Adenosine-5'-phosphosulfate (APS) as sulfate donor for assimilatory sulfate reduction in Rhodospirillum rubrum. Crude extracts of Rhodospirillum rubrum catalyzed the formation of acid-volatile radioactivity from (35S) sulfate, (35S) adenosine-5'-phosphosulfate, and (35S) 3'-phosphoadenosine-5'-phosphosulfate. An enzyme fraction similar to APS-sulfotransferases from plant sources was purified 228-fold from Rhodospirillum rubrum. It is suggested here that this enzyme is specific for adenosine-5'-phosphosulfate, because the purified enzyme fraction metabolized adenosine-5'-phosphosulfate; 3'-phosphoadenosine-5'-phosphosulfate, however, only at a rate of 1/10 of that with adenosine-5'-phosphosulfate. Further, the reaction with 3'-phosphoadenosine-5'-phosphosulfate was inhibited with 3'-phosphoadenosine-5'-phosphate whereas this nucleotide had no effect on the reaction with adenosine-5'-phosphosulfate. For this activity with adenosine-5'-phosphosulfate the name APS-sulfotransferase is suggested. This APS-sulfotransferase needs thiols for activity; good rates were obtained with either dithioerythritol or reduced glutathione; other thiols like cysteine, 2'-3'-dimercaptopropanol or mercaptoethanol are less effective. The electron donor methylviologen did not catalyze this reaction. The pH-optimum was about 9.0; the apparent Km for adenosine-5'phosphosulfate was determined to be 0.05 mM with this so far purified enzyme fraction. Enzyme activity was increased with K2SO4 and Na2SO4 and was inhibited by 5'-AMP. These properties are similar to assimilatory APS-sulfotransferases from spinach and Chlorella.


Allohydroxy-D-proline dehydrogenase. An inducible membrane-bound enzyme in Pseudomonas aeruginosa PA01. Growth of Pseudomonas aeruginosa PA01 on isomers of hydroxyproline induced the synthesis of an allohydroxy-D-proline dehydrogenase. The enzyme resembled the D-alanine dehydrogenase of this organism in its association with the particulate fraction and its linkage to oxygen through a cytochrome-containing respiratory chain, but differed from this and other bacterial D-amino acid dehydrogenases in its high substrate specificity and low Km.


Neurophysiologic and pathologic aspects of acute and chronic pain. Acute pain produced by disease or injury is the net effect of highly complex interactions of various neural systems and psychological factors. Through the interaction of the afferent systems and neocortical processes, the individual is provided perceptual information regarding location, magnitude, and spatial and temporal properties of the noxious stimulus that activates motivational tendencies toward escape or attack and permits analysis of multimodal information, past experience, and probability of outcome of different response strategies. In contrast, chronic pain is a malefic force that taxes the physical, emotional, and economic resources of the patient, his famiily, and society. Moreover, chronic pain is characterized by physiological affective and behavioral responses that are quite different than those of acute pain. The clinician must keep these differences in mind in order to provide patients with optimal relief of their pain.


[Ultrastructure of the parietal cells of the gastric mucosa in ulcer disease of the duodenum with a hyperacid syndrome (ultrastructurometric study)]. An electron-microscopy investigation and morphometric analysis were carried out in 30 specimens of bioptic material of the fundal part of the stomach in patients with duodenal ulcer with the hyperacid syndrome. At all levels of the Major gastric glands parietal cells with morphological signs of intensive secretory activity prevailed, which was one of the causes (apart from the increase in the mass of parietal cells) of considerable intensification of secretion of hydrochloric acid. It was suggested that a considerably increased (as compared with the norm) area of the surface of the crysts of mitochondria with simultaneous reduction of the coefficient of their fragmentation was the morphological substrate of a high functional activity of parietal cells of the gastric mucosa in the hyperacid syndrome.


Parkinson's disease.

Parkinson's disease. The classical clinical picture of Parkinson's disease is all too frequently encountered in the elderly patient. The modern classification of causes of Parkinson's disease lists 'idiopathic', and 'post-encephalitic forms', the latter being uncommon in this community.


[Carbohydrate-dependent lipidogenesis. Review of the literature and experimental study]. The literature on glycide-induced lipidogenesis is reviewed. This is an important question also because of its relation to the pathogenesis of pathological conditions such as ketoacidosis, obesity and hypertriglyceridaemia. Doubt is expressed concerning the interpretation of some experimental procedures used in determining the extent of lipidogenesis in man. The questions raised are fully discussed. Their solution is sought via an experiment on the rat involving the comparison of tissue and plasma lipid metabolites values following infusion of a labelled glucose bolus. Little correlation was noted, low plasma levels being found with high tissue radioactivity. It is emphasized that it is very difficult to ideate a valid experimental approach in order to investigate the extent of lipidogenesis in man in vivo: in this respect, the long term risk arising from the use of tracers with a very long half-life (e.g. 14C) in man must be carefully valuated.


Survival test of submersible life support systems. An experiment to validate predictions concerning submersible survivability was performed in December, 1975, by members of the Canadian Forces in the CF Submersible Lockout Vehicle SDL-1 in Halifax Harbour in water of 4 degrees C temperature at a depth of 40 ft. Data was collected relevant to the life support equipment to determine if it would operate for a simulated 6-h mission followed by a 24-h immobility period, at the end of which rescue was presumed to have occurred. Physiological data was collected from the submersible occupants in order to assess the degree of thermal stress experienced in this exercise. The experiment was terminated after a duration of approximately 25 h at 1 atm internal pressure due to exhaustion of two of the three on-board power supplies, causing the CO2 scrubbers to be inoperative and the CO2 content in the breathing gas to increase to toxic levels. Only two of the three submersible occupants experienced cold stress, one in the forward sphere and one in the aft sphere. At the end of 24 h, the core temperatures of both individuals had decreased by 0.5 degrees C and, during this time, skin temperatures, particularly of the extremities, had steadily and slowly decreased. Neither individual was hypothermic, but it was considered likely that after a 3-d exposure, at least two of the crew members would have had core temperatures of 35 degrees C or lower, assuming that CO2 poisoning had not occurred earlier.


Systematic desensitization of a phobia for flying with the use of suggestion. This case study describes the successful treatment of a phobia for flying in an active-duty United States Marine helicopter crew chief by using systematic desensitization. Systematic desensitization is briefly described as a treatment technique which incorporates relaxation exercises paired with the visualization of anxiety-producing images--in this case, images associated with flying. The patient and the therapist constructed a hierarchy of 32 items which were then rank ordered from least to most anxiety producing with the first item being "Ordering parts to repair a helicopter" and the last item being "A routine helicopter landing". Treatment consisted of having the patient visualize the scenes from the list while achieving a state of deep relaxation. The patient was unique in that he possessed an extraordinary ability to experience olfactory, tactile, auditory, and kinesthetic cues as well as vivid visualizations when imagining the items. The patient moved to the next item on the list only after he could feel relaxed while visualizing the previously anxiety-arousing scene of the preceding item. After progressing through the entire list, the patient initiated an actual helicopter flight. The patient participated in a 6-month follow-up helicopter flight with the therapist and experienced an appropriate, manageable level of anxiety. The presentation concludes with a brief discussion concerning the protential usefulness of routinely incorporating relaxation exercises into flight training as a preventative approach to anxiety- and stress-related illness frequently found in aviators.


The intracellular pH and the buffer capacity of the heart. The intracellular buffering (delta HCO3-/delta pH) in rats is lowest in skeletal muscle, intermediate in the diaphragm and highest in cardiac muscle. In the hypertrophic heart due to aortic constriction or to the Goldblatt mechanism the buffer value increases markedly during the first weeks, but returns to normal or even subnormal values during stable hypertrophy. There is an electrochemical disequilibrium for hydrogen ions caused by an active transport of ions across the cell membrane, which is possibly correlated to the myocardial activity.


Lymphocytotoxic antibodies. HLA antigen associations, disease associations, and family studies. Lymphocytotoxic antibodies (LCTAB) were sought in sera of patients with rheumatic diseases and in family members. Patients with SLE and cutaneous necrotizing venulitis and family members of JRA patients had an increased frequency of LCTAB; JRA patients and family members of SLE patients did not. The only association between LCTAB and the HLA phenotype of persons with LCTAB was a decreased frequency of LCTAB in individuals with HLA-B27.


Determination of flunitrazepam, desmethylflunitrazepam and clonazepam in plasma by gas liquid chromatography with an internal standard. A method is developed for gas-chromatographic determination of flunitrazepam, desmethylflunitrazepam and clonazepam in plasma, using methylclonazepam as internal standard. Following extraction of the benzodiazepines and their hydrolysis, the benzophenones are analyzed by gas-chromatography, with a glass column filled with 3% OV-225 on Gas Chrom Q and a 63Ni electron capture detector. The technique has a good selectivity. Its limit of sensitivity is about 0.5 ng/ml of plasma. It is suitable for investigations on pharmacokinetics and bioavailability as well as for monitoring the correlation between plasma level and therapeutic efficacy and toxicology.


[Studies on the reactability of chlordiazepoxide to its N-nitrosos derivative in physiological conditions (author's transl)]. It is shown that chlordiazepoxide readily reacts with sodium nitrite in an aqueous HCl solution as well as in human gastric juice to N-nitrosochlordiazepoxide with a high yield (up to 75%). For this reaction the pH optimum has been determined to be 1.4. After oral application of chlordiazepoxide alone or in combination with nitrite to rats, the drug as well as its N-nitroso derivative were recovered from serum and analysed by high-pressure liquid chromatography.


Serum gonadotropins in the rat after prenatal damage to the testes by busulfan and their reaction to cryptorchidism, castration and administration of testosterone. Adult male rats prenatally treated with 1,4-bis-(methanesulfonyloxy)-butane (busulfan), which damages the gonocytes, undergo cryptorchidization, castration or testosterone treatment and the FSH, LH and testosterone levels in serum are measured. Basal values of these hormones do not differ from those of controls. After cryptorchidization FSH levels rise but the LH levels do not. Following orchidectomy both hormone levels increase significantly. Application of testosterone in various doses increases FSH in normal animals but not in busulfan treated rats 24 h post infectionem. FSH and LH levels 7 days after administration do not differ from controls. According to the hypothesis that the FSH release depends on an inhibin from the testes, these experiments show evidence for inhibin being produced only in the Sertoli cells.


Double-blind cross-over clinical comparison of two 2'-chloro benzodiazepines: 7-chloro-5-(2-chlorophenyl)-1,3-dihydro-2H-1,4-benzodiazepin-2-one (chlordesmethyldiazepam) versus 7-chloro-5-(o-chlorophenyl)-1,3-dihydro-3-hydroxy-2H-1,4-benzodiazepin-2-one (lorazepam) in neurotic anxiety. A group of 20 female neurotic inpatients has been treated with 7-chloro-5-(2-chlorophenyl)-1,3-dihydro-2H-1,4-benzodiazepin-2-one-(chlordesmethyldiazepan)- 7-chloro-5(o-chlorophenyl)-1,3-dihydro-3-hydroxy-2H-1,4-benzodiazepin-2-one (lorazepam) according to a double-blind cross-over design. For each drug clinical evaluations were performed by means of Hamilton's rating scale for anxiety states and of Overall and Gorham's brief psychiatric rating scale, at the beginning, after the first week and at the end of the two-week period of treatment, in opposite sequence. A statistically greater efficacy of chlordesmethyldiazepam in  comparison to lorazepam was observed. Results are discussed with regard to benzodiazepine structure-activity relationships.


[Psychic drugs: predicting therapeutic effects and doses by test models with normal subjects (author's transl)]. Pharmacopsychology is concerned with the effects of drugs on "psychic" processes. It is attempted to demonstrate approaches which meet the criteria of scientific methodology. These approaches are based upon observation of somatic and/or behavioral changes. The steps from "naive" observation to the generation of objective data consist mainly of strict control of the observational situation and of technical refinement of the observation methods. Two kinds of pharmacopsychological models in healthy volunteers are described: models of normal and models of abnormal behaviour. Their relevance for the prediction of therapeutic drug effects and dosage is discussed. Possible new approaches are demonstrated: a) utilization of feedback form clinical investigation to control the relevance of pharmacopsychological models and, b) the concept of symptomatic volunteers, who are shown to be more suitable models than non-selected subjects.


Destruction of endogenous low density lipoprotein in incubated intima. The effect of incubation on the content of endogenous intact plasma lipoprotein (LP) has been examined in minced samples of normal intima and lesions from 38 patients. Both the electrophoretically mobile and the immobilized LP fractions decreased on incubation, and the rate of destruction was proportional to LP concentration (r=0.832, p less than 0.001). Mincing the intima with EDTA before incubation increased the rate of destruction about 4-fold in fibrous lesions but not in lesions containing numerous fat-filled cells. The destruction of LP was highly dependent on pH; the rate was highest below pH  5.5 and destruction was almost completely inhibited above pH 6.4. In standard cathepsin assays haemoglobin substrate was hydrolysed at a rate comparable to the rate of destruction of LP. The results suggest that LP may be degraded by a lysosomal cathepsin in intima.


Effect of prostaglandins E1 and F1alpha on the activities of cholesteryl ester synthetase and cholesteryl ester hydrolases of pigeon aorta in vitro. The in vitro effects of prostaglandins E1 and F1alpha on the activity of cholesteryl ester synthetase and cholesteryl ester hydrolase activities of the pigeon aorta were examined. It was found that prostaglandin E1 markedly inhibited the cholesteryl ester hydrolase activity in the supernatant fraction and slightly  inhibited the cholesteryl ester synthetase activity. Prostaglandin F1alpha, however, modestly stimulated the cholesteryl ester hydrolase activity both in the microsomal and in the supernatant fraction of the aorta. These observations strongly warrant further studies on the role of prostaglandins in atherogenesis.


Comparative beta-adrenoceptor blocking effects and pharmacokinetics of penbutolol and propranolol in man. 1 The beta-adrenoceptor blocking effects of penbutolol were compared with those of propranolol and a placebo in a double-blind trial involving six healthy volunteers. 2 Heart rate (HR), systolic blood pressure (SBP) and peak expiratory flow rate (PEFR) were measured at rest and during vigorous exercise before and at intervals up to 7 h after oral administration of the drugs. In addition, plasma renin activity (PRA) at rest and plasma levels of penbutolol and propranolol were determined. 3 Penbutolol proved to be a non-cardioselective beta-adrenoceptor blocking drug, antagonizing exercise-induced tachycardia, reducing exercise-induced increase in PEFR and decreasing PRA. The beta-adrenolytic potency of penbutolol was shown to be four-fold that of propranolol but the duration of its effect was similar. 4 The peak plasma level of penbutolol was reached 1 h after administration and its half-life was 4.5 h. 5 Comparison of plasma levels and biological activity of penbutolol revealed that after oral administration this drug is transformed into an active metabolite in man.


Measurement of thioridazine in blood and urine. 1 Thioridazine can be specifically, simply, and reliably measured in plasma and urine by gas chromatography using hexane extraction and prochlorperazine as internal standard; fluorimetry is non-specific. 2 The method can also measure thioridazine ring sulphoxide, and mesoridazine-plus-sulphoridazine (M/S). 3 After single doses plasma sometimes shows M/S in addition to thioridazine itself; it always does so on continued treatment. There is great individual variation in both components, and evidence of changes in metabolism during the early weeks. 4 Urinary excretion may be influenced by pH, but between pH 6.0-7.0 about 1% of the daily dose appears in 24 h urine as the following: free thioridazine in microng quantities, M/S and ring sulphoxide each in mg amounts. 5 Patients attain steady state conditions, although plasma levels rise considerably after each dose and settle again in about 10 h. After chronic treatment is stopped to half-life is at about 30 h. 6 Plasma levels cannot be related to therapeutic response when this is slow, as in schizophrenia, but interpretations are complicated by the production of clinically active metabolites, and by plasma protein binding.


The metabolism, distribution and elimination of chlorphentermine in man. 1 A gas-liquid chromatography procedure for the determination of chlorphentermine (I), N-hydroxychlorphentermine (II) and alpha,alpha-dimethyl-alpha-nitro-beta-(4-chlorophenyl)ethane (IV) in urine has been developed. Also methods are reported to determine conjugated II and the total N-oxidized metabolites of I, i.e. II, conjugated II, alpha,alpha-dimethyl-alpha-nitroso-beta-(4-chlorophenyl)ethane (III) and IV in urine. 2 The synthesis of alpha,alpha-dimethyl-alpha-nitroso-beta-(4-chlorophenyl)ethane (III) and its properties are reported. 3 The kinetics of urinary excretion of I and its metabolic products after the oral administration of I to a human subject on separate occasions have been studied. Under normal conditions of urinary pH, metabolism by N-oxidation was the main elimination route of I; acidifying the urine increased the urinary excretion of unchanged I at the expense of the N-oxidized products. 4 The importance of the N-oxidation metabolic route in the distribution of chlorphentermine (I) in man is discussed.


A repeated dose comparison of dichloralphenazone, flunitrazepam and amylobarbitone sodium on some aspects of sleep and early morning behaviour in normal subjects. 1 Seven normal subjects were given three different hypnotics (flunitrazepam 1 mg, amylobarbitone sodium 100 mg and dichloralphenazone 1300 mg) for four consecutive nights each. 2 All three substances improved subjective assessment of the ease of getting to sleep. Flunitrazepam was rated as better than eithr dichloralphenazone or amylobarbitone sodium in this respect. 3 The perceived quality of induced sleep was not altered by any of the preparations. 4 There was a disturbance of the subjective ratings of getting to sleep following cessation of treatment with dichloralphenazone, giving tentative support to the existence of a 'rebound' effect. 5 Dichloralphenazone produced an impairment in psychomotor performance as measured on a complex reaction time test following four nights medication with the drug.


Localized scleroderma-like lesions after bone marrow transplantation in man. A chronic graft versus host reaction. Localized scleroderma-like skin lesions which developed in two children, from 8 to 10 months after successful bone marrow transplantation for aplastic anaemia, showed histopathological features resembling those of scleroderma. This finding, like the animal models described in the literature, provides additional support for the auto-immune nature of scleroderma.


Proton nuclear magnetic resonance study of the effect of pH on tRNA structure. The low-field 220-MHz proton nuclear magnetic resonance (NMR) spectra of four tRNA molecules, Escherichia coli tRNAPhe, tRNA1Val, and tRNAfMet1, and yeast tRNAPhe, at neutral and mildly acidic pH are compared. We find a net increase in the number of resonances contributing to the -9.9-ppm peak (downfield from sodium 4,4-dimethyl-4-silapentanesulfonate) in three of these tRNAs at pH 6, while tRNAfMet1 does not clearly exhibit this behavior. The increase in intensity at this resonance position is half-completed at pH 6.2 in the case of yeast tRNAPhe. An alteration at the 5'-phosphate terminus is not involved, since removal of the terminal phosphate does not affect the gain in intensity at -9.9 ppm. Based on a survey of the tertiary interactions in the four molecules, assuming that they possess tertiary structures like that of yeast tRNAPhe at neutral pH, we tentatively attribute this altered resonance in E. coli and yeast tRNAPhe to the protonation of the N3 of the adenine residue at position 9 which results in the stabilization of the tertiary triple A23-U12-A9. This intepretation is supported by model studies on the lowfield proton NMR spectrum of AN oligomers at acid pH, which reveal an exchanging proton resonance at -9.4 ppm if the chain length N greater than or equal to 6.


Sodium-dependent methyl 1-thio-beta-D-galactopyranoside transport in membrane vesicles isolated from Salmonella typhimurium. Membrane vesicles isolated from Salmonella typhimurium G-30 grown in the presence of melibiose catalyze methyl 1-thio-beta-D-galactopyranoside (TMG) transport in the presence of sodium or lithium, as shown initially with intact cells by Stock and Roseman (Stock, J., and Roseman, S. (1971), Biochem. Biophys. Res. Commun. 44, 132). TMG-dependent sodium uptake is also observed, but only when a potassium diffusion potential (interior negative) is induced across the vesicle membrane. Cation-dependent TMG accumulation varies with the electrochemical gradient of protons generated as a result of D-lactate oxidation, and the vesicles catalyze D-lactate-dependent sodium efflux in a manner which is consistent with the operation of a proton-sodium exchange mechanism. Although the stoichiometry between sodium and TMG appears to be 1:1 when transport is induced by a potassium diffusion potential, evidence is presented which indicates that the relationship may exceed unity under certain conditions. The results are explained in terms of a model in which TMG-sodium (lithium) symport is driven by an electrochemical gradient of protons which functions to maintain a low intravesicular sodium or lithium concentration through proton--sodium (lithium) antiport.


Determination of the rate-limiting steps and chemical mechanism of fructokinase by isotope exchange, isotope partitioning, and pH studies. Isotope exchange studies show that beef liver fructokinase has a random kinetic mechanism in which release of fructose from the enzyme is slower than that catalytic reaction. The stickiness of fructose in the presence of MgATP is confirmed by isotope partition studies, which show it to be released 0.53 times as fast as V1/Et in the presence, and 80--130 times as fast in the absence of MgATP. Fructose-1-P release from it binary complex is not at all rate limiting in the forward direction since no exchange of MgADP back into MgATP could be observed during the forward reaction. Failure to find any isotope effect by the equilibrium perturbation method with [1-18O]fructose (upper limit, 1.003, shows that P--O bond cleavage or formation is not rate limiting. The pH profiles for the forward reaction show a group (probably carboxyl with pK 5.7-6.0 and deltaHion = 0) that must be ionized and a group (perhaps lysine, with pK 9--10, and deltaHion 5-9 kcal/mol) which must be protonated for activity. The profile for the back reaction shows only a group with pK 5.5--6 that must be protonated for activity. A chemical mechanism is proposed in which a carboxyl group on the enzyme accepts a proton from the 1-hydroxyl of fructose during the forward reaction and donates it back during the reverse reaction.


Carbon-13 nuclear magnetic resonance probe of active-site ionizations in human carbonic anhydrase B. Human carbonic anhydrase B (HCAB), prepared by a new affinity chromatography procedure, was carboxymethylated exclusively at NT of its active-site histidine-200 using 90% [1-13C]bromoacetate. The 13C nuclear magnetic resonance signal of the covalently attached carboxylate was easily detected over the natural abundance background due to the other carbonyl and carboxyl carbons of this 29 000 molecular weight zinc metalloenzyme. Its chemical shift proved very sensitive to the presence of inhibitors in the active site and to variations in pH. Two perturbing groups with pKa values of 6.0 and 9.2 were assigned to the modified histidine-200 itself and the zinc-bound water ligand, respectively, making use of 13C NMR titration data on Nr- and Nr-carboxymethyl-L-histidine model compounds. The results rule out histidine-200 as the critical  group whose ionization controls the catalytic activity. They also strongly suggest an interaction of the carboxylate of the carboxymethyl group with either the zinc or its water ligand around pH 8, possibly explaining the basis for the major differences between HCAB and CmHCAB.


Comparison of solubility properties of alpha-paramyosin, beta-paramyosin and acid-extracted paramyosin. The solubility properties of paramyosin in the zones of pH and ionic strength in which aggregation occurs were initially studied using preparations isolated by a method originally described by Bailey (Bailey, K. (1956), Pubbl. Stn. Zool. Napoli 29, 26). Other preparations yielding apparently different protein components have been described by Hodge (Hodge, A.J. (1952), Proc. Natl. Acad. Sci., U.S.A. 38, 850) using acid conditions, and Stafford and Yphantis (Stafford, W.F., AND Yphantis, D. (1972), Biochem. Biophys. Res. Commun. 49, 848) have identified alpha-, beta-, and gamma-paramyosin using various times and temperatures of extraction with or without ethylenediaminetetraacetic acid. We have found that acid-extracted paramyosin is very similar if not identical to alpha-paramyosin, but that both acid and alpha forms differ considerably from beta- and gamma-paramyosin. Beta-Paramyosin precipitates abruptly from solution in narrow zone of pH below neutrality, and increases in ionic strength shift the zone of precipitation toward lower pH values. In contrast, both acid and alpha-paramyosin show gradual aggregation with changing pH at lowerionic strength (less than 0.3) but sharp transitions similar to beta-paramyosin at higher ionic strength (greater than 0.3). Transitions were also found at lower pH (ca. 4.0) which were not mirror images of transitions at higher pH (ca. 7.0). Viscosity measurements show that acid extracted paramyosin is close in behavior to a native extract obtained by extraction in mild, nondenaturing media containing mixed antibiotics. Each of these extracts differed considerably from beta-paramyosin. Mild, nonhydrolytic procedures employed by others to remove small, noncovalent bonded components or to separate protein complexes were not effective in converting alpha- to beta-paramyosin. Comparison of extraction procedures strongly supports the suggestion of Stafford and Yphantis that beta- and gamma-paramyosin are hydrolytic products of alpha-paramyosin and that the proteases responsible may be of bacterial origin.


Cotransport of phosphate and sodium by yeast. Phosphate uptake by yeast at pH 7.2 is mediated by two mechanisms, one of which has a Km of 30 micronM and is independent of sodium, and a sodium-dependent mechanism with a Km of 0.6 micronM, both Km values with respect to monovalent phosphate. The sodium-dependent mechanism has two sites with affinity for Na+, with affinity constants of 0.04 and 29 mM. Also lithium enhances phosphate uptake; the affinity constants for lithium are 0.3 and 36 mM. Other alkali ions do not stimulate phosphate uptake at pH 7.2. Ribidium has no effect on the stimulation of phosphate uptake by sodium. Phosphate and arsenate enhance sodium uptake at pH 7.2. The Km of this stimulation with regard to monovalent orthophosphate is about equal to that of the sodium-dependent phosphate uptake. The properties of the cation binding sites of the phosphate uptake mechanism and those of the phosphate-dependent cation transport mechanism have been compared. The existence of a separate sodium-phosphate cotransport system is proposed.


Intracellular distribution of ribonuclease activity during erythroid cell development. Five ribonuclease activities, separable by polyacrylamide gel electrophoresis, have been detected in erythroid bone marrow cells from anaemic rabbits. Their intracellular distribution has been investigated and compared with that of the ribonucleases in reticulocytes. Both the acid and alkaline ribonuclease activities of reticulocytes are much lower (30--50 fold) than those of bone marrow erythroid cells. The most marked decrease in enzyme activity occurs in the fractions containing ribosomes and mitochondria plus lysosomes. In these subcellular organelles there was also a qualitative change in the ribonuclease electrophoretic pattern, whereas the cytosol enzymes of marrow erythroid cells and reticulocytes remained largely unchanged. Several ribonucleases released from reticulocyte membranes with urea were similar to those present in the lysosomal plus mitochondrial fraction, as shown by detection of enzyme activity after polyacrylamide gel electrophoresis. The decline in ribonuclease activity was found to begin in the orthochromatic cells, which have a highly condensed nucleus and are no longer active in DNA and RNA synthesis, and to coincide with a decrease in acid phosphatase activity and loss of lysosomes.


The essential activated carboxyl group of inorganic pyrophosphatase. 1. A carboxyl group of high reactivity has been found in inorganic pyrophosphatase (pyrophosphate phosphohydrolase, EC 3.6.1.1) from yeast. This group interacts with agents which react neither with carboxyl groups of low molecular weight compounds nor with other carboxyl groups of the protein. 2. The reaction of this activated carboxyl group with inorganic phosphate, hydroxylamine, N-methyl- and O-methylhydroxylamines, and glycine methyl ester has been studied. 3. Homoserine and homoserine lactone were found in the hydrolyzate of phosphorylated and NaBH4-reduced pyrophosphatase, indicating that an aspartyl residue is phosphorylated. 4. Hydroxylamine and other nucleophilic agents cause inactivation of pyrophosphatase as a result of interaction with a carboxyl group. Both diaminobutyric and diaminopropionic acids were seen in the acid hydrolyzate of the protein treated with hydroxylamine and subjected to rearrangement in the presence of carbodiimide. 5. The ways in which the activation of a carboxyl group in the enzyme is achieved and the presumed mechanism of action of inorganic pyrophosphatase are discussed.


The stereospecificity of nitrate reductase for hydrogen removal from reduced pyridine nucleotides. The stereospecificity of the hydrogen removal from reduced pyridine nucleotides catalyzed by nitrate reductase (NADH : nitrate oxidoreductase, EC 1.6.6.1, and NAD(P)H : nitrate oxidoreductase, EC 1.6.6.2) was investigated. A high degree of enzyme purification was required to obtain conclusive results. Improvements are described for the purification of nitrate reductase from Chlorella fusca and from spinach (Spinacea oleracea, L.) leaves. The latter enzyme is shown to contain a cytochrome. With highly purified nitrate reductase preparations from Cl. fusca, Neurospora crassa, Rhodotorula glutinis and spinach leaves the stereospecificity of the reaction was determined to be predominantly of the A-type in all cases.


Possible regulation of thiamine diphosphatase activity in rat brain microsomes by lipids. The effects of various treatments, which affect membrane structure, on microsomal thiamine diphosphatase and thiamine triphosphatase activities of rat brain, were examined. The treatment of micorosomes at alkaline pH caused a 2-fold activation of the thiamine diphosphatase, this being related to a change in membrane structure which was evidenced by a decrease of the turbidity of the microsomal suspension. Repeated freezing and thawing after hypo-osmotic treatment also increased the activity of microsomal thiamine diphosphatase. In addition, the thiamine diphosphatase activity was enhanced by treatment of the microsomes with phospholipase C or acetone. This lipid depletion resulted in a marked reduction in the apparent Km value of the thiamine diphosphatase with a corresponding loss in heat stability of the enzyme. We found further that brain thiamine diphosphatase was solubilized by Triton X-100. This decreased the phospholipid content in the preparation, but did not affect the apparent Km value and heat stability of the enzyme. In contrast with thiamine diphosphatase, thiamine triphosphatase was inactivated by treatment at alkaline pH or with acetone. However, treatment with phospholipase C did not affect the activity of thiamine triphosphatase.


Inhibition of alkaline phosphatase by sialic acid. The interaction of human organ alkaline phosphatases (orthophosphoric-monoester phosphohydrolases (alkaline optimum), EC 3.1.3.1) with sugars was studied. Hexosamines, N-acetylneuraminic acid (NANA or sialic acid), N-acetylmuramic acid and N-acetylglycolylneuraminic acid inhibited human organ alkaline phosphatase activities. Of these, sialic acid was the most effective inhibitor. The pH profiles for the enzymes in the absence and presence of sialic acid were similar. The sialic acid - enzyme complex was more heat stable than the free enzyme between 20 and 45 degrees C. Lineweaver-Burk plots of 1/v versus 1/S at various concentrations of sialic acid showed intersecting straight lines indicating that the mechanism of inhibition was a mixed type. The Ki value obtained from the plots of 1/v versus the square of sialic acid concentration was 0.07 mM for the hepatic, sialidase-treated hepatic, and intestinal alkaline phosphatases. The respective Hill coefficients varied somewhat with the alkaline phosphatase isoenzyme. Hyperbolic curves were obtained when the percentage of remaining activity was plotted against the substrate concentration at different concentrations of sialic acid. The Hill coefficient was lowered in the presence of sialic acid. The sialidase-treated hepatic enzymes used gave the most effective conversion. Partial denaturation of the enzyme with urea, or pronase digestion had a little if any effect on the sialic acid inhibition with constant time.


Some properties of human liver acid alpha-glucosidase. 1. Albumin activates human liver acid alpha-glucosidase (alpha-D-glucoside hydrolase, EC 3.2.1.20). From the Arrhenius plot, pH-dependence and Lineweaver-Burk plots it can be concluded that this activation is not only due to stabilisation of the enzyme, but also influences the enzymatic activity. It is proposed that for optimal functioning human liver acid alpha-glucosidase needs a protein environment. 2. Glycogen has a competitive inhibitory effect on the hydrolysis of 4-methylumbelliferyl-alpha-D-glucopyranoside, in contrast to maltose which exhibits a non-competitive type of inhibition. It is concluded that two catalytic sites exist, one for glycogen and one for maltose, while both sites influence each other. With glycogen as substrate a break in the Arrhenius plot is found. This is not the case when maltose is used as substrate. 3. The effect of antibody raised against human liver acid alpha-glucosidase on the activity of human liver acid alpha-glucosidase is studied. No corss-reacting material could be demonstrated in the liver of a patient with glycogen storage disease Type II (M. Pompe, acid alpha-glucosidase deficiency).


Lysosomal phospholipase A activities of rat ovarian tissue. 1.1. Lysosome-enriched fractions were prepared by differential centrifugation of homogenates of luteinized rats ovaries. Acid phospholipase A activities were characterized with [U-14C]diacyl-sn-glycero-3-phosphocholine and 1-palmitoyl-2-[9,10-3H]- or [1-14C]oleoyl-sn-glycero-3-phosphocholine as substrates. Acid phospholipase A1 activity had properties similar to other hydrolases of lysosomal origin; subcellular distribution, latency and acidic pH optimum. Acid phospholipase A2 activity with similar characteristics was also tentatively identified. We were unable to exclude the possibility that the combined action of phospholipase A1 and lysophospholipase contributed to the release of acyl moieties from the 2-position of the synthetic substrates. 2. Lysophospholipase activity was present in the lysosome-enriched fractions. This activity had an alkaline pH optimum. 3. Phospholipase A1 and A2 activities solubilized from lysosome fractions by freeze-thawing were inhibited by Ca2+ and slightly activated by EDTA. A Ca2+- stimulated phospholipase A2 activity, with an alkaline pH optimum, remained in the particulate residue of freeze-thawed lysosome preparations. This activity is believed to represent mitochondrial contamination. 4. Activities of acid phospholipase A, as well as other acid hydrolases, increased approx. 1.5-fold between 1 and 4 days following induction of luteinizatin, suggesting a hormonal influence on lysosomal enzyme activities.


Phospholipase A in pulmonary secretions of patients with alveolar proteinosis. This report presents evidence for the presence of phospholipase A2 (EC 3.1.1.4) activity in the insoluble pulmonary secretions of patients with alveolar proteinosis. The enzyme activity has a pH optimum between 7.5 and 8.5 and is stimulated by deoxycholate and Ca2+.


Specific phospholipid requirement for activity of the purified respiratory chain NADH dehydrogenase of Escherichia coli. The highly purified respiratory chain NADH dehydrogenase (EC 1.6.99.3) of Escherichia coli is inactive in the absence of detergent or phospholipid. Triton X-100 is the detergent that gives optimal activity, but the Triton X-100-activated enzyme is stimulated an additional 2-fold by E. coli phospholipids. Phosphatidylglycerol and diphosphatidylglycerol are the most effective lipid activators. The activated complex prepared with diphosphatidylglycerol is stable, whereas that with phosphatidylglycerol loses activity rapidly. Maximum activation by phospholipids occurs after preincubation at 0 degrees C and at pH 7. Triton X-100 is required at low concentrations for lipid activation, but high concentrations interfere with the activation. When the enzyme is optimally activated by phospholipids, it may be additionally activated 2-fold by spermidine, but not by magnesium. In contrast, the Triton X-100-activated form of the enzyme is stimulated by several divalent cations, without specificity. Thus, the most stable, active form of the purified NADH dehydrogenase is generated in the presence of diphosphatidylglycerol and spermidine.


Effect of hydrophobic probes on the higher structure of D-amino acid oxidase. 1. The holoenzyme of D-amino acid oxidase [D-amino acid: O2 oxidoreductase (deaminating), EC 1.4.3.3] was found to combine with 1-anilinonaphthalene-8-sulfonate without liberation of its coenzyme, FAD. No energy transfer interaction was found to occur between the bound dye and FAD of the holoenzyme. On the other hand, when the apoenzyme was bound to the dye and then to FAD, energy transfer interaction between the bound dye and bound FAD was observed. In both cases, the dye competes with the substrate, D-alanine. It is concluded that the dye bound to the holoenzyme is oriented in such a special manner that the mutual orientation factor between the dye and FAD becomes very small in magnitude. 2. When the apoenzyme combined with the dye, the monomer-dimer equilibrium of the apoenzyme shifted towards the dimer. On the other hand, 4-monobenzoylamido-4'-aminostilbene-2,2'-disulfonate combined with the apoenzyme to induce monomerization.


The interaction of histone H3 with histone H4 and with other histones studied by 19F nuclear magnetic resonance. The behaviour, upon variations in ionic strength, pH and temperature of 19F nuclear nuclear magnetic resonance signals of the trifluoroacetonylated derivative of histone H3 is compared with those of the H3-H4 complex and of the Hv fraction (an equimolar mixture of H2A, H2B, H3 and h4). The line width of the 19F-labelled histone H3 signals increases with ionic strength or pH, an effect consistent with aggregation of the protein. In the case of H3-H4 complex or Hv the line width decreases at intermediate ionic strengths (0.1-0.25 M NaCl). This effect is interpreted as the consequence of the formation of a well defined structure with ionic strength. At high salt concentrations the line width increases as a consequence of the final rigid quaternary structure or of the formation of higher aggregates.


The denaturation of beta-lactoglobulin-A at pH 2. The denaturation of beta-lactoglobulin-A by heat and guanidine hydrochloride at pH 2 has been investigated. The effect of ethylene glycol on the thermal denaturation at this pH has also been studied. The conditions of the experiments have been chosen so as to eliminate complications arising out of disulfide interchange, changes in the degree of association of the protein during denaturation, and intermolecular aggregation. The physical parameters characterizing the denatured states of the protein which are produced by heat and guanidine hydrochloride have been determined. The thermodynamic parameters for these transitions have been estimated using a two-state hypothesis in each case. Both the physical and thermodynamic parameters indicate that the heat-denatured state of beta-lactoglobulin-A retains about 15-20% of residual structure which is destroyed on adding guanidine hydrochloride.


[Influence of pH on the steady state kinetics of electron transfer through the cytochrome chain of submitochondrial particles. Kinetic model for regulating the activity of carriers by the local concentration of hydrogen ions in the membrane]. The kinetic parameters of the submitochondrial particles cytochrome chain obtained from steady-state kinetics were studied for pH dependence. The life-times of the activated states (tau) for cytochrome pairs b leads to c1 and a leads to a3 are shown to bear dissimilar dependence on pH of the medium, while for cytochrome pairs c1 leads to c and c leads to a they display practically no pH dependence at all. The rate constants of the non-activated state (alphai-kiCo) decreased for the pair b leads to c1 and increased for a leads to a3 with the increase of pH from 6.5 to 8.5. The apparent pK values obtained therefrom were 7.2 and 8.9, respectively. A kinetic model is proposed suggesting that local pH in the mitochondrial membrane, dependent on the rate of electron transfer, may be a controlling factor for the ratio of activated and non-activated carrier states. The model is in good consistence with the experimental dependences of k'i on V and the pH dependences of alpha2 for b leads to c1 and a leads to a3. It also gives a qualitative prediction for the pH dependences of the ordinate intercepts of the straight lines in l/(k'i--alphai) vs. l/V plots. The rate constants for the diffusion of hydrogen and hydroxyl ions in the membrane are estimated on the basis of our kinetic data to be 10(4)--10(5) s-1 and 10(2)--10(3) s-i, respectively.


[Properties of alpha-chymotrypsin and cis-cinnamoylchymotrypsin in free, covalently bound to a carrier and microcapsulated states]. alpha-Chymotrypsin and its light-sensitive derivative, cis-cinnamoylchymotrypsin, free and immobilysed (by means of microcapsulation, covalent binding with sepharose 4B and p-aminobenzyl cellulose) are studied. Effect of pH on the hydrolysis rate of N-alpha-acetyl-L-tyrosine ethyl ester by different immobilized enzyme preparations, their acylation with isomeric mixture of cis- and trans-cinnamoylimidazole, reactivation of stable cis-derivatives of different enzyme samples and thermal stability of native and microcapsulated enzyme preparations are investigated.


[Role of low-molecular weight factors isolated from Methanobacillus kuzneceovii in the activating effect of visible light on methane formation and reduction of pyridine nucleotides]. Four low molecular weight fluorescent factors are isolated from thermophylic methaneproducing facteria Methanobacterium kuzneceovii. These factors are: the factor 340; the factor 420 (both reduced and oxidized); and a flavine derivative, comprising together with proteins a biochemical system, which reduces pyridine nucleotides in dark (at 55degrees C) and in the light (at 21degrees C).


The effect of birth on the maturation of hepatic cytochrome(s) P-450 mono-oxygenase and tyrosine aminotransferase activities in the guinea pig. Mixed coat guinea pigs delivered by Cesarian section 5 days before term were compared to spontaneously delivered full-term animals with respect to the postnatal maturation of hepatic mono-oxygenase activity in vitro toward aniline and p-chloro-N-methylaniline. Tyrosine aminotransferase acitivity was studied in the same preparations as a positive control for birth-related phenomena. Mono-oxygenase activities toward both sustrates increased significantly in both premature and full-term animals during the first 3 postnatal days and approached adult values 72 h after birth. The maturation of tyrosine aminotransferase activity occured in a qualitatively similar fashion. The mechanism through which birth initiates the maturation of drug oxidative capacity is unresolved; the gestational age at which competence to respond to the event of birth is acquired, remains undefined.


Human cytomegalovirus. A review of developments between 1970 and 1976. I. Clinical developments. Cytomegalovirus (CMV), like other members of the herpesviruses, is widely distributed in human populations where the frequency of seropositive individuals is influenced by such factors as age, sex and socio-economic conditions. While this virus causes such diseases as mononucleosis, it is also implicated in autoimmune phenomena and plays an adverse role renal and bone marrow transplantation. Perhaps the most menacing aspect of CMV is the role which it play in congenital malformations resulting from  in utero or neo-natal infection. Indeed, this virus is one of the few which can be transmitted vertically. The above mentioned clinical aspects of CMV are discussed here in light of recent clinically studies. The second part of this review will be devoted to the great mass of information recently acquired through experimentation.


The effects of adrenergic agonists and blockers on antigen-induced DNA synthesis in vitro. The effects of both alpha and beta adrenergic agonists and blockers on antigen-stimulated DNA synthesis in human lymphocyte cultures were studied. Results show that antigen-induced responses were enhanced by the presence of the beta blockers, propranolol and dichloroisoproterenol, and that this effect appeared to be specific blocking of the lymphocyte beta receptor since D (+) propranolol, a compound devoid of such acitvity, has no effect. Similarly, and alpha adrenergic agonist such as norepinephrine enhanced lymphocyte responsiveness.


[Lactic acid as a  factor in altering vascular reactivity]. It was shown that the constrictor responses to the electrostimulation of the isolated vessel segments were replaced by dilatation when the drugs were contained in the lactic acid solution. The capacity to constrictor effects was not lost completely, but these effects appeared in repsonse to the intensive stimulation. Changes in the character of these reactions to the electrostimulation were connected with the changes in the pH of the solution, as well as with the specific action of lactic acid.


[Graft vs host reaction in reciprocal combinations of mouse strains differing by the H-2 complex of histocompatibility]. Intraperitoneal transplantation of 0.5 x 10(7), 1 x 10(7) or 2 x 10(7) spleen cells from the C57BL mice to newborn CBA recipients induced an acute form or runt disease which resulted in the death of 43%, 86% or 95% of the recipient mice, respectively, in the course of 2--3 weeks after the cell transfer. Preliminary immunization of C57BL donors with CBA isoantigens led to a marked enhancement, and immunization with foreign antigens (sheep red blood cells)--to weakening the reaction. In reverse combination of mouse strains the runt disease was 4--5 times less severe and no "preimmunization effect" occurred. In C57BL leads to CBA combination the reaction was accompanied by proliferation of pyroninophilic mononuclears and follicular destruction, while in the CBA leads to C57BL combination-by the retardation of their growth.


Blood pressure and heart rate and withdrawal of antihypertensive drugs. The immediate effects on heart rate and blood pressure of withdrawing antihypertensive drugs were studied over three-day periods in 26 patients. Four groups of drugs were studied. After withdrawal all patients taking clonidine showed a considerable increase in heart rate and blood pressure with intense ectopic activity. Patients taking postganglionic neurone-blocking drugs showed a similar but less pronounced reaction with increased ventricular ectopic activity. No alarming reactions were seen after withdrawal of methyldopa or beta-blocking drugs. Methyldopa and, especially, beta-blocking drugs are less likely to produce withdrawal reactions than clonidine or the postganglionic neurone-blocking drugs, and patients taking these drugs are therefore less likely to suffer violent reactions if they forget to take their tablets.


[Blood gas measurements and automatization (author's transl)]. The Corning 175, a fully automatic new blood-gas analyzer, was tested and compared with manual analyzers (Eschweiler-Radiometer). The precision was evaluated by means of tonometered bloods and precision gas mixtures. With the Corning 175 four parameters are measured: pH, PCO2, PO2, barometric pressure, and four computed: HCO3-, BE, oxygen content, oxygen saturation. These are automatically corrected for patient hemoglobin and temperature. A microprocessor controls the sequencing of all operations during the calibrations and the measurements and continuously checks the performances of the electrodes and the entire system. The operator part is limited to the introduction of blood. The advantages and the fiability of the automatism in clinical routine are described.


Transmitter histochemistry of the rat olfactory bulb. I. Immunohistochemical localization of monoamine synthesizing enzymes. Support for intrabulbar, periglomerular dopamine neurons. The rat olfactory bulb was studied at the light and electron microscopic level with the indirect immunofluorescence technique and the unlabelled antibody enzyme method (PAP-technique), respectively. Antibodies to all 4 enzymes in the catecholamine synthesis were used. In the principal bulb the first two enzymes, tyrosine hydroxylase (TH) and DOPA decarboxylase (DDC), but not dopamine-beta-hydroxylase (DBH), were present in a proportion of periglomerular cell bodies and dendrites indicating that these neurons synthesize dopamine (DA). This amine may therefore be released as a transmitter substance at some of the intraglomerular dendrodendritic synapses which periglomerular cells form with the mitral cells. There is evidence to suggest that some periglomerular cells use GABA as their transmitter. Thus, a morphologically and physiologically homogenous population of neurons can be subdivided on the basis of transmitter histochemical criteria. There was an impression of more DDC-positive than TH-positive fibers in the glomeruli. Such presumably DDC-positive, but TH-negative processes may represent 5-hydroxytryptamine (5-HT) nerve terminals. DBH-positive fibers were seen in the granular, external plexiform, and very rarely, in the glomerular layers, probably representing noradrenaline (NA) nerve terminals ascending from the lower brain stem. Weakly fluorescent DDC-positive fibers may represent nerve terminals of ascending 5-HT neurons. No phenylethanolamine-N-methyltransferase (PNMT)-positive neurons were observed.


Striatal lesions with kainic acid: neurochemical characteristics. Stereotaxic injection of 2.5 microng of kainic acid, a rigid analogue of glutamate into the rat striatum caused a 70% reduction in the striatum of the cholinergic parameters, choline acetyltransferase, acetylcholine and synaptosomal uptake of choline and a similar reduction in the GABAergic parameters, glutamic acid decarboxylase, psi-aminobutyric acid (GABA) and synaptosomal uptake of GABA. In contrast, the striatal content of dopamine and the synaptosomal uptake of dopamine were unchanged, and the activity of tyrosine hydroxylase was significantly increased. Significant changes in the activity of neurotransmitter synthesizing enzymes were demonstrable within 6h after injection of 2.5 microng of kainic acid and maximal effects occurred at 48h; the activities of choline acetyltransferase and glutamic acid decarboxylase remained depressed up to 21 days after injection. The kinetic characteristics of striatal tyrosine hydroxylase were altered 48h after injection with a two-fold increase in the Vmax for tyrosine and a three-fold reduction in Km for the pteridine cofactor. In contrast to the effects of kainic acid, the injection of copper sulfate, a non-specific toxin, caused a proportionate reduction in the dopaminergic as well as the cholinergic and GABAergic presynaptic markers. The kainate lesion caused an 85% decrement in the activity of dopamine-sensitive adenylate cyclase, a 40% reduction in the specific binding of [3H]quinuclidinyl benzilate and a 195% increase in the specific binding of [3H]GABA in the striatum. The morphology of the kainate injected striatum was markedly altered with nearly a complete loss of intrinsic neurons, increased number of glial cells but intact internal capsule fibers. Intracerebral injection of nanomolar quantities of kainic acid appears to cause degeneration of neurons with cell bodies near the injection site while sparing axons terminating in or passing through the region.


Methylamine dehydrogenase from the obligate methylotroph Methylomonas methylovora. An obligate methyltroph Methylomonas methylovora oxidized methylamine, formaldehyde, and formate. Enzymes oxidizing these substrates were detected in a cell-free system. Phenazine methosulfate-linked methylamine dehydrogenase was purified 21-fold. The enzyme had optimum activity at pH 7.5 and was stable at 60 degrees C for 5 min. The enzyme activity was inhibited by parachloromercuric benzoate, isonicotinic acid hydrazide, mercuric chloride, and sodium borate.


Partial purification and some properties of a neutral sulfhydryl and an acid proteinase from Entamoeba histolytica. The partial purification of two intracellular proteinases from the protozoan parasite Entamoeba histolytica is reported. One of these enzymes is an acid proteinase exhibiting maximum activity at pH 3.5 (hemoglobin substrate), is little affected by a range of inhibitors or activators, and is presumed to be similar to cathepsin D. Also present is a neutral proteinase exhibiting optimum activity at pH 6.0 (azocasein) but only poorly hydrolyzing either hemoglobin or serum albumen. This latter enzyme displayed no metal ion requirement, but was markedly inhibited by thiol-blocking agents and activated by free sulhydryl-containing compounds.


Lawnmower injuries in children.

Lawnmower injuries in children. Over an 8-year period 31 children in the Winnipeg area were admitted to hospital with lawnmower injuries. The accidents were always preventable and were caused by patient or parent carelessness. The wounds were severe, often resulting in amputation of a portion of an extremity. Infection was common; prophylaxis with a broad-spectrum antibiotic and human antitetanus globulin is recommended. Radiography, débridement of the wound with thorough exploration under general anesthesia, and defatting and reapplication of degloved skin should be done initially, then the wound should be reinspected and redressed at 2 to 3 days, and finally closed or skin grafted at 6 to 8 days. Children must be educated about the dangers of lawnmowers and prevented from riding on tractor mowers. The grass catcher should be used and the path cleared of missiles. Hand controls should be mounted on the mower's handle and the blade should be kept sharp so that the machine's speed can be reduced.


An endo-(1 leads to 3)-beta-D-glucanase from Mucor hiemalis. An endo-(1 leads to 3)-beta-D-glucanase (EC 3.2.1.6), isolated from the culture filtrate of the fungus Mucor hiemalis, was purified by ammonium sulfate fractionation, gel filtration, and column chromatography on O-(carboxymethyl)cellulose. The optimum pH, optimum temperature, and Km value of the enzyme were pH 5.0, 50 degrees, and 0.048%, respectively. The enzyme was strongly inactivated by Pb2+, Cu2+, and Hg2+ ions and also inhibited by Zn2+ and Fe3+ ions. The enzyme was specific for laminaran and the action pattern of the enzyme was of the endo-type. The molecular weight of the enzyme, as determined by gel filtration, was 30,000.


Effect of streptovaricin on the incorporation of uridine into cellular and viral RNA. Poxvirus replication is inhibited by streptovaricin. The most readily observed effect is the inhibition of incorporation of [3H]uridine into viral mRNA, suggesting an inhibition of RNA synthesis. Streptovaricin also inhibits the incorporation of [3H]uridine into cellular RNA but not as severely as viral RNA. On the other hand, [3H]uridine incorporation into the RNA of Semliki Forest virus (SFV), which contains a positive strand RNA genome, does not seem to be inhibited by streptovaricin. The inhibitory effect of streptovaricin is completely reversible after removal of the inhibitor. In addition to inhibiting RNA synthesis, streptovaricin also may inhibit the methylation of cellular RNA. Viral RNA is stable in the presence of streptovaricin.


[Control of pituitary secretion of melanotropin in an anuran amphibian by thyrotropin releasing factor (TRH). Study in vitro]. Intermediate lobes from Rana esculenta pituitary glands were continuously superfused for 7 hrs at 28 degrees C with amphibian culture medium. alpha-MSH release was measured by use of a sensitive double antibody radio-immunoassay system. alpha-MSH secretion was inhibited by low temperatures. A large increase in alpha-MSH release was observed when Thyrotropin Releasing Hormone (TRH) at doses ranging from 10(-9) to 10(-7) molar was added to the superfusion medium. Since large amounts of TRH are to be found in the hypothalamus of amphibians but have no effect over pituitary TSH secretion, the action of TRH over alpha-MSH release may have a physiological significance.


[Spontaneous phospholipase A activity of rat pancreatic homogenates]. Rat pancreas presents a spontaneous phospholipase A activity which appears before trypsin activation at optimal pH 6.5. The responsible enzyme is independent of pancreatic prophospholipase A, as can be seen through experiments done in the presence of trypsin inhibitors. On the other hand, this enzyme is distinct from excretory phospholipase which is more active and whose optimal pH is 8.8. Thermostability and insensibility of spontaneously active phospholipase A to DFP differentiate it from lipase, carboxyl-esterhydrolase and lysophospholipase, respectively.


Studies on alkaline phosphatase isoenzymes. Relation to gamma-glutamyltransferase and lactate dehydrogenase isoenzymes. Gamma-Glutamyltransferase (GT) and isoenzymes of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) have been studied in 282 cases with increased S-ALP and in 18 chronic alcoholics with normal routine liver tests. There was a high degree of correlation between S-GT and the bile (alpha 1) and liver (alpha 2) fractions of S-ALP. Fractionation of alkaline phosphatases sometimes yielded clinical information, which could not be obtained by determinations of S-ALP and S-GT only. The presence of alpha 1-ALP and increased S-GT appeared to be more sensitive indicators of ethanol-induced liver involvement than other liver tests, including LDH-5/LDH-4 ratios.


[Incidence of Arbovirus isolated from mosquitoes in the forest region of South Cameroun, region of Yaoundé ]. The authors give results of a survey of arboviruses isolated from mosquitoes in the neighborhood of Yaoundé, Southern Cameroon, from 1964 to 1972. During this period of nine years 127.520 female mosquitoes including at least 72 species were collected and processed for attempt of virus isolation. 55 strains belonging to 12 viral types were recovered from at least 19 species of mosquitoes. The appearance of different viruses over the years was quite intermittent. A possible reason for this is the need for viruses to move in search of susceptible vertebrate hosts. There was not definite evidence of vector host specificity except likely for Anopheles gambiae s. l. as a vector of Tataguine. However, among the more infected species, some ones show by their host preferences they could be true vectors of the viruses isolated from them.


Comparison of human membrane-bound neutral arylamidases from small intestine, lung, kidney, liver and placenta. Human membrane-bound neutral arylamidases were solubilized from small intestinal mucosa, lung, kidney, liver and placenta with trypsin. These five membrane-bound neutral arylamidases were identified by polyacrylamide gel-disc electrophoresis. The heat sensitivity of each enzyme was in the order, liver and placenta greater than kidney greater than lung greater than small intestine. This order correlates with that of electrophoretic mobility, except for the placental membrane-bound neutral arylamidase. Five membrane-bound neutral arylamidases have the same molecular weight, 240 000, as estimated by Sephadex G-200 gel filtration. The five membrane-bound neutral arylamidase have very similar KM values (8.7 x 10(-5) M towards L-alanyl-beta-naphthylamide), optimal pH values, hydrolysis ratios towards L-alanyl-beta-naphthylamide and L-leucyl-beta-naphthylamide, and sensitivities of inhibition by chelators or amino acids. These results suggest that the multiple forms of membrane-bound neutral arylamidase found in five different human organs are organ-specific isoenzymes.


Anti-tumour immunity by normal allogeneic blood transfusion in rat. Growth of transplantable KMT-17 tumour in syngeneic WKA/MK rats was inhibited by i.v. preimmunization with whole blood from normal rats of allogeneic strains. The inhibitory effect was also observed in rats immunized with allogeneic white blood cells alone. The strength of the inhibitory effect mainly depended on the the strain of donor rat used; blood from Donryu and Kyoto strain rats produced the strongest inhibition, blood from Tokyo and Fischer strain rats produced moderate inhibition to the syngeneic tumour growth. Blood from ACI/N strain rats did not produce the inhibition. The mechanism of blood transfusion in inhibiting tumour growth is not yet clear. However, it seems that GVH reaction does not play an important role in the mechanism of the inhibition effect, because the effect was obtained by immunization with mitomycin C-treated allogeneic white blood cells and also by the immunization effect may be due to nonspecific active immunization. Significance of blood transfusion with special reference to clinical immunogtherapy of cancer is discussed.


The immune reponse to type III pneumococcal polysaccharide in mice with malaria. The immune response of BALB/c mice to type III pneumococcal polysaccharide (SIII), as measured by splenic PFC, was abolished at the height of an acute self-limiting attack of malaria caused by the murine plasmodium P. yoelii, over a wide range of antigen doses. The response to antigen, given at various times after clinical recovery, gradually reappeared, but did not reach normal levels until 12 weeks after the injection of the parasite. A second injection of P. yoelii given 1 hr before SIII caused a moderate degree of depression, although in this case the plasmodium does not multiply. In chronic malaria the response to SIII was also very poor. Short term under-nourishment was found to reduce only slightly the response to SIII.


Beta-2 adrenoceptor blocking activity of penbutolol and propranolol at very low doses. The relative potency of penbutolol, a new beta adrenergic receptor blocking agent, was compared with propranolol by a four-point assay on healthy male subjects. A dose-response relationship to intravenous doses of propranolol in the microgram range was obtained during a steady state of infusion of epinephrine. Two subjects underwent the entire assay according to the Latin square design and four others each underwent one schedule of design on different days. The potency of penbutolol thus assessed was consistent with reported results. This study emphasizes the importance of intersubject variation and differential receptor sensitivity in individuals. The use of epinephrine as a beta receptor adrenergic agonist for evaluation of selective beta adrenergic receptor blocking activities is discussed.


Cardiovascular effects of total hip placement in man. With observations on the effects of methylmethacrylate on the isolated rabbit heart. The cardiovascular effects of total hip placement were evaluated in 10 surgical patients, aged 55 to 82, while receiving fluroxene-N2O-O2 anesthesia. The anesthetic regimen caused mild cardiovascular depression. The placement of the acrylic cement into the acetabulum and femoral shaft also induced mild cardiovascular depression, but these changes were not significant at p less than 0.05. In one 67-yr-old woman, there were significant reductions of cardiac output and stroke volume 2 min after the insertion of acrylic into the femoral shaft, despite careful replacement of intravascular loss and careful anesthetic management. Methylmethacrylate (1 X 10(-6) to 1 X 10(-4), v/v) was administered to 24 isolated perfused rabbits hearts. These concentrations of methylmetacrylate are of the same order as measurable blood levels in surgical patients. There was a dose-dependent depression of left ventricular dP/dt correlated with a depression of the spontaneous heart rate. When the bradycardia was prevented by electrically pacing the hearts or the administration of atropine, the depressed dP/dt rose to control levels. Reduction in myocardial temperature and heart rate by means of reduction in perfusate temperature of the isolated hearts reduced the myocardial depressant effect of methylmethacrylate.


Erythrocyte adenosine triphosphate and 2,3-diphosphoglycerate after human renal transplantation: dissociation from hypophosphataemia. 1. Erythrocyte 2,3-diphosphoglycerate (2,3-DPG) and adenosine triphosphate (ATP) were determined in normal individuals, uraemic patients on chronic haemodialysis and patients who underwent renal transplantation, and correlated with plasma phosphate and arterial blood pH. 2. Significant increases in the 2,3-DPG and ATP content were found in the uraemic patients and these persisted after transplantation in spite of marked hypophosphataemia. 3. No correlation was established with plasma phosphate for either of the compounds but 2,3-DPG had a significant correlation with arterial blood pH. 4. Normal values for ATP and 2,3-DPG were observed in post-transplant patients with normal haematological values. The high amounts of erythrocyte 2,3-DPG and ATP in the early post-transplant period are independent of the circulating concentration of inorganic phosphate, and might represent the response of erythrocyte glycolysis to changing arterial blood pH.


A study of the in vitro and in vivo interaction of barium sulphate particles with gastro-intestinal mucus. The in vivo interaction of various radiopaques with mucus was investigated by a determination of the pH-mobility curves of the treated particles. It was found that mucus covered the radiopaque particles both in vivo and in vitro. This interaction was not affected by the nature of the radiopaque or by the state of the mucous membrane. The changed state of the mucus was modified by the diet of the rat from which it was derived. The results are discussed with reference to the technique used in the administration of radiopaques.


In vitro efficacy of some drugs  on promastigotes of Leishmania donovani. Sodium stibogluconate, metronidazole and dehydroemetine Roche were tested for their in vitro action on the mobility, morphology and survival of promastigotes of Leishmania donovani. At low concentrations, sodium stibogluconate, the drug of choice for kala azar treatment in Iraq proved to be poorly  effective in the prevention of promastigotes growth in vitro. On the other hand, both metronidazole and dehydroemetine Roche at low concentrations were highly effective in the prevention of promastigotes of L. donovani growth in vitro. 3.51 X 10(2), 9.34 and 3.13 mmol of sodium stibogluconate, metronidazole and dehydroemetine, respectively, were required to show the same effect in vitro.


[The influence of surgery on serum enzyme changes]. Gamma-GT, which appears as a sensitive indicator in the enzyme pattern during postoperative examination, is especially recommended (besides the GOT and GPT enzymes) in detecting and watching the course of hepatocellular lesions. Distinction between parenchymatous and obstructive icterus can in some cases be made at an early stage by comparison with AP. Distinction between increases in biliary and ossal AP activity is possible with the help of Gamma-GT. Where liver metastases are suspected clinically, increased Gamma-GT activity makes metastization into the liver appear likely.


Spontaneous variability of the hemoglobin affinity for oxygen. In order to evaluate the accuracy of our method of P50 determination, we measured P50 and 2,3-DPG on different days on several members of our laboratory staff and were surprised to find significant variation. Initially, we indicted our methodology but subsequent data suggest that significant P50 variability occurs, not correlated to alterations in 2,3-DPG or red cell pH. The largest variability on different days. 8.5 torr, occurred in a laboratory staff member studied 7 days apart. The P50 determinations were made by fixing PO2 by tonometry and determining oxyhemoglobin saturation by Van Slyke analysis. In contrast to our in vivo data, serial studies over 5-20 days performed upon 3 samples of stored CPD bank blood demonstrated the expected progressive decline in both 2,3-DPG and P50.


Tardive dyskinesia: relationship with a primary affective disorder. We reviewed all Mayo Clinic case histories in which a diagnosis of tardive dyskinesia or dyskinesia might have been recorded during the years 1965 through 1973  and interviewed 18 consecutive patients in the Department of Psychiatry and Psychology. Among the histories and patients, we found a high incidence of primary affective disorders. Four of the five  men had a history of chronic alcohol abuse and symptoms of depression. We recommend that people who have primary affective disorders and chronic alcohol abuse with depression should be given antipsychotic medication, stimulants, or diazepam only after extremely careful consideration.


Oxygen availability during hypothermic cardiopulmonary bypass. Oxygen availability during cardiopulmonary bypass was assessed in 22 patients under hypothermic and relatively normothermic conditions. The patients were divided into two groups, 17 of whom received ACD blood and 5, CPD blood. The mean P50 for all patients fell from a preoperative value of 25.9 +/- 2.4 (SD) to 15.6 +/- 2.1 during hypothermia confirming a leftward shift of the oxyhemoglobin dissociation curve. Oxygen uptake, calculated from a-v oxygen content differences (avDO2) and flow, was significantly lower during hypothermic bypass (65 +/- 27 ml/min) than during rewarming (121 +/- 41 ml/min). The increase in oxygen affinity during hypothermia was influenced also by changes in acid base and 2,3-DPG concentrations, the changes being similar in both the ACD and CPD groups of patients. During rewarming, however, oxygen availability was increased in the CPD group presumably from significantly increased 2,3-DPG concentrations. A "functional" value of hemoglobin, based upon the effects of the shift of the oxyhemoglobin dissociation curve and, therefore, reflecting the true capacity of hemoglobin to unload oxygen at the tissue level, was calculated. During the hypothermic phase of bypass, this functional hemoglobin was only 4.2 g/100 ml blood, suggesting that, in spite of reduced metabolic demands, oxygenation reserves are minimal.


Multi-clinic cross-over comparison of triazolam (Halcion) and placebo in the treatment of co-existing insomnia and anxiety in anxious out-patients. In this multi-clinic double-blind cross-over designed study, forty-five anxious and insomniac out-patients received either triazolam (Halcion) 0.25 mg or placebo at bedtime. After seven days the medications were crossed over. The dosage was doubled after nights 2 and 9 if good sleep was not produced. Three patients dropped out, two on placebo (one for side effects and one for lack of efficacy) and one on triazolam (due to misunderstanding instructions). Analysis of sleep questionnaire data showed superiority of triazolam to placebo (p less .001) in all parameters. The incidence of dreams was not affected. Analysis of antianxiety efficacy showed that triazolam was superior to placebo (p less than .001) in all parameters, both physician's and self ratings. Thus, either triazolam or a good night's sleep significantly relieved the patient's anxiety. No side effects or abnormal laboratory values of significance were attributable to triazolam.


Interactions with antiepileptic drugs.

Interactions with antiepileptic drugs. Most of the currently available antiepileptic drugs have a low therapeutic ratio and therefore a drug interaction causing elevation of the serum level of one of these compounds can readily lead to drug intoxication. Phenytoin, in particular, is vulnerable because its metabolism is dose-related and at therapeutic serum levels the enzyme system involved in its degradation is easily inhibited by concurrent drug administration. As multiple drug therapy has traditionally been practised in the management of epilepsy, clinically important interactions are common. Furthermore, most of the drugs used in the treatment of major epilepsies are potent inducers of hepatic microsomal enzymes and can therefore stimulate the metabolism of concurrently-administered drugs to such an extent that they may be rendered ineffective. The use of one drug alone is recommended, where possible, in the management of epilepsy.


[Sugar studies at Turku].

[Sugar studies at Turku]. This survey concerns the essential findings in two clinical trials. The first study involved almost complete substitution of sucrose (S) by fructose (F) or xylitol (X). After 2 years the mean DMFS-increment was 7.2 in the S-group, 3.8 in the F-group, and 0.0 in the X-group. The second study comprised partial substitution, the effects of a S- or X-containing chewing gum being compared during 1 year. The subjects consumed 4.0 chewing gums per day in the S-group and correspondingly 4.5 in the X-group. In the S-group the caries incidence assessed independently by clinical and radiographical means was significantly higher than in the X-group. It is concluded that the metabolic studies indicate the relative safety of perorally administered xylitol at the present dosage levels. In view on the findings in the chewing gum study it is suggested that the non- and anti-cariogenic properties of xylitol principally depend on its lack of suitability for microbial metabolism and the physico-chemical effects in plaque and saliva brought about through low and repeated dosage.


The effects of dopamine on renin release in vitro. To examine the direct effects of dopamine on renin release, the in vitro rat kidney slice system, devoid of hemodynamic and humoral effects, was chosen. In the presence of an antioxidant, ascorbic acid (6 X 10(-4)M), a significant dose-related stimulation of renin release was observed with addition of 10(-5)M and higher concentrations of dopamine. When the monoamine oxidase inhibitor, pheniprazine (1 X 10(-5)M) was added, significant, dose-related stimulation of renin release was observed with 10(-8)M and higher concentrations of dopamine. Dopamine-induced renin release was not inhibited by the presence of the alpha-adrenergic antagonist, phentolamine (9 X 10(-4)M), the dopaminergic antagonist, haloperidol (5 X 10(-5)M) or the neural uptake inhibitor, cocaine (1 X 10(-5)M). However, the presence of the beta-adrenergic antagonist, propranolol (2 X 10(-4)M) completely inhibited dopamine-induced renin release. These studies indicate that dopamine can directly stimulate renin release in the absence of effects of hemodynamic factors, alterations in sodium metabolism or release of endogenous adrenergic agents. Further, this direct effect of dopamine on renin release appears to be mediated by an agonistic effect on the juxtaglomerular beta receptor rather than by the presence of a specific dopaminergic receptor for renin release.


Diagnosis of bilateral abdominal cryptorchidism by laparoscopy. The authors report a case of abdominal retention of both testes, where the exact diagnosis was made by laparoscopy. They suggest the usefulness of laparoscopy also in urology.


An approach to the toxicology of combustion products of materials. Physiological and behavioral (conditioned avoidance) responses of male Long-Evans rats were determined during exposure to combustion products produced on thermal degradation of three different polymeric materials. Arterial blood samples were obtained for determination of carboxyhemoglobin (COHb) and acid/base status. Material A produced a syndrome of carbon monoxide (CO)-induced anoxia, the severity of which was a function of the mass of material degraded. Material B produced grand mal seizures despite COHb levels of less than 10%. Material C produced metabolic acidosis and a mild degree of CO-induced anoxia. Loss of avoidance responses occurred at significantly lower COHb levels for materials B and C in comparison to CO alone. Using responses to COHb as a reference, it was possible to detect the presence of other toxicants present in combustion products. Compounds found in smoke in very low concentrations may have a high degree of biological activity and be responsible for impairment of survival responses. We have labeled these compounds "limiting" toxicants. They constitute a significant hazard, which is added to that of CO and anoxia.


Development of hepatic acetyl coenzyme A carboxylase in hormone-treated chicks. It has been suggested that the carbohydrate-rich diet of chicks after hatching is responsible for the emergence of hepatic enzymes involved in lipogenesis; the injection of glucose to newly hatched chicks gives rise to an appreciable elvation on the activities of acetyl coenzyme A carboxylase and fatty acid synthetase. The present study shows that during the first hours after hatching, there is a natural elevation of glycemia which parallels the increase in acetyl coenzyme A carboxylase activity. However, the administration of hormones which alter the blood glucose levels considerably (insulin, tolbutamide, glucagon and hydrocortisone) did not influence the enzyme activity. The administration of thyroxine, estradiol and cyclic AMP, was also without effect. These results do not support the theory that the increased amount of blood glucose is the natural effector of the induction acetyl coenzyme A carboxylase. They also show that different lipogenic enzymes are not regulated via the same 'operon' since thyroxine or glucagon which alter the level of some enzymes on this pathway did not modify that of the acetyl coenzyme A carboxylase.


The energy metabolism of the leukocyte. IX. Changes in the concentration of the coenzymes NAD, NADH, NADP, and NADPH in polymorphonuclear leukocytes during phagocytosis of Staphylococcus albus and due to the action of phospholipase C. The determination of the coenzymes NAD+, NADH, NADP+ and NADPH, by the use of a method of enzymatic cycling, demonstrates that the enzymes responsible for the stimulations found during the phagocytosis of Staphylococcus albus are NADH and NADPH oxidase of human leukocytes and NADPH oxidase in the case of guinea pig leukocytes. The effects of serum, of the bacterial strain used and of phospholipase C are also discussed.


Tissue distribution and blood levels of gamma-glutamyl transferase in the horse. In the horse, gamma-glutamyl transferase (GGT) was found to be mainly located in the kidneys, liver and pancreas. As renal lesions are followed by a urinary escape of enzyme, it can be assumed that if there are raised serum enzyme levels then the source will be chiefly from the liver and pancreas. In the blood, GGT can be measured either in plasma or serum. Its mean level in 58 horses was 12 U/L. This level was not affected by moderate dilution or slight haemolysis and its activity was only slightly decreased by storage at--30 degrees C. The relative hepatic specificity of this enzyme and its easy measurement make it a potentially very useful measure of liver dysfunction in the horse.


An isolation procedure for the native alpha chain of bovine hemoglobin. A study of the functional properties of this chain and its hybrid with the human beta chain. In this paper we present a procedure for the isolation of the native bovine alpha chain. The method is based on affinity chromatography. The results show that the ligand-binding properties of the bovine alpha chain are almost identical to those of the human alpha chain. The hybrid alphaB2 betaH2 prepared by mixing bovine alpha chains and human beta chains shows ligand binding properties similar to those of human hemoglobin and different from those of bovine hemoglobin.


Uridine phosphorylase from Escherichia coli. Physical and chemical characterization. Uridine phosphorylase from Escherichia coli has been purified to homogeneity. The enzyme was found to have a molecular weight of 176000 and to consist of 8 probably identical subunits with molecular weights of 22000. These numbers were determined from equilibrium centrifugations in the analytical ultracentrifuge, from dodecylsulphate gel electrophoresis and from amino acid analysis. Moreover the following physico-chemical constants were determined: s020,w = 8.2 x 10(-13) s, upsilon2 = 0.751 cm3/g, A1%280 (1 cm) = 6.73 and a specific activity of 183 units/mg towards uridine. The enzyme shows some activity towards deoxyuridine and thymidine. The activity is not impaired through substitution by bromo, fluoro or methyl groups in the 5-position of the uracil base, but no enzymatic activity is observed when cytosine base is used in the nucleoside substrate.


The exchange reaction of peptides R-D-alanyl-D-alanine with D-[14C]alanine to R-D-alanyl-D-[14C]alanine and D-alanine, catalysed by the membranes of Streptococcus faecalis ATCC 9790. Under alkaline conditions, the membrane-bound DD-carboxypeptidase of Streptococcus faecalis ATCC 9790 catalyses exchange reactions in which the X-L-R3-D-Ala moiety of peptides of the type X-L-R3-D-Ala-D-Ala is transferred to simple amino compounds such as D-alanine, glycine and glycyl-glycine. The enzyme system is unable, however, to catalyse complex reactions that would simulate the natural transpeptidation reaction.


The phosphoenolpyruvate-dependent phosphotransferase system of Staphylococcus aureus. 3. 1H and 31P nuclear-magnetic-resonance studies on the phosphocarrier protein HPr; tyrosine titration and denaturation studies. The phosphocarrier protein HPr has been investigated by proton nuclear magnetic resonance (NMR) at 270 MHz in order to evaluate structural properties of the whole molecule and its active site. The titration behaviour of the three tyrosines of the HPr protein was analysed by monitoring the chemical shifts of the aromatic proton resonances of these residues as a function of pH. It was found that the HPr protein contains a lot of slowly exchanging NH backbone protons which suggested a relatively rigid secondary structure of the protein molecule itself although it contains no disulfide bridges. The HPr protein shows a sharp reversible denaturation behaviour at alkaline pH values. Between pH 10.8 and 11.1 two C-2 proton resonance peaks for the single histidine residue could be observed together with abrupt changes in the aromatic and aliphatic absorption region of the HPr protein which are due to chemical exchange processes. The NMR spectrum of the HPr protein is only changed a little upon raising the temperature from 14 degrees C to 70 degrees C. At 76 degrees C all resonances in the spectrum broaden and almost disappear. This process is irreversible.


Stability in vitro of methylproscillaridin. The in vitro stability of methylproscillaridin has been compared with that of proscillaridin, the activities of the glycosides being assayed by the 86Rb-technique. After incubation in gastric juice at pH 1,2, and 3, the activity half life of each glycoside was proportional to pH and was approximately 0.25 h, 2.5 h, and 25 h, respectively. The inactivation rate in pure hydrochloric acid at pH 2 did not differ from that in gastric juice of the same pH. The glycosides were stable in bile and enteric juice. In faeces, methylproscillaridin was stable and proscillaridin was inactivated with a half life of 32 h. It is concluded that the difference in biological availability between the two glycosides cannot be explained by differences in gastrointestinal stability.


The effect of LSD and 2-bromo LSD on the striatal DOPA accumulation after decarboxylase inhibition in rats. LSD and BOL (0.125-0.5 mg/kg) were equipotent in increasing the in vivo tyrosine hydroxylation in the striatum as measured by the accumulation of DOPA after inhibition of neuronal decarboxylase. However, with 2--4 mg/kg doses, the maximum effect of BOL was larger than that of LSD. LSD and BOL antagonized the apomorphine-induced decrease of DOPA accumulation, without affecting the haloperidol-induced increase. LSD like apomorphine inhibited the increase of DOPA accumulation seen after reserpine, cerebral hemisection and after gamma-butyrolactone (GBL). The effect of apomorphine in rats given GBL was blocked by haloperidol, but not by BOL and promethazine, whereas that of LSD was inhibited by haloperidol, BOL, and promethazine. These findings suggest that LSD and BOL directly affect nigro-neostriatal dopamine neurons. LSD therefore appears to be a partial agonist and BOL a pure antagonist at dopamine autoreceptors. It is proposed in addition that LSD activates and BOL blocks 5-HT receptors that control DOPA formation.


Vascular beta-adrenoceptor stimulating properties of phenylephrine. In pithed rats pretreated with propranolol changes occurred in the slopes of dose--pressor response curves to i.v. phenylephrine and noradrenaline but not to LD 3098, a pure alpha-adrenoceptor stimulant. In dogs, cats, and spontaneously hypertensive rats pretreated with an alpha-adrenoceptor blocker, i.v. phenylephrine induced vasodilatory responses which were antagonised by propranolol. These results indicate that phenylephrine, in addition to its known beta1-agonistic properties, stimulates the vascular beta2-adrenoceptors.


Subpopulations of T-lymphocytes in Peyer's patches: sensitivity to antilymphocyte serum and adult thymectomy. Anti-lymphocyte (ALS) treatment or adult thymectomy of the donor have been shown to depress respectively the cell proliferation and the cytotoxicity in the graft-versus-host (GVH) reaction. A quantitative assay and the histological criteria of the GVH reaction have been used to demonstrate that all the known subpopulations of T-lymphocytes involved in the GVH reaction are present in the Peyer's patches as well as in the spleen and mesenteric lymph nodes in the rat.


Correlation between serum dopamine-beta-hydroxylase activity and dopamine-beta-hydroxylase and tyrosine hydroxylase activities in central and peripheral adrenergic neurons and adrenal glands. Serum dopamine-beta-hydroxylase activity in spontaneously hypertensive rats and Wistar-Kyoto rats had a positive correlation with dopamine-beta-hydroxylase and tyrosine hydroxylase activities in mesenteric vessels, vas deferens, and adrenal glands at 14-16 weeks of age, a negative correlation with dopamine-beta-hydroxylase activity in locus coeruleus at 3 weeks and 14-16 weeks of age, and a positive correlation with tyrosine hydroxylase activity only at 3 weeks of age, but not at 14-16 weeks of age.


Specific metabolic pathway in vitro of pinazepam and diazepam by liver microsomal enzymes of different animal species. The metabolic pathway of Pinazepam and Diazepam in vitro was studied with rat, guinea pig and dog liver microsomes using a chromatographic and spectrophotometric technique. Two main pathways were observed, N1-dealkylation and C3-hydroxylation. N1-dealkylation was shown to be the predominant reaction for Pinazepam in all the animal species studied, while  C3-hydroxylation was the major metabolic pathway for Diazepam in the rat. No oxazepam was found when Pinazepam and Diazepam were incubated with liver microsomes.


[Mechanism of the action of apressin under conditions of high temperature and physical exertion]. In experiments on rats subject to study were regularities governing the metabolism of glycogen, milk, pyruvic acid, phosphocreatine in the liver tissues, cortex, musculus gastrocnemius and in the myocardium, as well as of acids in the peripheral blood at 40 degrees C of outside temperature, 95 per cent humidity and during swimming. Investigations also covered preventive application of apressin in rats. The influence of stress was found to produce metabolic changes in the organism of rats. In hypoxia apressin tended to prolong the life-span of the test animals by as much as 30 per cent, to keep intact metabolism in the musculus gastrocnemius and in the myocardium, but, when used in a dose of 0.05 mg/kg, it failed to avert disorders in the cortex tissues.


[Sulfaphenazole binding by p-450 cytochrome of liver microsomes of rats and its effect of cytochrome-p-450-reductase]. Tests set up "in vitro" on the rats' liver microsomes showed that sulfaphenazol, which in an inhibitor of the mixed type N-and O-demethylation, is bound as a substrate of the II type (aniline type). It attenuates spectral changes caused by aniline and vice versa. Sulfaphenazol inhibits the activity of the ADPH cytochrome-P-450-reductase.


Regulation of gastric acid secretion.

Regulation of gastric acid secretion. The advent of the H2-histamine-receptor antagonists has given new life to the old hypothesis that histamine might be the final common chemical mediator of acid secretion. The available evidence, however, does not prove this hypothesis but does confer on histamine a role in the regulation of acid secretion in normal physiology. Evidence is mounting that, in addition to its stimulatory action, the vagus may play an inhibitory role in acid secretion and gastrin release. Our concepts of the gastric phase of acid secretion have been extended by the discovery of cross distension reflexes in the stomach: the pyloro-oxyntic reflex for acid secretion and the oxyntopyloric reflex for gastrin release. In addition, digested protein has been shown to stimulate directly the oxyntic gland mucosa, but the evidence is against a role for this mechanism in the intact stomach. The hormone(s) responsible for the intestinal phase have not been isolated but the physiological characteristics of entero-oxyntin (a nongastrin, enteric substance that acts on the oxyntic cell) have been defined. Gastric inhibitory polypeptide is an excellent candidate for the entero-gastrone released by fat, but whether it is the sole enterogastrone released is yet to be determined.


Localization of hypophysiotropic neurohormones by assay of sections from various brain areas. The results of studies of the localization of the hypothalamic hypophysiotropic factors based on their direct determination in sections or nuclear punches are described. Luteinizing hormone-releasing hormone was found in high concentrations in the median eminence-arcuate nucleus complex, in lower concentrations in the mediobasal zone of the preoptic area. In addition to these hypothalamic sites, it is present in all four periventricular organs, especially in the organum vasculosum laminae terminalis. Thyrotropin releasing hormone has a widespread distribution. High concentrations are in the median eminence, arcuate nucleus, dorsomedial nucleus, and anterior part of the ventromedial nucleus. Lower concentrations are in several other structures of the hypothalamus, preoptic area and septum, and low but measurable quantities are found in most of the structures of the brain. Somatostatin is also present in most structures of the central nervous system, with highest concentrations in the median eminence, arcuate nucleus, ventromedial nucleus and periventricular nucleus. There are indications that the ventromedial nucleus or its immediate vicinity contains growth hormone releasing factor. Prolactin releasing activity was present in the median eminence and mediobasal parts of the anterior hypothalamus, whereas prolactin inhibitory activity was in the dorsolateral parts of the anterior hypothalamus and/or preoptic area.


Lysis of periadnexal adhesions for correction of infertility. A series of patients is presented in whom lysis of periadnexal adhesions was carried out for correction of infertility. These 35 couples had been infertile for at least 1 year prior to surgery. Seventy-seven per cent had been trying to conceive for more than 18 months. Following diagnostic evaluation, periadnexal adhesions were found to be the sole cause of infertility in 83% of cases. Subsequent to surgery, 63% of the patients conceived, 82% within 18 months, and 57% gave birth to at least one viable child. There can be no doubt that periadnexal adhesions represent true pathology. Although often seemingly insignificant in character, at the macroscopic level, they appear to play a major role at the microscopic level in impairing ovum pickup by the fallopian tube. Gynecologists should recognize the importance of these structures as mediators of a condition of relative sterility. Thus, despite the demonstration of tubal patency, the function of the rather delicate fimbria may be compromised by periadnexal adhesions, conglutinations, and hydatids so that only a fraction of those eggs that leave the ovary at ovulation reach the interior of the fallopian tube.


Double-blind placebo-controlled efficacy study of ketazolam (U-28,774). The safety and efficacy of ketazolam (15 mg capsules) was compared to placebo in seventy-nine out-patients suffering from psychoneurotic anxiety, moderate or worse in severity. A flexible dosage range of 15-75 mg was used in this double-blind study lasting twenty-eight days. The average optimum therapeutic dose of ketazolam was 46-9 mg administered as a once-day dose at bedtime. Ketazolam was found to be significantly better than placebo in alleviating anxiety and its concomitant symptomatology as measured by the Hamilton Anxiety Rating Scale, three Physician's Global Impressions, two Patient's Global Impressions, and three Target Symptoms. Fifteen patients dropped from the placebo group before completion of the study, and two withdrew from the ketazolam group. The patients receiving ketazolam experienced a greater reduction in symptomatology throughout the study when compared to the placebo group. Side-effects experienced by the ketazolam patients were less than, or equal to, the placebo patients. No deleterious side-effects occurred. No differences between the two groups were found for vital signs, EKG's, laboratory tests, or physical examinations.


Hypnotic efficacy of triazolam and methyprylon ininsomniac in-patients. The hypnotic effects of a new triazolobenzodiazepine, triazolam (Halcion) 0-5 mg and methyprylon 300 mg was compared in twenty oncologic in-patient volunteers with insomnia using the preference technique. On the first night of the two-night trial, or methyprylon was given on a double-blind basis and on the second night the patients received the alternate medication. Following each trial night the patients were interviewed in regard to their sleep. Of the seventeen patients who completed the study, eleven patients preferred triazolam, three preferred methyprylon and three had no preference (p=0-057). Analysis of the various sleep parameters showed that triazolam helped the patients sleep more than methyprylon (p=0-13), induced more rapid sleep onset (p=0-003), gave a longer duration of sleep (p=0-013). The treatment was considered a success if the patient went to sleep in thirty minutes or less and slept for at least six hours. Triazolam was more successful than methyprylon in this respect (p=0-012). There were no side-effects reported on either of the drugs.


Intermediate use of triazolam: a sleep laboratory study. The present study was undertaken to evaluate the intermediate effects of triazolam 0-5 mg on the sleep of insomniac patients. The results showed that triazolam is effective in inducing and maintaining sleep without producing any major effects on sleep staging. Triazolam did not lead to significant subjective complaints or any deterioration in the physical condition of the patients. The effectiveness of traizolam did not vary over a two-week period.