blood: a comparison of blood in glass bottles and plastic bags. ACD blood was preserved in glass bottles with or without aeration and in plastic bags in air or nitrogen gas at 4.5 degrees C. The blood was examined for physical and chemical changes of erythrocyte membrane resistance, hemoglobin in the plasma, the viscosity of the blood, pH of plasma, and ATP and 2,3-DPG content of erythrocytes. The blood preserved in plastic bags showed less changes than blood in glass bottles. The presence of air or nitrogen gas in blood seems to increase the pH perhaps by elimination of carbon dioxide (CO2) which in turn causes the different rates of glycolysis in the erythrocytes.


[Metabolic disorders in the acute stage of cardiac infarction]. The level of lactates and pyruvates in arterial blood of 35 patients with myocardial infarction without shock (17-without heart insufficiency and 18-with manifested but in various degrees) is higher as compared with the level of 18 healthy subject and the elevation is better manifested in the patients with cardiac insufficiency. The highest level of lactates and pyruvates is reached in the first 48 hours of the onset of the disease, after which they decrease progressively and  in patients without cardiac insufficiency is normalized by the 5-6th day and in those with insufficiency-remains elevated. Slight to moderate correlation exists between the level of lactates and BE and PaCO2. Lacate and pyruvate level reoresents a more sensitive index for the existence of distrubances in the tissue metabolism as compared with the indices of alkaline-acid equilibtrium.


[Integrated stimulation test with releasing hormones on healthy men and on patients with various forms of male hypogonadism]. Ninety subjects were examined; 20 healthy males, 20 patients with primary hypogonadism, 30 patients with Klinefelter's syndrome and 20 patients with secondary hypogonadism. The stimulating test with 200 mug LH-RH and 200 mug TTH was carried out tp all subjects examined, applied in two successive o.v. injections. The plasma as regards the levels of TTH, prolactin, FSH, LH, ACTH and STH is investigated every 0,20, 30, 60 and 90 minutes. The levels of tbe mentioned hormones were determined as well as the normal respond reaction. The basic LH and FSH levels are considerabty elevated in patients with primary hypogonadism and Klinefelter's syndrome and the respond reaction to stimulation is normal or diminished. A strict correlation between the number of X chromosomes and the level of FSH and LH was established in the patients with Klinefelter's syndrome. With respect to TTH, the basic, level in patients with primary hypogonadism in normal in 13 patients and pathological deviations were found in 7 patients. A strict correlation between TTH level and the number of X-chromosomes exists, Prolactin level in both groups is decreased. In patients with secondary hypogonadism the basic level of LH and ESH is decreased or at the lower limits of the norm. The respond reaction is absent. Both TTH and prolactin levels are pathologically changed as well as the respond reactions to stimulation depending on the etiology of the primary endocrine distrubance that had led to hypoginadism development.


Effect of aqueous extract of Eugenia caryophyllus on brain acetylcholine esterase in rats. The effect of aqueous extract of Eugenia caryophyllus on brain acetylcholine esterase (AChE) activity in rats was studied. Results indicate that the aqueous extract reduced the hydrolysis of acetylcholine (ACh) by AChE. This reducing effect was not due to the acidic nature of the extract and suggests that Eugenia caryophyllus contains some water soluble substance(s) with anti-choline esterase activity.


Properties of proteolytic enzymes isolated from a thermophilic strain of Micromonospora vulgaris 42. Two proteolytic enzymes, protease A and protease B, were isolated in homogeneous state from the cultural broth of the thermophilic actinomycete Micromonospora vulgaris 42. Their physicochemical properties were studied, i.e., molecular weight (50 000 for protease A and 30 000 for protease B), amino acid composition, N-terminal amino acids (phenylalanine for protease A and alanine for protease B). The specificity of the action of these enzymes was assayed by splitting the B chain of oxidized insulin. Both enzymes are neutral proteases of the thermolysine type.


Histone-like protein fractions of Kluyveromyces fragilis and their relation to the cell cycle. Protein fractions were obtained from Kluyveromyces fragilis by pH titration, a technique used for histone extraction, following the inclusion of a distilled water extraction stage, and the fractions partially characterized. The inclusion of a distilled water step resulted in a tenfold purification of the fraction obtained at pH 2.20 as compared with pH titration alone. In synchronous cultures induced with 2'-deoxyadenosine or prepared by selection this fraction displayed a stepwise accumulation, and doubled in quantity at a point about one third of a cycle after cell division. The fraction obtained at pH 1.35 resembled calf thymus f2a2 histone in its extraction properties and amino acid composition, and also showed a possible stepwise accumulation. Other fractions appear to accumulate exponentially.


[Malabsorption caused by HBsAg-positive panareteritis nodosa]. A 36 year old man with panarteriitis nodosa (PAN) presented over a certain period of time meinly with gastrointestinal symptoms. He was HBsAg positive and a "reactive" hepatitis with histologically demonstrated. Arterial microaneurysms and stenoses were detected by angiography in the liver, kidney and small intestine. These findings were confirmed by autopsy. Vascular occlusions had caused infarctions of the small intestine as well as necroses and ulcerations of the large bowel. Chronic ischemia is considered as cause of the patient's malabsorption-syndrome. It is suggested, that cachexia of PAN may be partly due to malabsorption.


[Maldescensus testis (author's transl)]. During the last years numerous publications about clinical, endocrinologic, histologic, and animal experimental examinations for maldescensus have been made intending to find the most favourable time of therapy for obtaining fertility later on. Since the results partially differ considerably and even the findings in their prospective meaning are not easy to interpret, the right time is still under dispute, particularly because it is hardly possible to determine the limitation of inborn dysgenesis against acquired degenerative changes caused by a dystop position. The unanimous opinion exists for the scrotal position of the testicles to be repaired at least until puberty.


[The transfer of drugs into the amniotic fluid]. On the basis of the present literature and our own results a review will be given about the importance of the physico-chemical properties of drugs and of different factors of biological system "mother, fetus and amniotic fluid" for the passage of drugs into amniotic fluid. The amniotic fluid is relieved as a possible distribution volume for drugs.


Alkaline ribonuclease from rye germ cytosol. 1. Alkaline ribonuclease (pH optimum 7.6) was isolated from rye (Secale cereale L) germ cytosol and partially purified; the preparation was devoid of other nucleolytic activities. 2. The enzyme is a typical endonuclease hydrolysing all phosphodiester bonds in RNA, yielding ultimately purine and pyrimidine nucleoside 2',3'-cyclic phosphates and the corresponding 3'-phosphates. Upon extensive digestion of synthetic polyribonucleotides, pyrimidine, but not purine, nucleoside 3'-phosphates are formed. The enzyme does not hydrolyse synthetic purine cyclic nucleotides. 3. The enzyme does not depolymerize double-stranded complexes of poly(A) and poly(U). 4. Susceptibility to photooxidation and inhibition by 2-hydroxy-5-nitrobenzyl bromide and N-bromosuccinimide implies the involvement of tryptophan residue in the active centre of the enzyme.


[Fatal kidney diseases due to analogesic abuse in Switzerland]. As part of an investigation into death by poisoning in Switzerland we were able to examine the register of deaths at the Swiss Federal Bureau of Statistics. All the medical reports which named analgesic nephropathy as being the basic illness and the main cause of death have been taken into consideration in this work. 331 deaths occurred in the five year period 1967-1971, and were studied as closely as the documents allowed. The earliest cases of death arose in the thirties, the maximum number however occurring between the age of 60 and 70. Between the onset of kidney damage and the actual death elapses a period of about 10 years. The ratio from woman to man is almost 2,8:1. The occurrence is particularly frequent among childless housewives. The victims are people of widely different occupations and social classes. The analgesic nephropathy resulting from addiction and leading to death is most frequently encountered among the population of the cities, and the German Swiss Cantons, as well as among people of protestant faith. The shocking number of 331 deaths in five years (in 1973 there were a further 79 cases) bears witness to the need for adequate preventive measures.


Inhibition by alanine of AMP-deaminase from rabbit skeletal muscle. 1. Alanine inhibits rabbit muscle AMP-deaminase while aspartate, histidine and glutamate are ineffective. 2. The degree and type of inhibition of AMP-deaminase by alanine depend on pH; at pH 6.5 alanine behaves like an allosteric effector exerting a negative heterotropic effect. At pH 7.0 the inhibition is non-competitive, Ki being as high as 19 mM. 3. The probable significance of the effect of alanine on AMP-deaminase in muscle metabolism is discussed.


[Drugs and their way of application in juvenile drug abuse. A dimension-analytical study of consumption habits]. In a sample of 382 heavy users mode and extent of drug abuse were inquired. Factor analysis of the data gave three factors named "i.v. applicated illegal drugs", "oral applicated illegal drugs" and "oral applicated medicaments" for short. Results show that attitudes of the drug users towards their drug uses are strongly influenced by the mode of application and the definitions of the drug laws, whereas pharmacological aspects and subjective effects have hardly any demonstrable importance.


Regulation of aconitate hydratase activity from rat kidney cortex by bicarbonate. 1. The increase in pH value and bicarbonate concentration stimulated citrate synthesis from pyruvate and malate, inhibiting simultaneously conversion of isocitrate to citrate. 2. Bicarbonate inhibited competitively the activity of aconitate hydratase, probably binding with the two active sites of the enzyme. The Ki values for the cytoplasmic and mitochondrial enzyme were, respectively, 27 and 38 mM. The pH optimum for both forms of the enzyme in Tris-HCl buffer was in the range 7.8-8.6, and in bicarbonate buffer varied from 7.2 to 8.0, depending on the form of the enzyme and the substrate used. 3. Only free, completely dissociated citrate anion acts as a substrate for aconitate hydratase. 4. The role of aconitate hydratase as a factor controlling the rate of citrate metabolism in kidney in metabolic alkalosis is discussed.


Trypsin inhibitors in bovine lung and pancreas during development. 1. The activity of trypsin inhibitors in lung and pancreas increased with age of the animals. 2. In foetal lung the activity of the basic Kunitz-type inhibitor was 1% that, and in calf lung 5-30% that, found in adult animals. 3. In foetal pancreas the basic Kunitz-type inhibitor was not detected, and in calf pancreas its activity was much lower than in adult animals. The activity of the acidic Kazal-type inhibitor in foetal pancreas was significantly lower than in calf or adult animal.


Inhibition of Ca++-stimulated respiration by uncouplers. In a phosphate medium at pH 6.6 low concentrations of uncouplers such as p-trifluoromethoxyphenylhydrazone carbonylcyanide and 2,4-dinitrophenol inhibit the oxidation of beta-hydroxybutyrate and succinate, when added during Ca++-accumulation. The inhibition depends on the amount of accumulated Ca++, and is released by N,N,N',N'-tetramethyl-p-phenylendiamine plus ascorbate as substrate. Under identical conditions the uncouplers have no inhibitory effect when added to mitochondria during state 3 respiration or during accumulation of Sr++. Inhibition of respiration by the decrease of transmembranal succinate transport or by accumulation of oxaloacetate can be excluded. It is suggested that accumulation of Ca++ in the presence of phosphate induces structural alteration of the mitochondrial membrane, which on the one hand changes the accessibility or sensitivity of dehydrogenases to uncouplers and causes leakage of accumulated Ca++ on the other.


[Inhibition of the malic enzyme from Acinetobacter calcoaceticus by NADPH and NADH]. The malic enzyme enriched from Acinetobacter calcoaceticus is inhibited by NADPH and NADH. The inhibition afforded by the reduced coenzymes is not affected by NAD+, AMP and 3'.5'-AMP. Against L-malate, NADPH inhibits the enzyme in a noncompetitive linear fashion (Ki = 1.5 x 10(-4) M), against NADP+, competitively linearly (Ki = 5.0 x 10(-5) M). While NADPH acted as a product inhibitor, NADH seems to be an allosteric effector of the malic enzyme, because with L-malate as the variable substrate in the double reciprocal plot, a nonlinear curve is obtained.


[Oxygen binding of hemoglobin following covalent fixation in the deoxy- and oxy- conformation]. Deoxyhemoglobin (deoxyHB) and oxyhemoglobin (HbO2) were covalently fixed to BrCN-activated Sephadex G-200. At pH 6, the oxygen semi-saturation pressure for the deoxyHb coupling product was 14.1, and for the HbO2 coupling product, 7.2 mm Hg. The alkaline Bohr effect delta was calculated to be - 0.55 and - 0.4 respectively. The Hill coefficients n are for both Hb derivatives between 1.4 and 1.5, independently of pH (for free Hb the respective values are pO2 50% = 18.2 mm Hg, delta = -0.55 and n = 2.5). Non-crosslinked dextran and Sephadex G-200 have no influence upon the affinity of free Hb to oxygen and upon cooperativity. As a reason for the varying oxygen-binding properties for the two Hb derivatives it is assumed that the amino group of valin alpha 1 is involved in the HbO2 fixation. In deoxyHb, this group is not probably converted. The reduced Hill coefficients and enhanced oxygen affinity are assumed to be due to impairment of the inter-chain contacts, to restrained cooperative mobility, and heterogeneity of the coupling products.


[Intracellular protein breakdown. VI. Isolation, properties and biological significance of cathepsin D from rat liver]. The preparation and properties of cathepsin D from rat liver are reported. The enzyme is an endopeptidase of lysosomal origin. The molecular weight was estimated to be 49000 by sodium-dodecylsulfate electrophoresis. We did not find any dissociation into subunits under reducing conditions, in contrast to some other authors. We found the enzyme to occur in at least 4 forms with the isoelectric points 5.87, 5.65, 5.41 and 5.13. Strong -SH-blocking reagents inhibit the activity, but the most powerful and specific inhibitor was pepstatin (Ki=38 nM). The substrate specificity is discussed. There was no proof for any zymogen activation in a great number of experiments. Since the cathepsins B1, B3 and L obviously seem to play the major role in the intracellular protein breakdown within the rat liver, the main task of cathepsin D is the degradation of extracellular proteins in this organ.


[Intracellular protein breakdown. VII. Cathepsin L and H; two new proteinases from rat liver lysosomes]. Some properties (molecular weight, pI, temperature stability, action of selected inhibitors, substrate specificity and pH-activity dependence) of two not yet known cathepsins from rat liver lysosomes are compared with the properties of the known cathepsin B1. Cathepsin L is a thiolproteinase, has a molecular weight of 23--24000 and a pI of 5,8--6,1. By disc electrophoresis and isoelectric focusing there appear several protein bands which all have enzymatic activity. Leupeptin behaves as a strong inhibitor. The pH-optimum for digestion of proteins is close to 5,0. Cathepsin L does not hydrolyse esters and splits synthetic low molecular substrates only to a low degree. Cathepsin L stored in presence of glutathion and EDTA in liquid nitrogen kept its activity for some months. Cathepsin H is an aminopeptidase as well as an endopeptidase. An enzyme with these bifunctional properties was detected up to now only in E. coli but not in animal cells. Cathepsin H is a thiol-enzyme with a molecular weight of 28000 and a pI of 7,1. Strong inhibitors are leucyl-chlormethan and SH-blocking substances. Leupeptin shows only a weak inhibitory effect to this enzyme compared to its action on cathepsins L and B1. The pH-optimum for hydrolysis of all substrates is 6.0. Cathepsin H splits proteins, amino acid derivatives and selected N-protected amino acid derivatives. Cathepsin H compared to cathepsin L and B1 is quite temperature stable.


[Influence of glutathione on the catalytic properties of leucine aminopeptidase]. 1. Leucine aminopeptidase does not catalyze the hydrolysis of glutathione. 2. Glutathione inhibits the hydrolysis of the substrates leucine hydrazide and leucine-p-nitroanilide by leucine aminopeptidase. 3. By means of kinetic experiments the type of the inhibition has been determined as noncompetitive. The inhibition constant Ki for the Mg2+-activated enzyme is five times higher than for the non-activated enzyme. 4. The degree of inhibition caused by glutathione depends on the pH value indicating a competition between glutathione and OH- ions. Mg2+-activated enzyme is invariably inhibited in the investigated pH range of 7.2 to 9.8. 5. A preincubation of the enzyme with glutathione changes the degree of activity enhancement by metal ions.


Binding of tritiated methylated luteinizing hormone to bovine corpus luteum receptors. The binding of luteinizing hormone (LH) to cow corpora lutea homogenates was studied using a tritium labelled hormone obtained by reductive methylation. The KD observed was 0.9 10(-10) M and the number of sites was the equivalent of 0.4 10(-15) moles per mg of wet tissue. The influence of the pH and temperature was examined. HCG and LH produced the same binding inhibition properties of the derivative used for labelling LH were similar to those of native LH. The inhibitory activity of the subunits was extremely low (alpha-LH: 4%, beta-LH: 1%). No significant inhibition was observed in the case of FSH or prolactin.


A descriptive and comparative study of oral health in 8-year-old Swedish children. The aim of the study was to assess the oral health of a group of children, and to compare the findings with the results from an investigation performed 4 years earlier in children of the same age and living in the same area. The material consisted of 149 randomly selected 8-year-old children. The study included examination of caries and the state of the gingival conditions, enamel hypoplasia and opacities, supragingival calculus and extrinsic stains. Determinations were also made of the rate of secretion, pH and buffer effect of stimulated saliva. Socio-economic data and anamnestic dental data were noted. The investigation showed a decrease in caries frequency in the primary teeth, but not in the permanent teeth, during the four-year-period. As in the former investigation, regression analyses showed a negative correlation between the level of education of the parents and caries indices, and that the variation of the gingival indices could be explained by socio-economic factors. In addition, a negative correlation was found between the caries index for primary teeth and the rate of secretion of the saliva and the presence of supragingival calculus.


Inorganic pyrophosphatase in isolated enamel organ and odontoblasts from the rat incisor. The inorganic pyrophosphatase (PPiase) activity was determined by a colorimetric method in the odontoblasts and the parts of the enamel organ related to enamel matrix formation and enamel maturation. The effects on PPi hydrolysis by EDTA, R 8231, urea and heat treatment were found to be almost identical to those reported for nonspecific alkaline phosphatase (APase) in the same tissues. The Mg2+ activation curve for PPiase was also similar. Like those of APase, these characteristics of PPiase activity were identical in the three locations studied. It is suggested that the close similarity in the properties of PPiase and APase is due to activity of the same enzyme, a concept which is in agreement with recent biochemical and histochemical studies of calcification.


Dissolution rate of cadmium from dental gold solder alloys. The dissolution rate of cadmium from six different dental gold solder alloys was determined in an in vitro potentiostatic study. The measurements were made in the potential range 740--880 mV and attempts have been made to extrapolate the results to a potential region that might in reality occur in the oral cavity. For comparison the dissolution rates of copper and zinc were also determined. Electrochemically the most interesting quantity was the logarithm of the dissolution rate because it is a linear function of the applied potential. This linear relationship was given by the Tafel equation. The dissolution rate of cadmium was shown to be rather small even under circumstances which may be said to represent very unfavourable conditions within the oral cavity.


Preliminary report: parenteral Lorazepam in induced epileptic states in man. The anticonvulsive effect of Lorazepam -- one of the newer diazepines -- has been tested in 10 patients with induced epileptic conditions under continuous polygraphic control. The quantitative analysis of the paroxysmal discharges -- i.e. the evolution of the "paroxysmal index" -- shows that 5 mg IV Lorazepam normalize the tracings in 2.3 minutes. Parenteral Lorazepam inhibits pharmacologically-induced electroencephalographic abnormalities in man. Its action is slightly delayed. No effect on cardiac or respiratory function, or muscle tone was noted.


The correlation between salivary peroxidase activity, salivary flow rate, and the oxidation-reduction potentials of human saliva and dental plaque suspensions. 18 subjects, 9 males and 9 females, were examined regarding salivary oxidation-reduction potential, salivary flow rate, salivary peroxidase activity, oxidation-reduction potential of dental plaque samples, and dental health. Both the peroxidase activity, expressed as the salivary lactoperoxidase, and the salivary oxidation-reduction potential increased with increasing salivary flow rate. The variation of these variables was obviously due to changes in salivary flow rate during the day. The remarkably slight differences in peroxidase activities, oxidation-reduction potentials and salivary flow rate in this study did not have any marked correlation with the clinical recordings of the test groups.


Rapid diagnosis of Enterobacteriaceae. I. Detection of bacterial glycosidases. The paper describes a number of tests for the rapid detection of glycosidases including alpha-glucosidase, beta-glucosidase, beta-glucuronidase, beta-xylosidase and alpha-fucosidase. The methods use heavy suspensions of viable but non-multiplying bacteria in a buffered solution of a chromogenic substrate. The results of the tests are readable within 4 h. The application of these tests to a collection of 633 strains of Enterobacteriaceae and Vibrionaceae demonstrates that some of the tests may be valuable additions to the present tests available for the identification of bacteria belonging to these families. beta-glucuronidase activity was observed only in strains of the Escherichia-Shigella group. 97 per cent of the Escherichia strains possessed beta-glucuronidase activity. beta-xylosidase activity was almost completely restricted to strains of the Klebsiella-Enterobacter group in addition to Yersinia strains. None of the strains possessed alpha-fucosidase activity.


Proteolytic degradation of staphylococcal alpha-toxin. Staphylococcal alpha-toxin of mol.wt. 39,000 was degraded at an alkaline pH by staphylococcal extracellular proteases resulting in the formation of three relatively stable intermediates with mol.wt. 27,500, 23,500 and 12,000. The intermediate with mol.wt. 27,500 which existed in two charged forms, was isolated by column chromatography and found to be non-haemolytic. Furthermore, it could be obtained by proteolysis of alpha-toxin (mol.wt. 39,000) with chymotrypsin in low concentrations. This intermediate was further degraded by trypsin to the protein with mol.wt. 23,500 and 12,000.


Glycerol transport in human red cells.

Glycerol transport in human red cells. The kinetics of 14C-glycerol exchange was studied in human red cells. Glycerol appeared to be transported by two mechanisms: (i) by facilitated diffusion with permeability depending on glycerol concentration, and (ii) by an unspecific pathway, presumably representing the diffusion of individual glycerol molecules through the membrane with permeability independent of glycerol concentration. The latter permeability was 8 X 10(-8) cm/s at 20 degrees C, it was independent of pH, and had an activation energy of 25 kcal/mol. The facilitated transport of glycerol was completely inhibited by Cu++, and the activation energy was low, about 10 kcal/mol. The transport system was competitively inhibited by H+, reacting with at least three hydrogen analogue, as well as dimethylsulfoxide (a hydrogen bonding molecule with no structural resemblance to glycerol), inhibited glycerol transport competitively. Steins "dimerizer hypothesis" was revised according to our findings. A kinetic scheme describing the reactions of a transport controlling site with glycerol is presented in the Appendix. It is demonstrated in the article that the scheme accounts for out experimental results.


Significance of the extracellular bicarbonate buffer system to anaerobic glycolysis in hypoxic muscle. 1. The influence of the carbon dioxide-bicarbonate buffer system on anaerobic energy production during severe hypoxia was studied in isolated right hemidiaphragms of rats.--2. When the tissue was incubated in a Ringer solution containing 25 mM HCO-3 aerated with 7% CO2 in N2 at pH 7.4, the lactate production and lactate content of the tissue increased.--3. At an extracellular (tissue bath) pH OF 6.9 the lactate production was stimulated when carbon dioxide and bicarbonate were changed to 19% and 25 mM, respectively. This stimulatory effect disappeared when these values were lowered to 7% and 7 mM.--4. At pH 7.4 the stimulatory effect of the carbon dioxide-bicarbonate system persisted when the buffer value was lowered from 60 to 3 mM by changing the system from an open (i.e. continuous gas equilibration) to a closed one (i.e. without any gas phase). Decreasing the glucose in the media from 22 to 0 mM reduced the lactate production and abolished the stimulatory effect of the carbon dioxide--bicarbonate system.--5. There was no direct effect of this system on the glycolytic enzymes (i.e. lactate production and activity of phosphofructokinase of homogenates).


Influence of adipose tissue blood flow on the lipolytic response to circulating noradrenaline at normal and reduced pH. Hypercapnic acidosis (pH 7.0) inhibits the lipolytic response of canine subcutaneous adipose tissue to i.v. infused noradrenaline (NA) by 80 per cent or more. The response to sympathetic nerve stimulation, on the other hand, is only reduced by 10-40 per cent during acidosis. The fate of intravenously infused 3H-labelled NA (0.35 ug X kg-1 X min-1 for 30 min) was not significantly altered by acidosis. The rate of disappearance of unmetabolized NA from the arterial plasma after an infusion was the same at pH 7.4 and 7.0 and the calculated increase in circulating NA during infusions was 4 ng/ml at both pH:s. I.v. infusion of Na increases adipose tissue blood flow, an effect which is attenuated by acidosis. There was a significant correlation (p less than 0.001) between adipose tissue blood flow and the lipolytic response at normal pH. Preventing the NA-induced increase in blood flow by constant flow perfusion reduced the lipolytic response at normal pH. The degree of inhibition by acidosis of the lipolytic response to i.v. NA was significantly reduced (from 79 to 56 per cent, p less than 0.05) when the adipose tissue was perfused at constant flow. These data suggest that adipose tissue blood flow is important in determining the lipolytic response to i.v. NA, probably by influencing the delivery of NA to the tissue. The marked inhibition by acidosis of lipolysis due to i.v. infused NA therefore appears to be the combined effect of a direct antilipolytic effect of acidosis and a decreased delivery of NA to the adipose tissue due to the attenuated blood flow response.


Contrast media with and without calcium for cardioangiography in children. The possible beneficial effects of added calcium ions to contrast media to diminish myocardial toxicity was evaluated in 93 cardiac catheterizations with cardioangiography, performed in 50 children and adolescents. Two contrast media with equal iodine content (370 mg/ml) and similar physical properties were used: Isopawue-370, containing 0.34 mg calcium/ml, and Renografin-76, not containing calcium. It is concluded that addition of calcium to contrast media for cardioangiography in children and adolescents does not alter their myocardial, fluid or electrolyte effects when injected selectively into the cardiac chambers or great vessels.


Influence of the cation on the side-effects of urographic contrast media. The incidence of side-effects produced by meglumine diatrizoate and meglumine-, meglumine-calcium-, and sodium-calcium-magnesium metrizoate, was compared in 800 urographies. Patients older than 60 years seem to have less side-effects than younger patients. The incidence of sensation of warmth seems to be higher in patients who have had previous urographies, compared to those who are examined for the first time. The incidence of this effect is higher for the sodium-calcium-magnesium salt of metrizoate, than for other salts of metrizoate.


Regulation of lysine biosynthesis in Escherichia coli K12. A general survey of the regulation in lysine biosynthesis in Escherichia coli K12 is presented. No polygenic operon exists for the genes that code for enzymes of the lysine biosynthetic pathway. Lysyl-tRNA is not directly involved as a co-repressor in the pathway. Different regulation mechanisms must exist for the different enzymes. In the case of the last enzyme, diaminopimelate decarboxylase, its synthesis is induced in vivo by the lysine-sensitive aspartokinase under its non-inhibited allosteric conformation.


Regulation of tyrosine and phenylalanine biosynthesis in Salmonella. Several types of 4-fluorophenylalanine resistant mutants were isolated. In one type of mutant DAHP synthetase (tyr) and prephenate dehydrogenase were coordinately derepressed. The mutation was linked to aroF and tyrA and was cis- dominant by merodiploid analysis, thus confirming that it is an operator constitutive mutation (tyrOc). A second type of mutation showed highly elevated levels of tyrosine pathway enzymes which were not repressed by L-tyrosine. It was unlinked to tyrA and aroF, and was trans-recessive in merodiploids. These properties were attributed to a mutation in a regulator gene, tyrR (linked to pyr F), that resulted in altered or non-functional aporepressor. Hence tyrO, tyrA, and aroF constitute an operon regulated by tyrR. In a third type of mutation chorismate mutase P-prephenate dehydratase was highly elevated. It was not linked to pheA, was located in the 95--100 min region of the Salmonella chromosome, and was recessive to the wild type gene in merodiploids. A mutation was, therefore, indicated in a regulatory gene, pheR, which specified an aporepressor for regulating pheA. DAHP synthetase (phe), specified by aroG, was not regulated by pheR, but was derepressed in one of the tyrR mutants, suggesting that as in Escherichia coli tyrR may regulate DAHP synthetase(phe) and DAHP synthetase (tyr) with the same aporepressor. A novel mutation in chorismate mutase is described.


Blocking of olive oil induced plasma protein escape from the intestinal circulation by histamine antagonists and by a diamine oxidase releasing agent. Earlier studies have shown that feeding of olive oil to rats substantially increased the plasma protein in the intestinal lymph. The possibility of histamine mediating this response was examined. The plasma protein escape from intestinal circulation after olive oil feeding was measured in rats in terms of the amount of Evans Blue labelled plasma protein found in the intestinal lymph. Animals treated with histamine antagonists (H1-receptor antagonist pyrilamine, 16-22 mg/kg i.p., plus H2-receptor antagonist Burimamide, 12-16 mg/kg i.p.) did not show an increase in the quantity of lymphatic plasma protein. Heparin pretreatment which releases the histaminolytic enzyme, diamine oxidase, into the interstitial space also prevented the increased accumulation of labelled plasma protein in the lymph after olive oil ingestion. Based on these observations, histamine appears to act on the intestinal microcirculation during olive oil absorption and allows larger quantities of plasma proteins to leave the intestinal circulation.


Composition of the Zaysan virion.

Composition of the Zaysan virion. The sedimentation coefficient of Zaysan virion ribonucleic acid (RNA), estimated by sucrose density gradient centrifugation, was 42 S. Pancreatic ribonuclease digested the viral RNA to acid-soluble fragments. Two viral structural proteins with apparent molecular weight of 50,000 and 30,000 daltons were identified by polyacrylamide gel electrophoresis. A dense, RNA-rich particle, containing the smaller polypeptide, was isolated after mild detergent treatment of Zaysan virus.


Morphological, cytological and biological observations on viruses isolated from patients with subacute thyroiditis de Quervain. New data on viruses isolated from patients with subacute thyroiditis de Quervain are reported. Characteristic morphological, cytological, some physico-chemical and biological features of the isolated viruses are described. A possible role of these viruses in human and animal health disorders is discussed. The isolated viruses remain unclassified so far.


Interfering activity of virulent and attenuated influenza virus strains. The interfering activity of influenza virus variants A/Hong Kong/1/68 (H3N202), A/Victoria/35/72(H3N2-3), B/14/55 and B/USSR/69 differing in the level of their reactogenicity for adults and children was studied. An inverse relationship was established between reactogenicity of the strains and their interfering activity in the resistant chick embryo cell (CEC) cultures. Virulent strains did not interfere with vesicular stomatitis virus. Vaccine strains used for commercial live influenza vaccine safe for adults but reactogenic in children were intermediate and showed moderate interfering activity. The highest capacity for interference was demonstrated in cold-adapted thermosensitive variants non-pathogenic for both adults and children. The interfering activity of the attenuated strains increased progressively with increasing inocula.


Interference stimulated by tick-borne encephalitis virus: influence of divalent cations. The effects of divalent cations (barium, cobalt, calcium, copper, magnesium, zinc and iron) on the interference of tick-borne encephalitis (TE) virus with vaccinia virus in Detroit-6 cell cultures were investigated. An increase in interference was shown in the presence of cobalt, zinc, copper, and magnesium. Barium and iron displayed a marked inhibitory action. But an increase in interference was not parallel with an increased production of interferon.


Autoantibodies in sera of influenza patients. Sera of the influenza patients and healthy controls were tested for some types of autoantibody (SMA, ANA, ABBA, AMA). They were detected in 83.8% of the patients' sera and in 16.6% of controls. SMA were present in 77.4%, ANA in 54.8%, and ABBA in 16.1% of the patient's sera. AMA were not detected. A majority of the sera contained more than one autoantibody type. The possible mechanisms of induction of the autoantibodies in virus infection and their possible role in disease are briefly discussed.


Latent pseudorabies virus infection in pigs. Latent infection with the TOP strain of pseudorabies virus (PRV) was established in 6 weeks old piglets. Infectious virus was found in oropharyngeal swabs till day 10 post infection (p.i.); later on, attempts to detect the virus remained unsuccessful. However, PRV could be isolated by explantation of tonsils, cervical lymph nodes, nasal mucosa and gasserian ganglia. These tissues were removed between 160 and 181 days p.i., during cultivation, infectious PRV was released into the culture fluid from the 3rd to the 11th day of the explantation. PRV antigen was seen by immunofluorescence only in explants coming from gasserian ganglia. It was localized in both neurons and satellite cells. In piglets given hydrocortisone before explantation, the number of virus-producing explants was not enhanced as compared to that of virus-producing explants from untreated animals.


Experimental live tick-borne encephalitis vaccine (Labgat E5 "14" virus clone): volunteers 1 and 2 years after single-dose immunization. Groups of volunteers given intramuscularly 5 or 6.5 dex or perorally 6.5 dex newborn mouse icLD50 of the plaquesegregated "14" clone of the Langat E5 virus strain (tick-borne encephalitis comples) were followed for periods of 12 and 24-27 months. Circulating specific virus neutralizing antibodies persisted in them in the absence of apparent reaction as evidenced by clinical, electroencephalographic and cerebrospinal fluid findings.


Experimental infection of Macaca mulatta monkeys with Lednice (Yaba 1) virus. Macaca mulatta monkeys were inoculated with Lednice (Yaba 1) virus strain 6118. The animals developed no clinical signs and the body temperature and values of complete haematological examination remained within normal limits. In spite of positive immunofluorescence in regional lymph nodes, no virus could be recovered from them by passages in mice. Viraemia was not demonstrated. The antibody response was directly proportional to the amount of virus inoculated, i.e. it was significant after a higher virus dose or after repeated inoculation. The results obtained are discussed in respect of and in comparison with serological findings in man.


A stable antibody-sensitized erythrocyte diagnostic preparation for detection of rhinovirus type 17 and its antibody. A diagnostic preparation was obtained by sensitization of formalinized sheep erythrocytes with antibody to rhinovirus type 17. This preparation was agglutinated in the presence of relatively high concentrations of type 17 but not other types of rhinovirus. It could also be used for detection of antibody to rhinovirus type 17 in the antigen neutralization test, the antibody levels determined in this way being similar to those found by conventional virus neutralization tests in cell cultures.


Soldado virus from Ornithodoros (Alectorobius) maritimus (Ixodoidea: Argasidae) infesting herring gull nests on Puffin Island, Northern Wales. Three strains of Soldado (SOL) virus (Hughes serogroup) were isolated from nymphal and adult Ornithodoros (Alectorobius) maritimus Vermeil et Marguet collected in and near nests of the Herring Gull, Larus a. argentatus Pontoppidan, on Puffin Island, northern Wales. Reciprocal complement fixation (CF) titration results demonstrated recovered virus strains to be SOL virus and antigenically distinct from other Hughes serogroup members. All isolates killed mice and guinea pigs, and 1-2 day old domestic chicks when inoculated intracerebrally.


Infection of Hela cells with a virulent and an attenuated strain of pseudorabies virus studied by electron microscopy. Hollow-core particles, forming crystals in nuclei, prevailed in HeLa cells infected with an attenuated strain of pseudorabies virus (PRV). After infection with a virulent PRV strain, the cells contained mainly fully infectious dense-core particles. These findings might explain the lower susceptibility of HeLa and some other human cells to infection with attenuated strains of PRV as compared to virulent strains.


Respiratory syncytial virus-specific RNA synthesis in primary monkey kidney cell cultures. Primary rhesus monkey kidney (MK) cell cultures were inoculated with respiratory syncytial virus and treated or untreated with actinomycin D before pulse labeling with uridine-5-3H. The virus-specific RNA synthesis was noted at its peak in the nucleoplasm and possibly less so in the cytoplasm of infected cells. At 48 and 72 hours post-inoculation (p.i.), small fractions of available cells were synthesising virus-specific RNA with labeling index of 15% and 18% respectively. By 48 hours p.i. syncytia started appearing and a higher grain count in the cytoplasm of actinomycin D-treated infected cells was noted.


Cultivation of foot and mouth disease virus in bovine leukocyte cultures for use in the complement fixation test. Bovine leukocyte cultures achieved good growth after 72 hours of cultivation. Foot-and mouth disease (FMD) virus type 0 multiplied in such cultures to a titre of 10(6) TCID50/0.1 ml. The cell culture-grown virus was found to be suitable for the complement fixation test (CFT) after purification and concentration with calcium phosphate.


Non-infectious tick-borne encephalitis antigen. After formalin treatment of sucrose-acetone antigens, prepared from different tick-borne encephalitis (TBE) virus strains, their haemagglutination (HA) activity was preserved or even higher than that of untreated antigens. Formalin-treated TBE antigen was non-infectious and thus may be recommended for routine work to avoid laboratory infections.


Molecular weight analysis of the polypeptides of cholera phage PL 163/10 by polyacrylamide gel electrophoresis. Cholera phage PL 163/10, belonging to Mukerjee's Group I, was purified by alternate cycles of low and high speed centrifugation. Gel electrophoresis revealed the presence of four polypeptide chains of respective molecular weights of 10,370+/-515 (A), 30,000+/-1,303 (B), 40,000+/-1,049(C) and 64,000+/-2,433 (D) daltons. Electrophoresis of the sample alkylated with iodoacetic acid resolved the presence of only one polypeptide chain of an average molecular weight of 10,310+/-565 daltons. The polypeptides B, C and D could be interpreted as trimer, tetramer and hexamer respectively of the polypeptide A, which was the basic structural protein of this phage.


Investigation on the biological activity of fowl plague virus ribonucleoprotein. Fowl plague virus (FPV) ribonucleoprotein (RNP) bands in sucrose density gradient in a heterogeneous peak with sedimentation coefficients from 45 to 70 S, whereas in cesium chloride gradient it has a homogeneous density of 1.33-1.34 g/cm3. FPV RNP contains 7.4-8% RNA. Upon inoculation of chick embryo cell cultures. FPV RNP shows no infectivity, does not induce virus-specific protein synthesis and does not participate in complementation or recombination interactions with ts mutants of FPV. The biological activity of FPV RNP demonstrable under certain experimental conditions is due to admixture of undestroyed virions and is completely eliminated by treatment of the preparation with gamma-globulin fraction of antiserum to FPV haemagglutinin, but not with antiserum to RNP proteins.


Interaction of plasma membranes with influenza virus. VI. The possible role of the adenylate cyclase system. The amounts of released soluble (s) antigen of influenza A/WSN virus were increased when the virus was allowed to interact with isolated plasma membranes in a medium containing substances enhancing the level of adenosine 3',5' cyclic monophosphate (c'AMP) or activating the enzyme adenylate cyclase. By contrast, less s-antigen was released upon addition to the incubation medium of foetal calf serum or calf serum proteins which activate c'AMP phosphodiesterase and thus decrease the level of c'AMP. Changes in the amount of released s-antigen were parallelled by changes in the activities of membrane Ca-adenosine triphosphatase and creatine phosphokinase.


Virion heat-sensitivity of adenovirus type 5 temperature-sensitive mutants and interferon induction. Kinetics of interferon induction with wild type adenovirus 5, and three temperature-sensitive mutants, ts 1, ts 18 and ts 19 were examined in chick embryo cells (CEC) at permissive (31 degrees C) and non-permissive (38 degrees C) temperatures. Interferon levels reached a maximum four days after infection with the wild type and all ts mutants at 31 degrees C. However, at 38 degrees C, ts 18 and ts 19 failed to induce interferon, while wild type and ts 1 induced normal levels which reached a maximum by three days after infection. Striking differences in virion heat sensitivities between wild type, ts 1, ts 18 and ts 19 at both 50 degrees C and 52 degrees C were detected. These results suggested that ts 18 and ts 19 might each have a mutation in a gene coding for a virion structural component. The production of infectious progeny by shift-up experiments on Hela cells showed that mutant ts 18 was affected immediately after shift-up at all times throughout the growth cycle. On the other hand, interferon production in CEC by mutant ts 18 was only affected by shift-up during the first five hours of incubation, which indicated involvement of an early gene function in elliciting interferon production.


Ts mutant of NWS influenza virus: behaviour in vitro and in vivo. After passaging the NWS influenza virus at increasing temperatures in the hamster embryo fibroblast (HEF) cell line, a temperature-resistant (tr) mutant which grew well at 39 degrees C was isolated. Attempts were made to isolate temperature-sensitive (ts) mutants from the tr virus. After growth in the presence of 5-fluorouracil, 233 virus clones were isolated. Among these only one was incapable of replicating at 39 degrees C. The physiological studies, the shift-up experiment and the electron microscopic investigation suggested that the genetic defect responsible for its ts character influenced a late function of the virus genome. In the lungs of intranasally infected mice, the ts mutant replicated to a lesser extent than the tr virus. During its growth in vivo, the ts virus exhibited a high degree of genetic stability.


Replication dynamics of tick-borne encephalitis virus in, and glycolytic activity of, human diploid cells. Tick-borne encephalitis (TBE) virus (Western subtype) strains replicated very rapidly in human diploid embryonic lung (HDEL) cells, namely within the first 24 hours of infection. Greater amounts of both intra- and extracellular virus were produced after inoculation of a virulent than of an attenuated strain. With the virulent strain, more infectious virus was found in the medium than in the cells: the opposite was true for the attenuated strain. No cytopathic effect was observed during the period studied. The rates of glucose uptake and lactate production by cells were markedly influenced by the infection. Lactate production was retarded by both virus strains to a comparable degree. Glucose uptake was markedly increased, appreciably more so after infection by the virulent than the attenuated strain. A method for differentiating between attenuated and virulent strains based on the determination of glucose conthe medium was developed.


Affinity chromatography of mouse interferon: a modified purification procedure utilizing specifically purified antibodies. Interferon preparations of a high degree of purity were obtained by a one-step procedure using affinity chromatography on specifically purified immunoadsorbent. The procedure consisted of binding interferon harvested from serum-free medium and purified by Zn-acetate precipitation and SP-Sephadex chromatography to CNBr-activated Sepharose 4B (Column No. 1). In the next step, antiinterferon globulin was purified by affinity chromatography on Column No. 1 with the bound interferon. In this way, antibodies against purified interferon, which were free from non-antibody components, were obtained. The purified antibodies were then coupled to CNBr-activated Sepharose 4B forming Column No. 2. The latter had a binding capacity of 125 800 mouse interferon units per 4.8 ml of gel. This capacity was not altered during an 8-month period of use. The gel was capable to bind interferons obtained from fibroblasts and leukocytes and, partially, from serum. The resulting purified, products were similar, i.e. they were not influenced by volume, interferon activity, or purity of the starting material. The electrophoretic profiles of the products had a similar shape irrespective of the origin of the starting material.


Use of cross-linking in studying the structure of RNA tumour viruses. Treatment of intact avian myeloblastosis virus (AMV) with dimethyl suberimidate dihydrochloride (DMS), a cross-linking agent specific for amino groups, was found to result in progressive cross-linking among viral proteins, as revealed by polyacrylamide gel electrophoresis (PAGE) in the presence of sodium dodecyl sulphate (SDS). Free viral proteins were not cross-linked. The cross-linked protein complex with an apparent molecular weight of 50,000 daltons was studied in detail.


Eyach--an arthropod-borne virus related to Colorado tick fever virus in the Federal Republic of Germany. From Ixodes ricinus ticks collected in a tick-borne encephalitis focus in Baden-Württemberg, an agent pathogenic exclusively for suckling mice was isolated. Obviously a virus, it passed membrane filters with a pore width of 200 nm. It was resistant to ether and sodium deoxycholate, but not to chloroform. In the complement fixation test it showed a close, and in the neutralisation test a more one-sided relationship to Colorado tick fever virus.


Effect of adamantane derivatives on the activity of orthomyxovirus RNA-dependent RNA polymerase. The effect of several adamantane derivatives on the activity of virion-associated RNA-dependent RNA polymerase of fowl plague virus (FPV) and influenza B virus was studied in vitro. Some of the derivatives inhibited the activity of the polymerase by 60 per cent. A correlation was established between the previously demonstrated capacity of these inhibitors to suppress orthomyxovirus reproduction in vivo and their ability to reduce the activity of virion-associated RNA-dependent RNA polymerase in vitro.


Increased resistance of an interferon-resistant cell subline to the toxic effect of double-stranded RNA. The toxicity of double-stranded (ds) f2-phage RNA for interferon-treated L cells and an interferon-resistant subline of L cells (CVS cells) was compared. The interferon-resistant subline proved to be about 20 times more resistant to ds RNA than the parent L cells. The degree of toxicity of ds RNA was dependent on the concentration of interferon to which the cells were exposed.


[Some aspects of phylogenetic development of cellular immune reaction in vertebrates]. The division of the immune system into humoral and cell-mediated immunity is established in vertebrates. Moreover single manifestations of cell-mediated immunity are observed in invertebrates, too. In protochordates, e.g., a genetically fixed ability to recognize "Self" and "Not-Self" could already be demonstrated, but the interpretation of this phenomenon as a preliminary stage of cell-mediated immunity in vertebrates is not established yet. Especially by the investigation of the transplantation tolerance, for example between mammalia and inferior vertebrates, essential results as to the development of the immune system in vertebrates could be obtained. It is quite sure that the experimental findings obtained till now are not sufficient for giming a complete picture of the phylogenesis of immunologic reactivity. But they very unanimously support the conception that nearly all of the nowadays living representatives of the vertebrates dispose of cell-mediated immunity, which is equal in its characteristic features. And the small lymphocyte takes the central part in the cell-mediated immune reactions of the vertebrates, and the complexity of the lymphoid organs, which is increasing in the course of phylogenesis leads to a quantitative but not to a qualitative change.