Effect of ultraviolet radiation on pinocytosis in Amoeba proteua. Ultraviolet (UV) irradiation (4 000-10 000 erg X mm(-2) decreased membrane potential and input resistance of Amoeba proteus and induced formation of pinocytotic channels. Submaximal pinocytosis induced by UV light was additive to pinocytosis induced by K+ or Na+ and stimulated in the presence of EGTA. It was not inhibited by the presence of La+++ or by pretreatment with dibucaine. In these respects and with respect to optimum pH and pCa, UV induced pinocytosis. Accumulation of K+ in the amoeba membrane after a dose of radiation may explain the similarity between pinocytosis induced by UV light and potassium salts. Ca++ present during the period of irradiation inhibited the effect of UV light. Instead Ca++ applied after irradiation (1-20 mM) increased channel formation. This effect was stimulated the presence of local anesthetic drugs. It is suggested that high doses of UV light may induce channel formation by releasing Ca++ from the cell membrane into the cell (UV induced pinocytosis). Ca++ may be released at the moment of absorption of UV light in the membrane as well as during the period of depolarization which follows irradiation. Low doses of UV light may permit extracellular Ca++ to enter the cell and stimulate channel formation (calcium induced pinocytosis). Dithiotreitol (1 mM) applied after irradiation depressed both UV and calcium induced pinocytosis so these may be the result of the same structural change which involves the formation of disulphide bonds in the membrane.


An in vitro-formed protamine-heparin complex as a model for a two-compartment store for biogenic amines. The capacity of an in vitro-formed protamine-heparin complex (PHC) to store inorganic cations and biogenic amines was investigated. The PHC behaves like a two-compartment storage system. One compartment corresponds to the terminal free carboxyl groups of the protamine moiety and has the characteristics of a cation exchanger, with the ability to bind inorganic cations and biogenic amines in a reversible and rather unslective manner. The cations and biogenic amines therefore compete for and displace each other from the common ionic binding sites. The binding sites in the other compartment, corresponding mainly to the carboxyl groups of the heparin moiety, are only unmasked at high ionic concentrations and show a specific affinity for biogenic amines. The storage of amines in this compartment of the PHC is reversible but is dependent not only on simple ionic binding but evidently also on other attractive forces, such as dipole and hydrogen bonding.


The local effect of pH changes in the cerebrospinal fluid on the ventrolateral areas of medulla oblongata and on the spinal cord surface on the activity of cardiac and vertebral sympathetic nerves. Application of acid cerebrospinal fluid at pH 6.8 to the ventral surface of the medulla oblongata in cats increased the arterial blood pressure and the activity of the vertebral nerve and enhanced respiratory modulation of the vertebral and cardiac nerves. Alkaline cerebrospinal fluid at pH 7.7 applied to the ventral surface of the medulla oblongata increased the arterial blood pressure without changes in the sympathetic nerve activity. When the spinal cord was superfussed with artificial acid cerebrospinal fluid at the Th1 level the activity of the vertebral nerve and cardiac nerve increased and the arterial blood pressure rose. The authors discuss the possibility of regulation of the activity of sympathetic neurons in the medulla and spinal cord by means of direct action of pH and pCO2 on the neurons, or through an indirect effect mediated by superficial chemoreceptors of the medulla oblongata. The role of local changes of the brain blood flow in the mechanism of these results is discussed as well.


Oxygen uptake and tissue oxygen tension during adrenergic stimulation in canine subcutaneous adipose tissue. The effect of sympathetic nerve stimulation (NS) and injected noradenaline (NA) or isoprenaline (Iso) on PVO2, VO2 and PtO2 was studied in isolated canine subcutaneous adipose tissue. These effects were compared to those produced by mechanical blood flow reduction (clamping). Resting VO2 measured 13.0+/- 2.3 mumol X min-1 X 100 g-1. When blood flow was reduced by 20% or less there was no significant change of VO2. Reducing blood flow to 50% of control or less by NS caused a parallel reduction in VO2, while clamping reduced VO2 significantly less. NA gave effects similar to those of NS. After NS or NA there was a period of hyperemia and increased oxygen extraction which more than compensated for the decrease in VO2 during vasoconstriction. Such a net increase in VO2 was not produced by clamping. Control PtO2 averaged 29+/-2 mmHg. NA reduced it by 70% and clamping to the same blood flow level only by 14% (p less than 0.01). Thus, a mere reduction in blood flow has little effect on PtO2, while blood flow reduction combined with redistribution of blood flow and an increased oxygen deman can lead to tissue hypoxia.


Beta2-adrenoceptors facilitating noradrenaline secretion from human vasoconstrictor nerves. Isolated biopsy specimens of human peripheral arteries and veins, preincubated with 3H-(-)- noradrenaline (NA) to label the neural stores of NA, were used to study the Beta-adrenoceptors previously found to increase the secretion of 3H-NA evoked by electrical field stimulation of the adrenergic nerves of this tissue. The increase in nerve stimulation induced secretion of 3H-NA caused by 0.04 muM isoprenaline was prevented by 1 muM propranolol. This beta-blocking drug by itself slightly but significantly depressed the secretion of 3H-NA caused by nerve stimulation in the absence of isoprenaline. While the secretion of 3H-NA was not affected by known beta1-agonists, it was dose-dependently and reversibly increased by two different beta2-agonists. The effect of isoprenaline on 3H-NA secretion was not altered by a selective beta1-antagonist, but strongly reduced or abolished by a beta2-blocking drug. The results indicate that the beta-adrenoceptors involved in the control of NA secretion from the vasoconstrictor nerves of human omental blood vessels are only to a minimal extent stimulated by NA secreted from the nerves, and therefore do probably not mainly serve to mediate local positive feedback control of transmitter secretion; the receptors appear to be beta2 in nature.


Vagal reflexes in the bronchoconstriction occurring after induced intravascular platelet aggregation. Intravascular platelet aggregation induced in cats by i.v. infusions of collagen caused a transient increase in pulmonary vascular resistance (PVR) and in non-elastic pulmonary resistance (RL), and a transient decrease in dynamic lung compliance (dyn CL). PVR,dyn CL and platelet aggregation after collagen infusions were unaffected by bilateral cervical vagotomy and atropinization, wheras these procedures reduced the post-infusion rise in RL by about 50 per cent. The administration of indomethacin inhibited platelet aggregation as well as bronchoconstriction after collagen infusion. The present investigation indicates that intravascular platelet aggregation will cause reflex bronchoconstriction mediated by vagal efferent fibres.


Clinical features in poisonings by tricyclic antidepressants with special reference to the ECG. Clinical variables, and especially their relation to the ECG, have been studied in 153 cases of poisonings by tricyclic antidepressants (TCA). The mean age of the patients was 34 years. Amitriptyline poisoning accounted for 112 (73%) of the cases and the mean dose ingested was about 1 000 mg. Coma was present in 87 patients (57%) and on admission 40 (26%) had a systolic blood pressure (BP) below 100 mmHg. The systolic BP on admission was significantly lower (p less than 0.001) and the heart rate (HR) higher (p less than 0.001) than when the patients left the ward. Apart from an increased HR (greater than or equal to 90 beats/min), which was present in 73% of the cases, the most characteristic ECG change was a QRS prolongation (greater than or equal to 0.11 sec), this being found in 42% of the cases. About the same proportion displayed a QT prolongation and 28% had a prolonged PQ time. The mean of the QRS times was 0.11 sec. Unlike the QT time, the QRS time was not correlated to HR. Statistical analysis of the material with regard to clinical variables (dose of TCA, BP, coma duration, etc.) showed that the QRS time was closely related to the severity of poisoning. Five patients (3) died, all of whom already on admission demonstrated advanced ECG changes with arrhythmias and a mean QRS time of 0.19 sec. Excluding dibenzepine poisonings (4 cases, all fatal), the mortality rate was 0.7%. The importance of high initial preparedness for cardiac complications is pointed out, as is the value of the QRS time as a guide to the severity of poisoning.


The mechanism of action of glycosidases. The factors that may contribute to the rate enhancement observed with enzymatic versus non-enzymatic hydrolysis of glycosides are discussed. The nature of the active site as deduced from labelling studies with beta-glucosidases is described. A two-step mechanism involving either an enzyme stabilized glycosyl ion or a covalent glycosyl-enzyme intermediate is proposed. Experiments with a beta-glucosidase from almonds show that even with 2-deoxy glucosides with good leaving groups as aglycon which are hydrolyzed 1000 times more slowly than the corresponding glucosides, the deglucosylation step is faster than the cleavage of the glycosidic bond.


Properties of hydroxysteroid oxidoreductase isolated from yeast. Yeasts can advantageously be utilized for the production of the 17beta-hydroxy-derivative, from 3-methoxy-8,14-seco-1,3,5(10),9(11)-estratetraene-14,17-dione (14,17-dione) while 14alpha-hydroxy and 14alpha,17beta-dihydroxy-derivatives are also formed. The biochemical properties of yeasts' enzymes responsible for the formation of the two monohydroxy-derivatives have been studied in detail. In the cell-free extract of Saccharomyces the presence of two hydroxysteroid oxidoreductases could be detected. The first enzyme forms 3beta,17beta-dihydroxy-derivative from 5alpha-androstane-317-dione. This enzyme is responsible for the formation of 17beta-hydroxy-derivative from 14,17-dione. The second enzyme forms 3alpha-hydroxy-derivative from 5beta-androstanedione as well as 14alpha-hydroxy-derivative from its 14,17-dione. The cofactor of both enzymes is pyridine nucleotide. The two enzymes possessing different properties can selectively be inhibited.


Properties of delta5-3beta-hydroxysteroid oxidoreductase isolated from Streptomyces griseocarneus. Delta5-3beta-hydroxysteroid oxidoreductase was extracted in magnesium-containing Tris buffer from sonicated Streptomyces griseocarneus cells. The enzyme was partially purified (150 X) by ion exchange chromatography and gel filtration following (NH4)2SO4 fractionation. Upon gel filtration on Sephadex G-75 to G-200, the greatest part of the activity gave a peak in the fractionation range. The enzyme obtained from the gel yielded small enzyme molecules on repeated chromatography. A molecular weight of 32 to 36 000 was calculated for the activity appearing in the fractionation range of Sephadex G-75 to G-200. The enzyme is highly specific for the irreversible oxidation of the 3beta-hydroxyl group in steroids with a trans-anellated A : B ring system with either C5 or C6 double bond. Delta5-3-ketosteroids are converted into delta5-3-ketosteroids at a high rate, but the isomerase activity cannot be separated from the oxidoreductase activity either by chromatography or by selective heat inactivation. NAD, NADP, FMN or FAD did not influence the activity, but the enzyme is inactive in the absence of molecular oxygen.


Soil microflora of the rhizosphere of plants from several habitats in the botanical garden in Poznań. The abundance and activity of certain groups of soil microorganisms were estimated in the rhizosphere of 13 plants from four different habitats at several dates. An additional study was made of the rhizosphere microflora of Ledum palustre from a peat-bog. Numbers of proteolytic, nitrate assimilating, denitrifying, and cellulolytic bacteria were estimated. The intensity of ammonification, nitrification and Clostridium growth was estimated. The studies have revealed that the abundance and the activity of all the groups of bacteria studied depended on the habitat. Within one habitat, however, the influence of the plant was sometimes more pronounced than that of the habitat itself, particularly on the abundance of proteolytic bacteria (Fig. 1). The date of sampling had very little effect on the abundance of all the bacterial groups studied except the cellulolytic bacteria.


Kinetic studies on citric acid production by Aspergillus niger. II. The two-stage process. A two-stage process of submerged citric acid fermentation with replacement of growth medium by fermentation medium has been worked out. The optimum composition of mineral nutrients and pH of the fermentation medium of the second stage of the process were determined. An addition of 0.5 g/l of NH4NO3 as nitrogen source and 0.1 g/l of MgSO4-7H2O as magnesium source ensured effective conversion of sucrose to citric acid. An addition to KH2PO4, on the other hand, was definitely unfavourable as it considerably reduced the product yield. The medium for the second stage of fermentation should be acidified to about pH 2.2, while the water used for washing the mycelium from the remains of the growth medium should have a pH of 2.5--3.5. Under these conditions, with an initial sucrose concentration of 100 g/l, after 132 hr fermentation at 26 degrees up to 90 g/l of citric acid was obtained, which corresponds to a productivity of over 16 g/l. day. The highest activity for citric acid formation was found in three- or four-day-old mycelium.


L-Glutamate-glyoxylate aminotransferase in Lactobacillus plantarum. Glutamate-glyoxylate aminotransferase which mediates the reaction of glyoxylic acid with glutamic acid to yield glycine and alpha-oxoglutaric acid has been isolated and purified 84-fold from extracts of Lactobacillus plantarum. Purified enzyme requires the addition of pyridoxal phosphate and magnesium ions for its activity. The molecular weight of the enzyme estimated by Sepharose 4B gel filtration amounts to 37.000. Micaelis constants for glyoxylate and glutamate are corresponding to 6.25 X 10(-3) M and 2.75 X 10(-3) M, respectively. Optimal pH in phosphate and veronal buffers is 8.0 and optimal temperature 35--37 degrees C.


Inhibition of Newcastle disease virus replication by 6-azauridine. II. Combination of 6-azauridine and adenine derivatives. Twenty-five metabolites (purines, pyrimidines, nucleosides and nucleosides) were tested for their simultaneous action with 6-azauridine (AzUrd) in inhibition of Newcastle disease virus (NDV) replication. With the exception of deoxyadenosine and cyclic AMP all natural adenine derivatives exerted a synergic effect with AzUrd like ATP. Glutamine in combination with AzUrd did not inhibit NDV replication. The inhibitory effect of the combination of AzUrd and adenine derivatives was reversible by guanosine, uridine and cytidine but not by orotic acid or orotidylic acid.


Some biological activities of rabbit anti-interferon serum. After prolonged immunization of rabbits with a semipurified mouse interferon preparation in Freund's incomplete or Al-Span-Oil adjuvant, a specific interferon-neutralizing immunoglobulin was obtained from antiserum with a capacity of neutralizing about 49000 mouse interferon units per ml. The specific activity of the antiserum and immunoglobulin was confirmed in tests in which the interaction of antibodies with the cell surface was ruled out. The antiserum (and the immunoglobulin) neutralized both the antiviral and the cell-growth inhibitory activities of interferon. The "slow" and the "fast" fractions of purified interferon preparations were equally sensitive to the neutralizing effect of antibodies. On the other hand, the reaction of heat-inactivated interferon with the antiserum did not diminish the neutralizing activity of the latter, suggesting a destruction of interferon antigenic sites.


Interferon-producing capacity of human tonsil cells and properties of interferon produced by these cells. Tonsils excised from human beings for chronic tonsillitis have been shown to be an accessible and sufficiently rich source of human lymphocytes. Tonsil cells produced interferon as intensively as blood leukocytes and the properties of this interferon were similar to those of leukocyte interferon. The optimal conditions for interferon production by tonsil cells were established.


Diminished virucidal activity of kethoxal against vesicular stomatitis virus pretreated with guanidinating reagent and proteases. The reactivity of vesicular stomatitis virus (VSV) with kethoxal can be appreciably altered by treatment with 1-guanyl-3, 5-dimethyl pyrazole nitrate (GDMP) and proteolytic enzymes. Pretreatment of purified VSV with GDMP or proteolytic enzymes markedly reduced the effectiveness of kethoxal as a virucide. The rate of neutralizability of GDMP- and trypsin-treated viruses by specific antiserum differed from that of controls.


Levels of immunoglobulins and antibodies to haemaglutinin and neuraminidase of influenza virus in nasal secretions after natural infection. Nasal washings (NW) from 16 influenza patients in the course of an epidemic in November and December, 1974 were examined for the presence of influenza virus, immunoglobulins (Ig) and titres of haemagglutination inhibiting (HI) and neuraminidase inhibiting (NI) antibodies. Influenza virus identical with A/Port Chalmers/1/73 (H3N2), increased levels of IgA and occasionally IgG, and specific antibodies were detected in the NW. The dynamics of HI and NI antibody formation did not differ substantially, but there were individual differences in titres and persistence of antibodies. Convalescent sera always contained increased levels of HI and NI antibodies. In some cases, the titres of antibody to viral ribonucleoprotein did not increase.


Associated seroconversions to respiratory viruses in volunteers with experimental influenza infection. Serological examinations of 573 volunteers with mild experimental influenza infection and 86 volunteers of a control group hospitalized in a special clinic revealed a significant rise in the titre of antibodies (seroconversion) not only to influenza A or B viruses used for the experimental infection but in 23.3 to 29.8% of cases also to other respiratory viruses. Based on a number of arguments, associated seroconversions are interpreted as due to mixed or sequential infections of different aetiology.


Different allotypic susceptibility of mice to Rickett-sia tsutsugamushi. Some lines of mice and random-bred mice show different susceptibility to infection with Rickettsia tsutsugamushi, displayed mainly in the development of overt infection. The latter is most markedly manifested in random-bred and Swiss mice, causing high death rates and massive accumulation of rickettsiae in the peritoneum. Black mice respond to inoculation with asymptomatic infection with no microscopically detectable rickettsiae. The infection is accompanied in either case by immunological response and persistence of the agent in mice. One of the mechanisms of resistance to R. tsutsugamushi in some mouse lines is due to phagocytosis at the inoculation site of the main mass of rickettsiae by macrophages which digest the agent and retain their viability.


Cytological investigation of Rickettsia tsutsugamushi infection of mice with different allotypic susceptibility to the agent. Cytological and cytochemical studies on Swiss and random-bred white mice susceptible to Rickettsia tsutsugamushi infection as well as resistant C57 Black 6 and CBA mice revealed a clear-cut relationship between the severity of infection and the pattern of macrophage reaction. In highly susceptible mice the fatal infection was accompanied by death of macrophages and necrotisation of the peritoneal exudate cells. The resistant mice showed no clinical signs but developed an intensive macrophage reaction, and the main mass of the inoculated rickettsiae died at the inoculation site, the macrophages remaining viable. Phagocytosis and digestion of rickettsiae were accompanied by markedly increased activities of hydrolytic and oxidizing-reducing enzymes.


Characterization of an endotoxic lipopolysaccharide from Coxiella burnetii. Phase I Coxiella burnetii antigen isolated by phenol extraction from purified suspensions of C. burnetii in phase I is a complex lipopolysaccharide (LPS) molecule containing substances typical of the bacterial LPS. Some endotoxic properties of this C. burnetii LPS, namely pyrogenicity and skin epinephrine reaction in rabbits, hypothermia in white rats, lethal effect on chicken embryos or on actinomycin-D-treated mice are similar to those of LPS isolated from other Gram-negative bacteria.


Electron microscopy of virus particles in ultrathin frozen sections. A procedure with the use of polyethylene glycol (PEG) 600 to support biological materials for ultracryotomy was developed. Developing and mature virions were demonstrated by electron microscopy in negatively stained frozen sections of vaccinia virus-infected human diploid cells and of herpes simplex virus (HSV) type 1 infected primary rabbit kidney cells. Approximate localisation of the virus particles in the cells was possible.


Interferon pretreatment primes interferon production by human adenovirus in chick embryo cells. Pretreatment of chick embryo cells (CEC) with homologous interferon enhanced the subsequent interferon production induced by human adenovirus. This priming effect of interferon pretreatment was also demonstrable when trypsin-treated virus was used for the induction of interferon.


Enhancement of interferon induction in mice by polycationic modified polypeptides. Combined intraperitoneal treatment of mice with poly I: C and a polycationic modified polypeptide (poly-DMAE-glutamine) was investigated. It was established that the presence of appropriate amounts of poly-DMAE-glutamine produced markedly enhanced serum interferon levels as compared to those produced by poly I: C alone. Similar combinations injected into mice produced full protection against lethal doses of mouse-virulent Semliki forest virus.


Inhibition of Newcastle disease virus replication by 6-azauridine. I. Inefficacy of purified uridine kinase, effect of adenosine-5'-triphosphate. As distinct from cell-free extracts prepared from tumour cells, partially purified uridine kinase prepared from the same cells was not effective in 6-azauridine (AzUrd) inhibition of Newcastle disease virus (NDV) replication. This showed that uridine kinase was not the effective component of cell-free extracts. Adenosine-5'-triphosphate (ATP) was found to exert a synergic effect in combination with AzUrd in the inhibition of NDV replication.


Effect of vagus nerve stimulation upon excitability of the canine ventricle. Role of sympathetic-parasympathetic interactions. The effect of vagus nerve stimulation on ventricular excitability was studied in 28 dogs under various conditions of adrenergic neural tone. Strength-interval curves were delineated from the apex of the right ventricular endocardium with a transvenous bipolar catheter. Vagus nerve stimulation in both closed chest and open chest dogs shifted the strength-interval curve 6 to 8 msec later into electrical diastole (P less than 0.001). Left stellate ganglion stimulation shifted the strength-interval curve 9 to 11 msec earlier into diastole (P less than 0.001). The effect of simultaneous left stellate ganglion and vagus nerve stimulation was not significantly different from that of left stellate ganglion stimulation alone. The influence of vagus nerve stimulation on the strength-interval curve under basal conditions was abolished by acute beta adrenergic blockade with propranolol. It is concluded that vagus nerve stimulation affects ventricular excitability as well as vulnerability by opposing the effects of sympathetic neural tone.


An automated flavin adenine dinucleotide-dependent glutathione reductase assay for assessing riboflavin nutriture. An automated AutoAnalyzer method using 5:5'-dithiobis-2-nitrobenzoic acid is described for determining whole blood glutathione reductase (BGR) activity and for measuring in vitro activation of BGR with flavin adenine dinucleotide (FAD). BGR activity is expressed as mumoles glutathione regenerated from oxidized glutathione per ml of whole blood (WB) or per g of hemoglobin. The stimulatory effect of FAD on BGR activity divided by the activity without FAD determined the activity coefficient (AC). We found that NADPH and oxidized glutathione assay concentrations of 0.100 mmole/liter and 0.250 mmole/liter, respectively, in 0.1 mole/liter phosphate buffer, pH 7.4, gave consistent results when WB, before assay, was diluted 20-fold. WB samples to be stored are initially diluted 10-fold with distilled water and frozen. Prior to assay, two aliquots of the sample are diluted 2-fold, one aliquot with distilled water and another with 46 mumole/liter FAD. With sample and manifold dilutions the assay FAD concentrations is 1.0 mumole/liter: assay concentrations greater than 5.0 mumole FAD/liter were shown to be inhibitory. We examined blood samples from 617 children in the age range 6 to 60 months and determined the normal AC range to be between 1.00 and 1.35. Six weaned rats (23 days of age), maintained on a riboflavin-deficient diet, showed a mean AC of 1.23, 1.54, 2.02, and 2.41 at 23, 26, 30, and 36 days of age, respectively. Six control rats maintained an AC of 1.23 +/- 0.05 (SD) during the same period.


Accidental poisoning with psychotropic drugs in children. Seventy-seven (0.24%) of 32,005 admissions to the Massachusetts General Hospital pediatric service during the period 1962 to 1973 were due to accidental poisoning. In 27 cases, mostly involving children less than 6 years of age, psychotropic drugs were implicated. These included sedative-hypnotics in six cases, phenytoin in two, major tranquilizers in five, antidepressants in three, stimulants or hallucinogens in three, and drug mixtures in eight. Toxicologic analyses contributed little to diagnosis and initial management. Except for one child who ingested ferrous sulfate, no patient was seriously intoxicated, and all recovered rapidly without sequelae. Although referral of serious poisoning cases to another hospital may have biased the results, the findings suggest that accidental psychotropic drug poisoning is not a major source of childhood morbidity.


[Mediators of the allergic reaction. Slow reacting substance (SRS-A)]. The slow reacting substance of anaphylaxis (SRS-A) belongs to a group of substances which produce a slow progressive and sustained contraction of some smooth muscles. It is released by the interaction of the antigen with certain antibodies; in humans through the interaction with the IgE or reagine. The SRS-A is a heat-labile sustance, chemically unstable, of an acid character, with a low molecular weight. It is not destroyed by the action of proteolytic enzymes. Its molecular structure has not yet been elucidated. It is not found accumulated in the cells but synthesized and released by some white cells mainly by sensitized mast cells and polymorphonuclear leukocytes after challenging with the specific antigen. The SRS-A is a powerful pharmacodynamic agent, it produces contraction of the bronchial smooth muscle in doses of nanograms. It probably plays a predominant role in the physiopathology of asthma. The chain of chemical reactions elicited by antigen-antibody interaction does not end with the release of SRS-A and the other mediators (histamine, eosinophil chemotactic factor of anaphylaxis, ECF-A), on the contrary, these mediators especially SRS-A induce the release of prostaglandins of type E (PGE1 and PGE2) which produce bronchodilatation and inhibit the release of SRS-A itself, perhaps being a selfregulating mechanism. The PGF2a, on the other hand, produces bronchoconstriction. The release of SRS-A is also inhibited by the action of diethyl-carbamazine and especially by sodium chromoglicate and compound AH-7725. From the biochemical point of view it is found that the antigen Igells, a serine esterase, initiating several chemical reactions whose consequence is a decrease in the cAMP concentration. This reduction in the cAMP intracellular level is followed by synthesis and excretion of the SRS-A as well as by the aggregation of the microtubules and excretion of the stored histamine. The PGE (1 and 2) acting on one type of membrane receptor and the beta-agonist catecholamines on another produce a common phenomenon: the activation of the adenylcyclase whcih produces the increase of the concentration of cAMP and inhibits the release of mediators of the anapylactic reaction. The parasympathetic system through its chemical mediator acethylcholine, by a mechanism in whcih adenycyclase is not involved is also capable of stimulating the release of histamine and SRS-A. Something similar happens with PGF2a. In conclusion, self-regulatory mechansims for the release of mediators of the anaphylactic reactions may exist. The "perpetuation" of an asthamtic reaction would signify a failure of these self-regulatory mechanisms due to, for example, to a temporary block of the beta-adrenergic receptors, overstimulation of the alfa-receptors or insufficient production of PGE or a transformation of the PGE in PGF.


Polyarteritis in children.

Polyarteritis in children. Polyarteritis was diagnosed in three girls, 9 to 10 years old, by kidney and skin biopsies. They were treated with a combination of prednisone (1.5 to 2 mg/kg) and cyclophosphamide (2 mg/kg) for up to 12 months. The illness was severe in all three, complicated by hypertension, seizures, pulmonary infiltrates, renal failure, or hallucinations. All three patients are alive and well with no or minimal residual symptoms two to three years after therapy was discontinued. The treatment with corticosteroids or with a combination of steroids and immunosuppressive drugs seems to improve the prognosis of polyarteritis considerably.


Changes in mucosal and venous histamine concentrations during instillation of ethanol in the canine stomach. Histamine concentrations in the gastric mucosa, gastric vein blood, and gastric contents were measured after instillation of 300 ml of 40%, 20% and 12.5%, vol/vol, ethanol intragastrically for 30 min in anesthetized dogs. The mean histamine concentration in the gastric mucosa, gastric vein blood, and gastric contents in dogs treated with 40% ethanol was significantly higher than in dogs treated with sodium chloride (NaCl). Serial studies of the gastric vein blood of the ethanol-treated dogs should significant elevation of histamine concentrations for 2 hr after instillation of 40% ethanol, but less increases at 20 and 30 min after instillation of 12.5% and 20%, respectively. No change was observed after administration of 50% glucose and 25% mannitol. Morphologically, the gastric mucosa of the ethanol-treated dogs was edematous, congested and hemorrhagic. The severity of these changes increased with the concentration used. The mucosa of the dogs treated with sodium chloride, glucose, or mannitol was normal. The factors that may underlie the increase in histamine concentrations in the gastric vein blood and the possible relation of such an increase to the morphologic alterations are discussed.


Organic anions induce colinic secretion. Experiments were conducted by instilling test solutions into the cleaned colon of anesthetized rats. Isotonic test solutions at an acid (2.9% or neutral (7.0) pH and containing acetic acid or acetate ions were used. At the lower pH, acetic acid was absorbed well absorbed and the colon absorbed fluid. The secretory effect could not be reproduced by hydrochloric acid solutions at pH 2.9. Fluid secretion induced by acetic acid was associated with discharge of mucus from goblet cells. The interference of colonic absorption by acetic acid might be important in some diarrheal diseases.


Residues in antibiotic preparations, i: scanning electron microscopic studies of surface topography. The surface characteristics of residues obtained from several commercial antibiotic products were studied using scanning electron microscopy. The photomicrographs showed the presence of particulates possessing crystalline or amorphous properties with particle sizes ranging from 1 mum to several mum. Small, well-defined, granular crystalline particulates seemed to predominate in residues of products which were processed  under optimum pH conditions. Coalescing or aggregating of small crystalline particles (1-10mum) to form larger masses was interpreted as a sign of instability of the antibiotic. Most of the penicillin and semisynthetic penicillins produced residues that were characterized as amorphous, flaky, bulky and of no distinct shape. Such structures were attributed to the drying of gelatinous particulates. The possible sources and causes of the occurrence of these particulates are discussed. It appears that in many of the products the particulates are product-related rather than process-related, and subtle degradation may be involved. Advice is given to practitioners regarding the preparation, storage and administration of these products.


Residues in antibiotic preparations, ii: effect of pH on the nature and level of particulate matter in sodium cephalothin intravenous solutions. The effect of pH on the extent and nature of particulate contamination in sodium cephalothin intravenous solutions was studied. The extent of particulate contamination was determined by microscopic and automatic electronic counting procedures, and the physical nature of the particles was determined using scanning electron microscopy. At low pH values (i.e., 4.9--5.9), an amorphous residue was predominant, while at higher pH values (6.9--7.8), the gelatinous nature of the residue disappeared and small crystalline particulates of varying sizes were predominant. The level of particulate contamination (particles over 10 mum in size) decreased with an increase in pH to a minimum level between pH 7 and 8. Advice is given to practitioners regarding the preparation of sodium cephalothin solutions.


Compatibility of penicillin and ascorbic acid injection. The stability of Potassium Penicillin G, USP, when mixed with Ascorbic Acid Injection, USP, in 5% Dextrose Injection, USP, was studied. The change in concentration over an eight-hour period of potassium penicillin G in the admixture was determined by the hydroxylamine colorimetric assay method and the microbiological assay method. The stability of penicillin was not adversely affected by the presence of sodium ascorbate. Reports of incompatibilities between penicillin and ascorbic acid are a function of pH rather than a characteristic of the ascorbate ion. Articles reporting studies involving ascorbic acid should specify whether the work refers to the use of ascorbic acid or Ascorbic Acid Injection, USP. Confusion in the literature could be reduced by changing the official title from Ascorbic Acid Injection to Sodium Ascorbate Injection.


Self-injurious behavior in schizophrenic and retarded children. Self-injurious behavior  is a problem with some children who are primarily nonverval and low-functioning. This behavior has resulted in management difficulties far out of proportion to its incidence. In the present paper, we have considered possible operant and respondent paradigms instrumental in the acquisition and maintenance of several different topographies of self-injurious behavior. Support for these paradigms was gathered from existing epidemiological literature dealing with humans and primates and from the literature concerned with treatment of self-injurious behavior. Immediate outcome and results of subsequent follow-up were presented as a function of type of intervention, the nature of positive reinforcement utilized, and the topography of the self-injurious response involved. Implications were drawn for future research and treatment.


Fetal acid-base balance. I. Interdependence of maternal and fetal PCO2 and bicarbonate concentration. This paper presents the results of measurements of the acid-base and gaseous status of maternal and fetal capillary blood at the time of amniotomy, before the onset of labor. The correlation coefficient between maternal and fetal bicarbonate concentration is 0.6, and that between maternal and fetal PCO2 is 0.31. From the results obtained, as well as from a survey of other work in the field, it is proposed that the diffusion of carbon dioxide across the placenta could not explain some of the findings and that bicarbonate concentrations equilibrate across the placenta.


Breathing patterns before death in fetal lambs. Continuous observations in 16 fetal lambs over many days before death in utero from hypoxia, asphyxia, infection, and other causes have shown that death is preceded by variable changes in breathing movements. There was always a prolonged period of apnea succeeded by continuous abnormal breathing in six lambs and by gasping or brief episodes of abnormal breathing in the remainder. These abnormal patterns are described as a warning of what may be encountered in the human fetus.


The evaluation of continuous fetal heart rate monitoring in high-risk pregnancy. Intrapartum electronic fetal heart rate monitoring of the high-risk obstetric patient is thought to improve the perinatal outcome. A prospective randomized study of 483 high-risk obstetric patients in labor was carried out comparing the effectiveness of electronic fetal monitoring with auscultation of fetal heart tones. The infant outcome was measured by neonatal death, Apgar scores, cord blood gases, and neonatal nursery morbidity. There were no differences in the infant outcomes in any measured category between the electronically monitored group and the auscultated group. The cesarean section rate was markedly increased in the monitored group (16.5 vs. 6.8 per cent in the auscultated patients). The presumptive benefits of electronic fetal monitoring for improving fetal outcome were not found in this study.


Evaluation of redox state of isolated perfused rat lung. The metabolic responsiveness of lung tissue to inhibition of oxidative metabolism was determined by measurement of the redox state of the isolated perfused and ventilated rat lung. Changes in redox state were evaluated by fluorescence from the lung surface at wavelengths suitable for reduced pyridine nucleotides and by measurement of the ratios of redox couples in rapidly frozen lung tissue. Maximal change of redox state was observed during ventilation with carbon monoxide; surface fluorescence increased 6.6%, lactate/pyruvate increased 5.8 times, glycerol 3-P/dihydroxyacetone-P increased fourfold and glutamate/alpha-ketoglutarate doubled. KCN infusion resulted in similar changes. Hypoxia produced with N2 ventilation resulted in less than maximal changes in redox couple ratios until alveolar PO2 was reduced below 0.1 mmHg. Redox changes observed during infusion of 0.5 mM aminoxyacetic acid suggested that maintenance of cytoplasmic redox state depended on functioning of a malate-aspartate "shuttle." The isolated perfused lung appears suitable to study factors controlling pulmonary parenchymal oxidative metabolism. The results emphasize the need for ventilation with CO to establish intracellular anoxia.


Altered hepatic glycogen metabolism and glucoregulatory hormones during sepsis. Levels of glucose, insulin, and glucagon in portal vein plasma and of liver glycogen and cyclic AMP and activities of glycogen synthase and phosphorylase in liver were assayed in control (CONT) rats and rats infected (INF) with Diplococcus pneumoniae. In INF rats compared with CONT rats, insulin and glucagon levels were higher (8,12,24 h). Activity of synthase I was lower (8, 12, 24 h) and of phosphorylase higher (12 and 24 h) in INF rats. Cyclic AMP levels were higher in INF rats at 12 and 24 h. Total synthase activity was lower in INF rats at 24 h. Glucose given intravenously increased glycogen less in INF than in CONT rats and activated synthase and inactivated phosphorylase in all animals except at 24 h in INF rats. However, in situ perfusion of the livers at 24 h with glucose in buffer decreased phosphorylase activities in all animals and increased synthase I activities in CONT but not INF rats.


Histidine decarboxylase-mediated histamine synthesis in glomeruli from rat kidneys. Enzymatic histamine synthesis by renal glomeruli of the rat has been examined. Assays of the partially purified enzyme demonstrated a pH optimum of 6.2, a Michaelis-Menten constant of 2.4 X 10(-4) histidine, and lack of potentiation by benzene. These data thus indicate that renal glomeruli contain the histidine-specific histidine decarboxylase.


Effects of temperature transients on gas exchange and acid-base status of turtles. Pulmonary ventilation (VE), O2 consumption (VO2), and CO2 production (VCO2) were measured continuously on each of 10 turtles, Pseudemys scripta elegans, at 20 degrees C, during and for 1 h after heating to 30 degrees C and during and for 1 h after cooling to 20 degrees C. In seven of the animals, arterial blood was sampled at the three temperature plateaus. Ventilatory ratios (VE/VO2 and VE/VCO2) and metabolic rate adjusted promptly to temperature change, stabilizing at values similar to those observed previously in turtles following 1 day or more at each temperature. Likewise, mean blood pH and PCO2 values conformed both in absolute values and in temperature-dependence to data previously obtained from animals exposed to the various temperatures for longer time periods or from turtle blood thermally equilibrated in vitro. Total plasma [CO2] did not change significantly, suggesting that steady-state CO2 exchange prevailed throughout the experiment. In accordance with this, R(VCO2/VO2) did not change significantly during the 20-30 degrees C transition; however, R rose during the 30-20 degrees C transition, suggesting possible hyperventilation at this stage. We conclude that the respiratory control of blood acid-base status adjusts rapidly to temperature change in the turtle and its adjustment minimizes disturbance to CO2 balance.


Direct effects of various catecholamines on liver circulation in dogs. As measured by electromagnetic blood flow transducers, direct infusion of epinephrine, norepinephrine, and dopamine into the portal vein (PV) produced a 40-50% decrease in hepatic arterial (HA) blood flow; isoproterenol increased HA flow by about 69%. No changes in PV flow or pressure were observed. Direct HA infusion of the vasoconstrictors decreased HA flow by amounts comparable to those occurring after PV infusion. However, HA infusion of isoproterenol increased HA flow only 15% suggesting a difference in beta-receptor population in the two vessels. When infused directly into the superior mesenteric artery (SMA), epinephrine and norepinephrine reduced SMA flow by about 45% and PV flow by 20-25%; HA flow increased 6-8%. Infusion of isoproterenol and dopamine into SMA increased SMA flow by 115% and 206% and PV flow by 60% and 70%, respectively, whereas HA flow decreased by 25% and 50%. Portal vein pressure increased less than 3 mmHg. Alpha- and beta-receptor blockade of the liver did not change significantly the alterations in hepatic arterial blood flow that were secondary to changes in portal venous blood flow. It is likely that regulation of hepatic arterial flow resides in mechanisms located within the liver sinusoids.


Clinical response and plasma levels: effect of dose, dosage schedules, and drug interactions on plasma chlorpromazine levels. Plasma chlorpromazine (CPZ) levels of 50 psychotic inpatients were measured by gas liquid chromatography; the clinical progress of 29 of these patients with acute psychoses was also assessed. CPZ levels of 50-300 ng/ml were usually associated with clinical improvement; there was also a relationship between CPZ levels and increases in certain symptoms. The 50-300 ng/ml level was best attained by doses of 400-800 mg/day. Trihexyphenidyl decreased plasma CPZ by a mean of 44.7% in 12 of 15 patients. A single 400-800-mg dose of CPZ at bedtime produced steady states equal to or better than those achieved with multiple doses. Those patients who failed to attain CPZ levels of more than 70 ng/ml despite doses of 400-1000 mg/day were receiving lithium throughout the study and had discharge diagnoses of manic-depressive psychosis, manic type, and schizo-affective schizophrenia--a finding with implications for future research.


Accidental hypothermia: core rewarming with partial bypass. Three patients with profound hypothermia were treated by rewarming on partial bypass. Two surivived and have normal mental and metabolic functions. The resuscitation of the hypothermic patient should be approached with enthusiasm since the outcome is often much better than expected from initial vital signs and neurologic examination. To avoid ventricular fibrillation the patient should be handled gently and an effort should be made to keep the patient well oxygenated and the pH normal. Blood gases should be measured often and corrected for temperature. The potassium concentration and hydration status of the patient should also be monitored closely. The rewarming of profoundly hypothermic patients can readily be accomplished with a pump oxygenator and heat exchanger. The indications for this method are not established from our small experience and the few cases reported in the literature. Certainly ventricular fibrillation is a compelling indication. Patients with frozen extremities might also benefit from this method since theoretically tissue salvage would be increased. Finally, those patients who do not respond rapidly to external rewarming may be at less risk of ventricular fibrillation if rewarmed on bypass.


Premedicant drugs and gastric juice pH and volume in pediatric patients. The effects of premedication on gastric juice volume and pH were evaluated in five groups of 206 pediatric patients undergoing elective surgical procedures: Group 1 (Control) received no premedication; Group 2 was given morphine sulfate and pentobarbital as premedicants. The other groups received, in addition to morphine and pentobarbital, atropine (Group 3), scopolamine (Group 4), or glycopyrrolate (Group 5). After endotracheal intubation, gastric aspirates were examined for volume, pH and color. Neither premedication with morphine and pentobarbital nor addition of atropine or scopolamine to the premedication significantly altered volume. In patients treated with glycopyrrolate, volume was reduced to less than a third of that of patients in Group 1 (P less than 0.001), and the percentage of pH's higher than 2.5 was significantly greater than in other groups. The incidences of unobtainable samples and samples with pH's higher than 2.5 were greatest with atropine (32.0 per cent, P less than 0.05) and glycopyrrolate (58.1 per cent, P less than 0.01). In 60 per cent of the bile-stained specimens, pH's were below 2.5. It is concluded that because of its selective inhibitory effect on gastric acid secretions, glycopyrrolate appears superior to other anticholinergic drugs. The reduction of gastric juice volume and acidity produced by glycopyrrolate would have important clinical implications in case of accidental aspiration. It is also concluded that bile staining of gastric contents is not a reliable indicator of gastric juice pH.


Arterial hypoxemia caused by intravenous ketamine. Ketamine given IV in a dose of 2 mg/kg caused a significant reduction in Pao2 in 7 patients spontaneously breathing with an unassisted airway. Under the same conditions, in 7 patients, ketamine (2 mg/kg IV) preceded by diazepam (0.2 mg/kg IV) also caused a reduction in Pao2 not significantly different from that caused by ketamine. In some patients, alarmingly low levels of Pao2 ( less than or equal to 40 torr) were seen following ketamine administration. Based on these findings, the authors recommend that O2 and ventilatory assistance accompany ketamine given IV for anesthesia.


Clinical investigation of a new intravenous anesthetic--etoxadrol hydrochloride (CL-1848; U-37862A). Twenty-eight patients were anesthetized with etoxadrol as primary agent. The anesthesia produced was characterized by profound analgesia and amnesia, while pharyngeal and laryngeal reflexes, as well as swallowing and lid reflexes, remained active. Systolic, diastolic, and pulse pressure were slightly increased, with associated tachycardia and tachypnea. A dose of 0.75 mg/kg produced anesthesia for an average of 26 (14 to 53) minutes. Alternating nystagmus was present for several hours and associated with dreams and/or visions that were pleasing to most patients. Six patients, however, had unpleasant dreams for up to 24 hours. One patient given an excessive dose (4.65 mg/kg) was cataleptic, amnesic, and analgesic for 6 days. The occurrence of unpleasant dreams and aberrations in over 20% of the patients suggests that the drug probably has little usefulness in anesthesia. However, the extreme safety of the drug (an LD50 equal to some 20 to 40 times the ED50) and the prolonged analgesia justified clinical testing. There was no evidence of metabolic or systemic organ system change from any of the clinical laboratory studies.


The effect of enflurane, isoflurane, fluroxene, methoxyflurane and diethyl ether anesthesia on ouabain tolerance in the dog. Digitalis tolerance in dogs anesthetized with enflurane, isoflurane, fluroxene, methoxyflurane, and diethyl ether was compared with that in dogs anesthetized with pentobarbital. Ouabain dosage needed to cause ventricular tachycardia was significantly higher than that of pentobarbital with all agents except fluroxene, as was the LD50. The relative potency of these anesthetics in converting ouabain-induced ventricular tachycardia to sinus rhythm, in order of descending effectiveness, was: diethyl ether, methoxyflurane, enflurane, fluroxene, isoflurane, pentobarbital.


An evaluation of memory under regional anesthesia with IV lorazepam as a premedicant. Forty male volunteer patients undergoing regional anesthesia were evaluated for alertness and memory in a double-blind study which compared IV lorazepam (4 mg) plus IM meperidine (50 mg) with IV placebo plus IM meperidine (50 mg) as premedicants. The data indicate no significant retrograde effects. Significant differences at all measurement points from 20 minutes to 3 hours following IV drug administration indicated greater sedation and less recall and recognition of events and stimuli (auditory and visual) with the combination of lorazepam and meperidine than with placebo and meperidine. A significant number of patients over age 40 were judged to be excessively sedated after administration of lorazepam. However, respiration, blood pressure, and pulse did not appear to be differentially affected by lorazepam and there were no adverse changes even in patients considered to be oversedated.


[2 cases of chylothorax caused by catheterization of left large venous trunks]. The authors present two cases of almost certain catheterization of the thoracic duct, by infusion catheters placed in the large veins at the base of the neck (once in the left internal jugular, once in the left subclavian). Their clinical manifestaztions are the existence of a rapidly progressive pleural effusion, apart from the abnormal course of the catheter. Removal of the catheter is sufficient to lead to the disappearance of the symptomatology, so that it is difficult to speak of an accident as such, but rather of an incident. These incidents could be avoided by systematic radiography from the time that the catheter is placed in position or the positioning of the latter under control by brilliance amplifier.


[Complications of lignocaine].

[Complications of lignocaine]. In three cases, following cardiac surgery operations, the appearance of a sudden loss of consciousness followed by an epileptic - like crisis was recorded. The accidents have a relationship with an administration of lignocaine in the form of a perfusion. Description of the observations and discussion of the toxicity of the product.


[Use of a new long acting local anesthetic, etidocaine in anesthesia-resuscitation (Duranest)]. Our experiment covered 123 patients who were given this new preparation with long action for various types of local and regional anesthesia. In 9 cases of peridural administration for several days, the total doses were very high and tolerance excellent. Results were good for all local and regional anesthesia for which this product was used: - Good local and general tolerance - Short latency period - Duration constantly and regularly prolonged: an average of 3 hours for non-adrenalin forms and 5 hours for adrenalin forms.


[The MA 1 B respirator. Apropos of an experiment involving 91,300 hours of function]. Description of the working principle and the characteristics of the Bennet MA 1 B respirator, which is of the release of volume type with electro-mechanical functioning. Study of the advantages and drawbacks. During an experiment involving 91,300 hours of utilization of 21 sets of apparatus, 12 breakdowns concerning a mechanical or electrical component were recorded. No incident in the functioning of the electronic part of the system of control or of surveillance was noted. The reliability of the apparatus appears on the whole to be satisfactory, taking into account its remarkable working posibilities.


[Enzyme system and coenzymes involved in the energy metabolism of leukocytes. Function and metabolism of polymorphonuclear neutrophils]. Mitochondria may be isolated from various types of leukocyte (neutrophil polymorphs and lymphocytes from human blood, neutrophil polymorphs and macrophages from peritoneal exudates of the guinea pig) after destruction by heparin of the cell membrane. This procedure is very simple and less traumatic for these subcellular structures than the usual mechanical procedures. The enzyme activities of the respiratory chain and oxygen consumption may be measured in these mitochondrial preparations. The oxygen consumption is determined using oxyhemoglobin which serves both as oxygen donor, as in the respiratory system in vivo, and as indicator of the reaction at 435.8 nm. The integrity of the mitochondria may be demonstrated by determination of the "acceptor control index", the existence of ADP phosphorylation coupled with oxygen consumption (phosphorylating oxidation) was proved in all the cells studied even if the ADP/O ratio can only be calculated for certain of them (lymphocytes, macrophages). In these cases, the ratios obtained are close to theoretical values whatever the oxidation substrate used. The mitochondria of leukemic cells have a higher oxidation activity than the corresponding reference cells. Determination of leukocyte coenzymes by enzyme cycling (NAD, NADH, NADP, NADPH) showed the following facts: -- Generally, the NAD concentrations remain constant, those of NADH increase whilst those of NADP and NADPH fall during incubation of neutrophil polymorphs in Dulbecco's medium. -- The metabolic changes observed during S. albi heat-induced endocytosis are in favour of simultaneous stimulation of NADH oxidase and NADPH oxidase in human polymorphs, and of NADPH oxidase in the corresponding cells of peritoneal exudates in guinea pigs.


[Use of the Cessna 206 for medical evacuations]. The CESSNA 206 aircraft belonging to the National Police Force has been used to effect medical evacuations. After describing the specifications of this aircraft and underlining its advantages (speed, autonomy, cost price) and its disadvantages (very small cabin, necessity of prepared landing grounds, sound level) compared with the helicopter generally used in similar circumstances, the authors conclude that this new aircraft could be used to advantage for evacuating medium-serious cases over distances greater than 200 kilometer provided that the cabin was modified. Even more than with the helicopter, the pilot must remain the judge concerning the technical flying possibilities of the aircraft, which is not pressurized and which has no defrosting equipment.


[Incidence of deep venous thromboses in traumatology and methods of early detection]. In this work carried out at the Traumatology Centre in Strasbourg, out of 227 patients operated for a fracture situated between the pelvis and the tibial plateau, the authors endeavoured to show a thrombosis of the lower limbs. The means of exploration included in addition to the clinical and biological blood tests an isotopic, rheographic and phlebographic surveillance. The latter three ways of exploration are particularly viewed in this study, during which it was possible to demonstrate 44 p. 100 thrombo-embolic manifestations of which 38 p. 100 were detected only by the isotopic method, the phlebographic check for its part confirming 80 p. 100 of the clinically undeclared cases of thrombosis.


Detection of mutant hemoglobins with altered affinity for oxygen. A simplified technique. The detection of high- or low-affinity hemoglobins in subjects with polycythemia or anemia is difficult for most physicians because of the requirement for special equipment to do oxygen-hemoglobin dissociation curves. Measurement of the pH, oxygen tension, and oxygen saturation of antecubital venous blood with instruments present in most clinical chemistry laboratories permits an estimate of the strength of oxygen binding to hemoglobin. An equation can be used to convert the venous oxygen tension (standardized to pH 7.4) and the oxygen saturation to the P50 of the oxygen-hemoglobin dissociation curve on which the observed point falls. The data indicate that this method is a reliable initial step in the identification of a hemoglobin with abnormal affinity for oxygen and may be applied to population studies, since reliable results are obtained with venous blood stored at 4 degrees C for up to 24 hours.


[Post-traumatic fat embolism of the hemorrhagic pulmonary edema type]. The authors present a case of post-traumatic fat embolus, which was remarkable by the very haemorrhagic nature of the pulmonary oedema, which healed without sequel.


Antigenemia in fulminant pneumococcemia. Two asplenic patients with fulminant pneumococcemia developed purpura and coagulopathy. Levels of C3 and C4 in the serum were low. Both patients had high levels of circulating capsular polysaccharide, and one patient had visible diplococci on a smear of the peripheral blood. Pneumococcal group C polysaccharide (C-substance) was detected in the serum of one of the patients. Possible pathogenetic relation of circulating pneumococcal antigens to the development of coagulopathy in pneumococcemia is discussed.


[Acute tamponade: complication of resuscitation (cardiac perforation by a subclavian catheter)]. The authors report a case of acute tamponnade as result of the collection under pressure of a lipid solution in the pericardium following perforation of the right atrium by a sub-clavian catheter. Diagnosis and treatment were practically siumultaneous. Aspiration of the lipid solution (450 ml) via the catheter confirmed the diagnosis and successfully treated the problem. With regard to the case, the following are discussed: - diagnostic features of acute tamponnade (acetiological, clinical, heemodynamic, electrocardiographic and radiological signs); - the physiopathological mechanism of tamponnade with changes in intra-pericardiac and ventricular pressure/volume cures; - finally, the management of tamponnade.


RNA-protein interactions in alfalfa mosaic virus. Particles of the bottom component of alfalfa mosaic virus have a compact bacilliform structure at neutral pH. When the pH is raised to 8.3 the structure unfolds. Since the particle weight does not change it is concluded that the protein subunits remain attached to the RNA. The particle weight of the spheroidal top component a of the virus is halved when the particles unfold at pH 8.3. This can be explained by the fact that these particles contain two RNA molecules of identical size. Free RNA molecules of alfalfa mosaic virus are able to withdraw protein subunits from intact particles of the virus. It is demonstrated that per RNA molecule there are a few sites with a high affinity for coat protein. Possibly these are the sites where the coat protein plays its role in activating the genome.


Human enolase isozymes: electrophoretic and biochemical evidence for three loci. 1. Four major enolase isozymes have been identified in human tissues and are referred to as L, M, 'intermediate' and 'fast'. The M isozyme is the major form found in skeletal muscle and heart extracts and the L isozyme the major form found in extracts of liver and most other tissues. The 'intermediate' and 'fast' isozymes are most active in brain but are observed as weak components in most other tissues including heart but are not seen in skeletal muscle. It was observed that during fetal development of heart and skeletal muscle the L form declines in activity while the M form increases in activity. 2. The kinetic properties, heat stabilities and molecular sizes of the main enolase isozymes have been compared. Although the isozymes share many features in common, the 'fast' isozyme is more stable when subjected to heat treatment than either the L or M isozymes. Further, the 'fast' isozyme retains its dimeric structure and activity in the absence of magnesium ions while the L and M isozymes dissociate and lose activity. The 'intermediate' isozyme has properties which are intermediate to those of the L and 'fast' isozymes. 3. The 'intermediate' isozyme can be partially dissociated to equal quantities of L and 'fast' isozymes by storage at room temperature or by freezing and thawing in the presence of 2 M-NaCl. Conversely, mixtures of L with 'fast' and M with 'fast' give rise to an 'intermediate' isozyme after freezing and thawing. 4. Evidence derived from this study has led to the suggestion that three separate gene loci are involved in the determination of human enolase. It is proposed that one of these, ENO1, determines the L isozyme which is the homodimer alphaalpha; another locus, ENO2, determines the 'fast' isozyme which is the homodiner betabeta; and the third locus, ENO3, determines the M isozyme which is the homodimer gammagamma. The 'intermediate' isozyme seen as a strong component in brain and as a weak component in most other tissues is thought to be the heterodimer alphabeta. In heart however it is probably mainly betagamma.


Adenylate kinases in man: evidence for a third locus. The tissue distribution of the adenylate kinase isozymes in man has been examined using various substrates. The isozymes attributable to the AK1 and AK2 loci were identified, and an additional set of isozymes probably attributable to a third locus was also found. This locus has been provisionally designated AK3. The AK3 isozymes show activity with either GTP + AMP or ITP + AMP but do not show activity with ATP + AMP. They also differ from the AK1 and AK2 isozymes in electrophoretic mobility and from the AK1 isozymes in being resistant to silver inhibition. They are similar in molecular size to the AK1 isozymes whereas the AK2 isozymes are apparently larger. The AK3 isozymes evidently correspond to the enzyme nucleosidetriphosphate-adenylate kinase (2.7.4.10). Somatic cell hybrid studies indicate that the AK3 locus is not syntenic with that of AK2 (chromosome 1). The AK3 locus is however, probably syntenic with the AK1 locus, on chromosome 9. Genetically determined variation of AK3 has not been seen in a survey of about 80 individuals.


[Criteria for evaluating calcium carbonate from the point of view of chlortetracycline biosynthesis]. Calcium carbonate is added to fermentation media in biosynthesis of tetracyclines for providing definite pH values and binding tetracycline into insoluble complexes. Seven different samples were studied with respect to their physical properties, such as the microscopic size of the particles, their form, capacity for agglomeration, specific volume, rate of the particle precipitation and chemical properties, such as purity, buffer capacity, effect on the medium pH before and after sterilization. The above properties were studied in comparison with activity chlortetracycline biosynthesis. Microfine calcium carbonate proved to be the best from the point of view of productivity of Str. aureofaciens. With its use the activity of the culture fluid increased by 20 per cent as compared to the other samples. The titration curve of the sample had the lowest bend.


[Study of the penicillin amidase from E. coli. An ultrasonic method of studying the ph- and temperature-conformational transitions in the active center of the enzyme]. pH and temperature conformation transitions in the active center of penicillin amidase i.e. penicillinamidohydrolase E.C. 3.5.I.II were investigated by means of the kinetic method and a new ultrasonic method. It was shown that the catalytic activity of the enzyme was controlled by 2 ionogenic groups with pK 6.1 and 10.2. The study of penicillinamidase by means of the ultrasonic method showed that the ionogenic group with pK 10 was responsible for maintaining the catalytically active conformation of the enzyme active center. Investigation of the temperature relation between the kinetic parameters of the enzymatic hydrolysis of benzylpenicillin catalyzed by penicillin amidase and the data on the effect of ultrasound on the enzyme showed that the enzyme was subjected to the temperature conformation transiton. The temperature and thermodynamic parameters of the conformation transition were determinded (T=318 degrees K, delta H=81 kcal/mole and delta S=255 e.u.). The structure of the active center of the enzyme is discussed on the basis of the data obtained.


[Chromatographic control of the process of rifamycin B isolation and purification]. A method of chromatography in a thin layer of Silica Cel No. 2 was developed for rifamicin B, rifamicin complex and some admixtures. Extracts of rifamicin B in an organic solvent, aqueous buffer reextracts and dry preparations of rifamicin B were analyzed with the above method. A half-quantitative chromatographic procedure for estimation of the main admixture in dry preparations was proposed.


Regulatory mast cells. I Suppressive action of their products on an in vitro primary immune reaction. Products of mast cell degranulation, as well as histamine and serotonin, were added to a Mishell and Dutton preparation for in vitro primary immunisation (induction of IgM antibody formation) to sheep or horse red blood cells. Degranulation products were either obatined beforehand by reacting passively sensitised mast cells with the corresponding antigen (unrelated to or identical with the in vitro immunising antigen) or liberated into the culture medium where mast cells actively sensitised to the in vitro immunising antigen had been added. A 46 to 72 % reduction of direct (IgM) plaque forming cells was observed in all cases. This reduction was prevented by anti-histamine. The responsible mediators were active in the 0 to 24 hour period after antigen introduction. The anaphylactic degranulating antibodies triggering this inhibitory activity were found to be thermolabile in one experiment. An in vivo-induced mast cell degranulation led to a reduced formation of plaque forming cells. The enhancing and immunoregulatory activity of anaphylactic mouse antibodies is therefore tentatively and at least partially attributed to their capacity to degranulate mast cells after contact with the antigen.


[Relationship between the antibacterial activity and the acceptor properties of certain penicillins]. Antibacterial activity of 2 natural and 12 semisynthetic penicillins against 5 strains of grampositive bacteria was determined and quantum chemical estimation of their molecules was performed with the Hukkel method. The data were indicative of the fact that antibacterial activity of the penicillins was connected with the acceptor properties of their molecules.


[Effect of different factors on the rate of benzylpenicillin conversion in an aqueous system-butylacetate system]. It was shown that the mass-transfer coefficient of non-dissociated benzylpenicillin during extraction was more than 1000 times higher than that during reextraction. For the latter it was found that the aqueous phase exerted limiting resistance during mass-transfer of benzylpenicillin. The effect of a number of factors on the kinetics was shown. These factors were the following: hydrodynamic conditions, temperature, pH of the aqueous phase.


[Regulation of tetracycline biosynthesis by controlling the growth of the producer]. Regulation of the rate growth of Act. aureofaciens in batch fermentation by maintaining the concentrations of phosphorus, ammonium nitrogen, glucose and pH values at the levels favourable for intensive growth at the beginning of the process and after accumulation of the biomass at the levels optimal for retarded growth of the organism resulted in significant prolongation of the period of intensive antibiotic production, i.e. intensification of the fermentation process. Microscopic investigation of the organism development under conditions of regulated fermentation revealed the presence of significant amounts of free peripheral highly basophilic hyphae for a prolonged period of time. The hyphae possessed a capacity for growth and intensive metabolism unlike the control culture which was liable to early autolysis.


[Study of the biosynthesis of a carbohydrate fragment of rubomycin]. When the rubomycin-producing organism was grown in the presence of an evenly labeled C14-glucose and various non-labeled carbon sources, the carbohydrate part of the antibiotic also became evently labeled. The activity of I carbon atom of rubomycin sugar grown against the background of non-labeled glycerol was almost 3 times higher than the respective value of the aglycone activity. C14-acetate, C14-propionate, C14-methionine and daunosamine were not incorporated. The data are indicative of the fact that there was no splitting of the glucose carbon skeleton during conversion of glucose into amino sugar.


[Physicochemical properties of 7-phenylacetamidodesacetoxy-cephalosporanic acid]. 7-phenylacetamidodesacetoxycephalosporanic acid was prepared by transformation of benzylpenicillin. The acid was subjected to potentiometric titration and investigation of its electrophoretic mobility at wide pH ranges. The data of the potentiometric titration and electrophoresis were used for calculation of the acid ionization constant. Minimum solubility of the acid was determined and the curve of its solubility at wide pH ranges was estimated.


Beta-glucosidase of Trichoderma: its biosynthesis and role in saccharification of cellulose. The extracellular beta-glucosidase of Trichoderma viride generally is present in low levels when the organism is cultured on cellulose because it is inactivated under the acid conditions which develop in the medium while the other enzymes of the cellulase complex are more stable. With the appropriate pH control, inactivation of beta-glucosidase is prevented and the activity of this enzyme increases during growth. In the saccharification of crystalline cellulose, or of cellulose at low concentrations, much of the glucose produced is the result of the cleavage of cellobiose by beta-glucosidase. However when high concentrations (10%) of pretreated cellulose are saccharified, significant quantities of glucose are produced by action of enzymes other than beta-glucosidase.


Growth of Fusarium moniliforme on carob aqueous extract and nutritional evaluation of its biomass. Fusarium moniliforme was cultured semicontinuously on a carob medium in a 14-liter fermentor (8.5-liter working volume). The growth medium provided 2.4% carob sugar, 0.72% NH4H2PO4, and 0.03% MgSO4-7H2O. The biomass harvest was 8.8 g/liter per day. Ninety percent of the sugars were consumed, and the pH dropped from 5.9 to about 3.7. The crude protein (N X 6.25) of the spray-dried mycelium was 380 g/kg, 300 g/kg for the true protein (Lowry), and 4.8 g/kg for the (Folin-Denis) tannic acid. The mycelium was evaluated nutritionally with the weanling rat as experimental animal. The protein efficiency ratio and net protein utilization values for the unsupplemented mycelium were 1.15 and 0.42, respectively, and for the mycelium supplemented with DL-methionine (5 g/kg) they were 2.31 and 0.72, respectively. No growth depression was observed in the experimental rats, and on dissection of the carcasses the internal organs were found to be normal.


Incorporation of proline into prodigiosin by a Put mutant of Serratia marcesens. A Put mutant of Serratia marcescens, deficient in proline oxidase and therefore unable to degrade proline, was used to assay for an enzymatic reaction responsible for incorporation of proline into prodigiosin. The reaction had a pH optimum of 7.5 and a Km of 1.1 X 10(-4) M at 27 C. At temperatures above 27 C, the velocity of the reaction decreased with increasing temperature and little activity was detected at 42 C. Activity of the enzyme was directly proportional to the quantity of pigment formed and was inhibited by thioproline, a substrate analog. These data suggested the presence of a unique and specific enzyme in the biosynthetic pathway for prodigiosin.


Enzymatic conversion of sterigmatocystin into aflatoxin B1 by cell-free extracts of Aspergillus parasiticus. A cell-free extract, prepared from Aspergillus parasiticus ATCC 15517 grown in synthetic medium, was active in converting [14C]sterigmatocystin into aflatoxin B1 in the presence of reduced nicotinamide adenine dinucleotide phosphate. The activity was demonstrated by the time course of conversion and the linear dependence of the yield of product on enzyme concentrations. Optimum activity was obtained at pH 7.5 to 7.8 at 27 C. The results confirm sterigmatocystin as a biogenetic precursor of aflatoxin B1. Techniques were developed for enzymatic studies on aflatoxin biosynthesis.


Cord gamma glutamyl transpeptidase activity and neonatal jaundice. gamma Glutamyl transpeptidase (GGT) activity was measured in normal neonates and in maternal serum post partum. Levels were above the normal adult range (35I U/1) in all neonates and a significant correlation was bound between enzyme activity and bilirubin levels on day 7(P less than 0-005). The mean bilirubin level on days 4 and 7 was higher in babies with cord values less than 90 IU/1. In certain circumstances increased plasma GGT activity may serve as an index of enzyme induction. However, our results suggest that raised levels in the neonate may reflect hepatic microsomal damage with subsequent impairment of bilirubin conjugation. Further evaluative studies of cord GGT activity in neonates at risk, with a view to early prophylactic or therapeutic measures, are indicated.


Cardiorespiratory response to feeding in newborn infants. Milk feeds were given through indwelling nasogastric tubes to 14 infants with respiratory distress. Similar cardiorespiratory disturbances were observed when the infants were fed (5 ml/kg per feed) human milk, cow's mild, or distilled water. Pao2 fell after a feed but recovered to the prefeed  value at 30 minutes, at which time Paco2 had fallen and the respiratory rate had increased. No changes in pH, heart rate, or blood pressure were observed. Portal sinus pressures rose after feeding in association with an increase in central venous pressure. In contrast, when the ill infants were fed human milk at a volume of 2-5 ml/kg per feed no consistent changes in any of the measurements were found. These studies suggested that the cardiorespiratory effects were related to volume displacement resulting from feeds being introduced into the stomach. The relation of the increase in central venous pressure and the magnitude and direction of shunting in infants with the respiratory distress syndrome is uncertain. Adverse effects may be avoided by giving smaller, and therefore even more frequent, feeds.


Clinical experience with the Teflo disposable membrane oxygenator. A simple, inexpensive, highly efficient disposable membrane oxygenator with low priming volume and a microporous membrane recently has become available. Animal and clinical investigations of its use have been most satisfactory, and clinical experience now has been extended to include 285 patients. Its primary advantage has been the ability to control oxygenation and carbon dioxide separately. Disadvantages have included the somewhat increased complexity of the system as compared with bubble oxygenator systems, the necessity of converting pumps for its use, and excessive condensation of water vapor in the gas phase of the oxygenator unless certain precautions are followed.


Hypothermia and rewarming by peritoneal dialysis and temperature-controlled inhalate. In 12 rabbits hypothermia and rewarming were induced with temperature-controlled circulating peritoneal dialysis in combination with temperature-controlled hypoxic and hypercapnic gas mixtures. The average cooling time necessary for the esophageal temperature to decrease from 37.7 degrees +/- 0.7 to 20.6 degrees +/- 1.0 degrees C was 81 +/- 34 minutes with a range of 41 to 150 minutes. The average warming time for esophageal temperature to increase from 20.6 degrees +/- 1.0 degrees C to 35.2 degrees +/- 1.8 degrees C was 90 +/- 35 minutes. Time of cooling was related to the proportions of inspired carbon dioxide and oxygen. In contrast to surface and bypass methods, esophageal and muscular temperatures agreed very closely, suggesting an absence of regional temperature gradients.


Angioplastic repair of a ruptured pulmonary artery aneurysm. A 28-year-old woman had a ruptured solitary pulmonary arterial aneurysm which was successfully treated by pericardial patch graft. The etiology of the aneurysm could not be determined although the patient had had several episodes of blunt chest trauma in the past. Unusual aspects of this case include: location of the aneurysm in the intermediate portion of the left pulmonary artery within the major intralobar fissure, intrapleural rupture, preoperative diagnosis by pulmonary angiography, and an incidentally discovered histopathological abnormality of the aneurysm itself for which no satisfactory explanation has been found.


Suppression of renin release by timolol. The beta-adrenergic blocking agent, timolol, administered to resting rabbits as an i.v. bolus (0.125 mg/kg) sustained by a 2-hr infusion at 0.0625 mg/kg/hr, caused significant depression of plasma renin activity (PRA) to 49% of the control level. Significant correlations emerged between the fall in mean blood pressure and changes in both heart rate and PRA. Timolol also antagonized isoprenaline-induced renin release. In anaesthetized normal rats, timolol (0.2 mg/kg i.p.) suppressed mean plasma renin concentration (PRC) to 16% of the pre-treatment value. Furthermore, the mean PRC of normal rats, bled immediately after decapitation, to avoid stimulating renin secretion, was reduced by 55% one hr after i.p. injection of timolol. The potency of timolol in this respect was 8 times that of dl-propranolol. Thus, in rabbits and rats, timolol effectively depresses both basal and stimulated plasma renin levels.


Effect of sodium acetate infusion on renal function in the dog. Sodium acetate (AC) is routinely used in dialysis solutions in hemodialysis units as it provides a ready source of fixed base. The renal dynamics of the buffer salt are not well known. Anesthetized dogs received i.v. infusions of AC at a rate of 40 to 600 muEq/kg/min. Urinary AC and bicarbonate excretion was measured and correlated with PAH and creatinine clearance and urinary electrolyte excretion. AC appeared in the urine as quickly as it was detected in the plasma. Its excretion gradually increased and tended to parallel bicarbonate excretion. PAH clearance increased with the advent of AC infusion, plasma pH and bicarbonate increase as AC is introduced. AC infusion caused an immediate drop in blood pressure. The systolic blood pressure returned to control in 3-5 min; there was a more gradual return in diastolic pressure. We conclude that AC is filtered and excreted into the urine at low plasma levels and has a low renal threshold.


Effects of intravenous anesthetics on continuous avoidance behavior in the rat. The actions of ketamine, etoxadrol and thiopental on continuous avoidance behavior in the rat were evaluated following i.v. administration. Ketamine and thiopental exerted only a depressant effect on avoidance behavior which was attributable to the anesthetic properties of the drugs. In contrast, biphasic activity was displayed with etoxadrol; avoidance responding was increased at low doses and decreased at high doses. A persistent rebound stimulation of responding occurred upon recovery from the anesthetic effect of high doses of etoxadrol. The effects of these drugs on continuous avoidance behavior do not correlate with previously reported effects on the disposition of brain monaoamines.


Drugs and PGO waves in the lateral geniculate body of the curarized cat. V. Miscellaneous compounds. Synopsis of the role of central neurotransmitters on PGO wave activity. In the last part of this series we have studied the effects of various drugs on ponto-geniculo-occipital (PGO) waves induced by the benzoquinolizine derivative, Ro 4-1284 (PGO(1284)), and by the inhibitor of trypotophan hydroxylase, p-chlorophenylalanine (PGO(PCPA)), and continuously recorder and counted in the lateral geniculate bodies (LGB) of unanaesthetized and immobilizedcats. The major aim of this study was to test the specificity of drug-induced alterations of the PGO wave activity suggested by the previous investigations. Hypnotics-sedatives of different classes had no significant effects in doses that did not markedly alter the electrical background activity in the LGB. A notable exception was gamma-hydroxybutyric acid which increased the density of PGO(1284) and PGO(PCPA). A number of neuroleptics were found inactive; sulpiride surprisingly decreased the density of PGO(1284). Bulbocapnine had a similar effect. Convulsants in subconvulsive doses did not uniformly affect PGO waves; while pentetrazole had no consistent effect, strychnine decreased and picrotoxin increased the density of PGO(1284). High doses of morphine, methadone and meperidine decreased the PGO(1284). Ethanol was inactive even in high doses. Caffeine and mefexamide reduced the density of PGO(1284). Mepiprazol was the most potent depressant of PGO(1284, probably by inhibiting the uptake of 5-HT. Mescaline was a weak depressor of PGO(1284). p-Chloromethamphetamine induced PGO waves in untreated cats less consitently than did PCPA. Amantadine reduced the amplitude of PGO waves due to a central antinicotinic action. The results of this study and of the whole series suggested a tentative scheme of the generation and modulation of PGO waves, in which the hypothetical roles and sites of action of four central neurotransmitters are included.


Nitrogen fixation by hydrogen-utilizing bacteria. Seventeen strains of nitrogen-fixing bacteria, isolated from different habitats on hydrogen and carbon dioxide as well as on other substrates, morphologically resembled each other. All strains, including Mycobacterium flavum 301, grew autotrophically with hydrogen. The isolate strain 6 was sensitive to oxygen when dependent on N2 as nitrogen source, a consequence of the sensitivity of its nitrogenase towards oxygen. At the same time, strain 6 was sensitive to hydrogen when growing autotrophically on N2 as nitrogen source, but hydrogen did not affect acetylene reduction by these cells.


Isolation of a moderate halophilic ammonia-oxidizing bacterium, Nitrosococcus mobilis nov. sp. An ammonia-oxidizing bacterium was isolated from a sample of brackish water (North Sea, Harbour of Husum). It is a motile large coccus 1.5-1.7 mum in diameter. The extensive cytomembrane system occurring as flattened vesicles in the peripheral region of the cytoplasm and as intrusions into the center of the cytoplasm is to be emphasized as a characteristic mark of identification. The lithoautotrophically growing bacterium turned out to be an obligate halophile. Because of its physiological and morphological properties, we assigned it to the genus Nitrosococcus and propose the name Nitrosococcus mobilis.


An ammonia-oxidizing bacterium, Nitrosovibrio tenuis nov. gen. nov. sp. An ammonia-oxidizing, autotroph growing, slender, curved rod was isolated from the soil of Hawaii. It is well distinguishable from any other nitrifying bacteria thus far described by their morphology. The cells are 1.1-3.0 mum long and 0.3-0.4 mum wide. They are motile by means of 1-4 subpolar to lateral flagella. In contrast to most of the ammonia-oxidizing bacteria the isolated vibrio is void of an extensive cytomembrane system. To categorize this not yet described species we propose to create the new genus Nitrosovibrio and to classify the isolated strain as Nitrosovibrio tenuis.


Properties of heterocysts isolated with colloidal silica. A method is described for the isolation of heterocysts that are virtually free of contaminating cell debris after sonication of aerobically grown Anabaena 7120. Isolated heterocysts reduced acetylene in a light-dependent process in the absence of exogenously provided ATP; heterocysts supplied with ATP and Na2S2O4 reduced acetylene slowly in the dark but still showed a marked light activation. Nitrogenase activity was greatest in fractions containing intact heterocysts. Up to 13% of the activity of the intact filaments was accounted for in the isolated heterocyst preparation. Isolated heterocysts took up O2 in a light-independent process; O2 uptake with added NADP+ was enhanced by pyruvate, isocitrate and intermediates of the oxidative pentose pathway.


Nitrogenase activity in cultured Rhizobium sp. strain 32H1: nutritional and physical considerations. Nutritional and physical conditions affecting nitrogenase activity in the strain of "cowpea" rhizobia, 32H1, were examined using cultures grown on agar medium. Arabinose in the basic medium (CS7) could be replaced by ribose, xylose, or glycerol, but mannitol, glucose, sucrose, or galactose only supported low nitrogenase (C2H2 reduction) activity. Succinate could be replaced by pyruvate, fumarate, malate, or 2-oxoglutarate, but without any carboxylic acid, nitrogenase activity was low or undetectable unless a high level of arabinose was provided. Inositol was not essential. Several nitrogen sources could replace glutamine including glutamate, urea, (NH4)2SO4 and asparagine. The maximum nitrogenase activity of cultures grown in air at 30 degrees C was observed under assay conditions of pO2=0.20-0.25 atm and 30 degrees C incubation. Greatest activity occurred after a period of rapid bacterial growth, when viable cell count was relatively constant. Compared with results obtained on the CS7 medium, nitrogenase activity could be substantially increased and/or sustained for longer periods of time by using 12.5 MM succinate and 100 mM arabinose, by increasing phosphate concentration from 2 to 30-50 mM, or by culturing the bacteria at 25 degrees C.


Ribulose biophosphate carboxylase from Thiobacillus A2. Its purification and properties. Ribulose bisphosphate carboxylase (EC 4.1.1.39) from Thiobacillus A2 has been purified to homogeneity on the basis of polyacrylamide gel electrophoresis and U.V. analysis during sedimentation velocity studies. The enzyme had an optimum pH of about 8.2 with Tris-HCl buffers. The molecular weight was about 521000 with an Srel. of 16.9. Km for RuBP was 122 muM, for total "CO2" it was 4.17 mM, and for Mg2+ 20.0 muM. The absolute requirement for a divalent cation was satisfied by Mg2+ which was replaceable to a certain extent by Mn2+. Activity was not significantly affected by SO(2-4), SO(2-3), or S(2)O(2-3) at 1.0 mM. At this concentration S(2-) caused a 27% stimulation. All mercurials tested were inhibitory. pHMB was the most potent causing about 60% inhibition at 0.04 mM. This inhibition was reversible by low concentrations of cysteine. Cyanide was also inhibitory. Its mode of inhibition with respect to RuBP was un-competitive and with a Ki of 20 muM. Lost activity could be restored partially by GSH or Cu2+. Although azide at the concentration tested had no significant effect on enzyme activity, 2, 4-dinitrophenol at 1.0 mM caused 91% inhibition. Finally, activity was also affected by energy charge.


[Activity and properties of alkaline phosphatase in the plasma and various organs (kidney, liver, small intestine mucosa, bone) of the swine]. There was a high activity of alkaline phosphatase in the blood plasma of piglets during the first few days of live; enzyme obtained at this time had high heat stability and was readily inhibited by L-phenylalanine (5 mM). The enzyme in blood was inhibited to a greater extent than alkaline phosphatase from intestinal mucosa. With increasing age there was a fall in heat stability and in the ease with that the enzyme could be inhibited by phenylalanine. The proportion of alkaline phosphatase derived from bone and present in blood plasma increased with increasing age. Two isoenzymes were detected in liver, kidney, lung, intestinal mucosa and endometrial mucosa by electrophoresis in polyacrylamide gel. Heat lability and inhibition by phenylalanine were good criteria for differentiating different types of alkaline phosphatase in pigs. In the case of alkaline phosphatase in blood plasma, disodium phenylphosphate was split more readily than p-nitrophenyl phosphate and very much more readily than phenolphthalein diphosphate and beta-glycerophosphate.


Glycogenolysis in the heart and skeletal muscle during stimulation and blocking of cholinergic receptors. III. Action of acetylcholine under conditions of sympathicolysis. Changes in enzymatic activities in phosphorolytic and hydrolytic glycogenolysis in the working heart and in resting skeletal muscle after intravenous injection of sympathicolytic compounds were studied in vitro. Reserpine, alderline and ligation of the adrenals inhibited the influence of acetylcholine on phosphorylase, Reserpine alone or ligation of the adrenals caused an increase in hydrolytic activity similar to that produced by compounds acting on the ganglia. This confirms the concept according to which the adrenergic system (beta-receptor) inhibits hydrolytic activity during glycogenolysis.


[Morphologic (bioptic) conferences: experience with organization, tasks and results]. The article deals with the experience in organization of practical conferences in Moscow (The Central Institute of Advanced Medical Training) and in Lvov. The conferences enable pathoanatomists of various institutions to obtain consultation concerning bioptic material, to exchange view with regard for any individual observation. This new form of a collective considaration of complex biopsies should be regarded as one of important methods of the work of pathoanatomists. Some problems associated with interpretation of bioptic specimens, avoidance of errors in the process of an investigation of histological preparations, as well as with the role of the conferences in the upgrading professional qualities of pathoanatomists are discussed.


Biomedical aspects of oxygen regulator performance: I. Static characteristics. Static performance characteristics of current-inventory USAF oxygen regulators were evaluated with the use of a specialized regulator test stand. Outlet suction pressures, flows, positive pressures, and delivered oxygen dilutions were monitored and recorded as functions of operational altitudes. General findings indicated that: 1) excessive oxygen addition occurs in all models, especially at low cabin altitudes; 2) positive pressure schedules generally conform to specifications; 3) negative suction pressures for most regulators are less than -2.54 cm H2O. The validity of static evaluation is discussed and data interpretation is considered with respect to biomedical compatibility emphasizing maintenance of crewmember physiological sufficiency.


Biomedical aspects of oxygen regulator performance: II. dynamic characteristics. Several oxygen regulators were quantitatively assessed during interface with dynamic respiratory simulation and human user breathing. Continuous measurements of regulator flow, delivered pressure, and delivered oxygenation was provided during variable tidal volumes and breathing frequencies as accomplished initially with breathing simulation and subsequently with human users. The regulators demonstrated large sensitivity to flow demand and the characteristics of the inspiratory flow pulse. Large variations in outlet suction pressure and delivered oxygen concentration accompanied the variable breathing modes of human subjects. The inability of oxygen regulators to consistently provide required oxygen/air delivery is discussed with respect ot inadequate performance specifications currently used.


Short-chain fatty acid synthesis in brain. Subcellular localization and changes during development. Acetyl-CoA synthase (EC 6.2.1.1), Propionyl-CoA synthase (EC 6.2.1.-) and butyryl-CoA synthase (EC 6.2.1.2) were measured in subcellular fractions prepared by primary and density-gradient fractionation from adult rat brain by a method resulting in recoveries close to 100%. Most of the activity of the three enzymes was recovered in the crude mitochondrial fraction. On subfractionation of this crude mitochondrial fraction with continuous sucrose density gradients, most of the activity of the three enzymes was found at a higher density than NAD+-isocitrate dehydrogenase and at about the same density as glutamate dehydrogenase, confirming earlier reported data for acetyl-CoA synthase. The finding that propionyl-CoA synthase and butyryl-CoA synthase had about the same distribution in the gradients as acetyl-CoA synthase adds support to the hypothesis that mitochondria involved in the metabolism of these short-chain fatty acids (all three of which have been shown to result in a rapid and high labelling of glutamine in vivo) form a distinct subpopulation of the total mitochondrial population. The three synthase activities were found to differ from each other in their rate of change and their subcellular localization during rat brain development. This, in combination with the observation that in gradients of adult brain preparations the three activities did not completely overlap, suggests that the three synthase activities are not present in the same proportion to each other in the same subpopulation (s) of mitochondria in the brain.


Proton translocation and the respiratory nitrate reductase of Escherichia coli. Stoicheometries and rates of proton translocation associated with respiratory reduction of NO3- have been measured for spheroplasts of Escherichia coli grown anaerobically in the presence of NO3-. Observed stoicheiometries [leads to H+/NO3- ratio; P. Mitchell (1966) Chemiosmotic Coupling in Oxidative and Photosynthetic Phosphorylation, Glynn Research, Bodmin] were approx. 4 for L-malate oxidation and approx. 2 for succinate, D-lactate and glycerol oxidation. Measurements of the leads to H+/2e- ratio with formate as the reductant and oxygen or NO3- as the oxidant were complicated by pH changes associated with formate uptake and CO2 formation. Nevertheless, it was possible to conclude that the site of formate oxidation is on the inner aspect of the cytoplasmic membrane, that the leads to H+/O ratio for formate oxidation is approx. 4, and that the leads to H+/NO3- ratio is greater than 2. Measurements of the rate of NO3- penetration into osmotically sensitive spheroplasts demonstrated an electrogenic entry of NO3- anion. The permeability coefficient for nitrate entry at 30 degrees C was between 10(-9) and 10(-10) cm- s(-1). The calculated rate of nitrate entry at the concentration typically used for the assay of nitrate reductase (EC 1.7.99.4) activity was about 0.1% of that required to support the observed rate of nitrate reduction by reduced Benzyl Viologen. Measurements of the distribution of nitrate between the intracellular and extracellular spaces of a haem-less mutant, de-repressed for nitrate reductase but unable to reduce nitrate by the respiratory chain, showed that, irrespective of the presence or the absence of added glucose, nitrate was not concentrated intracellularly. Osmotic-swelling experiments showed that the rate of diffusion of azid anion across the cytoplasmic membrane is relatively low in comparison with the fast diffusion of hydrazoic acid. The inhibitory effect of azide on nitrate reductase was not altered by treatments that modify pH gradients across the cytoplasmic membrane. It is concluded that the nitrate-reducing azide-sensitive site of nitrate reductase is located on the outer aspect of the cytoplasmic membrane. The consequences of this location for mechanisms of proton translocation driven by nitrate reduction are discussed, and lead to the proposal that the nitrate reductase of the cytoplasmic membrane is vectorial, reducing nitrate on the outer aspect of the membrane with 2H+ and 2e- that have crossed from the inner aspect of the membrane.


Subcellular structure of bovine thyroid gland. A study on bovine thyroid membranes by buoyant-density-gradient centrifugation in a B-XIV zonal rotor. A combined mitochondrial and light mitochondrial fraction and a microsomal fraction were isolated from bovine thyroid gland and fractionated further in a B-XIV zonal rotor. A density gradient ranging from 20 to 50% (w/w) sucrose was used. The rotor was operated for 3 h at 45 000 rev./min. All manipulations were performed at 4 degrees C and at pH 7.4. 2. Membranous material was recovered in two zones: zone I, containing microsomal material derived from both smooth endoplasmic reticulum and plasma membranes and probably also from other smooth membranes; zone II, containing material from rough endoplasmic reticulum. 3. Increasing the pH of the medium up to 8.6, or the addition of Mg2+ to the medium resulted in the formation of a single zone at intermediate densities (aggregation of membranes?). An analogous effect was obtained after treatment with Pb (NO3) 2. 4. In the presence of heparin (50 i.u./ml) the bulk of the membranes was found in zone I. This was due to the release of ribosomes from the rough endoplasmic reticulum.