Intracellular pH transients in rat diaphragm muscle measured with DMO. Changes of the intracellular pH of rat diaphragm muscle were monitored at 30-min intervals with the weak acid DMO (5,5-dimethyl-2,4-oxazolidinedione). Transferring the muscle from a CO2-containing to a CO2-free solution caused intracellular pH (pHi) to rise by an average of 0.18 during the first 30 min and then to level off at a slightly lower value over the next 60-90 min. Transferring the muscle from a CO2-free to a CO2-containing solution caused pHi to fall by 0.18 during the first 30 min and then to recover by 0.05 over the next 90 min. Subsequent return to the CO2-free solution caused pHi to overshoot the control value by 0.10. Both the recovery and the overshoot can be accounted for by an acid-extruding pump. Intracellular acid loading with 118 mM DMO similarly caused pHi to fall initially, to recover slowly during the acid loading, and then to overshoot the control pHi on removal of the acid load. In the absence of HCO3-/CO2, acid extrusion was reduced by about a fifth. SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid) had no effect. The absence of either Na+ or Cl- from HCO3-/CO2- free solution reduced acid extrusion by about a half.


Infection rates of Aedes triseriatus following ingestion of La Crosse virus by the larvae. Infection rates ranged from 0-2.1% in adults of Aedes triseriaus reared from groups of larvae that had ingested La Crosse (LAC) virus (Clifornia encephalitis group) at dosages of 7.0-8.3 log 10 SMICLD50/ml. Females form orally infected larvae transmitted the virus to suckling mice. Larvae that devoured carcasses of transovarially infected larvae containing 3.0 log 10 SMICLD 50/ml failed to become infected. Ingestion by larvae of infected carcasses appears, therefore, to be unimportant as a method of horizontal amplification of LAC virus.


[Effects of diazepam and flunitrazepam on the undesired postoperative side-effects of ketamine anaesthesia (author's transl)]. 150 patients undergoing minor gynaecological procedures (curretages) were anesthetized with ketamine and either diazepam flunitrazepam or placebo in order to establish the best combination for the prevention of postoperative psychotomimetic effects. From observations, up to 24 h after operation, it was found and statistically proven that the combination of ketamine/flunitrazepam as compared to the combinations ketamine/diazepam and ketamine/placebo led to a remarkable reduction of psychotomimetic side reactions.


[The use of naloxone in neuroleptoanalgesia]. Naloxone was used in 20 patients divided into two series: series A consisted of 10 adults with an average age of 50.6 years (+/- 12.03) and series B 10 children with an average age of 8.5 years (+/- 5.16). Naloxone was given in the treatment of postoperative respiratory depression related to persistence of morphine impregnation, the patients having received either fentanyl (mean dose 0.04 mg/kg/h) or dextromoramide (mean dose 1.15 mg/kg/h). The mean dose of naloxone was 0.26 mg +/- 0.10, i.e. 3.9 microgram/kg in series A, and 0.13 mg +/- 0.11, i.e. 5.3 microgram/kg in series B. In both series, study of ventilatory function showed correction and stabilisation of the various parameters (F/min, Vt and V) up to 180 minutes after the injection. Recovery was rapid in both groups (7 to 10 min) and of good quality. Whilst it was accompanied in a number of cases by the recurrence of pain, the latter never required specific relief. The administration of naloxone was associated with an increase in heart rate (non-significant) at 10 min in series A and 30 min in series B. Apart an episode of nausea in one case of series A, no disagreeable side effects were observed.


[Antagonistic effects of naloxone against morphinomimetic agents. Apropos of 10 cases in gynecologic surgery]. The object of this study was the use of naloxone to correct hypoventilation related to the use of morphinomimetics without suppressing the analgesic effect of these agents. The study involved ten patients undergoing gynaecological surgery under neuroleptanalgesia and who at the end of surgery had hypoventilation due to the use of fentanyl (average dose : 4.87 microgram/kg/h) or phenoperidine (average dose : 48.7 microgram/kg/h). Naloxone was administered intravenously in an average dose of 1.37 microgram/kg (in one or two injections) followed by an intramuscular injection of an average of 0.73 microgram/kg. Under these conditions, respiratory depression was completely corrected in all cases, the effect being durable. Good analgesia was retained and there was a normal return to consciousness without undesirable effects.


[Electrophysiologic aspects of the effects of etomidate in man]. Recordings were electroencephalographical (EEG with spectral analysis), polygraphical (EMG, EKG, breathing). Visual and Auditory Cerebral Evoked Responses were also recorded. The depth of narcosis was quantified upon EEG data. The reactivity during the narcosis and the mechanisms of action of Etomidate on the "non specific structures" are discussed. Cautions should be taken with the use of tomidate in anesthesiology with epileptic and arteriopathic patients.


[Effect of etomidate on cerebral blood flow and oxygen metabolism in man]. The effects of Etomidate on cerebral blood flow (C.B.F.) and cerebral oxygen consumption (C.02 C.) were studied in 13 patients in a neuroradiology department. Induction of anaesthesia was ensured using a standard dose of 15 mg of Etomidate, followed by a constant rate infusion (2.8 mg/kg/hour). Seven subjects were considered to be normal whilst six were suffering from tumour pathology. Under the influence of Etomidate, C.B.F. in the normal subjects decreased by 34 p. 100 and C.02 C. by 46.7 p. 100. The relationship between C.B.F. and arterial PC01 was studied under the influence of Etomidate: cerebral vasoactivity to C02 was maintained. A special study was made of focal changes in flow in patients with tumour pathology. Under the influence of Etomidate, there were zones of adaptation of flow, indicative of vasoparalysis situated opposite the tumour.


[Hemodynamic effects of etomidate].

[Hemodynamic effects of etomidate]. A study of the haemodynamic effects of an injection of 0.3 mg/kg of Etomidate was carried out in 11 multiple trauma patients under constant controlled ventilation. At the 3 rd minute after injection there was a slight, but significant, increase in cardiac output and in the systolic index of the left ventricle, persisting at the 20th minute. It may be concluded that the injection of a dose of 0.3 mg/kg has no harmful haemodynamic effects in this type of patient.


[Constant flow anesthesia using a combination of etomidate and fentanyl]. In 35 orthopaedic surgery patients (33 adults and 2 children), anaesthesia was obtained using an association of etomidate and fentanyl. Induction was obtained by injection from a syringe of a mixture of etomidate and fentanyl in an average dose of 21 mg for etomidate and 0,08 mg for fentanyl. Anaesthesia was maintained by constant dose of etomidate was 1.29 mg/kg/h and 4.96 microgram/kg/h for fentanyl. The mixture used for both induction and maintenance contained 1.3 mg of etomidate and 5 mg of fentanyl per ml. The results, value, indications and contraindications of this technique are described.


[Choice of a local anesthetic in obstetrical analgesia]. A local anaesthetic may be selected from amongst the two major groups of drugs of this type: amino-alcoyl-esters and amino-acylamides. In practice, three substances are available or will shortly be available in France: lignocaine, etidocaine and bupivacaine, thus limiting the choice. This choice is guided by the toxicity of the substance, its clinical effects and 1st side effects. 1-Toxicity depends upon the nature of the substance, the dose necessary to obtain analgesia and to prolong it for a sufficient time. 2-Amongst the clinical effects which should be noted in the choice of a local anaesthetic are the latent period before analgesia is obtained, the degree of sensory block, the degree of motor block and the duration of action. The three substances available are compared in relations to these parameters and in the light of the specific requirements of obstetric analgesia. The value of the addiction of adrenalin is discussed. 3-The side effects are those of the local anesthetic itself and of its possible association with adrenalin. On the basis of the various criteria mentioned, it would seem that the analgesic of choice for obstetric analgesia is bupivacaine used with or without adrenalin.


[Effects of peridural analgesia on labor]. A review of the effects during labour of epidural analgesia on dilatation, expulsion and delivery of the placenta. Uncertainties in knowledge of the precise organisation of uterine innervation and of the intrinsic innervation of the organ render the study of changes in uterine contraction and in cervical resistances difficult. At circulating levels higher than those found in clinical practice, local anaesthetics have a definite oxytocic e-fect, local anaesthetics have a definite oxytoci effect, whilst adrenalin is a powerfult tocolytic. By contrast, the indirect effects related to neurolysis in extradural analgesia are negligeable. Cervico-segmentary resistances are decreased. In practice, there is no increase in the period of dilatation in the absence of any haemodynamic disturbances and if mixtures containing adrenalin are avoided. Expulsion is favourised by muscular relaxation but impaired by the lack of voluntary expulsive effort, hence the choice of a local anaesthetic with the lowest possible motor effect. Delivery of the placenta, which involves only uterine contraction, which is not affected, may be accelerated by oxytocics.


[The use of Marcaine in obstetrical analgesia]. Bupivacaine (Marcaine) is being used increasingly in obstetrics for epidural analgesia, by virtue of the good sensory block obtained and the minimum of side-effects on the mother and newborn infant. At a concentration of less than 0.5 p. 100, analgesia is excellent with a minimal effect on motor fibres. Side-effects and toxicity are limited by the use of fractionated doses from the beginning of labour or of a single dose during or at the end of labour calculated in relation to the effect sought. Any action on the newborn infant would appear to be exceptional, if dose recommendations are respected, the high percentage of Marcaine bound to proteins limiting its transplacental passage.


[Experience of labor and obstetrical analgesia]. A prospective study was undertaken at Strasbourg of the assessment of the patients of the techniques used by the obstetric team and of the general atmosphere in which labour and delivery took place. A questionnair of 115 items was analysed by computer using a Pastis-Pascal programme (see attached). The first results, concerning the analysis of 100 cases and taking into account approximately ten variables, gave the following findings: -the intolerable pain factor is related to the period of dilatation; -the analgesic or amnesic factor in the mother influences the satisfaction she experiences in relation to her child; -demand for epidural anaesthesia appeared the same, whether the woman had been prepared by a psychoprophylactic method or not; -psychoprophylactic preparation would not appear to influence the assessment of pain; -women having an epidural anaesthetic never considered their labour to have been intolerable. Discriminant analysis with study of correlations is not possible at present. Given the number of variables to be considered, the population analysed is inadequate. A multicentre study would improve the performance of this research. Our questionnaire is available to any teams who might wish to use it.


[The organization of an obstetric center in relation to obstetrical analgesia]. In order that continous epidural anaesthesia may be given safely in an obstetric centre, the permanent presence of an anaesthetist is necessary. Obstetricians and midwives must be familiar with the management of labour using this method and team spirit involving continous cooperation between the anaesthetist, obstetrician and midwives is essential. Training related to epidural anaesthesia is necessary  for the members of the obsteric centre, as well as education of the patients.


[Economic and social problems posed by peridural anesthesia and obstetrical analgesia]. It is impossible not to note the small number of epidural anaesthetics given for obstetrics in France. The rarity of centres where it is offered by a team of specially trained anaesthetists and obstetricians contrasts with the increase in demand from the patients themselves. Study of the number of epidural anaesthetics predictable in obstetrics according to the criteria of indication indicates that the procedure could be used in as many as 30 to 50 per cent of deliveries. This implies a need for an increase in the number of anaesthetists, i.e. an increase not so much in the total number as in those trained in obstetrics with experience of epidural anaesthesia. Similar changes are necessary in hospital organisation, in such a way that the place of the anaesthetist in the maternity unit is recognised officially, this recognition being strengthened by routine out-patient consultation with all patients during their pregnancy. The economic aspect of a possible increase in the number of epidural anaesthetics carried out in France and in the number of anaesthetists performing the procedure is envisaged in terms of salaries in the public hospital system, costing of the procedure by the Social Security and the true cost of epidural anaesthesia.


[Study of a new respiratory analeptic: injectable almatrine. Apropos of 22 cases of acute hypoventilation: respiratory activity and electroencephalographic control]. 2620 is a respiratory analeptic which acts by stimulation of peripheral chemoreceptors. It was administered intravenously in 22 patients in acute respiratory insufficiency. Doses during the first hour varied between 0.7 and 3 mg/kg. A careful study of the effects on respiration showed a significant increase in minute ventilation without any proportional increase in frequency for doses of between 1.7 and 3 mg/kg. Continuous recording of the electroencephalogram over a period of one hour revealed no convulsant effect even in particularly vulnerable patients. In seven patients, tracings showed a change in the sense of increased consciousness which corresponded with the clinical course of the diseases responsible for the state of hypoventilation.


[Apropos of complications following the use of methylmethacrylate. Possible prevention by betamethasone]. The aim of this study was to determine whether the prior injection of betamethasone was capable of preventing blood pressure changes and the fall in arterial pO2 when following the fixation of total hip protheses with methyl metacrylate. It involved 54 patients in whom systolic, diastolic and mean blood pressure (BP), arterial pO2, pCO2 and heart rate were noted at different times: T1, 15 minutes after the beginning of the operation; T2 and T3, one minute and five minutes after application of the cement; T4, at the end of the operation. There was no significant difference between the various measurements at times T1, T2, and T3 as far as blood pressure and heart rate were concerned. By contrast, there was a fall in pO2 at times T2 and T2 in relation to times T1 and T4. This fall was significant (p is less than 0.01) but never theless remained moderate. The possible influence of betamethasone is discussed.


[Comparative study of venous contamination with low FiO2 and with pure oxygen in disease-induced pulmonary edema]. A comparative study of the venous contamination (Qs/Qt = (Cc'O2 - CaO2) / (Cc'O2 - C-vO2) under FIO2 = 0.45 - 0.6 and under FIO2 = 1, was realised in 9 patients with lesions of pulmonary oedema. Increasing the FIO2 lead to an increase in the venous contamination three times (average increase was 36 p. 100 of the initial value), to no change two times and a decrease four times (average decrease of 34 p. 100). The values of the venous contamination under average FIO2 are statistically lower in the group where it rises on increasing the FIO2 than in that in which it falls or remains unchanged (p is less than 0.005).


[Biological tests as indirect indicators of the activity of drug metabolizing enzymes. Use of glucaric acid and gamma-glutamyltransferase]. Laboratory tests are increasingly important in the surveillance of drug treatments. People in charge of clinical laboratory science must now become aware of a new development, the use of as indicators of the activity of drug-metabolizing enzymes, especially of the induction of such enzymes. Ideally, the activity of such enzymes should be measured in the tissues. In man, however, easier indirect methods of application are now most often used : measurement of antipyrine half-life, urinary and plasma drug metabolites, protein and specific enzyme levels in palsma, changes in endogenous substrates and endogenous metabolites, and enzymes in circulating blood cells. Of all these, two examinations are now widely performed, those for urinary glucaric acid and gamma-glutamyltrasferase. The value and limitations of these two tests are discussed.


[Gamma-glutamyltransferase. Human pathology and animal biology]. Gamma-glutamyltransferase (GGT, EC 2.3.2.2) is in man and animals mainly a renal enzyme. It is also present in non-negligeable quantities in the liver except in certain rodents such as the rat. In the cells, the cytosoluble fraction of the enzyme is very low and most of the enzyme is linked to the microsomal fraction in vitro. This fraction permits its use in the diagnosis of liver and biliary disease by measurement of its serum activity and that of renal disease by measurement of its urinary activity. In biological research, the prospects offered by GGT both in human and animal cancers, and in pharmacology and toxicology makes it probably one of the most appreciated biological tests in the exploration of liver, biliary and renal function.


[Neutral leucinamidasic activity of human serum. Semiautomatic method of analysis]. Half automated method for determining a L-leucinamide splitting enzymatic activity in human sera. In normal and pathological human sera, two distinct L-leucinamide splitting enzymatic activities have been demonstrated. One of them has an optimal activity at pH 9 (alcaline leucine amidase activity) and shows the most properties of a classic leucinaminopeptidase (E.C. 3.4.11.1). The other (neutral leucine amidase activity) has an optimal activity at pH : 7,5--7,8 and is not activated by Mg2+ ions. In the present work a semi-automated method permitting the determination of the "neutral leucine amidase activity" is presented. The mean of the reference values for normal human sera are established to 31,54 mUI/ml, and the upper normal limit is 48 mUI/ml. The neutral leucine amidase activity is studied in pathological sera comparatively with two other aminopeptidase activities : "alcaline leucine amidase activity", and "leucine-arylamidase". Our study shows that in pathological sera, the neutral leucine amidase activity" varies often without any correlation with those parameters.


Safety of therapy for allergic symptoms during pregnancy. We undertook a literature review to document whether certain therapeutic measures could be considered safe during pregnancy in the allergic patient. With the possible exception of brompheniramine, the commonly used antihistamine drugs appear to be safe during pregnancy. The bronchodilators ephedrine and theophylline also appear to be safe, as does cromolyn. Corticosteroids do not appear to have adverse effects in pregnancy beyond those well recognized in nonpregnant patients. Because side effects are reduced when steroids are administered as aerosols in the nose or lung, these preparations seem well suited for use in pregnancy. The safety of allergic immunotherapy has been confirmed. For asthma, annual influenza vaccination is indicated.


Bilateral central retinal artery occlusions in polyarteritis nodosa. The third reported case of bilateral central retinal artery occlusion associated with polyarteritis nodosa is presented. An alternative etiology of the frequently observed association of polyarteritis nodosa with optic atrophy is suggested. A plan of therapy is recommended.


beta-Blockers and the eye: an overview.

beta-Blockers and the eye: an overview. Some beta-adrenoceptor blocking drugs are exciting new ocular hypotensive compounds. Unlike pilocarpine, the mainstay of glaucoma treatment for the last 100 years, beta-blocking agents do not contract the pupil nor interfere with vision even in patients with central lens opacities. They also do not cause spasm of the ciliary muscle producing transient myopia and disturbance of accommodation. Paradoxically, one of these agents, timolol, causes a fall in intraocular pressure when administered locally even in patients responding to beta-agonists with a reduction in intraocular pressure. Concomitant administration of timolol with epinephrine seems to enhance its ocular hypotensive effect in many patients. The exact mode of action of beta-blockers in reducing intraocular pressure is unknown but appears to primarily reduce aqueous production. This new class of drugs seems destined to play an important role in the treatment as well as in the understanding of glaucoma.


Athlete exertion injuries.

Athlete exertion injuries. Over a period of three years 829 cases of greater than or equal to 16-year-old athlete exertion injuries and syndromes were collected. There were 75 women and 754 men in the series. About 90% of the athletes had been training regularly for more than two years, and 75% of them trained 6 times a week or more. Approximately 52% of the injuries occurred in track and field athletics, about 17% in ball events, 13.6% in skiing, 7.4% in orienteering, and 4.7% in power events. Other sports were associated with fewer exertion injuries. 28.7% of the conditions occurred in the knee, 17% in the ankle, foot and heel, 14.8% in the leg, 8.2% in the back and trunk, 8.1% in the thigh, 7.4% in the achilles tendon. The rest were in the shoulder, neck and upper extremities. In 92% of the patients conservative treatment and rest were used. Only 8% of the cases were treated surgically.


A short review of techniques for the localisation of gamma-glutamyl transferase isoenzymes after electrophoresis. The development of a sensitive method for the localisation of gamma-glutamyl transferase isoenzymes after electrophoresis on various media is described. Recommendations on final conditions are given for optimal concentrations of substrate, acceptor and buffer, and for pH of the reaction mixture. The reaction conditions are compared with other isoenzyme localisation techniques previously reported.


Butanol extraction of serum and urinary gamma-glutamyltransferase and its application in clinical diagnosis. Gamma-glutamyltransferase (gamma-GT) activity was measured in serum and urine before and after extraction with n-butanol. Residual gamma-GT activity after extraction of normal serum with butanol was more than 75%, whereas for normal urine the figure was less than 68%. The loss of activity from urine was not affected by prior dialysis, and the stabilisation which resulted from adding bovine serum albumin at a concentration of 40 g/l did not approach the residual activity with normal serum. In patients with a variety of renal diseases, residual gamma-GT activity in the urine after butanol extraction was inversely correlated with the creatinine clearance. Butanol extraction was performed on serum samples from 182 patients with a variety of diseases. Eighty-one per cent of patients with elevated gamma-GT activities caused by hepatobiliary disease had an increased loss of activity after butanol extraction. By contrast, only 34% of patients with increases gamma-GT activities in whom there was no clinical or other biochemical evidence of hepatic disease, had increased loss of gamma-GT activity after butanol. The reasons for differences between urinary and serum gamma-GT in response to butanol, and the implications in interpreting serum gamma-GT activities are discussed.


Anomalous behaviour of control sera in automated versions of the Kind and King alkaline phosphatase method. Differences in alkaline phosphatase activity of some commerical control sera were observed when comparing manual with AutoAnalyzer versions of the Kind and King procedure. This phenomenon was found to be associated with sera boosted with exogenous alkaline phosphatase and to be owing mainly to inadequate provision of buffer in some automated methods. It is suggested that a modification to the SMA procedure may lessen the problem, but the constraints of the system necessitate a compromise. There was better precision after the modification had been introduced. Awareness of this phenomenon is of paramount importance if commerical sera are to be used in any way in the calibration of automated systems for alkaline phosphatase assay.


A comparison of two methods of quality control of pH, Pco2, and Po2 measurements. pH, Pco2, and Po2 measurements have been made over a three-week period on General Diagnostics Blood GAS Control Ampoules and on a tonometered buffer system. The latter seems adequate as a precision and accuracy control of pH, Pco2, and Po2 measurements in a single solution. The former seems adequate as a precision control of these measurements, but, on the particular batch provided, mean values need to be determined within the customer's laboratory.


A comparison between metoprolol and pindolol in the treatment of essential hypertension. The antihypertensive effects on metoprolol and pindolol were compared in 50 patients with essential hypertension belonging to WHO stage 1 or 2. After four weeks of placebo treatment the patients were randomly allocated to treatment with metoprolol or pindolol. During the initial dose-titration period of six weeks the dose was individually adjusted, aiming at a diastolic blood pressure in the seated position less than 95 mm Hg. The maximum possible dosage for metoprolol was 50, 100 and 150 mg b.i.d. and for pindolol 5, 10 and 15 mg b.i.d. Thereafter followed a fixed-dose period of 20 weeks. Six patients did not complete the study. At the end of the study satisfactory blood pressure control, defined as a diastolic blood pressure in the seated position less than 95 mm Hg, was achieved in 15 of the 20 patients (75%) in the metroprolol group and in 13 out of 24 patients (54%) in the pindolol group. The average reduction in diastolic blood pressure was more pronounced with metoprolol than with pindolol, while no difference was found between the two beta-blockers with regard to the reduction of the systolic blood pressure. Generally, metoprolol and pindolol were both well tolerated by the patients.


Penbutolol, a new non-selective beta-adrenergic blocking compound in the treatment of hypertension. A comparision with propranolol. The antihypertensive effect of a new non-selective beta-adrenergic blocking compound, penbutolol (40 mg b.i.d.), was compared with propranolol (160 mg b.i.d.) in a series of 20 hypertensive patients using a double-blind cross-over scheme. Both compounds reduced both the supine and the standing blood pressure significantly. The magnitude of the responses did not differ significantly and both compounds evoked a commensurate decrease in heart rate. No significant side-effects were noted and a series of laboratory tests did not disclose any biochemical changes.


Response of left ventricular myocardial perfusion and cavity size to beta-blockade by acebutolol. The effect of beta-blockade by acebutolol on global and regional myocardial perfusion (133Xenon wash-out) was studied in 10 patients with coronary artery disease. Another group of 10 similar patients was used to study the effect of acebutolol on left ventricular cavity size (metal markers--spot film camera). Global perfusion responses roughly paralleled the changes in rate-pressure variable which decreased in 8 patients and increased in 2 who had spontaneous angina pectoris. Regional perfusion decreased more in areas distal to less than 75% stenoses than in those distal to less than 75% stenoses (29 vs 12%; p = 0.10 less than 0.20). Left ventricular asynergy did not modify the response, nor did the presence or absence of collateral vessels. No evidence was found to support the thesis that beta-blockade may evoke a redistribution in perfusion which favours the potentially ischaemic areas of myocardium. Left ventricular cavity size remained unchanged after acebutolol, a cardioselective beta-blocking compound with some degree of agonist activity.


Further characterization of bound measles-specific IgG eluted from SSPE brains. Tissue residues from 3 SSPE brains extracted at neutral pH were thoroughly washed with phosphate-buffered saline and further extracted at acidic pH. The low-pH extracts had significant measles antibody titers, and their measles-specific IgG content ranged from 40 to 60% of total IgG. Agarose-gel electrophoretic analysis of the extracts showed oligoclonal IgG bands, most of which disappeared after suitable absorption with measles virus, demonstrating the presence of restricted measles antibody population. Ouchterlony analysis showed a reaction of identity between bound IgG and IgG extracted at neutral pH. Thus IgG extracted at low and neutral pHs is remarkably similar.


Extracellular accumulation of a new amino acid, O-2-hydroxypropylhomoserine, from 1,2-propanediol by flavobacterium rigense. During an investigation of microorganisms utilizing petrochemicals, a strain identified as Flavobacterium rigense was found to accumulate a new amino acid in a medium containing 1,2-propanediol as the sole carbon source. Cultural conditions for the accumulation of the product were investigated, and as a result, the yield was increased to 2.8 mg/ml after a 5-day incubation in a medium containing 8% 1,2-propanediol. The pure amino acid was isolated, and its structure was investigated. Elemental analysis and infrared, nuclear magnetic resonance, and mass spectral analyses indicated that the amino acids is O-2-hydroxypropylhomoserine.


Viral aggregation: quantitation and kinetics of the aggregation of poliovirus and reovirus. The aggregation of poliovirus and reovirus was followed in buffers at various pH values by means of a single particle analysis (SPA) test. The SPA test used here was modified from the original test reported earlier to prevent disaggregation of virus clumps from invalidating the results. The modified SPA test demonstrated that the efficiency of aggregation, which is a measure of the percentage of collisions which are effective in producing an aggregate, may vary widely depending on the conditions in which the virus is placed. The modified SPA test was also used to demonstrate that the kinetic features of viral aggregation follow the classical laws of colloid particle aggregation, which in turn are solely dependent upon diffusion of the particles as caused by brownian motion.


Viral aggregation: effects of salts on the aggregation of poliovirus and reovirus at low pH. As a first step toward the understanding of virus particle interactions in water, we have used the modified single particle analysis test to follow the aggregation of poliovirus and reovirus as induced by low pH in suspensions containing varying amounts of dissolved salts. Salts composed of mono-, di-, and trivalent cations and mono- and divalent anions were tested for their ability to reduce or increase the aggregation of these viruses in relation to that obtained by low pH alone. Mono- and divalent cations in concentrations covering those in natural waters were generally found to cause a decrease in aggregation, with the divalent cations having a much greater effectiveness than the monovalent cations. Trivalent ions (Al3+), in micromolar concentrations, were found to cause aggregation over that at low pH alone. Anions, whether monovalent or divalent, had little ability to produce inhibition of viral aggregation, and thus the overall effects were due almost exclusively to the cation. This was true regardless of whether the overall charge on the virus particle was positive or negative, as determined by the relation between the isoelectric point and the pH at which the tests were carried out. Thus, whereas virus particles conform to classical colloid theory in many respects, there are specific exceptions which must be taken into account in the design of any experiment in which viral aggregation is a factor.


Simple method for the isolation of astaxanthin from the basidiomycetous yeast Phaffia rhodozyma. A method is described for the quantitative and, possibly, large-scale extraction of astaxanthin from the yeast Phaffia rhodozyma. The method utilizes extracellular enzymes produced by the bacterium Bacillus circulans WL-12, which partially digests the yeast cell wall and renders the carotenoid pigments extractable by acetone or ethanol. Complete recovery of astaxanthin from heat-killed P. rhodozyma cells was obtained after growing B. circulans WL-12 on these yeast cells for 26 h and then extracting the yeast-bacterium mixture with acetone. A bacteria-free lytic system, which gave quantitative extraction of astaxanthin from P. rhodozyma, was obtained by concentrating the culture broth from the growth of B. circulans WL-12 on P. rhodozyma cells. Hydrolytic enzyme activities detected in this concentrate included beta-(1 leads to 3)-glucanase, beta-(1 leads to 6)-glucanase, alpha-(1 leads to 3)-glucanase, xylanase, and chitinase. The lytic system was found to work most efficiently at pH 6.5 and with low concentrations of yeast.


Sulfide oxidation by spheroplasts of Thiobacillus ferrooxidans. Thiobacillus ferrooxidans is an acidophilic organism important to metal leaching of low-grade ores. The aforementioned importance is related to the ability of the bacterium to oxidize reduced iron and sulfur, principally found in nature as pyrite (FeS2). The present study dealt with sulfide oxidation at low pH values and the involvement of the cell envelope in the process of the inorganic oxidations. Sulfide oxidation was noted in spheroplasts of T. ferrooxidans prepared by enzymatic and chemical treatments and partially purified by differential centrifugation. No enzyme activities were noted in membrane fractions containing enrichments of lipopolysaccharide symbolic of outer membrane material or in membrane vesicles containing (or associated with) higher levels of proteins. Results to date indicate that in an acid milieu the envelope structure containing both the outer membrane and the intact inner cytoplasmic membrane is required for sulfide oxidation.


Induction of colonial growth and replica plating of the white rot basidiomycete Phanaerochaete chrysosporium. A medium containing L-sorbose and sodium deoxycholate was devised which induces colonial growth of Phanaerochaete chrysosporium in the presence of a variety of supplements. The restricted size and heavy conidial production of the colonies permits high plating densities and the use of a replica-plating technique for copying the pattern of colony growth.


Preliminary data on antiserotonin effects of oxatomide, a novel anti-allergic compound. Oxatomide or R 35443, 1-(3-[4-(diphenylmethyl)-1-piperazinyl]propyl)-1,3-dihydro--2H-benzimidazol-2-one, a compound with potent antihistaminic and anti-anaphylactic activities, shows specific antiserotonin effects on isolated caudal arteries of the rat. In vivo it is a powerful antagonist of bronchospasm induced by serotonin and by histamine; it has no anticholinergic effects and does not affect blood pressure in spontaneously hypertensive rats. These antiserotonin properties of oxatomide may contribute significantly to its anti-allergic activity.


Discriminable effects of antihistamine drugs. Rats were trained to discriminate drug vs. no-drug in a shock-escape T-maze task; a right turn was required when rats were drugged and a left turn when undrugged. The three antihistamine drugs pyrilamine, dimenhydrinate (Dramamine) and diphenhydramine (Benadryl) were discriminated after an average of 20 training sessions indicating that their effects were only moderately discriminable. After criterion performance was achieved under the training conditions, substitution tests were conducted during which rats received a novel drug and were then run in the maze with both goals accessible. Rats trained with antihistamine drugs made drug choices during tests with other antihistamines. Rats trained with other types of drugs made no-drug choices during tests with antihistamines. The results suggest that the antihistamines share a discriminable effect which is relatively unique to that class of drugs.


Clobazam. A 1.5 benzodiazepine derivative: effects upon human psychomotor performance under different levels of task reinforcement. Two dose levels of clobazam, a benzodiazepine derivative, were compared to placebo for their effects upon psychomotor performance in an acute dose daytime study with normals. An acute dose of 10 mg clobazam significantly reduced response latencies in the low reinforcement condition of a psychomotor performance task but not in the high reinforcement condition. However 20 mg clobazam did not produce any significant changes in performance under either low or high reinforcement. Response speed changes generally correlated positively with trait anziety and neuroticism scores at both doses of clobazam.


Studies on the inhibition by beta adrenergic receptor agonists of cyclic AMP phosphodiesterase activity of rat heart. Isoproterenol (10(-5) and 10(-4)M) inhibited a low affinity but not a high affinity form of rat heart cyclic AMP phosphodiesterase. The concentrations of isoproterenol required to produce inhibition of the isolated enzyme were 10,000 to 100,000 fold larger than those required to produce a positive chronotropic response in the isolated atria. Another beta adrenergic receptor agonist, soterenol, had no effect on any of the isolated forms of the enzyme. Theophylline produced inhibition of low and high affinity forms of phosphodiesterase at the same concentrations required to produce a positive chronotropic response in the isolated atria. Results from two experimental models failed to reveal any circumstances under which a contribution to the positive chronotropic response could result from isoproterenol-induced inhibition of cyclic AMP phosphodiesterase.


The role of vanadium in green plants. IV. Influence on the formation of delta-aminolevulinic acid in Chlorella. In a series of experiments, it is demonstrated that the trace element vanadium (4.10(-7) g-at/1 as NH4VO3) has a considerable positive influence on the synthesis of delta-aminolevulinic acid(delta-ALA) in the autotrophically growing green alga Chlorella pyrenoidosa, the effect being visible by an enhanced output of the amino acid into the culture medium in presence of levulinic acid (LA). The level of intracellularly accumulated delta-ALA, however, is not changed in presence of the metal. The V-effect on exogenous found delta-ALA is suppressed, when LA is added to the nutrient medium at low pH (pH5), although V-uptake into the algal cells is not disturbed by LA. As demonstrated in culture media with various nitrogen sources (urea, partially hydrolized urea, ammonium salts), the development of the pH during the cultivation time is important for the presentation of the V-effect on delta-ALA. It is suggested that vanadium acts as a catalyst in the conversion of 4,5-dioxovaleric acid to delta-ALA by transamination.


[Endocrine data in cryptorchism].

[Endocrine data in cryptorchism]. Endocrine evaluation with LH-RH (0.1 mg/m2) and chorionic gonadotrophin (HCG 3 X 1,500 I.U.) in 154 cryptorchid boys aged 1 month to 15 years showed a decrease of LH pituitary secretion and Leydig-cells response to HCG in prepubertal and early pubertal patients. These deficiencies were positively correlated. Partial and at least transient descent of cryptorchid testis or testes has been obtained in 87 of 265 patients treated with HCG (3 to 9 X 1,500 I.U.). Plasma testosterone after HCG 3 X 1,500 I.U. was less increased in patients whose cryptorchid testis or testes descended after 9 X 1,500 I.U. than in those whose testes remained undescended. These data suggest that a partial, early and transient deficiency of pituitary LH secretion may be responsible for testicular maldescent in part of cryptorchid boys.


Standard and long-acting depot neuroleptics in chronic schizophrenics: an 18-month open multicentric study. The overall objective of this 18-month open study was to compare standard neuroleptics and long-acting depot neuroleptics following the current psychiatric practice in order to determine the best therapy. Thirty French psychiatrists from 15 different wards participated in this experiment. One hundred eighty-one chronic schizophrenic patients were randomly assigned to receive one of the following three treatments: standard neuroleptics, pipotiazine palmitate, or fluphenazine decanoate. Criteria used for evaluation were an overall clinical evaluation by a psychiatrist, a Brief Psychiatric Rating Scale, and a Nurse's Observation Scale for Inpatient Evaluation. No significant difference (P greater than .05) was observed between the three groups in drug effectiveness or tolerance.


Vestibular responses in schizophrenia.

Vestibular responses in schizophrenia. In a study of vestibular responses to caloric stimulation that controlled opportunity for fixation and state of alertness, we evaluated previous findings of diminished nystagmus in schizophrenia. We failed to replicate earlier reports in these respects: (1) None of the psychotic patient groups, when compared with normal controls, showed lower response intensity, latency, or culmination time of the nystagmic response. (2) The schizophrenic groups did not manifest a prevalence of clinically significant asymmetry. We did, however, observe that chronic deteriorated schizophrenics and recent schizophrenics have significantly greater dysrhythmic responses. This diminished orderliness of nystagmus may explain previous reports of absent or diminished nystagmus in the schizophrenics. The results are not compatible with peripheral vestibular disease in schizophrenia, but they may reflect state-related phenomena consistent with disturbances in alertness, which are not necessarily voluntary or motivational in origin.


[Proteolytic activities in mature human placenta. II. Neutral peptidases and PZ-peptidase (author's transl)]. Continuing our contributions on proteases of human placenta we enriched some unidentified neutral peptidases by chromatographying freeze dried placenta extracts on CM-Sephadex, followed by preparative isoelectric focusing and chromatography on Ultrogel. By this procedure two Acetyl-aminoacyl-p-nitranilidases, three Ala-Ala-p-nitranilidases, one Benzyloxycarbonyl-Ala-Ala-p-nitranilidase, one Carbobenzoxy-Gly-Pro-Arg-p-nitranilidase and one PZ-peptidase could be characterized in greater detail. Here molecular weights, isoelectric points and pH-optima of these enzymes are reported.


Concept of paraneurons.

Concept of paraneurons. Paraneurons are those cells which have not been designated as neurons but are recognized as closely related to neurons on the basis of their fine structure (possession of neurosecretion-like and synaptic vesicle-like granules/vacuoles), metabolism (production of neurosecretion- and neurotransmitter-like substances) and origin (evidenced or proposed neuroectodermal). Some paraneurons are mainly endocrine in function, while others are sensory. Some should be called interneurons. There are graduations between these types of paraneurons; also a definite boundary between neurons and paraneurons can not be determined. Different types of paraneurons (together with some related neurons) are shown in Figure 1. Comments and discussions were given on the neuron-like characteristics of the individual paraneurons.


Neuro-epithelial bodies in the lung of the rat and the mouse. Histochemical and ultrastructural examinations were performed on neuroepithelial bodies (N. E. B.) in the lung of the mouse and rat. The N. E. B. were identified as specialized groups of pale, columnar cells. They were located throughout the intrapulmonary airway. These cells displayed some special cytochemical properties also seen in the APUD (Amine Precursors Uptake and Decarboxylation) endocrine system, such as cytoplasmic argyrophilia and the capability of selective uptake of amine precursors. Ultrastructrally the N. E. B. were composed of a kind of granulated cell which had concentrations of specific cored vesicles in the basal cytoplasm. It seemed likely that the cells released the contents of the cored vesicles into the extracellular space by exocytosis. These cells made extensive and intimate contact with the mitochondria-rich nerve terminals which occurred mainly at the level of the supranuclear portions of the granulated cells. In this contact area, the membrane of the nerve terminals were apposed to that of the granulated cells with a consistent gap of about 20 nm. Among these membrane appositions, some membrane thickenings were observed in which cored vesicles were closely associated in the granulated cell cytoplasm. From these observations, it was conceived that the lung N. E. B. function both as receptor and endocrine organs and that their specific cored vesicles may have an intimate correlation with these dual functions.


Cimetidine, carbenoxolone sodium, and antacids for the prevention of experimental stress ulcers. Cimetidine, an H2-receptor, antagonist, and carbenoxolone sodium, a drug used to stimulate gastric mucus synthesis and increase the life span of gastric epithelial cells, were studied to determine their possible prophylactic effect on the formation of stress ulcers, gastric transmucosal potential difference and  gastric pH in rats, using a restraint stress model. The number of ulcers substantially decreased in the rat groups receiving either cimetidine or carbenoxolone pretreatment for two or more days before stress. When the drugs were used in combination, there was an additive effect, since there was a significant decrease in ulcer formation even after half a day of pretreatment. When antacids were used, there was also decreased ulcer formation with half a day of pretreatment. Gastric pH was significantly increased from control levels in all the groups treated with antacid, cimetidine and carbenoxolone. Both cimetidine and carbenoxolone, either alone or in combination, prevented the decrease in potential difference that was produced in the untreated stressed rats. This study suggests that either cimetidine or carbenoxolone, and particularly their combination, may be a useful prophylactic regimen for the prevention of stress ulceration in patients subjected to severe stress.


[The effect of pelleting cereal straw with NaOH-addition on rumen fermentation in cows]. Experiments with rumen-fistulated dairy cows were performed to study the effect of NaOH-addition to cereal pellets. Compared to straw-concentrate rations in which the straw pellets were free of NaOH, cereal straw pellets containing 2.4 kg NaOH per 100 kg straw caused the rumenal pH to go up from 6.21 to 6.47, the cellulotlytic activity to rise by some 20%, and the passage turnover rate of dry matter to increase from 0,039 to 0,056. The concentrations of volatile fatty acids and the fermentation patterns were not influenced by the NaOH-content of the pellets. It is concluded that NaOH-addition during straw pelleting has a favourable effect on digestion in the bovine rumen.


[On the toxicology of carbromal. III. Role of active metabolites in humans acutely poisoned with carbromal-containing sedatives (author's transl)]. Carbromal is metabolized extensively in humans. The major metabolites known to date are bromoethylbutyramide, ethylbutyrylurea and inorganic bromide. After ingestion of a therapeutic dose of 1.0 g carbromal (4.2 mmoles) by four healthy volunteers highest concentrations in serum were found to be for carbromal 30 mumoles/l, for bromoethylbutyramide up to 20 mumoles/l and for ethylbutyrylurea 2--3 mumoles/l. In patients acutely poisoned by carbromal-containing sedatives serum concentrations measured were in the range of 200 mumoles/l carbromal, 350 mumoles/l bromoethylbutyramide and 50 mumoles/l ethylbutyrylurea. These patients were comatose, apneic, had isoelectric encephalographic records and decreased body temperature. The degree of central nervous depression as judged by clinical signs was found to correlate with the serum concentrations of carbromal and of bromoethylbutyramide. Pharmacological activity and acute toxicity of carbromal and its two metabolites were examined in rats and compared with the activity of phenobarbitone. For intraperitoneal injection LD-50 values were found to be for carbromal 1.8 mmoles/kg, for bromoethylbutyramide 1.5 mmoles/kg, for ethylbutyrylurea 5.0 mmoles/kg and for phenobarbitone 0.9 mmoles/kg. Carbromal and bromoethylbutyramide severely decreased body temperature. The relative narcotic activity was estimated to be for carbromal = 100; bromoethylbutyramide = 66; ethylbutyrylurea = 33; phenobarbitone = 100. The anticonvulsive activity against pentetrazol-induced generalized seizures was nearly identical for carbromal, bromoethylbutyramide and phenobarbitone. Anticonvulsant activity of ethylbutyrylurea was two to three times less than that of carbromal. Inorganic bromide was found to increase the narcotic activity of carbromal and of bromoethylbutyramide. The findings show that the clinical signs of central nervous system depression seen in patients acutely poisoned with carbromal are caused mainly by unchanged carbromal and by its metabolite bromoethylbutyramide.


Hepatotoxic effect of thioacetamide (TAM) on NADP-linked enzymes, aminotransferases and glutamate dehydrogenase. NADP-linked dehydrogenases, glucose-6-P dehydrogenase (G 6PDH) 6-P gluconate dehydrogenase (6 PGDH), isocitrate dehydrogenase (ICDH), malate dehydrogenase decarboxylating (ME) and aminotransferases GOT and GPT were analyzed in the soluble fraction of blood free homogenates. Glutmate dehydrogenase (GDH) was assayed in the mitochondrial fraction. TAM was i.p. administered to male albino rats (50 mg/kg/day) for 28 days. enzyme activities were determined as described by Bermeyer 1965 (Methods Enzymatic Analysis. Verlag Chemie. Acad. Press).


Changes in liver tyrosine aminotransferase after acute and chronic administration of morphine in the rat. Acute administration of Morphine (20 mg/kg/s.c.) in the rat results in a rise of liver tyrosine aminotransferase (TAT) expressed as mumoles of p-hydroxyphenylpyruvate/100 mg/h. With chronic administration, a tolerance develops to this enzymatic effect. TAT induction is not evident in pregnant rats, given the narcotic, in which enzyme levels are already initially high. After delivery TAT returns to normal levels and it is possible to show both induction and tolerance developing to morphine. Enzyme activity in fetal livers is much lower than that of adult animals: after maternal administration of morphine only a modest TAT increase is seen which is not, however, statistically significant. TAT activity is fully evident in livers of offspring, with much higher mean levels in newborn rats from morphine-treated animals, as a possible consequence of morphine deprivation. In this latter group of newborn rats narcotic administration causes TAT activity to return to levels as high as those of naive animals. On the other hand, morphine administration to the prole of naive rats results in an induction of liver TAT.


[Morphometric and electron microscopic study of the small intestine following total orthotopical auto- and allotransplantation]. Total orthotopic autotransplantation of the small intestine was performed in 22 dogs, allotransplantation--in 13 dogs. Changes in the wall of the graft resulted from the effect of both nonspecific (ischaemia, blockage of lymph outflow, denervation) and specific factors connected with the reaction of tissue incompatibility. Changes caused by the nonspecific factors are demonstrated as lymphostasis, edema of the wall in the graft, atrophy of the villi with epithelial flattening, hyperplasy of the intestinal gland, macrophagal and plasmocytic reaction in the mucous lamina propria, drop in villous-cryptic coefficient and reduction of the absorbing surface of the villi. At autotransplantation, structural-functional changes in the intestine undergo three stages: the stage of acute morphological changes and pronounced functional disturbances (up to 1,5 months); the stage of relative structural stabilization and gradual functional restoration (up to 6 months); the stage of atrophic-hyperplastic changes in the wall and decrease in functional activity of the intestinal graft (followed-up for 15 months). At allotransplantation, depending on the degree of cellular reaction and vascular changes, as well as on the localization of the process in the wall of the graft, a "graft versus host" reaction and a "recipient versus graft" reaction is demonstrated. Peculiar rosettes of plasmocytes have been described as a possible index of the reaction "graft versus host".


[Morphological characteristics of acute respiratory tract diseases of mixed etiology in children]. The analysis of 30 etiologically deciphered (by the direct Coons method) autopsy observations of acute respiratory diseases in children under 8 is presented. Monoinfections were established only in 6 cases. Morphological examinations of respiratory organs in 24 cases of mixed acute respiratory diseases (associations of virus-bacteria and virus-Mycoplasma pneumoniae-diplostreptococcus) revealed marked circulatory disorders and desquamative, dystrophic, and inflammatory processes in segmental bronchi, bronchioles and alveoles. Morphometric studies demonstrated statistically significant differences in the cell composition of infiltrates of interalveolar septa in monoinfections and associated infections.


The effect of cryptorchidism on the quantitative histology, histochemistry and hydrolytic enzyme activity of the rat testis. Cryptorchidism of the mature rat testis led to degeneration of the seminiferous tubules and changes in enzyme patterns and activities. Spermatogenic stages 1-4, containing pachytene primary spermatocytes in late meiotic prophase, and stage 5, containing recently formed round spermatids, were damaged by 48 h. Within 96 h stages showed a loss of germinal cells into the lumen and this was almost complete by 192 h. Acid phosphatase showed increased histochemical activity in the basal area of the seminiferous tubule up to 96 h of cryptorchidism, and at 192 h much of the activity was located in large lipidcontaining bodies within the remaining seminiferous epithelium. Total and free biochemical acid phosphatase decreased during cryptorchidism in parallel with cell loss; there were no significant changes in total cathepsin D activity but free enzyme activity was increased throughout the experimental period indicating increased lability of lysosomes in the Sertoli cell. Lactate dehydrogenase activity was mainly tubular but  succinate dehydrogenase also showed interstitial activity. Lipoamide dehydrogenase (NADH) was found mainly in the interstitium. During cryptorchidism both lactate and succinate dehydrogenase activity decreased in the tubules parallel to the loss of germinal cells, whereas lipoamide dehydrogenase (NADH) activity increased in both interstitial and tubular areas. It is suggested that the initial lesion in the seminiferous epithelium, produced by cryptorchidism is in the Sertoli cell and that germ cell damage may result from reduced function of the Sertoli cell.


Rudimentary locus of Drosophila melanogaster: partial purification of a carbamylphosphate synthase--aspartate transcarbamylase--dihydroorotase complex. Glutamine-dependent CPSase, ATCase, and DHOase from Drosophila, the first three enzymes in pyrimidine biosynthesis, show coordinate variation in activity throughout development. The three activities were highest in first instar larvae and decreased as development proceeded. The three activities cosediment in sucrose gradients as a single peak with a relative sedimentation coefficient of approximately 30S. CPSase, ATCase, and DHOase copurify during (NH4)2SO4 fractionation and during DEAE-cellulose and hydroxylapatite chromatography.


Genetics and physiological expression of beta-hydroxy acid dehydrogenase in Drosophila. A mutant Hadnl was induced in Drosophila melanogaster and found to be deficient in beta-hydroxy acid dehydrogenase. This mutation was utilized to study the genetics and physiological expression of Had+ . Had+ was mapped to the X chromosome at 54.4 and seems to be the structural gene for the enzyme. Enzyme activity in male and female flies indicates that the gene shows both dosage compensation independent from dose effect and differential activity during ontogeny. Electrophoretic mobility data indicate that the enzyme is a dimer which forms by random association of subunits. The fact that the mutant shows no detrimental effect implies that the enzyme is dispensable, at least under laboratory conditions. The biological and technical implications of this gene--enzyme system are discussed.


Pharmacokinetics of fazadinium in man.

Pharmacokinetics of fazadinium in man. The disposition of fazadinium was studied in 10 patients with normal hepatic and renal function undergoing abdominal surgery. The serum and urinary concentrations were determined fluorimetrically. A single dose of fazadinium 1.5 mg kg-1 was administered i.v. The plasma decay curve for fazadinium was found to be diphasic and the data were interpreted according to a two-compartment open model. The half-lives of the distribution and elimination phases were 12.3 and 76.4 min respectively. The total apparent volume of distribution averaged 234 ml kg-1 and the plasma clearance 132 ml min-1. From these results it is suggested that fazadinium should have a shorter duration of action than other non-depolarizing agents because of the more rapid decline of the serum concentration corresponding mainly to the short half-life of the beta phase. This rapid elimination phase of fazadinium did not correspond to an extensive biotransformation because fazadinium was excreted almost unchanged in the urine and this excretion accounted for 50% of the injected dose within 24 h.


Pharmacokinetics of sulphamethoxazole in man: effects of urinary pH and urine flow on metabolism and renal excretion of sulphamethoxazole and its metabolite N4-acetylsulphamethoxazole. A high performance liquid chromatography method for the determination of sulphamethoxazole and its metabolite N4-acetylsulphamethoxazole is described. The renal excretion rate and cumulative renal excretion of sulphamethoxazole is markedly influenced by urinary pH. With constant urinary pH, the renal excretion rate and the renal clearance of sulphamethoxazole is dependent on the urine flow. The renal clearance of the metabolite N4-acetylsulphamethoxazole is not influenced by urinary pH or urine flow. No clear acetylator phenotype could be detected in the group of volunteers studied. The extent of acetylation depends on the amount of sulphamethoxazole available for acetylation, thus indirectly on the urine pH and flow.


The use of chlorazepate for premedication on the night before surgery. Chlorazepate, a long-acting benzodiazepine, allayed anxiety before operation, being superior to placebo in a double-blind study in which the drug was given the night before surgery. Long-term relief of apprehension before surgery by the administration of chlorazepate, combined with a narcotic analgesic when indicated, is advocated.


Comparative peripheral and coronary haemodynamic effects of rimiterol and isoprenaline. 1. Rimiterol and isoprenaline produced significant dose-related increases in cardiac output. 2. These changes in cardiac output were accompanied by increases in heart rate and myocardial oxygen consumption which were similar for each drug and dose-related. 3. Isoprenaline in contrast with rimiterol produced direct coronary vasodilation, i.e. coronary vasodilation in excess of that required to meet increases in myocardial oxygen demands. 4. It is suggested that the beta-adrenergic receptors in the human coronary vasculature are mainly of the beta1 type. 5. Rimiterol, because it does not produce direct coronary vasodilation may be preferable to isoprenaline in the treatment of low-cardiac output syndrome where there is regional myocardial ischaemia, since it would be less likely to produce a "coronary steal" effect.


13C nuclear magnetic resonance study of the cis-trans isomerism in X-Pro-Pro tripeptides. 13C nuclear magnetic resonance has been used to characterize quantitatively the cis-trans isomerism about both peptide bonds in the tripeptides Ser-Pro-Pro and Arg-Pro-Pro. Detailed pH titration data indicate that the configuration about both peptide bonds is closely linked to titration of the terminal carboxyl group and, to a lesser extent, to titration of the terminal amino group. The Pro2 C-3 resonance has been found particularly useful for interpretation due to its sensitivity to the isomerization about both peptide bonds. Analysis of the probabilities of the trans-trans, cic-cis, cis-trans, and trans-cis isomers in aqueous solution indicates a stability decrease in the order given. Similarities in the isomerization behavior of the two peptides indicate that side chain interactions involving the first residue have very little effect on the observed cis/trans ratios. The sensitivity of the cis/trans ratio to titration of the terminal amino group is most readily explained on the basis of an indirect effect on carbonyl-carbonyl repulsion.


Enzymatic basis for the Ca2+-induced cross-linking of membrane proteins in intact human erythrocytes. The accumulation of Ca2+ ions in intact human erythrocytes leads to the production of membrane protein polymers larger than spectrin. The polymer has a heterogeneous size distribution and is rich in gamma-glutamyl-epsilon-lysine cross-links. Isolation of this isodipeptide, in amounts as high as 6 mol/10(5) g of protein, confirms the idea [Lorand L., Weissmann, L.B., Epel, D.L., and Bruner-Lorand, J. (1976), Proc. Natl. Acad. Sci. U.S.A. 73, 4479] that the Ca2+-induced membrane protein polymerization is mediated by transglutaminase. Formation of the polymer in the intact cells is inhibited by the addition of small, water-soluble primary amines. Inasmuch as these amines are known to prevent the Ca2+-dependent loss of deformability of the membrane, it is suggested that transglutaminase-catalyzed cross-linking may be a biochemical cause of irreversible membrane stiffening.


Effects of substituents on the rates of deacylation of substituted benzoyl papains. Role of a carboxylate residue in the catalytic mechanism. The effect of ring substituents on the rates of deacylation of 8 meta- and para-substituted benzoyl papains was evaluated. The rate constants were found to depend upon a single ionizing group of pKa = 4.2--4.3, and to decrease by a factor of approximately 2.2 when measured in 94% D2O/H2O. The rates of deacylation are increased greatly by electron-withdrawing groups on the benzene ring. The Hammett rho value is 2.74 +/- 0.32. A plot of the rate constants for deacylation of the benzoyl papains against the corresponding constants for substituted benzoyl chymotrypsins generates a straight line of slope 1.0. This result suggests a very similar distribution of charge on the benzoyl moiety in the transition state for the two enzymes, which is interpreted in terms of the net charge of the transition state for the deacylation of nonspecific acyl papains being equal to--1 with the general base catalyzed assistance to the attack of water on the acyl enzyme being provided by the negatively charged Asp-158 rather than by the neutral Asn-175-His-159 hydrogen bond network. This result together with a survey of literature data suggests that the role of Asp-158 in papain catalysis has been underestimated. The evidence advanced to date in support of the proposition that an imidazolium-159-cysteine-25 thiolate ion pair exists in native papain is evaluated and considered to be insufficient to decide the issue.


The status of tyrosyl residues in a Formosan cobra cardiotoxin. Spectrophotometric titration of Formosan cobra cardiotoxin showed that two of the three tyrosyl residues were titrated freely with a normal apparent pKa of 9.6 whereas the remaining one ionized at pH above 11.0. Nitration of cardiotoxin in Tris . HCl buffer with tetranitromethane resulted in the selective nitration of tyrosine 11 and tyrosine 22. It also revealed that tyrosine 51 was the abnormal one in the spectrophotometric titration. Complete nitration occurred in the presence of 6.0 M guanidine hydrochloride. Compared with the conformation of native cardiotoxin, the peptide conformation of the partially nitrated cardiotoxin did not change significantly but the conformation of the completely nitrated cardiotoxin changed remarkably. The biological activity of cardiotoxin was indeed affected by nitration, but the immunological activity was nearly intact even when all the tyrosine residues were nitrated.


Spectroscopic studies on the tyrosine residues of canavalin. Canavalin is a tetramer with 6 tyrosines per subunit. In the work presented here, we have classified these tyrosines by their spectrophotometric and fluorometric pH titration and their ability to be quenched I-. Of the 6 residues, 2 were found to be exposed to the solvent. One (pK = 10.2) contributes 28% of the total fluorescence intensity; the second has a pK of 11.50, and a lower quantum yield, contributing only 16% of the total intensity. The remaining 4 residues (pK = 12.5, contributing 54% of fluorescence intensity) are buried; their titration is irreversible, requiring protein denaturation.


Proton NMR studies on soya bean lipoxygenase-1. The 270 MHz 1H-NMR spectrum of soya bean lipoxygenase-1 (linoleate: oxygen oxidoreductase, EC 1.13.11.12) was investigated at 298 K and at several pH values. A large fraction of the protein (50%) was found to be effectively random coil.


[Role of the electron acceptor properties of lysozyme and its substrate inhibitors in photosensitized electron transport in their complexes by the EPR method]. The nature of electron-acceptor groups in the system of lysozyme with its substrate-inhibitors has been studied in a wide range of pH values by the method of photosensitized electron transfer. In the lysozyme molecule disulphide bonds and peptide groups are the electron--acceptor groups. The nature of radicals in irradiated lysozyme depends on pH. At complex-formation of lysozyme with oligomeres of N-acetylglucosamine the electron transfer from the enzyme molecule to N-acetyl group of the substrate-inhibitor molecule is realized. Under conditions ruling out complex-formation of lysozyme with the inhibitors (N-acetylglucosamine and its dimer) the electrons are localized on disulphide bonds of the protein molecules at alkaline pH and at pH less than or equal to 3 the radicals are observed which are due to the remove of hydrogen atom from the Calpha-atom of the protein polypeptide chain.


Sympathetic and parasympathetic control of heart rate in unanaesthetized fetal and newborn lambs. Development of autonomic nervous control of basal heart rate was studied in unanesthetized fetal lambs (93 days to term) and newborn lambs (2--29 days), using atropine and/or propranolol blockade. Fetal lambs showed a progressive increase in parasympathetic restraint of heart rate; vagal influence in the newborn lamb was similar to the term fetus. Sympathetic stimulation of fetal heart rate declined toward term, possibly due to the strongly increasing parasympathetic influence. Sympathetic influence in the newborn was similar to the early-gestation fetus. Intrinsic heart rate was about 185 beats/min throughout the fetal and newborn life span studied. Thus changes in basal heart rate resulted from a different balance of the sympathetic and parasympathetic components of the autonomic nervous outflow.


[Structural changes in cell nuclei in the physiologic pH range]. Fluorescence of the suspension of cell nuclei from rat liver was investigated under neutral pH range. The pH decrease from 7 to 5 causes the quenching of protein fluorescence of nuclei and leads to an increase of the polarization degree of the fluorescence. Buffering properties of the nuclei in this range were detected. The use of 1-anilino-8-naphthalenesulfonate as the fluorescent probe permits to find the changes of nuclei surface properties with increasing the proton concentration in the solution. Circular dichroism spectra of nuclei were shown to depend on the pH solution too. It was suggested that structure state of the nuclei depends on the pH solution. The structural changes are reversible in the physiological pH range and were concluded to be in the envelope and chromatin of the nuclei. The role of histidine residues of nuclear proteins in the initiation of the structure changes was discussed. It was supposed that structure changes of the nuclei in the physiological pH range have an essential functional role.


Fundamental aspects of antimony thin film electrodes for pH measurement. This report concerns the investigation of the sensitivity, temperature dependence, accuracy, and the standard electrode potential EO of an antimony thin film pH electrode which was prepared with electron beam evaporation techniques. The air-formed oxide film on antimony thin film electrodes has been proved by both the cathodic reduction method and electron spectroscopy for chemical analysis (ESCA). The antimony thin film electrode responded rapidly to pH changes and its sensitivity was slightly changed depending on the buffer composition. The accuracy of this electrode was compared with that of the glass electrode. Temperature had some influence on the function of this electrode. The standard electrode potential of this electrode was discussed together with that of other forms of antimony electrodes. The structure and thickness of the surface oxide on antimony thin film electrodes was confirmed by cathodic reduction and ESCA. It was clear that the surface oxide governs the electrode reactions. Possible applications of the antimony thin film electrode are discussed stating some limitations in the use.


[Use of acetylcholinesterase reactivators and central cholinolytics in the treatment of experimental tetanus poisoning]. Experiments were conducted on rabbits with tetanus intoxication induced by the intravenous injection of a lethal dose of the toxin; a study was made of the therapeutic efficacy of the acetylcholinesterase reactivators--dipyroxim and isonitrosine, and also of the central cholinolytics--amizyl and diphacyl. In dose of 25 mg/kg dipyroxim produced no therapeutic effect, and in doses of 30--40 mg/kg caused the animal death. Amizyl and diphacil in a dose of 3--4 mg/kg caused elimination of tonic convulsions for 1 1/2--2 hours. As to isonitrosin--it produced the same effect for 4--5 hours. In case of combined administration of reactivators and cholinolytics convulsions were eliminated for 4--5 hours.


Prediction of wound sepsis following gastric operations. Gastric aspirates were obtained from 12 healthy volunteers, 49 patients with duodenal ulcer, 14 with gastric ulcer and 35 with gastric carcinoma. The mean total viable bacterial counts in these groups were as follows: volunteers 0, duodenal ulcer 3.8 X 10(1), gastric ulcer 6.95 X 10(4), carcinoma 1.9 X 10(7) organisms/ml. The incidence of wound sepsis in patients without antibiotic cover was; duodenal ulcer 17 per cent, gastric ulcer 38 per cent, carcinoma 56 per cent. Regardless of the underlying pathology, patients with counts greater than 5 X 10(6) organisms/ml in the gastric aspirate had a 93 per cent incidence of wound sepsis, compared with 16 per cent in patients with counts of less than 5 X 10(6) organisms/ml (P less than 0.001). In the group with high counts all except one of the wound infections were caused by organisms present in the stomach at the time of operation. There was a good correlation in the bacteriology of apirates obtained during preoperative endoscopy compared with operative nasogastric samples (n = 31) both for viable counts (r = 0.93) and for the counts of individual organisms. Therefore, preoperative endoscopy can be used to identify patients who are at risk of developing wound sepsis after gastric surgery.


Characteristics of Thiobacillus thioparus and its thiocyanate assimilation. Thiocyanate-assimilatig bacterium, TK 21, was isolated from activated sludge used for the treatment of thiocyanate contained in coke-oven liquor. This organism oxidized thiosulfate and elemental sulfur, causing a decrease of pH of the medium. These facts indicated that it belongs to the genus Thiobacillus. Potassium thiocyanate (0.5 g/l) was completely assimilated during 60 h. Thiosulfate inhibited the assimilation of thiocyanate but elemental sulfur did not. This bacterium did not evolve cyanide as its oxidation product after the decomposition of thiocyanate. The isoalted bacterium was identified as Thiobacillus thioparus. Examination of the composition of cellular fatty acid of three strains of T. thioparus showed that they prossessed 3-hydroxy fatty acid of C10 and C12; saturated straight chains of C10, C12, C15, C16, C17, and C18; monounsaturated straight chains of C16 and C18; and cyclopropane acid of C17.


Oxidation of gallium sulfides by Thiobacillus ferrooxidans. The bacterial oxidation of naturally occurring gallium-bearing chalcopyrite concentrate and a pure synthetic gallium (III) sulfide has been investigated at pH 1.8 and 35 degree C, using an active culture of Thiobacillus ferrooxidans. This oxidation process may proceed by direct or by indirect bacterial action. The highest dissolved gallium and copper concentrations were about 2.2 and 40.2 g/l, respectively. The order of the specific rate of oxygen uptake by T. ferrooxidans in approximately CuFES2 greater than or equal to gallium-bearing CuFeS2 greater than FeS2 greater than Cu2S greater than Cu2S greater than Ga2S3.


Cyclophosphamide-induced oncogenic transformation, chromosomal breakage, and sister chromatid exchange following microsomal activation. Cyclophosphamide, an extensively used cancer chemotherapeutic agent, requires metabolic activation through a mixed-function oxygenase system. The capacity of this agent to produce oncogenic transformation and chromosomal damage, including increases in sister chromatid exchanges, was investigated in cell culture with or without an exogenous liver metabolic activation system. No oncogenic transformation or chromosomal aberrations were produced by cyclophosphamide in the absence of metabolic activation, whereas significant transformation, chromosomal breaks, and increases in sister chromatid exchanges were observed when the activation system was incorporated into the assays. The oncogenic transformation and chromosomal changes were completely eliminated by removing glucose 6-phosphate and nicotinamide adenine dinucleotide phosphate from the metabolic generating system. These studies emphasize the necessity to incorporate some activation procedure into short-term assays used for evaluating the mutagenic and/or oncogenic potential of various chemicals.


Complexes of trivalent oxygenated phosphorus compounds with cytochrome P-450 and cytochrome P-420: the origin of double Soret spectra. Trivalent oxygenated phosphorus ligands include alkyl and aryl phosphites, (RO)3P, phosphonites, (RO)2PR, and phosphinites, ROPR2. All such compounds tested, with the exception of triphenyl phosphite, interact with ferrous cytochrome P-450 and its denatured form, cytochrome P-420, to produce complexes having two peaks in the Soret region of their optical difference spectra. Careful evaluation of these spectra indicate that they arise for different reasons for each of the two cytochromes. Clear evidence shows that cytochrome P-450 is not denatured by these ligands. The high affinity of these ligands for heme iron is indicated by small Ks values. The experimental results are used to substantiate a theory of the origin of microsomal double Soret spectra and the nature of the environments available for microsomal cytochromes P-450 and P-420.


The anaerobic dechlorination of trichlorofluoromethane by rat liver preparations in vitro. Incubation of trichlorofluoromethane with a liver microsomal fraction and an NADPH generating system under anaerobic conditions produced a metabolite dichlorofluoromethane, characterised by gas chromatography and mass spectrometry. The metabolic reaction was carried out by liver microsomes from the mouse, rabbit, hamster and rat and was increased by phenobarbitone pre-treatment. The formation of dichlorofluoromethane in vitro was enhanced by the addition of FMN, but partially inhibited by the presence of air, oxygen, SK&F 525-A, metyrapone and carbon tetrachloride and totally inhibited by carbon monoxide. The consumption of NADPH in the reaction was greater than could be accounted for by the production of dichlorofluoromethane indicating the possible formation of other metabolic products. It is suggested that trichlorofluoromethane interacts with the reduced form of cytochrome P-450 at the oxygen binding site and a possible mechanism for its subsequent reductive dechlorination is proposed.


A comparison of the effects of 5- and 6-hydroxydopamine on the isolated canine saphenous vein. We studied and compared the effects of 5- and 6-hydroxydopamine (5- or 6-OHDA) on venous tone and on adrenergic and nonadrenergic responses of the isolated canine saphenous vein. In the tissue bath, both 5- and 6-OHDA caused increases in tone and abolished adrenergic responses to transmural nerve stimulation. Effects of 6-OHDA were blocked by desmethylimipramine. 6-OHDA reduced norepinephrine levels of venous strips to 15% of those in control strips. Neither 5- nor 6-OHDA markedly altered peak contractile responses to norepinephrine but 6-OHDA prolonged the response to norepinephrine. Both compounds reduced responses to tyramine, with 6-OHDA producing a greater reduction than 5-OHDA. Large contractile responses to 5-OHDA were blocked only partially by treatment of strips with 6-OHDA, suggesting that contractile responses to 5-OHDA were mediated by direct and indirect actions. These results suggest differences in the actions of 5- and 6-OHDA on adrenergic terminals and smooth muscle in saphenous veins. The results of the experiments suggest that both 5- and 6-OHDA may be useful pharmacological interventions for the study of adrenergic mechanisms in isolated blood vessels.


Separation and analysis of arylsulfatase isoenzymes in body fluids of man. Soluble arylsulfatase (EC 3.1.6.1) is present in the body fluids of man in the form of two isoenzymes, arylsulfatase A and B, which reportedly are useful biochemical markers for certain types of malignancy. However, rapid assay of the individual isoenzymes is extremely difficult; procedures based on differential inhibition or activation of the isoenzymes in a mixture yield only semiquantitative results. A feature of these isoenzymes is their inhibition by some common anions (notably phosphate) at physiologic concentrations. The isoenzymes can be separated by anion-exchange chromatography, the B isoenzyme being eluted in the void volume and the A isoenzyme and the anionic inhibitors retarded. Lead is used to sequester phosphate, enabling measurement of A in the salt-eluted fraction. Using this technique, we have found significant elevations of B in the sera of patients with colorectal cancer. The potential of rapid, chromatographic separation coupled with continuous monitoring for arylsulfatase activity is discussed.


Acid beta-glucosidase and the diagnosis of Gaucher's disease in liver and spleen. Variable amounts of residual 4-methylumbelliferyl-beta-glucosidase activity gave rise to difficulties in the diagnosis of Gaucher's disease using spleen and particularly liver. Soluble and particulate components of beta-glucosidase, which interfere with the diagnosis, may be eliminated by preincubation of homogenates at pH 3.0 or with 100 mM sodium chloride at pH 4.0. After either of these treatments the optimum of acid beta-glucosidase was pH 4.5 and diagnosis of Gaucher's disease could be more easily made using homogenates of spleen and liver. There was residual acid beta-glucosidase activity in one of the Gaucher livers.


Methylmalonic/beta-hydroxy-n-valeric aciduria due to methylmalonyl-CoA mutase deficiency. A patient with methylmalonic and beta-hydroxy-n-valeric aciduria, apparently due to deficiency of methylmalonyl-CoA mutase, is described. The excretion of beta-hydroxy-n-valerate did not parallel that of beta-hydroxypropionate and methylmalonate but was observed, together with beta-keto-n-valerate, only during ketosis. beta-Hydroxy-n-valerate excretion thus correlates primarily not with the pool size of propionyl-CoA but with that of acetyl-CoA, and may occur during ketosis in any disorder causing accumulation of propionyl-CoA.


Glycogenosis type II: the infantile- and late-onset acid maltase deficiency observed in one family. The concurrence of the infantile and adult form of acid maltase deficiency in one family is presented. The muscle of both cases possesses residual activity. The glycogen amount in the infantile form is increased tremendously, while in the late-onset the amount of glycogen is normal.


Considerations on physiopathological approaches to the treatment of hypertension. 1. Two main physiopathological approaches to the treatment of hypertension are discussed, the first based on renin profiling and the second on haemodynamic measurements. 2. Reduction in plasma renin activity does not appear to be a clinically important mechanism in the hypotensive effect of beta-adrenergic blockers and of alpha-methyldopa. In particular, alpha-methyldopa has been found equally effective as a hypotensive agent both in normal renin and in low renin hypertensive patients. 3. Diuretics are certainly possessed of a renin-stimulating action, but in most patients this action does not obscure the hypotensive effect of these drugs. 4. Responders and non-responders to the hypotensive activity of beta-adrenergic blockers do not differ among them in their control haemodynamics.


alpha-Methyldopa and beta-blockers in hypertension--a comparison of their haemodynamic effects. 1. During acute injection of alpha-methyldopa there is a reduction in the blood pressure within a period of a few hours and a modest reduction in the cardiac output, sometimes due to a modest decrease in heart rate. Frequently the total peripheral resistance is decreased. In contrast, acute injection of beta-blockers induces a marked decrease in heart rate and cardiac output, a compensatory increase in the total peripheral resistance and no decrease in the blood pressure. 2. During long-term use alpha-methyldopa frequently reduces the total peripheral resistance and the reduction in cardiac output is usually modest, at rest as well as during exercise. The beta-blockers reduce the blood pressure, but induce a marked decrease in cardiac output and heart rate. The total peripheral resistance is little affected compared to the pre-treatment level.


Immunotherapy of human acute leukaemia.

Immunotherapy of human acute leukaemia. Conclusions are difficult to draw. In the six studies of immunotherapy of AML discussed, all the three employing BCG and cells showed a prolongation of survival and the major contributing factor to this prolongation of survival was extension of life after relapse. In the three studies using BCG alone only one shows a beneficial effect, but some more time must be allowed to elapse before this can be concluded with confidence.


Pamatolol: phase I evaluation of the pharmacodynamics of a cardioselective beta adrenoceptor blocking drug. A Phase I evaluation of a new adrenoceptor blocker, pamatolol, was performed in 10 healthy male volunteers. Minor reductions in standing and exercise and isoproterenol-induced increases in heart rate were observed with the 10-mg oral dose and appeared maximal with the 400-600 mg dose. The rate of decline of effect averaged 1.5% of the exercise heart rate/hr. Neither resting systolic time intervals nor post-exercise pulmonary function was affected by this dose range of pamatolol. Based on the latter responses and the isoproterenol dose response curve, we tentatively conclude that pamatolol is relatively cardioselective in man.


Assessing the subjective effects of stimulants in casual users. A methodology and preliminary results. In order to assess the subjective effects of nefopam, a new non-opiate analgesic, a study was designed using highly educated, young, middle-to-upper class subjects in a naturalistic setting. Results suggest that the design is capable of differentiating variations in subjective drug effects. On a number of measures, 10 mg of d-amphetamine, a modest dosage, could be distinguished from placebo, showing changes in the direction expected for stimulant drugs. Nefopam (90 mg), on the other hand, showed few differences from placebo or caffeine (300 mg). Nefopam appeared mildly dysphoric, rather than stimulant, in subjective effects.


Analytical subcellular fractionation studies on enterocytes from the jejunum and ileum of the rat and some properties of brush-border alkaline phosphatase. 1. Enterocytes, isolated from the proximal jejinum and distal ileum of the rat, were homogenized and their organelles separated by isopycnic centrifugation on continuous sucrose density gradients. The distributions of marker enzymes for the principal organelles, RNA and protein were determined in the sucrose gradients and related to the activities per entercocyte. 2. In the jejunum the modal equilibrium densities of the various organelles were: brush borders (1.20), lysosomes (1.20), peroxisomes (1.19), mitochondria (1.17) and basal-lateral membranes (1.13). The values were not significantly different in the ileum. The activities of brush-border enzymes, soluble and mitochondrial malate dehydrogenase, soluble and membrane-associated lactate dehydrogenase and particulate protein content, however, were greater in the jejunal than the ileal enterocytes. 3. Detergent exposed latent alkaline phosphatase activity in jejunal enterocytes and indicated that this enzyme is present not only in the brush border but also in the basal-lateral membrane and soluble fractions of the cell. 4. Isolated jejunal brush-border preprations showed latent activities of both alkaline phosphatase and gamma-glutamyltransferase whereas the activities of alpha-glucosidase and leucyl-beta-naphylamidase were not affected by detergent. Mechanical disruption of these preparations suggested the presence of two forms of alkaline phosphatase in the brush border and provides a technique to assess membrane fragility.


Urinary dopamine in man and rat: effects of inorganic salts on dopamine excretion. 1. Plasma and urine free dopamine (3,4-dihydroxyphenethylamine) were measured in six normal male volunteer subjects and the urinary clearance of dopamine was calculated for each subject. 2. The excretion rates for free dopamine in man were greater than could be explained by simple renal clearance. It was concluded that free dopamine must, therefore, be formed in the kidney. 3. Changes in urinary dopamine excretion were studied in four groups of rats initially maintained on low sodium diet and then given equimolar dietary supplements of NaCl, NaHCO3, KCl or NH4Cl, to study the specificity of the previously observed increase in dopamine excretion after increased dietary NaCl. 4. The mean dopamine excretion increased significantly in rats given NaCl, KCl and NH4Cl, whereas dopamine excretion decreased in those given NaHCO3. 5. The failure of dopamine excretion to rise in response to loading with NaHCO3 was unexpected, and argues against a simple effect of volume expansion by the sodium ion. The increase in dopamine excretion with KCl and NH4Cl showed that this response was not specific to the sodium ion.


Effect of varying PCO2 on intracellular pH and lactate consumption in the isolated perfused rat liver. 1. The effects of varying PCO2 on lactate uptake and intracellular pH (pHi) were studied in the isolated rat liver perfused with differing concentration of lactate. 2. In general, pHi and lactate uptake are inversely related to PCO2, and pHi and lactate uptake are directly related to each other, but the quantitative aspects and significance of these relationships vary with the availability of lactate. A model of hepatic lactate metabolism is proposed which may account for the quantitative variation. 3. The metabolism of lactate within the hepatocyte exerts a destabilizing effect on hepatocyte cell pH, in contrast to the buffering effect seen in predominantly glycolytic tissues. 4. An attempt is made to relate the findings to the disturbances of lactate metabolism in clinical respiratory failure.


Mechanism of the effect of varying PCO2 on gluconeogenesis from lactate in the perfused rat liver. 1. The effects of varying PCO2 on glucose output and the intracellular concentrations of lactate, pyruvate, phosphoenolpyruvate, 2-phosphoglycerate and 3-phosphoglycerate were studied in the isolated rat liver perfused with differing concentrations of lactate. 2. When the perfusate lactate concentration is above 1.5 mmol/l respiratory acidosis (simulated by high perfusate PCO2) inhibits gluconeogenesis from lactate, whereas respiratory alkalosis stimulates gluconeogenesis. 3. In general there were significant positive correlations between intracellular pH (pHi) and hepatocyte phosphoenolpyruvate, 2-phosphoglycerate and 3-phosphoglycerate concentrations, and negative correlations between pHi and lactate and pyruvate concentrations; there were usually significant correlations in the opposite  sense between these metabolites and log PCO2. 4. The results suggest that CO2 exerts an inhibitory effect on gluconeogenesis at a step between pyruvate and phosphoenolypruvate; however, this is not the only effect of CO2 on the gluconeogenic sequence. CO2 probably acts by changing pHi, but direct effects of CO2 and HCO-3 cannot be excluded. 5. Except at low lactate concentrations, nonionic diffusion probably does not play a major role in the entry of lactate into the hepatocyte.


[Crohn's disease: course and prognosis in 169 patients (author's transl)]. The most common initial symptoms in 169 patients with Crohn's disease, followed from 1956 to 1973, were diffuse abdominal pain, diarrhoea without any blood admixture, loss of weight, and fever. At the onset of the disease 84% of patients were aged 10-39 years. The most important complications (in descending order of frequency) were fistulae and abscesses, ileus, anaemia, and malabsorption. A breakdown into active and passive phases indicated that the younger the patient at onset of the disease, the more severe its course. The disease seemed to take a more favourable course when only the colon, but not the terminal ileum, was involved. Recurrence occurred more frequently after a short than after a long preoperative history. Recurrences were more frequent in patients under than over 31 years of age at their first operation.


Studies on the microsomal metabolism and binding of polybrominated biphenyls (PBBs). The metabolism of polybrominated biphenyls (PBBs) was studied in vitro by using rat liver microsomes in the presence of NADPH and atmospheric O2. Quantitative recoveries of all PBBs were obtained after incubations with control or 3-methylcholanthrene (MC) induced microsomes. Of the twelve major components, losses of only peaks 1 (2,4,5,2',5'-pentabromobiphenyl) and 3 (a hexabromobiphenyl) were observed following incubations with microsomes from phenobarbital (PB)- or PBBS- pretreated rats. Of seven structurally identified PBB components, only peak 1 has a bromine-free para position. Peaks 1, 2, and 5 all have two adjacent unsubstituted carbons, yet only peak 1 is metabolized. Of two dibromobiphenyl model compounds studied, the 2,2'-congener was very rapidly metabolized by PB-induced microsomes whereas its 4,4'-isomer was not. These results suggest that the presence of a free para position is required for the metabolism of brominated biphenyls. Of lesser importance appears to be the number of bromines or the availability of two adjacent unsubstituted carbons. In vivo evidence for the metabolism of peaks 1 and 3 was also provided by their drastically diminished levels in liver and milk extracts. When a 14C-PBB mixture consisting almost exclusively of peaks 4 (2,4,5,2',4',5'-hexabromobiphenyl) and 8 (2,3,4,5,2',4',5'-heptabromobiphenyl) was incubated with PB- or PBBs- induced microsomes and NADPH, only traces of radioactivity remained with the microsomes after extensive extraction. However, less radioactivity was bound to microsomes from MC pretreated or especially control rats. No radioactivity was bound to exogenous DNA included in similar microsomal incubations, regardless of the type of microsomes used. Under the same conditions, [3H]-benzo[a]pyrene metabolites were bound to DNA, and PBB-induced microsomes enhanced this binding more than six-fold.


Purification and properties of 5'-nucleotidase from lymphocyte plasma membranes. 5'-Nucleotidase is purified from lymphocyte plasma membranes by two affinity chromatographies. The first one, on Lens culinaris lectin-Sepharose 4B yields a fraction of twelve lectin-binding glycoproteins (lectin-receptor fraction). The second one on 5'-AMP-Sepharose 4B leads to pure enzyme. This enzyme is a glycoprotein with a molecular weight of 130 000; it gives a single band in polyacrylamide/dodecylsulfate electrophoresis and displays a very high specific activity (2500-3000 mumol Pih-1mg-1). Some properties of purified 5'-nucleotidase are similar to those of membrane-bound enzyme: substrate specificity, temperature dependence, effects of ions and SH-blocking reagents. Others are completely different for the two systems and these differences result from an interaction between the enzyme molecule and other Lens culinaris lectin binding proteins.


Purification of aspartate transcarbamylase from Drosophila melanogaster. A purification procedure is described by which aspartate transcarbamylase was obtained from cultured cells of Drosophila melanogaster as part of a high-molecular-weight enzyme complex. The complex is shown to contain several polypeptides. An antiserum directed against the complex enzyme inhibited in vitro the activity of aspartate transcarbamylase, carbamylphosphate synthetase and dihydro-orotase which were shown to copurify on a sucrose gradient and by gel electrophoresis. A fast preparation procedure using this antiserum yielded a 220 000-molecular-weight protein in addition to the polypeptides present in the complex. A purification procedure is also described to obtain aspartate transcarbamylase from second instar larvae of Drosophila. At this stage, the enzyme is not complexed with carbamylphosphate synthetase and dihydro-orotase but exhibits the same molecular weight as the aspartate transcarbamylase moiety found in the high-molecular-weight complex of cultured cells.


Identification of the functional ionic groups of papain by pH/rate profile analysis. The pH dependence of papain catalysis was analyzed by a scheme which evaluates the kinetic contribution of both protonated and unprotonated species of functional groups involved in catalysis. Kinetic measurements were made at constant pH, without buffers, by automatic titration. The rate-determining step for papain-catalyzed hydrolysis of alpha-N-benzoyl-L-arginine ethyl ester, determined by nucleophile competition, changed from acylation below pH 6.5 to mixed acylation-deacylation above pH 6.5. Kinetic analysis indicated that three prototropic groups governed the pH-specificity of alpha-N-benzoyl-L-arginine ethyl ester hydrolysis. These prototropic groups had pKa values of 4.8, 6.5 to 6.7, and 8.7. Theoretical treatment of the kinetics provided an excellent fit with the experimentally found profile when the contribution of all three prototropic groups was considered. Analysis showed that, in acid, the pathways of papain catalysis were functional with either two or three active-site protons. In base, a single functional ionic pathway is associated with an active site with only one proton. Pathways involving an unprotonated active site are catalytically inoperative in both acid and base. These results indicate that papain exhibits several catalytically functional ionic pathways. The results are discussed in terms of pKa assignments, and the mechanism of papain catalysis.


Hyperphenylalaninaemia due to dihydropteridine reductase deficiency. Two siblings with increased levels of serum phenylalanine were detected by newborn screening. The older sibling deteriorated neurologically and mentally, despite early dietary control, and died at the age of 6 1/2 years. In the younger sibling phenylalanine hydroxylase activity in liver tissue was normal. Further investigations revealed increased concentrations of biopterin derivatives in the blood, a low excretion of 5-hydroxyindole acetic acid in the urine, and a dihydropteridine reductase deficiency as the cause of hyperphenylalaninaemia. The parents of the siblings showed 50% of the normal dihydropteridine reductase activity in their fibroblasts grown in culture. Neurotransmitter therapy was started in the second child at the age of 6 months and this was followed by distinct neurological and mental improvement.


Effects of stored and fresh blood transfusions on postoperative pulmonary function. A clinical study on patients following extracorporeal circulation in association with aortic valve surgery. The effects on postoperative pulmonary function of fresh versus stored blood when priming the heart-lung machine were studied in 49 patients subjected to aortic valvular replacement. The onset of extracorporeal circulation was considered to mimic the conditions of massive blood transfusion. The patients receiving heparinized fresh blood required more oxygen in the respirator than those receiving stored blood, but still had a significantly lower arterial oxygen tension during the first and second postoperative days. The fresh blood patients also showed a tendency to require a larger amount of diuretics to maintain adequate urine output. Pre- and postoperative bleeding and changes in postoperative hematological variables did not differ in the two groups. The results imply that stored blood should be preferable when substituting blood losses.