Interrelationship between the dietary regulation of fatty acid synthesis and the fatty acyl-CoA desaturases. In this paper we present further evidence for the close control of fatty acid synthetase and stearoyl-CoA desaturase. Furthermore, we have established that whereas dietary palmitic acid may influence the activity of this desaturase but not of fatty acid synthetase, dietary linoleic acid appears to control both these enzymes. Finally, we have studied the influence of dietary fat and carbohydrate on the activities of the delta6 and delta5 desaturases. The former is only slightly affected by these dietary components. The delta5 desaturase activity is stimulated as the dietary fat content rises but is unaffected by dietary carbohydrate. The control of these enzymes is therefore independent of the control of fatty acid synthetase and stearoyl-CoA desaturase. From the data presented, the magnitude of the controlling effect of polyunsaturated fatty acids on fatty acid synthetase and stearoyl-CoA desaturase activity is determined and its relevance to lipogenesis in man based on daily intake of carbohydrate and linoleic acid is discussed.


Studies on the inhibition of the desaturases by cyclopropenoid fatty acids. Unwashed rat liver microsomes were used to study the inhibition of the delta6 and delta9 desaturases by cyclopropenoid fatty acids with the ring structure about the 9,10 or 6,7 carbon atoms. The 9,10 cyclopropenoid acid (sterculic acid) is shown to be an effective inhibitor of only delta9 desaturase and then only in the presence of MgCl2 and coenzyme A (presumably due to the formation of sterculoyl-CoA). Two 6,7 cyclopropenoid acids of different chain lengths showed no marked inhibition of either the delta6 or delta9 desaturase. By the use of [3H]-sterculic acid, it has been shown that under conditions of high inhibition of the delta9 desaturase the inhibitor is not covalently attached to the enzyme at any point. This disproves older ideas on the mechanism of inhibition that assumed reaction between the cyclopropenoid ring and sulphydryl groups on the enzymes.


Penicillin combinations against multi-resistant urinary pathogens as an alternative to gentamycin treatment. A total of 42 multi-resistant urinary pathogens, sensitive to gentamycin only, were investigated with regard to the effects of dicloxacillin and ampicillin or cloxacillin/flucloxacillin and amoxycillin in combination. A synergistic antibiotic effect with resulting low MIC levels was demonstrated for 81% of the strains. It was also found that a further increase in sensitivity might be achieved by adjustment of the pH level.


Pathway of glucose catabolism in Caulobacter crescentus. Glucose when present as a sole organic carbon source in a mineral salts medium is dissimilated by Caulobacter crescentus ATCC 15252 (strain CB-2) by the Entner-Doudoroff pathway throughout the culture cycle (exponential, transition, and stationary phase). Most of the available glucose that is present at the onset of exponential growth is assimilated by the cells during the transition phase or the period associated with stalk cell development. Swarmer cell development is minimized during this phase. During this same period the pH drops from 6.1 to 4.9 as a result of an abundant excretion of acetic acid. Simultaneously, poly-beta-hydroxybutyrate accumulates within the cells at an accelerated rate. An NADP-dependent glyceraldehyde-3-phosphate dehydrogenase is also present throughout the culture cycle which subsumes the presence of the subsequent enzymes of the Embden-Meyerhof-Parnas pathway in pyruvate formation. An operative tricarboxylic acid cycle is associated with cells throughout the culture cycle.


Changes in blood pressure and antihypertensive treatment in a community over four years. In 1971 persons aged 50 to 59 years in Albury were screened for hypertension and those with diastolic blood pressure equal to or over 110 mm Hg were notified. On screening of the sample in 1975, the proportion of persons receiving treatment had doubled to 22%. Most of those newly receiving treatment in 1975 had had diastolic blood pressures under 110 mm Hg in 1971. The mean systolic and diastolic pressures of the newly treated group had fallen by 13-7 and 9-1 mm Hg respectively to 153-5 and 91-6 mm Hg. The mean pressures of subjects already receiving treatment in 1971 and continuing it remained higher (163-1 mm Hg systolic and 100-9 mm Hg diastolic), despite their receiving relatively more medication. The latter group was composed of patients likely to have had more severe hypertension originally, and the importance of effective treatment for them is emphasized. The frequency with which subjects with mild hypertension are being treated demonstrates the urgent need to complete randomized controlled trials of treatment in this group.


[Influence of amino acids on synthesis of extracellular proteases by Actinomyces thermovulgaris]. Various amino acids had different effect on the synthesis of exocellular proteases by Actinomyces thermovulgaris T-54. Glutamine and arginine induced the synthesis, the effect depending on their concentration in the medium. Other amino acids inhibited the synthesis of proteases. The synthesis of alkaline and neutral proteases was not coordinated, and the ratio between these enzymes was not constant in different variants of the experiment. The maximum synthesis of alkaline proteases was found before the formation of submerged spores.


A comparison of the genetic and physical size of the streptomycin resistance locus in Pneumococcus. Polyacrylamide gel electrophoresis of the 30S ribosomal proteins derived from six streptomycin resistant strains indicates that each mutation alters the same ribosomal protein (str-r protein). Preliminary data utilizing SDS gels indicates that the str-r protein has a molecular weight between 10,000 and 20,000 daltons. No significant differences could be detected between the molecular weight of the str-r protein when it is derived either from a sensitive or from a resistant strain, including those derived from strains carrying multisite mutations of different genetic size. We have estimated the size of the multisite str-r mutations to be less than 30 base pairs. Two factor crosses with str-r markers in the trans position demonstrate recombination frequencies expected of closely linked, intragenic markers although cotransfer frequencies, of these same markers from the cis position, are very low. It is concluded that the cotransfer frequencies represent a marker effect and possible explanations are discussed. A reinterpretation of the genetic map of the pneumococcal str-r locus is presented.


[Ecologic features of cultures of the species Clostridium gürfelii]. The Clostridium cultures decomposing pectin were isolated from flax straw during flax retting. The activity was manifested only at high pH values, the optimum pH being 6.7--8.2. Such ecological adaptation of the cultures is determined by the properties of the pectolytic enzyme complex which they synthesize. The complex does not contained polygalacturonase (the optimum pH 5.0) but has a high activity of pectate transeliminase (the optimum pH 8.0--8.4). The cultures differ from Clostridium felsineum which possesses a high activity of polygalacturonase and therefore can decompose pectin in acid medium. By their morphological, cultural and physiological properties, the bacteria are classed as Clostridium güfelii (Prévot, 1966).


Organization of purpine degradation in the liver of a teleost (carp; Cyprinus carpio L.). A study of its subcellular distribution. Carp liver was fractionated by differential and density gradient centrifugation and assayed for enzymes of purine catabolism. While urate oxidase is an exclusively peroxisomal enzyme, only a very small percentage of the enzymes xanthine oxidase, allantoinase and allantoicase is associated with subcellular or ganelle fractions. There is no general purine catabolizing subcellular compartment. There is some but not yet conclusive evidence for the assumption that urate oxidase is a membrane bound enzyme.


Enzymatic synthesis of polyprenol monophosphate mannose in insects. The microsomal fraction of insects was found to contain an enzyme which transfers mannose from guanosine diphosphate mannose to an endogenous or exogenous insect lipid and to other acceptors such as dolichol monophosphate or ficaprenol monophosphate. This activity depended on the presence of Triton X-100 and magnesium ions, the optimal concentration of the latter being 10mM. The optimal temperature of the reaction was 25 degrees C and the maximal activity was obtained at pH 7.9. The mannolipid formed behaved as a monophosphodiester when chromatographed on DEAE-cellulose. Weak acid treatment of the product liberated mannose. Its behaviour both on thin layer and Sephadex G-150 chromatography would indicate the presence of a number of isoprenyl units similar to the dolichol and different from the ficaprenol derivative. Stability to phenol treatment indicated that the lipid fraction of the mannolipid is an alpha-saturated polyprenol phosphate similar to dolichol monophosphate.


A primer independent activity of rabbit muscle phosphorylase b. Rabbit muscle phosphorylase b was found to be capable of forming protein bound alpha-1,4 glucosyl chains upon incubation of the enzyme with appropriate concentrations of glucose-1-phosphate with no primer addition (unprimed synthesis). This activity would only be present in a small fraction of the total muscle phosphorylase b activity, as judged from the high concentrations of enzyme which are required to demonstrate the occurrence of unprimed synthesis. Polyacrylamide gel electrophoresis shows the presence of a phosphorylase isoenzyme capable of accepting glucosyl moieties, giving rise to a glucosylated protein enzymatically active in the chain lengthening of its own glucan.


Sucrose metabolism in green algae. I. The presence of sucrose synthetase and sucrose phosphate synthetase. The presence of sucrose synthetase and sucrose phosphate synthetase has been demonstrated in two species of green algae: Chlorella vulgaris and Scenedesmus obliquus. Partial purification from crude extracts allowed the determination of the kinetic constants of algae enzymes. They are very similar to the ones reported for enzymes from higher plants.


Trisialoganglioside synthesis by a chicken brain sialyltransferase. Comparative study with the similar reaction for the synthesis of disialoganglioside. An enzyme preparation from embryonic chicken brain catalyzes the transfer of sialic acid from CMP-N-acetylneuraminic acid to ceramide-Glc-Gal(NeuAc-NeuAc)-GalNAc-Gal (GDlb) to form ceramide-Glc-Gal(NeuAc-NeuAc)-GalNAc-Gal-NeuAc (GTlb). The sialyltransferase activity was measured during the development of the embryo, the subcellular distribution of this activity was determined and several kinetic properties of the reaction were examined. A comparative study with the similar reaction involved in the transfer of sialic acid to the terminal galactose in ceramide-Glc-Gal(NeuAc)-GalNAc-Gal (GMl) was made. The results obtained in this comparative study suggest that the transfer of sialic acid in both reactions is catalyzed by the same enzyme.


Porphyrin biosynthesis. Immobilized enzymes. IV. Studies on aminolaevulate dehydratase attached to Sepharose. Bovine liver aminolaevulate dehydratase (ALA-D) has been chemically attached to Sepharose 4B and its properties have been studied. The optimal conditions for coupling have been determined. It was found that the immobilized enzyme retained a significant percentage of the activity of the free enzyme. The coupling yield was rather high. The insolubilized enzyme requires both anaerobiosis and a thiol activator for maximal activity. It can be stored at 4 degrees C for long periods with little loss of activity and it can be repeatedly used without alteration of its enzymic capacity. Attachment of ALA-D to the gel has led to an enhanced thermal stability. pH optima of free and bound enzyme was the same while a small decrease in the Km of the matrix bonded ALA-D as compared to that of the soluble enzyme was observed. The use of the fixed-ALA-D for the preparation of PBG is described.


[Complications of pertussis immunization (author transl)]. 16 cases of neurological disease and/or death shortly after pertussis immunization are reported. Eight patients had convulsions, six with ensuing permanent defects. Severe polymyositis was observed in one case. Five infants died 12 h to 4 days after vaccination: two after acute encephalopathy and three in the form of a sudden unexpected death (SID). In two fatal cases the morphological changes in the brain corresponded to those of pertussis encephalopathy: neuronal degeneration in various parts of the cortex, especially in the region of the ammons horn, and in the cerebellum. There were no signs of inflammation. Three cases underwent forensic autopsy and death was attributed to bronchopulmonary infection. Complete neuropathological work-up was only done in one case, in which the brain was normal. The critique of episodical reports and the demand for prospective studies is appreciated. Knowledge of all possible forms of complications, however, is indispensable for future investigations. Polymyositis and SID have so far not been listed as abnormal reactions to immunization. The majority of our cases became known accidentally from hospital sheets or from discussions with collegues. For a detection of all possible cases a greater awareness of doctors for the problem of pertussis immunization appears necessary. Only another 23 cases have been reported to the health authorities of the state of Lower Saxony during the last 6 to 7 years. Of those, nine were either harmless reactions or diseases probably unrelated to vaccination. Two were cases of SID, 12 and 72 h after vaccination. It is concluded that only a minor proportion of possible complications is presently reported to the health authorities.


Amelioration of metabolic acidosis with fludrocortisone therapy in hyporeninemic hypoaldosteronism. In four patients with renal hyperchloremic acidosis and hyperkalemia, hyporeninemic hypoaldosteronism and chronic renal insufficiency (glomerular filtration rates of 13, 31, 35 and 44 ml per minute per 1.73 m2), prolonged administration of fludrocortisone increased urinary potassium and net acid excretion, corrected hyperkalemia and substantially ameliorated acidosis. Except in the patient with the lowest glomerular filtration rate, the increased net acid excretion was due mostly to increased ammonium excretion. Urine pH decreased initially in each patient, but in the three patients with the highest filtration rates, it increased subsequently as ammonium excretion increased, indicating that renal ammonia production increased. Urinary ammonium excretion correlated inversely with serum potassium concentration and did not decrease on discontinuation of therapy if hyperkalemia was prevented from recurring. In patients with renal acidosis and hyporeninemic hypoaldosteronism, administration of mineralocorticoid hormone can augment both renal hydrogen-ion secretion and, by correction of hyperkalemia, renal ammonia production, and thereby ameliorate metabolic acidosis.


The scientific foundations of clinical enzymology. Clinical enzymology is the study of the activity and properties of enzymes in specimens (usually of blood) taken from patients, as an aid to the diagnosis and understanding of disease. Research in many areas of enzymology is undertaken to strengthen the foundations on which the clinical interpretation of such enzyme measurements rests. Some aspects of this research are briefly described.


Modification of actins by phallotoxins.

Modification of actins by phallotoxins. Phallotoxins bind to filamentous actin (F-actin) from liver or rabbit muscle with Kdiss approximately 10(-8) M. By this combination the structure of F-actin is stabilized to such an extent that it will be resistant to the depolymerizing action of 0.6 M KI and of deoxyribonuclease I, to denaturation by heat (70 degrees C), and to local ruptures caused by ultrasonication or by cytochalasin B. The structural features of phallotoxins essential for affinity to F-actin are specific.


Characteristics of chick cerebral beta-adrenoceptors assessed by cyclic adenosine 3',5' monophosphate formation and [3H]-propranolol binding. Chick cerebral beta-adrenoceptors have been characterised by measurement of cyclic AMP accumulation in brain slices and assessment of the specific binding of [3H]-propranolol to cerebral membranes. The binding of [3H]-propranolol was inhibited by beta-adrenoceptor agonists and antagonists with affinities that correlated well with their ability to stimulate cyclic AMP formation or to antagonise the cyclic nucleotide accumulation induced by isoprenaline. The relative potencies of a number of drugs in several cerebral regions suggests that the receptors may be of the beta2 subtype. Regional distribution studies revealed the highest density of binding sites in the cerebellum and lowest in the optic lobes. However, the concentration of [3H]-propranolol that produced half-maximal specific binding was similar in all regions. Subcellular fractionation of cerebral hemisphere tissue demonstrated an enrichment of [3H]-propranolol binding sites in the synaptosomal and microsomal fractions. There are discrepancies between the topographical distribution of beta-adrenoceptor binding sites and the endogenous noradrenaline level in chick brain and between the number of binding sites and the intrinsic activity of beta-adrenoceptor mediated cyclic AMP formation.


On the optimal dosage of Pro-Leu-Gly-NH2 (MIF) in neuropharmacological tests and clinical use. Pro-Leu-Gly-NH2 (MIF) inhibits the tremor induced by oxotremorine. Objective measurement of this tremor permits the drawing of a dose-effect curve. The inhibitory effect of the peptide increases linearly with increasing doses until an optimum is reached (between 30 and 40 mg/kg i.p.). At still higher doses the peptide is inactive. The same phenomenon is observed with analogues of MIF. This finding may have important bearings on the interpretation of clinical and experimental data obtained with MIF.


[Use of an anxiolytic agent (lorazepam) in low doses in the treatment of anxiety states]. A nonblind clinical study was carried out by administering Lorazepam at a low dosage to patients with a light to moderate state of reactive anxiety. The drug was shown to be effective in the management of the target symptoms studied: anorexia, sleep induction, rhythm, and duration disturbances, depression, irritability and moodiness, fatigue, anxiety and tension, somatic anxiety, social adaptation. The results obtained are practically identical to those reported in the literature for higher doses. Moreover, low-dose Lorazepam was well tolerated and without undesired side-effects.


Injuries to the ankle and foot in athletics. Accurate diagnosis in ligament and ligament-bone injuries of the foot and ankle demands a knowledge of the anatomy and mechanics of the foot and ankle, a good history of the mechanism of injury, clinical examination including fingertip palpation and assessment of stability, and adequate x-ray studies, perferably in the deformed position, with stress films and arthrograms in selected cases. By making an accurate diagnosis, early proper treatment is instituted, the athlete is restored to health sooner, and, almost as important, the athlete and the coach get a more realistic prognosis regarding the end result and duration of disability.


Polyarteritis nodosa in older children.

Polyarteritis nodosa in older children. Polyarteritis in the older child is thought to be a rare disease. This study describes 11 children, 3 to 12 years of age, with polyarteritis seen over a five-year period. Fever, abdominal pain, hypertension, and leukocytosis were found in almost all. Renal disease occurred in eight. Examination of muscle, gut, or kidney tissue was an effective means of diagnosis. The pathological changes were the same as those seen in adults. There seemed to be an association between polyarteritis and group A streptococcal infection. Ten patients had a salutary response to high-dose prednisone administration.


Properties of the HCO-3-stimulated Mg2+ -ATPase activity in red cell membranes. The HCO-3-stimulated Mg2+ -ATPase activity in red cell ghost fragments was investigated. Increasing the HCO-3 concentration in the incubation medium resulted in increased ATPase activity. NaHCO3 appeared to be more effective than KHCO3 in this regard. The ATPase activities were slightly stimulated by increases in ionic strength and utilized ITP almost as readily as ATP. A Mg/ATP ratio of 1.0 and a pH of 7.6 yielded maximum activity. These properties are of interest since the present enzyme is the only unquestionable instance where a HCO-3 ATPase is located in the surface membrane of a cell.


Methylprednisolone pulse therapy in the treatment of polyarteritis nodosa. Two cases of lung granulomata associated with "crescentic" glomerulonephritis both in a clinical setting of polyarteritis nodosa were treated with high doses of intravenous methylprednisolone ("pulse therapy") in single injections of 30 mg/kg body-weight. In the first case rapidly progressive glomerulonephritis with 100% crescents was arrested and followed by improvement of renal function from a creatinine clearance of 5 ml/min to 30 ml/min; in the second case multiple lung granulomata of 8 months' standing, unresponsive to oral steroids, disappeared 8 days after treatment with high dosage intravenous methylprednisolone. The use of "pulse therapy" with methylprednisolone is advocated, not only in such cases where arteritis is known or suspected, but also in radidly progressive glomerulonephritis associated with crescents.


Cryoenzymology in mixed solvents without cosolvent effects on enzyme specific activity. Water-soluble polyelectrolytes in interaction with proteins are described. These polyelectrolytes make it possible to investigate enzyme-catalyzed reactions in cooled mixed solvents without the usual effects of their organic solvent component  on enzyme specific activity. The applicability of techniques developed is illustrated by results obtained on several systems. The possibility of an electrostatic "sorting out" of solvents and its potentialities in cryoenzymology are discussed.


A role for asparaginyl-tRNA in the regulation of asparagine synthetase in a mammalian cell line. The expression of asparagine synthetase activity [L-aspartate:ammonia ligase (AMP-forming), EC 6.3.1.1] in cultured Chinese hamster ovary (CHO) cells is regulated by asparagine. After transfer of CHO cells from an asparagine-supplemented medium to a medium lacking asparagine, activity increases 1.5- to 2-fold. If asparagine is added back to the medium, activity returns to control levels. To test the possible involvement of Asn-tRNAAsn in regulating the levels of asparagine synthetase, we have examined the levels of asparagine synthetase in a mutant of CHO cells containing a temperature-sensitive asparaginyl-tRNA synthetase [L-asparagine:tRNA ligase (AMP-forming), EC 6.1.1.22]. Under conditions of limited asparaginyl-tRNA synthetase activity in the mutant, there is a 2- to 3-fold increase in the level of asparagine synthetase activity. Under identical conditions, there is no change in asparagine synthetase activity in the wild type. This correlation between asparaginyl-tRNA synthetase activity and asparagine synthetase levels may be a consequence of a direct role of tRNAAsn in the regulation of the in vivo expression of the asparagine synthetase structural gene.


Isolation of a chloroplast N,N'-dicyclohexylcarbodiimide-binding proteolipid, active in proton translocation. The N,N'-dicyclohexylcarbodiimide-binding proteolipid from lettuce chloroplast membranes has been purified by a novel, rapid technique involving I-butanol extraction and ether precipitation. Reconstitution of this proteolipid into liposomes composed of chloroplast lipids and subsequent incorporation of bacteriorhodopsin resulted in the formation of liposomes exhibiting a light-dependent accumulation of protons. This accumulation was significantly enhanced upon addition of N,N'-dicyclohexylcarbodiimide at concentrations similar to those that inhibit chloroplast adenosinetriphosphatase activity. Radioactively labeled N,N'-dicyclohexylcarbodiimide was found to be incorporated essentially into the proteolipid of the reconstituted liposomes. These results suggest that the functional unit responsible for proton channeling in the chloroplast membrane has been isolated and reconstituted in the native state.


Lack of effect of chronically administered thyrotropin-releasing hormone (TRH) on regional rat brain tyrosine hydroxylase activity. Chronic treatment of adult male rats with TRH (1 or 10 mg/kg IP) for 5 or 9 days failed to alter the activity of tyrosine hydroxylase (TH), the enzyme regulating the rate-limiting step in catecholamine biosynthesis. In contrast, as previously described, chronic reserpine administration (0.5 mg/kg IP: 9 days) resulted in a significant rise in TH activity in midbrain, hypothalamus, pons-medulla and forebrain. These results suggest that the enhanced brain norepinephrine turnover reported to occur after treatment with TRH is not due to synthesis of new TH enzyme protein.


Phosphohistone and glycogen synthase D phosphatase activities in a liver glycogen pellet preparation. A liver glycogen pellet preparation previously found to contain synthase D phosphatase activity was shown to contain also phosphohistone phosphatase activity. Pellet phosphohistone phosphatase and synthase D phosphatase competed for the same substrates and appeared to be the same enzyme. ATP, a potent inhibitor, and G-6-P, a potent activator of the synthase phosphatase reaction, had little effect on the phosphohistone phosphatase reaction. These observations suggest that the ATP and G-6-P effects are relatively specific and are probably caused by binding to the synthase D substrate. The observed effects of NaCl and KCl were more complex. They stimulated phosphohistone phosphatase activity but strikingly inhibited synthase phosphatase activity. Sodium fluoride inhibited both reactions.


Antagonism of the analeptic activity of thyrotropin-releasing hormone (TRH) by agents which enhance GABA transmission. Administration of 10 mg/kg TRH to mice was found to reduce the sleep and hypothermia induced by 4.7 g/kg ethanol. However, TRH did not reduce the sleep of mice that were given gamma-hydroxybutyric acid (GHBA), baclophen, or aminooxyacetic acid (AOAA) in combination with 3 g/kg/ ethanol. TRH also failed to reverse the hypothermia induced by the combination of ethanol and baclophen or GHBA, and the characteristic neurological effects of TRH e.g. tremor, increased muscle tone, and increased respiratory rate were reduced. In addition, TRH-induced locomotor stimulation was prevented by pretreatment with small doses of the GABA-ergic agents, and while 30 mg/kg TRH reduced the hypothermia produced by large doses of the GABA-ergic drugs, it did not antagonize the locomotor retardation produced by baclophen or GHBA. A hypothesis that the analeptic effects of TRH may be medicated via an inhibition of GABA systems is discussed.


Benzodiazepines and discrimination behaviour: dissociation of response and sensory factors. Ten London pigeons were trained on a schedule which allowed concurrent measurement of motor and exteroceptive sensory changes. The task involved a conditional colour discrimination contingent upon the completion of a fixed interval schedule of responding. As a preliminary pharmacological study the effects of the benzodiazepines, chlordiazepoxide and flurazepam were investigated. Both drugs enhanced perseverative responding (after completion of the FI), and to a variable degree responding during the FI, although there was no evidence for an increase in responding during the inter-trial-intervals. On the other hand, no marked changes in discrimination performance were observed. It is concluded that the most significant effects of these benzodiazepines are on motor mechanisms.


Effect of apomorphine on the antinociceptive activity of morphine. Apomorphine pretreatment potentiated the analgesic effect of morphine in a dose-dependent manner both in rats and in mice measured by five different tests (writhing, hot plate, inflamed foot, tail-pinch and tail-flick procedures). Furthermore, apomorphine augmented the antinociceptive activity of morphine in tolerant animals as well. In morphine dependent mice the nalorphine precipitated jumping--a withdrawal symptom--was found inhibited by apomorphine treatment. The results are discussed in the light of the numerous but contradictory data available in the literature.


Assay and characterization of serum-stimulated lipolytic activity in homogenates of human adipose tissue. An assay of lipolytic activity in human adipose tissue is described, in which native homogenates of the adipose tissue yield the enzyme, as well as the triglyceride substrate, and the emulsifying phospholipids. The lipolytic activity in the presence of serum is characterized mainly as lipoprotein lipase activity by a pH-optimum of 8.0, by the fact that serum is necessary for full activity, and that it is inhibited by 1 M NaCl and by protamine. At serum concentrations of higher than 50% a marked inhibition of the lipolysis is observed. Noradrenaline, insulin, and heparin have no effect on the serum-stimulated lipolytic activity.


Oxygen affinity of duck blood determined by in vivo and in vitro technique. Half saturation partial pressure of O2,P50, was determined in domestic Muscovy ducks (Cairina moschata) both by in vivo and in vitro techniques. For in vivo determination, blood samples were drawn from the anesthetized, artificially ventilated animals and analyzed both for O2 content, Co2, and O2 partial pressure, Po2. O2 capacity was detemined in arterial samples during hyperoxic ventilation (arterial Po2 about 180 torr). P50 was calculated from measurements in venous blood samples (O2 saturation near 50%). For in vitro determinations, Co2 was measured in blood samples equilibrated with Po2 close to P50. No significant difference was found between P50 values determined by both techniques. At 41 C and pH 7.50, P50 averaged 41.7 torr when analyzed by in vivo technique and 41.4 torr when determined in vitro. The variability between animals was less than 1 torr (SD) and could be explained by the experimental error. The partially discordant literature data on P50 of duck blood are reviewed and critically discussed.


Inhibition of secretin release and pancreatic bicarbonate secretion by somatostatin infusion in man. Five healthy students were investigated on two different days with or without a constant infusion of somatostatin (500 microgram/h) into an arm vein a fluoroscopically placed Lagerlöf tube was used for the collection of gastric and duodenal juice. After 30-min basal period, 40 ml 100 mmol/l HCl was infused into the midpart of the duodenum over 5 min through a thin catheter attached to the tube. Plasma immunoreactive secretin was measured by radioimmunoassay employing 125I-labelled synthetic secretin, antibody against synthetic secretin, and standards prepared from pure natural porcine secretin. Secretin levels without somatostatin infusion were 4.6+/-0.7 pmol/l (mean+/-S.E.M.) basally and rose to a peak of 21.8+/-6.2 pmol/l after duodenal acidification (p less than 0.05) and with somatostatin infusion were 4.4+/-0.4 pmol/l basally and rose to a peak of 6.7+/-1.7 pmol/l (n.s.) after duodenal acidification. Pancreatic bicarbonate output increased from 8.0+/-2.5 mumol/min (mean+/-S.E.M.) to 283+/-44 mumol/min without somatostatin infusion (p less than 0.05) and from 6.7+/-2.1 mumol/min to 70+/-13 mumol/min somatostatin (p less than 0.05). This study shows that somatostatin (500 microgram/h can inhibit the release of secretin and the pancreatic bicarbonate secretion after duodenal acidification in man.


Gastro-oesophageal sphincter pressure, motility and acid clearing. A study of hiatus hernia patients and normal subjects and of the effect of a modified belsey MK IV repair on the results of the manometric and acid-clearing tests. Basal gastro-oesophageal sphincter pressure was recorded in 68 patients with symptomatic, radiologically verified sliding hiatus hernia, and in 37 healthy subjects. The diameter of the probe was 2.5 mm, and the flow rate 0.5 ml/min. Mean sphincter pressure was lower in patients (6 mm Hg) than in normal subjects (15 mm Hg). In 32 per cent of the patients sphincter pressure was within normal range (8-24 mm Hg), whereas in 68 per cent it was lower than in the normal subjects. There seemed to be no relationship between sphincter pressure and severity of symptoms. Oesophageal acid clearing was investigated in 57 of the patients and in 26 of the normal subjects. Normal subjects clear the bolus of acid in an average of 10 swallows (range 4-16). In patients the incidence of prolonged clearing was greater, but the acid-clearing ability did not seem to be related to the degree of severity of the symptoms. A manometric study was made of 45 patients and an acid-clearing study made of 40 patients, before and 3 months after a modified Belsey MK IV repair for hiatal hernia. Mean postoperative sphincter pressure was higher (10 mm Hg) than the preoperative mean (6 mm Hg), but was still lower than the normal mean. Significant changes in acid-clearing ability could not be demonstrated. The results are inconclusive with regard to the importance of oesophageal motility disturbances for the symptomatology and acid-clearing ability. Thirty-seven out of 45 patients became free of symptoms, and the rest - except 1 (relapse of hernia) - improved.


Enzymatic Determination of bile acids. the NADP-specific 7alpha-hydroxysteroid dehydrogenase from P. testosteroni (ATCC 11996). By use of the specific substrates, 7alpha-hydroxy-5beta-cholanic acid and 3alpha, 7alpha, 12alpha-trihydroxy-3,12-diacetyl-5beta-cholanic acid methyl ester, crude extracts of P. testosteroni grown on a steroid-containing medium have been shown to exhibit 7alpha-hydroxysteroid: NADP-oxidoreductase activity. The enzyme is highly specific for NADP. Both free and conjugated 7alpha-hydroxy bile acids can act as substrates, but those of low polarity (few hydroxyl groups) seem to be preferred, judging from initial reaction velocity studies. Optimal conditions appears to be at pH 8.5-9.5 and at 25 degress C. Free SH-groups are essential for maximum catalytic activity, since the enzyme is inhibited by SH-reacting substances such as p-chloromercuribenzoate and monoiodoacetic acid. Also the ketone-trapping agents hydrazine hydrate and semicarbazide act as inhibitors. Upon dilution, the storage stability is severely reduced, but this effect may be counteracted by the addition of glycerol at concentration of 20% or more. By gel filtration experiments on Sephadex G-100, the molecular weight was estimated to about 80,000.


Quantitative determination of serum bile acids using a 7alpha-hydroxysteroid dehydrogenase. A crude 7alpha-hydroxysteroid dehydrogenase isolate from P. testosteroni requires NADP specifically and provides a simple and specific means for the determination of 7alpha-hydroxy bile acids and their conjugates. A specific and reliable enzymatic-fluorimetric method for the quantitative measurement of the primary bile acids in serum is described. Overall recovery from normal and jaundiced serum averaged 85%. The mean concentration of serum 7alpha-hydroxy bile acids in fasting, healthy subjects was 2.5 mumoles/1 (S.D. 1.45 AND RANGE 1.2-6.3), averaging 73% (S.D. 11.5) of the total, measured as the 3alpha-hydroxy bile acid concentration. In parenchymatous liver disease and cholestatic hepatobiliary disorders, the proportion of 7alpha-hydroxy bile acids was significantly increased, mean 92.5% (S.D. 6.6)(p less than 0.001).


Effect of cimetidine on pentagastrin-stimulated gastric secretion in healthy man. Two series of experiments were carried out in three healthy volunteers. In one series increasing doses of cimetidine were tested on a constant background stimulation with 0.15 microgram-kg-1h-1 of pentagastrin. In the other series of experiments increasing doses of pentagastrin were given either alone or in combination with a fixed dose of cimetidine (1.2 mg-kg-1h-1). Pentagastrin and cimetidine were given as continuous intravenous infusions, the various doses tested on separate days. The inhibition of acid output in response to pentagastrin stimulation increased almost linearly with the log dose of cimetidine. Fifty per cent inhibition of the response to 0.15 microgram-kg-1h-1 of pentagastrin was achieved by about 0.6 mg-kg-1h-1 of cimetidine. 2.4 mg-kg-1h-1 of cimetidine gave on average 90% inhibition. The inhibition caused by cimetidine decreased to a minimum of 57% and could not be overcome by increasing the doses of pentagastrin. The results suggest a mixture of competitive and noncompetitive interaction. The effect of cimetidine on pepsin secretion was less regular. Generally, pepsin output was less inhibited than acid output.


Microcalorimetric studies of human platelet metabolism at rest. Influence of pH, temperature, cell concentration, preparation methods and storage. Heat production in human platelets has been measured under different conditions of pH, temp., cell concentration, preparation and storage time. The heat effect was found to increase linearly in the pH range 7.00-7.95, by 20% per pH unit. The temp. coefficient for the heat effect was determined to be Q10 = 2.0 for the temp. interval 32-42 degrees C. Heat production per cell was not significantly affected by variations in cell concentration, or by storage for several hours at room temp. When citrate replaced heparin as anticoagulant significantly higher heat effect values were found.


A deficient pyruvate kinase with an electrophoretically slow-moving component. A deficient erythrocyte pyruvate kinase observed in a patient with congenital non-spherocytic anaemia was characterized by the following properties: very low activity in haemolysates, decreased thermal stability, slightly increased urea denaturation, high affinity for PEP, poor FDP activation, normal ATP inhibition, decreased affinity for ADP, normal pH of optimal activity, and presence of an abnormal slow-moving component in this layer polyacrylamide gel electrophoresis. The patient was probably double heterozygous for two different deficient mutants of erythrocyte pyruvate kinase.


Asthma and urticaria during disodium cromoglycate treatment. A case report. A 10-year-old asthmatic boy began to suffer from urticarial rash and moderately severe bronchospasm after 8 weeks' treatment with disodium cromoglycate. Initially, DSCG had helped to control his asthmatic attacks, and steroid therapy could be discontinued. Inhalation provocation test with DSCG aerosol, 4 months after stopping DSCG treatment, showed an immediate-type 1 response and urticaria. A repeat provocation test, under antihistaminic cover , failed to produce similar response. When DSCG was withdrawn, urticaria vanished and the child remained symptom-free. Disodium cromoglycate is regarded as the possible aetiological agent.


[Effect of alclofenac on the prothrombin level in patients under treatment with anticoagulants]. Simultaneous administration of the anticoagulants acenocoumarol, phenprocoumon or chlorindion and the antirheumatic substance alclofenac in long term trials has no observable influence on the prothrombin time of stabilized patients. When the anticoagulant (acenocoumarol or phenprocoumon) and the alclofenac therapy are begun simultaneously, a variation of the dose is necessary. Either a one-third lower initial dosage can be given, or, after attaining the maximal anticoagulation effect on the prothrombin time (48 h after beginning acenocoumarol therapy, 72 h with phenprocoumarol), a lower daily dosage must be administered once, in comparison to the other group without alclofenac therapy. The maintenace dosage is not influenced by alclofenac.


An entomological survey on the mosquitoes of Wuvulu Island, Papua-New Guinea. An entomological survey in Wuvulu Island, Papua New Guinea, in August 1975 and 1976 shows the presence of six mosquito species: Aedes (S.) hebrideus, AE. (F.) notoscriptus, AE. (V.) lineatus, ? Ae. (L.) dasyorrhus, Culex pipiens fatigans and Armigeres breinli. The medical significance of these mosquitoes is discussed, with special reference to the problem of dengue virus transmission.


The effect of acid-stimulating and inhibiting drugs on the ultrastructure of gastric parietal cells in man. The ultrastructure of the human parietal cell at rest, after stimulation of gastric acid secretion by pentagastrin and after inhibition with propantheline or cimetidine, an H2-receptor antagonist, was studied. On stimulation there was an increase in the total secretory surface area of the gastric gland and canaliculus, with a concomitant decrease in the tubulovesicular system. On inhibition there was a decrease in the secretory surface area and a corresponding increase in the tubulovesicular system. These results are in agreement with other reports and support the hypothesis that during secretion of acid the tubulovesicular system increases the secretory surface by eversion or fusion of the vesicles and tubules with the plasmalemma. On inhibition there is thought to be reformation of the tubulovesicular system by pinocytosis. There is also ultrastructural evidence for the theory of membrane flow between the tubulovesicular system and the microvilli of the canaliculus and gastric lumen. Furthermore, there was an increase in lysosomal activity after the administration of the inhibiting drugs. It is suggested that the increased lysosomal activity could be involved in reducing the quantity of membrane after the membrane flow which occurs on gastric acid inhibition.


The effect of cyclizine hydrochloride on lower oesophageal sphincter pressure in man. Gastro-oesophageal reflux and pulmonary aspiration of acid gastric content remain significant causes of morbidity and mortality. A drug which increases lower oesophageal sphincter (LOS) tone would reduce this hazard. The effect of LOS function of intravenous cyclizine (25 mg), in half the recommended adult dose, was investigated in 8 volunteers. Cyclizine increased the LOS pressure by an average of 14,4 cm H2O (P less than 0,005). Cyclizine, like metoclopramide, has a desirable functional effect on the LOS. Both drugs are, in addition, potent anti-emetics. On the grounds of these pharmacological properties they are recommended in the preparation of patients for emergency surgery.


The role of adrenergic stimulation in the pathogenesis of pulmonary insufficiency. Recent studies have shown that epinephrine (E), norepinephrine (NE), and isoproterenol (I) caus'e substantial pulmonary shunting. This study was undertaken to determine the role of alpha (pulmonary vasconconstriction) and beta (pulmonary vasodilatation) adrenergic stimulation in the pathogenesis of pulmonary insufficiency. Forty-three mechanically ventilated, anesthetized dogs received infusions of E, alpha and beta stimulant; NE, a relatively pure alpha stimulant; I, a relatively pure beta stimulant; and dextran (D), a drug with no known adrenergic activity. The cardiac output and shunt measurements were determined simultaneously in the control period and four times during a 1 hour infusion. The quadratic equation of best fit was determined for each group for the relationship between the cardiac output and the shunt over a wide range of cardiac outputs, and statistical comparisons were made between all groups of the shunt at the same cardiac output. In all groups an increase in cardiac output was associated with an increase in the shunt, but the magnitude of the increase in the shunt differed significantly with each drug over the entire range, being smallest with D and becoming progressively larger with I, E, and NE, respectively. It is concluded that an increase in the pulmonary blood flow causes an increase in the pulmonary shunt, but that the magnitude of the increase is dependent on the specific pulmonary microcirculatory effects of each drug.


Dilemmas in the early diagnosis and treatment of multiple endocrine adenomatosis, type II. Fifteen patients with the diagnosis of multiple endocrine adenomatosis, type II, syndrome (MEA II) were reported from a single center to discuss the dilemmas of early detection and treatment of the adrenal medullary, thyroid, and parathyroid gland diseases. Ten patients came from three families. Three of the patients died, none in hypertensive crisis. Bilateral adrenal medullary disease was present in six patients. Five patients with proved pheochromocytoma had hypertension. All had diagnostic urinary catecholamine values. Nine normotensive patients without proved pheochromocytoma but in a high-risk category for adrenal medullary disease, have multiple suspicious urinary cathecholamines suggestive of adrenal medullary hyperplasia. Bilateral adrenalectomy is recommended for proved adrenal medullary disease in the MEA II syndrome. Medullary carcinoma of the thyroid gland was found in 13 patients and is believed to be present in two others. Five of the proved cases were occult, being discovered by elevation of pentagastrin-stimulated serum calcitonin levels, justifying total thyroidectomy. Parathyroid hyperplasia was found in three patients with preoperative hypercalcemia and in four others with preoperative normocalcemia. Conservative treatment of parathyroid gland hyperplasia in the MEA II syndrome is substantiated. Metachronous phenotypic expression of the syndrome components was significant.


Potentiation by the injection vehicle of the teratological action of acetazolamide in rats. The high alkalinity of the injection vehicle of certain parenteral solutions of acetazolamide produces necrosis of the skin upon sc injection. The possible modification of this effect on the teratogenicity of acetazolamide was examined. Acetazolamide in a vehicle of pH 10.5 produced 36.6% fetal malformations, in a vehicle of pH 8.7, 6.1%, and in neutral suspension, 11.8%. Adrenal medullectomy or phentolamine plus the high pH acetazolamide reduced the frequency to 23.2 and 24.4%, respectively. The teratogenicity of the low pH acetazolamide was increased by epinephrine to 64.2%. The frequency of hemimelia and micromelia, and of bilateral involvement, was greater in litters exposed to the high pH acetazolamide or the epinephrine-acetazolamide combinations, and was reduced by phentolamine or adrenal medullectomy. Neither the high pH vehicle nor epinephrine produced fetal defects in the absence of acetazolamide. The biological disposition of acetazolamide was not altered by any of the treatments. Reduction of uterine blood flow may be responsible for the potentiation of teratogenicity by the high pH vehicle.


Purification and properties of a neutral protease from soil bacterium WM 122. An extracellular protease SN 687, secreted by the soil bacterium isolate WM 122, has been purified by means of gel filtration, ammonium sulfate precipitation, DEAE-Sephadex and hydroxylapatite chromatography. Apparent homogeneity was ascertained by polyacrylamide gel electrophoresis. The protease was inactivated by ethylenediamine tetracetic acid (EDTA) but not by diisopropylfluorophosphate (DFP), and it was partially inhibited by serum inhibitors. SN 687 was shown to be of high specific activity against casein and fibrin, but it did not hydrolyze L-lysine-methyl ester dihydrochloride (LME), p-tosyl-L-arginine-methyl ester hydrochloride (TAME) and N-benzoyl-L-tyrosine-ethyl ester hydrochloride (BTEE) synthetic substrates. The optimum pH for hydrolysis of casein was 7.5 and the molecular weight, as determined by gel filtration, was 31,000.


Fetal suppressor cells. Their influence on the cell-mediated immune responses. The cell-mediated immune responses are significantly reduced in fetal and newborn mice, furthermore, spleen cells of these animals are able to suppress the immune responses of adult lymphocytes in a variety of situations. Newborn mice sensitized to picryl chloride within 24 hr after birth fail to develop contact sensitivity reaction when tested several weeks later, and fetal mice do not develop graft-versus-host reaction when given injections of parental lymphocytes. Spleen cells of fetal or newborn mice are able to suppress the passive transfer of contact sensitivity and the local graft-versus-host reaction elicited by immunized parental cells in F1 hybrid mice, and reduce significantly the severity of graft-versus-host reaction and mortality rate in cyclophosphamide-treated F1 recipients. In no experiment were thymus cells of either fetal or newborn mice found to be inhibitory. The possible mechanisms of action and biological significance of fetal suppressor cells are discussed.


Graft-versus-host reaction. Influence of genetic background in donor-recipient pairs incompatible for major histocompatibility complex (MHC). In two H-2b anti-H-2d but not in H-2b anti-H2k donor-recipient combinations, graft-versus-host reaction (GVHR) mortality was found to vary as a function of the host's genetic background; the same non-major histocompatibility complex genes and/or antigens which influence GVHR mortality do not influence the intensity of GVHR splenomegaly or the time of skin rejection. In contrast, the severity of GVHR mortality correlates with the intensity of stimulation in mixed lymphocyte culture. The roles of minor histocompatibility (H) antigens and Mls product are discussed.


[Immunosuppressive properties of vinblastine and colchicine]. The action of vinblastine and colchicine on the populations of antibody-forming cells, their precursors and stem cells using a model of primary immune response and in the transplantation system was studied using techniques by Jerne and Nordin (1963), Kennedy et al. (1965), and Till and McCulloch (1961). It was discovered that vinblastine and colchicine, at single administration 48 hours after immunization, inhibited immune response completely. The administration of colchicine and vinblastine to a donor gave an exponential decrease of in foci-forming cell number in a recipient in the system of syngenic transfer. But at the same time, vinblastine at small doses stimulated population of antibody-forming and stem cells, whereas at large doses it was not effective. Colchicine, in contrast, strongly inhibited both antibody-forming and haemopoietic cells in the recipient.


[Temporary characteristics of the process of stem cell inactivation by allogenic lymphocytes]. The time during which transplated lymphocytes block proliferation and differentiation of non-syngeic stem cells has been determined by retrasplantation of immuno-competent cells from one lethally irradiated recipient to another one. It was established that process of inactivation of CFU by allogeneic lymphocytes proceeds itwo stages. At the first stage, the colonization of recipient's tissues takes place. The colonization of tissues and processes of early recognition are completed during the first hours after transplantation of cell mixtures. At the second stage, the processes of redistribution of injected cells occur and a complete inactivation of stem cells take place. These events are completed in bone marrow and spleen 4-5 days after transplantation of cells mixture, possibly with the participation of lymphocytes sensibilized with the target-cells.


[Physicochemical characteristics of calcium binding with plasma membranes of rabbit skeletal muscles]. It is established that Ca2+ binding with rabbit skeletal muscles plasmatic membrane depends on this cation concentration in the incubating medium. pH, temperature, incubation period and presence of Mg ions in the medium affect the Ca2+-binding ability of sarcolemma. The entropic changes observed in sarcolemma Ca2+ binding are due to conformation reconstructions of the membrane components.


[Properties of chymotrypsin proteinase from Aspergillus oryzae]. Chymotrypsin-type proteinase is detected in the proteolytic system of Asp. oryzae. The action of it and chymotrypsin is shown to depend on formaldehyde. Hydrolysis of substrates, p-nitrophenyl acetate (p-NPA) and N-benzoyl-tyrosine methyl ether (BTME), by both preparations is almost the same. The obtained activity pH-optimum for the studied proteinase esterolytic activity is located in the alkaline zone as well as for crystalline chymotrypsin (substrate p-NPA). It concerns pH of both enzymes stability as well. The enzyme under study is relatively labile. At 50 degrees C there are only traces of the activity in the medium with p-NPG. Its considerable decrease is observed at 40 degrees C. This type activity is more stable on the substrate BTME. 10 min later it disappears completely in the enzymic preparation at a temperature of 60 degrees C at 40 degrees C it is 96.8%. For 24 h at 25 degrees C the activity lowers only by 8%. Crystalline chymotrypsin is stable under these conditions. DEAE-cellulose chromatography (different types of elution) detected multiple forms of proteinase differing in solubility chromatographic properties and specific activity when splitting the substrates p-NPA, BTME and casein.


Feasibility of localizing ectopic testes by 131I-labeled luteinizing hormone scintigraphy. A new method for localizing ectopic testes preoperatively was investigated in rats. Twenty hours after administration of iodine-131 luteinizing hormone the position of normal descended testes was easily identified by scintigraphy. Clinically, this technique may offer a noninvasive and practical means of locating testes undetected by palpation. The complexity and hazards of the surgical intervention to follow would thus be greatly reduced.


[Effect of acid anions and medium acidity on pH during a change of immunoglobulin G, peculiar to malignant growth, to water-insoluble state]. Affinity of an acid anion for proteins has a decisive effect on conditions for detecting using an acid medium immunoglobulin G peculiar to the malignant growth in blood serum of patients with cancer. Depending on the protein affinity of the anion of the acid used for the reaction immunoglobulin G appearing in blood of patients with cancer under equal conditions may be in a soluble state or change to a sediment. The presence of salts in the reaction mixture has no effect on conditions of the reaction performance. Exposition of blood sera to the acid medium before titration favours a more distinct detection of the studied protein in blood of patients with cancer.


[Incorporation in vivo of 14C-precursors into pyrimidine nucleotides of chicken liver and spleen tissues]. Maximum incorporation of the label after administration of 14C-bicarbonate to free pyrimidine nucleotides begins by the 60th minute. Radioactivity of cytidine nucleotides at all time points is four-eight times as low as that of uridine nucleotides. 2-14C-orotic acid is incorporated into acid-soluble nucleotides and RNA of the liver and spleen tissues. 60 min after administration of the isotope 20% of radioactivity was found in the liver and 0.3% in the spleen, in both tissues radioactivity of RNA is 20% of 14C-radioactivity in the acid-soluble fraction. The formation rate for cytidine nucleotides from uridine ones in the liver and spleen tissues is low, about 2-6% of uridine nucleotides radioactivity is found in them only, that may be due to a low activity of CTP-synthetase. The content of cytidine nucleotides in these organs is three-life times as low as compared to uridine nucleotides.


[Some mechanisms of carbohydrate metabolism regulation with NADP participation]. It is shown that in the presence of NADP in the myocardium extracts glucose-6-phosphate is transformed into fructose-6-phosphate, fructose-1,6-diphosphate and lactate. The intensity of this transformation is close to that when NAD is introduced. The addition of crystalline glucose-6-phosphate dehydrogenase to the  extracts is not accompanied by formation of lactate, but reduced NADP accumulates which oxidizes due to introduction of pyruvate or oxalacetate into the medium. This process is reconstructed in the system with pure enzymes (glucose-6-phosphate dehydrogenase, lactate dehydrogenase), that evidences for hydrogen transfer from reduced NADP to pyruvate, or to the oxalacetate without participation of transhydrogenase. The oxidation rate of the reduced coenzymes in the myocardium extracts depends on the medium pH: when pH increases, it lowers for NAD-H and rises for NAD-H. The NAD-H divided by NADP-H rate ratio at pH 7.0 for pyruvate is 11.5 and for oxalacetate is 6.5.


[Dissociation of Penicillium vitale catalase under the effect of urea and acid pH]. Conditions are studied for Penicillium vitale catalase dissociation into subunits under the effect of urea (0.7-8.5 M) and acid pH (5.0-2.0). In 8.0 M urea (pH 5.0) a molecule of the P. vitale catalase dissociates with formation of the components, the sedimentation coefficient of which is 2.4+/-0.2S, the molecular weight is 153000+/-2800. Dissociation at pH 2.2 in 4.0 M urea results in formation of components with the sedimentation coefficient 2.2+/-0.3 S. The catalase molecule dissociation under the mentioned conditions is accompanied by a loss in the enzyme activity. The results obtained show that dissociation of the P. vitale catalase molecule requires more rigid conditions than dissociation of the animal catalase. At the same time the catalase after dissociation in 8.0-8.5 M urea loses its ability to reassociation with a decrease in the urea concentration up to 0.08 M.


[Changes in enzyme activity induced by experimental aflatoxicosis in dairy cows]. The activities of GOT, GPT, LDH, gamma-glutamyltranspeptidase (gamma-GTP), alkaline phosphatase (AP), glutamate dehydrogenase (GLDH) and the concentrations of bilirubin in blood plasma after a single intraruminal application of aflatoxins were studied in four dairy cows. The maximum changes in the activities of the enzymes and the maximum bilirubin concentration in plasma were obtained in the first two to three days following the application of aflatoxins. The statistically significant increase of GOT activity, compared with activity before the application of aflatoxins, persisted until the 23rd day; in the case of LDH and GLDH the increase persisted until the 38th day from the application of aflatoxins. The activities of gamma-GTP and AP were slightly higher even on the 50th day. The increased concentration of bilirubin in plasma lasted until the 23rd day from aflatoxin application. The increased activities of enzymes testify to an impaired function of the liver, which is also proved by the specific enzymes GLDH, gamma-GTP, by increased bilirubin levels, and by histological changes known from literature. The evaluation of enzymatic activities and bilirubin concentration in plasma can make a valuable contribution to correct diagnosis of aflatoxicoses in cattle.


[Chronic metabolic acidosis in dairy cows]. Complex clinical and clinico-biochemical examination of the blood, urine and rumen liquor in a herd of dairy cows revealed chronical metabolic acidosis accompanied by rumen dysfunction and by a reduced butterfat content of milk. During the first examination of the acid-base state of the blood was almost at a standard level. An increased level of urea in blood plasma and a higher GOT transaminase activity testified to an excessive load on the liver. Urine pH was considerably deviated towards the acidic side and inorganic phosphorus was present in urine in a greater concentration. The pH of rumen liquor was slightly shifted towards alkalinity owing to the release of NH3 from urea in the food ration. The diagnosis--suspect chronical metabolic acidosis--was determined on the basis of the first examination. Chronical metabolic acidosis was definitely proved by the second examination when urea had been excluded from the feed ration. Repeated examinations revealed chronical metabolic acidosis which had originally been accompanied by a higher rumen liquor pH. On the basis of case histories and mechanisms of chronical acidosis, measures were proposed, resulting in an increase of the butterfat content of milk. Chronical metabolic disorders often follow a long-lasting latent pattern, manifesting themselves as a reduced milk yield and lower resistance; the clinical form of disease appears only at a later stage. The system of preventive diagnostics provides information on the changes in the composition of internal medium and of the faeces before a drop occurs in milk and fat production. These measures prevent metabolic disorders and high losses of produce which otherwise remain hidden for a long time.


[Significance of ruminal juice examination in the diagnosis of subclinical rumen dysfunction]. The suitability of the metabolic test of rumen liquor for the diagnosis of subclinical rumen dysfunction was tested. The test for the assessment of the metabolic profile of the rumen included: the determination of pH, total titration acidity in clinical units, lactic acid in mg%, fatty acids in mmol 1-1, ammonia N in mg%, urea N in mg% and the number of infusorians in 1 ml of rumen liquor. This test enabled, on the basis of the examination of rumen liquor, to determine not only the kind and form of the disease but also its cause, with an incomplete clinical syndrome. Therapeutic measures, either direct (in the rumen liquor) or indirect (adjusted monodiet) improved the health condition and provided optimum rumen fermentation of permanent character. The resuption of rumen fermentation also enabled an increased production of fatty acids which favourably influenced the content of butterfat in milk.


HLA-antibody-induced 14C-serotonin release from platelets. A methodological analysis. The prerequisites and kinetics of HLA-antibody-induced 14C-serotonin release from platelets were investigated with five HLA-specific antisera. By studying the influence of incubation time, temperature, pH, platelet count, stirring and extent of platelet labeling, optimal results were obtained after 30 (to 60) min of incubation at 37 degrees C, pH 7.5-8.0, under constant stirring (400 rpm). Optimal platelet counts varied between 160,000 and 400,000/microliter depending on the antiserum applied. A comparison of the amount of 14C-serotonin released by different antisera from washed platelets and from platelets in platelet-rich plasma suggests that plasma components--most likely complement--may additionally influence the results. Based on these experiments, a standardized microtechnique of HLA-antibody-induced 14C-serotonin release is recommended.


[Electrophoretic and kinetic properties of the erythrocyte glucose-6-phosphate dehydrogenase of patients with hemolytic anemia related to a deficit in the activity of that enzyme]. Partially purified glucose-6-phosphate dehydrogenase (G6PD) was studied in erythrocytes of patients with hereditary hemolytic anemia. Kinetic and electrophoretic properties of G6PD distinctly varied both in patients--homozygotes and in maternal heterozygotes. All the enzymes studied together with the decreased activity possessed the reduced Km value for NADP and G6P. Anomalous enzymes were shown to differ from normal ones by several patterns (pH-dependency, thermostability, % of the substrate analogues utilization, electrophoretic mobility etc). The mutant variant of G6PD was not described earlier. The anomalies variant was called "Kremenchug" to indicate the place of proband origination.


[Purification and several properties of thiamine pyrophosphokinase from rat liver]. Thiamin pyrophosphokinase (TPK) (EC 2.7.6.2) was isolated and purified from rat liver tissue ion in exchange chromatography, rechromatography and gel filtration. The enzyme was purified 3000-fold with yield of 12%, it was homogenous in polyacrylamide gel disc electrophoresis. TPK had two pH optima; Km value for thiamin was equal to 6-10(-6) M. Pyrithiamin was shown to be a competitive inhibitor with K1=3-10(-6) M and hydroxythiamin--an inhibitor of the mixed type with K1=1-10(-2) M. The relationship between the rate of the reaction and the substrate concentration (ATP-Mg2+) at the Mg2+/ATP ratio of 1:1 was described by S-like curve, which acquired a hyprebolic form in presence of an excess of Mg2+. Dependence of the reaction rate on Mg2+ concentration was also described by a sigmoid curve. The data obtained suggest that liver TPK was apparently an allosteric enzyme with quaternary structure.


[Immunochemical characteristics of hydrocortisone induced and hydrocortisone non-induced rat liver tyrosine aminotransferase isoenzymes]. Two isoenzymes of tyrosine aminotransferase (induced and noninduced by hydrocortisone were isolated from rat liver tissue and subjected to chromatographic purification. Specific antibodies with high titer were obtained after immunization of rabbits by the inducible isoenzyme. The antigenic properties of the obtained tyrosine aminotransferase isoenzymes were compared after double immunodiffusion in agar and immunoelectrophoresis. The purified induced and non-induced isoenzymes were shown to possess both similar and different antigenic determinants. The data obtained suggest that differences in antigenic properties of the tyrosine aminotransferase isoenzymes reflect those variations in their structure, which are expressed as various thermostability, sensitivity to proteolytic enzymes and substrate specificity.


Contamination of rat urine with gut flora using all-glass metabolism cages for collection of urine and faeces. In rat urine collected in all-glass metabolism cages, at least four strains of intestinal microflora were found: two types of E. coli, Enterobacter cloaceae and Proteus vulgaris. The number of bacteria of each strain increased with time. 2. The pH of the urine increased from 6-9 after 24 h to 8-95 after 120 h. The pH of the urine of neomycin-treated rats remained nearly constant over a period of two days. 3. Nitroreductase activity was present in the rat urine. Added p-nitrobenzoic acid was reduced within the first 24 h. Nitroreductase activity in the urine of neomycin-treated rats was significantly lower than in the urine of normal rats, during the second 24-h period only. 4. Collection of urine at -10 degrees prevented the consequences of contamination.


[Influence of the age of old primiparae on the perinatal infant morbidity and mortility (author's transl)]. 157 old primiparae (greater than 30 a) have been compared to a randomised control group of primiparae between the 16th and 30th year of age. In addition to the perinatal infant mortality rate, the number of children with malnutrition, the frequency of premature babies, the presence of a low Apgar score (less than or equal to 7) and pathological pH-values in the umbilical artery (pH less than or equal to 7,10) were cheked. Moreover the frequency of early neonatal morbidity (presence of one of the following criteria: 1 minute Apgar score less than or equal to 7, pH of the umbilical artery less than or equal to 7, 10, a transitoric or severe RDS and neurological complications during the newborn period) was investigated in both groups. In the group of the old primiparae a significantly higher perinatal infant mortality rate was found compared to the control group (3,8% versus 0,6%, p less than 0.05). The rate of small for date babies was remarcably higher than in the control group (14,6% versus 6,4%, p less than 0.05). There was no difference in the prematurity rate in both groups. The children of old primiparae had more frequently a low Apgar score (less than or equal to 7) than infants of the randomised control group (18,5% versus 11,5%), but the difference is statistically not significant. Furthermore no difference in the number of pathological pH values in the umbilical arteries (pH less than or equal to 7, 10) were found in the both groups. The children of old primiparae show a higher early neonatal morbidity rate compared to the control group (21.7% versus 14.0%); however, the difference is statistically not significant. According to our results the risk for the newborns is increased already with primiparae over 30 rather than 35 years of age.


[Pharmacokinetic of piracetam during labour influence to acid-base-status in maternal and fetal blood (author's transl)]. PIRACETAM concentrations in fetal and maternal blood were measured during the first and second stage of labor and the elimination in maternal and fetal blood was studied. Fetal heart-rate, pH- and base-excess of the maternal and fetal blood were investigated to evaluate the influence of PIRACETAM on the acid-base-status of mother and child. PIRACETAM was administered intravenously to 43 patients in a dosage 2 g, 4 g and 6 g. The concentration in the maternal and fetal plasma was measured by gaschromatography. Before and after the injection of PIRACETAM and at delivery, blood was sampled from the mother's earlobe and the umbilical artery and vein, respectively. The results were compared with a control group. There was an exponential fall of PIRACETAM concentration in maternal and fetal blood. Maternal and fetal elimination half-life of PIRACETAM was about 112-98 minutes and 200 minutes, respectively. A fairly good correlation between the concentration of PIRACETAM in maternal and fetal blood was found. The fetal PIRACETAM concentration was about 50% below the maternal values. Fetal heart-rate, maternal and fetal pH- and base-excess-values were not significantly altered following PIRACETAM infusion. It may be concluded, that there exists a transfer of PIRACETAM across the placental barrier. However, the cytoprotective effect of PIRACETAM, as described in animal observations and by investigations in human, could not be verified by the methods and technology used in this study.


Studies on in vitro survival of Isoparorchis hypselobagri. Adult parasites, Isoparorchis hypselobagri were maintained in vitro in four basic salt solutions, viz. Stephenson saline, AB saline, Tyrode's solution and Locke's solution. Survival was prolonged by adding sugars to Tyrode's solution. Of all the sugars, glucose was found most effective and parasites survived for 39 days in solution containing glucose, 6 days in galactose and sucrose, 10 days in fructose and lactose and 12 days in maltose. The parasites absorb carbohydrates through the cuticle. Ligated parasites survived for a longer period than unligated parasites. The pH 9 was considered as an optimum for maximum survival of the parasites Isoparorchis hypselobagri for 39 days.


[On the enzymatic breakdown of tripolyphosphate and diphosphate in comminuted meat. XI. Influence of heating and freezing (author's transl)]. The tripolyphosphatase and diphosphatase activities of comminuted bovine muscle increase with rising temperature until at about 40 degrees C a denaturation of the enzyme proteins occurs. Prae or post rigor frozen bovine muscle, which was stored at -18 degrees C, shows after thawing and mincing the same tripolyphosphatase activity as the non-frozen tissue post rigor. Freezing and thawing under the same conditions cause a reversible inhibition of the  diphosphatase activity of bovine muscle. The reactivation depends on the pH. This inhibition is probably identical with that occurring during storage of beef at -4 degrees C.


[Formation of dehydroalanine, lanthionine and lysinoalanine during heat treatment of beta-lactoglobuline A (author's transl)]. Solutions of beta-lactoglobulin A were heated at 97.5 degrees C at pH 6.9 and 9.8 80 min. The formation of dehydroalanine, lanthionine and lysinoalanine as well as changes in the levels of cysteine, cystine and the basic amino acids were recorded. Lanthionine (equal amounts of the 1- and the mesoform) was formed only in small amounts, while the lysinoalanine levels were rather high. Beside this, lysine was converted to unknown compounds. The main sources for the formation of dehydroalanine are the cystine moieties of the protein. Cysteine residues are first oxidized to cystine before beta-elimination occurs.


[Detection and quantitative estimation of 2,2-diethyl-4-pentenamide in a fatal case of poisoning (author's transl)]. Toxicological investivation in a fatal case of poisoning by Novo-DolestanR and DoromaR is presented with methods for identification and quantitative estimation of 2,2-diethyl-4-pentenamide especially. The findings confirm 2,2-diethyl-4-pentenamide as the main toxic agent in this case.


[Evaluation of congenital-cryptorchid testicles after surgical treatment (author's transl)]. The volume of 271 congenital-cryptorchid testicles which were subject to surgical treatment was compared with that of 128 descended testicles of patients of the same age. Undescended testicles, also after successful surgical treatment, remain smaller on volume than descended ones. By means of the hitting method of the point counting technique the proportion of all seminiferous tubules in the total volume of one gonad can be calculated. The seminiferous tubules of congenital-cryptorchid testicles, also after intrascrotal replacement, maintain an essentially smaller proportion concerning the total volume as in descended testicles. Among the numerous methods of surgical treatment preference must be given to those which are especially favourable to the parenchyma.


Accentuation of Staphylococcal hemolysis (CAMP-like test) by pneumococci. A CAMP-like phenomenon can be observed in Streptococcus pneumoniae. It is supposed to be caused likewise as in group B streptococci by a haemolysis accentuating factor. However, the form of the accentuated haemolysis is different from that in group B streptococci and similar to that in listeria.


A catalogue of isohormones of human growth hormone based on quantitative polyacrylamide gel electrophoresis. With the growing recognition that all polypeptide hormones exist in heterogeneous forms, i.e., isohormones, this study represents the initial development of a comprehensive catalogue of the various isohormone species of human growth hormone (hGH) in relation to molecular structure. The catalogue is based primarily on results obtained in quantitative polyacrylamide gel electrophoresis (PAGE) in a multiphasic buffer system optimized for characterization of hGH. Thus, hGH forms can be recognized in objective, numerical terms based on physicochemical parameters of molecular size and net charge. Isoelectric focusing in polyacrylamide gel; and PAGE in sodium dodecyl sulphate in the presence and absence of reducing agents supplement the quantitative PAGE criteria. Specific hGH immunoreactivity is necessary to completely distinguish hGH from hormones of similar size and charge properties (prolactin and chorionic somatomammotrophin). In the examination of eight standard hGH preparations obtained from four laboratories, all preparations exhibited multiple isohormone species, from two to five components per preparation. These identifiable components appear to represent the predominant species of a bidirectional continuum of hGH components, the continuum being in regard to both molecular size and charge. Within any given preparations, the continuum presents itself as a series of identifiable "charge isomers". Between preparations, in addition, there are size differences as well, which may relate to conformational changes (unfolding) in some preparations.


[Alteration of several indices of carbohydrate metabolism in the blood and cerebrospinal fluid following surgery on patients with intracranial aneurysms]. The content of the lactic acid (LA) and pyruvic acid (PA) was determined pre- and postoperatively in the arterial blood and CSF of 31 patients with intracranial aneurysms. A considerable elevation of the concentration of LA and PA was noted along with an increasing LA/PA in the CSF of patients with symptoms of ischaemic damage of the brain. During surgery under fluothane narcosis these patients also demonstrated a distinct elevation of the LA level along with an increasing LA/PA ratio. In the CSF the growing content of glycolysis metabolites was not accompanied by LA/PA changes. The employment of artificial hypotension had no important effect upon the changes of these parameters in the CSF.


[Behavior therapy and neurotic behavior]. The first part of the lecture presents three classical aspects of behavior therapy: therapeutic efficiency, utilisation of well defined methods, rapidity of treatment. Emphasis is brought to the importance of behavior analysis and to the dangers arisen from insufficient knowledge or ignorance of learning theories and fundamental principles of behavior analysis. One insists on experimental analysis which is the basis for methods of action having as consequence for the therapeut: controlling strategy of treatment and direct responsibility concerning success and failure. Then, actual limitations of behaviour therapy are described. The second part deals with one of the most important working hypothesis on human neurosis originating from laboratory and clinical research. Behavior therapy refuses to elaborate hypothetical deductive constructions, unless working hypotheses to be verified. Then the four essential clues to behavioural psychotherapy are formulated. The main methods of action presently utilized are presented: aversive methods, operational and systematic desensitization technics through reciprocal inhibition. Finally, some of the main criticism usually made on behaviour therapy are being discussed.


[Behavior therapy of fear of flying in professional pilots]. The theorical frame proposed by behaviour principles seems to have an explanatory value in the genesis of some troubles of traumatic origin. This is the case for fear of flying which can appear suddenly in airline personnel following accidents, troubles or problems while flying and which could bring a definitive incapacity of flying. The use of behaviour therapy, like systematic desensibilisation, is then being considered. It is illustrated by examples taken from the literature and by two clinical cases.


[Schultz autogenic training and galvanic skin response in the systematic desensitization of a compulsive phobia]. A patient suffering suicidal phobia and resistant to antidepressants as well as electroconvulsive therapy was submitted to Wolpe's systematic desensitization associated with Schult's autogenic training. Ten sessions were sufficient to cure the patient. A follow-up period now extends to one year, without relapse. The concomitent measure of skin galvanic response by an electrodermograph appears a privileged psychophysiological control of such a therapy, i.a. allowing for detection of the avoidance of the anxiogenic evocation by the patient.


Syntheses of virus-induced thymidine kinase and viral DNA in herpes simplex type 1 virus-infected chick embryo fibroblasts. Herpes simplex virus type 1, strain Kupka, did not replicate in chick embryo fibroblasts (CEF), but the infection was followed by the development of cytopathic changes. This effect could be abolished by UV irradiation of the virus. Virus-induced thymidine kinase was synthesized in the infected cells reaching a maximum level at 24 hours post infection (p.i.). In the presence of cytosine arabinoside, thymidine kinase synthesis was enhanced. This suggested that the late (post-replicative) viral function, which turns off thymidine kinase synthesis, was expressed in the infected CEF untreated with the drug. HSV type 1 laboratory strains Kupka and KOS were capable of inducing the synthesis of virus-specific DNA in CEF. But in CEF infected with fresh type 1 virus isolates replication of viral DNA was not observed.


Differential expression of D and R components of Epstein-Barr virus early antigen after superinfection and after induction with 5-iododeoxyuridine. Three non-productive human lymphoblastoid cell lines, NC-37, RAJI and L-1, either superinfected with Epstein-Barr (EB) virus from P3HR-1 cells or treated with 5-iododexyuridine (IUDR), were studied for the development of early antigen (EA) of EB virus. In all three superinfected cell lines, both the restricted (R) and diffuse (D) componets of EA were synthesized, with R-component formation preceding D-component synthesis. After IUDR treatment both components of EA were only formed in RAJI cells, while in NC37 and L-1 cells only the R-component was synthesized.


Production and some properties of neutralizing antigens of herpes simplex virus. Human diploid cells (LEP) infected with herpes simplex virus (HSV) type 1 were extracted by treatment with Nonidet P-40. The content of antigens reactive with neutralizing antibody in the clarified extract was measured by the blocking test. Influence of various factors on the production of the blocking antigens (BA) was tested. Highest BA titres in LEP cells were achieved at a multiplicity of infection (m.o.i.) of 0.5 TCD50/cell, medium 199 and 22 to 29 hours incubation at 37 degrees C. The use of resting cultures or the presence of cytosine arabinoside in medium reduced the BA production. The activity of BA was reduced by repeated freezing and thawing. The antigen was stable at -20 and -70 degrees C. One hour heating at 60 degrees C resulted in a marked decrease of the antigen titre. BA was resistant to formalin; formalin treatment did not influence its thermosensitivity.


Heterogeneity of pseudorabies virus studied by isoelectric separation. Heterogeneity of four lines derived from a single strain of pseudorabies virus (PRV) was studied by isoelectric separation. The individual lines differed in the distribution pattern of infectious fractions separated in the pH range from 3.5 to 10.0. All virus fractions of the virulent and attenuated lines were infectious, but some fractions of the highly attenuated avirulent line were devoid of infectivity. Infectivity of the fractions isolated from the individual PVR lines depended on the degree of attenuation; the higher degree of attenuation, the greater the differences in infectivity of the isolated fractions. The possible causes of the different distribution of infectious fractions obtained by isoelectric separation from virus lines differing in biological properties are discussed. The relationship between physico-chemical and biological properties of PRV is stressed along with the possibilities of applying isoelectric separation in narrow pH gradients.


Nucleic acid biosynthesis in rat embryo cells infected with X14 or H-1 parvovirus. Nucleic acid biosynthesis was studied in rat embryo cell (REC) cultures 48 hours after infection with X14 or H-1 parvovirus. The incorporation of 14C-formate and [6-(14C]-orotic acid into purines and pyrimidines of various was lowered after infection with these parvoviruses. 14C-Formate incorporation into acid-soluble thymine was greatly inhibited in H-1 virus-infected cells whereas it was slightly inhibited in X14 virus-infected cells. These results suggest that X14 virus-infected cells can carry out the biosynthesis of thymidylic acid utilizing some endogenous pyrimidine nucleotide (e.g. deoxycytidylic acid, via deoxyuridylic acid). In the infected cells, the nucleoplasmic RNA polymerase activity was strongly inhibited. This results suggests an interference by the two viruses with hosts RNA synthesis.


Properties of virions and ribonucleoproteins of Venezuelan equine encephalomyelitis virus. The physicochemical properties of Venezuelan equine encephalomyelitis (VEE) virus and of its ribonucleoprotein (RNP) were studied. Upon purification in a discontinuous or linear sucrose gradient, the losses of infectivity were small (25%), and 96% of cellular proteins were removed. The purified virus was homogeneous with respect of sedimentation rate (s20, w = 265 S). The CsCl gradient was unsuitable for purification of infectious virus because the latter was destroyed at high CsCl concentrations. Buoyant density of the virus in CsCl after formaldehyde fixation was 1.21--1.22 g/cm3. Treatment of the virus with 1% Nonidet P-40 proved to be the most effective method for isolation of the RNP. The structures thus obtained contained practically all of the viral RNA and about 20% of viral proteins, and were homogeneous with respect of sedimentation rate (153 S) and buoyant density (1.40--1.42 g/cm3 in CsCl after formaldehyde fixation). The RNPs were sensitive to ribonuclease.


Latent infection and measles meningoencephalitis in Syrian hamsters. A strain designated 42-Edmonston was isolated from the brain of a Syrian hamster 30 days after inoculation during the neonatal period with the reference "wild" measles virus strain Edmonston which produced a latent infection accompanied by production of measles antibody and insignificant pathomorphological changes. Intracerebral inoculation with the 42-Edmonston strain of newborn, one day-and 5-7-day-old Syrian hamsters resulted in the development of acute measles encephalitis with typical clinical symptoms and pathomorphological changes. It is suggested that selection and adaptation of a neurotropic measles virus variant probably had occurred in the course of virus persistence in the Syrian hamster brain.


Serodiagnosis of influenza by indirect haemagglutination test. Results of studies on serodiagnosis of influenza A and B by the indirect haemagglutination test (IHA) with lyophilized erythrocyte diagnostic preparations, highly stable upon storage, are presented. The diagnostic value of the IHA test was in good agreement with, that of virus neutralization tests.


Improved methods of influenza virus propagation. I. Enhancement of virus reproduction in cell cultures. Comparative studies on the reproduction of a set of influenza A and B virus strains in different cell cultures were carried out. A cultivation system yielding influenza virus with high infectivity, based on the use of roller cultures of bovine embryo kidney cells, of virus strains adapted to this culture, and of a maintenance medium of improved composition, was developed.


Improved methods of influenza virus propagation. II. Characteristics of cell culture and allantoic virus preparations. Regular reproduction of influenza A virus with infectious titres of 7.4--8.5 log EID50/0.2 ml was observed in roller bovine embryo kidney cell cultures. The haemagglutinating activity of the cell culture preparations was 8-33-fold lower than that of the allantoic preparations, the infectivity being similar. Cell culture preparations of influenza A/Victoria/35/72 and A /Leningrad/538/74 viruses were markedly immunogenic in laboratory animals and antigenically active in complement fixation (CF) test. The CF activity of liquid and lyophilized preparations remained unchanged for 12--13 months (the period of observation). The stability of infectivity of lyophilized cell culture virus preparations was similar to that of allantoic virus preparations.


Analysis of antibody response and immunoglobulins in sera of rabbits and guinea pigs immunized with Coxiella burnetii. Rabbit and guinea pig sera and their immunoglobulin fractions (IgM and IgG) were examined by complement-fixation (CF), microagglutination (MA), opsonization-phagocytosis (OP) and serum protection (SP) tests at intervals after immunization with live phase I and phase II Coxiella burnetii suspensions. In general, MA antibodies, but also decreased, earlier than CF antibodies. The anamnestic immune response was higher with lower primary doses. Both phase II and phase I CF and MA antibodies as well as phase I opsonins and protective antibodies were found in either immunoglobulin fraction depending on post-immunization intervals. At later intervals and especially after the second immunizing dose the levels of protective antibodies in whole sera, but mainly in the IgG fractions, showed a better agreement with those of phase I opsonins rather than of phase I CF and MA antibodies.


The role of the nucleus in the replication of measles virus. The replication of measles and SSPE viruses in enucleate BSC-1 cells was measured by plaque assay, and the synthesis of virus-specific antigen was measured by indirect immunofluorescence. Titres of infectious virus produced in enucleate cells at 24 hours post inoculation (p.i.) were consistently 100-fold less, and the number of enucleate cells containing virus antigen at 30 hours p.i. was 10-fold less, than in nucleate control cultures. Enucleate cells at this time had 30% of the protein synthetic capacity of control nucleate cells, and supported the growth of Semliki Forest virus to titres equivalent to nucleate cell production.


Antiviral effect of tebrofen.

Antiviral effect of tebrofen. Tebrofen inhibited the replication of western equine encephalomyelitis (WEE) virus in multicycle experiments in chick embryo cell cell cultures. The compound did not inhibit the replication of vaccinia and Newcastle disease viruses.


Effect of physical and chemical agents on the blue-green algal virus N-1. The stability of the blue-green algal virus N-1, infecting the blue-green alga Nostoc muscorum, against temperature, EDTA and pH was studied. Ninety per cent thermal inactivation of virus occured at 55 degrees C in 20 minutes with an average inactivation rate constant of 0.07 per minute; at 60 degrees C the virus was completely inactivated within 5 minutes. 10(-4) M EDTA inactivated 30 per cent of free virus in 60 minutes; 10(-3) M EDTA was completely lethal. Intracellular virus was comparatively resistant to EDTA. The virus was stable in the pH range from 6.7 to 10.5 and was inactivated at acidic pH.


Scanning and transmission electron microscopy of Rickettsia rickettsii propagated in cell culture. Scanning electron microscopy utilizing critical-point drying and transmission electron microscopy employing air-dried agar pseudoreplicas and critical-point dried carbon replicas were used to study the surface of Rickettsia rickettsii propagated in cell culture.


Adrenergic mechanisms in infection. III. alpha-and beta-receptor blocking agents in treatment. Raised epinephrine concentrations concentrations developed in the blood of mice near death after an intraperitoneal challenge with 10(9) Staphylococcus aureus or Escherichia coli. Both epinephrine levels were elevated in the urine near death. These challenges were infections and not intoxications, as they could be successfully treated with antibiotics. Using the same model infections, we studied the effect of alpha-and beta-receptor blocking agents and other adrenergically active compounds. The compounds alone were not toxic in the doses used. alpha-Blockers, such as phenoxybenzamine, dibenamine, and ergotamine and beta-blockers, such as butoxamine, timolol, and practolol, were ineffective. Phentolamine alone or propranolol alone showed some evidence of an ability to prolong life in S. aureus infections. Together they lengthened life significantly in S. aureus-challenged mice. In E. coli infections this beneficial effect was not noted. Mice pretreated with reserpine (7 mg/kg) and challenged with S. aureus lived 55% longer than saline-treated controls. Less benefit was noted in E. coli infections. Serontonin treatment was helpful in S. aureus infections and deleterious in E. coli infections.


Effects of pneumococcal infection on rat liver microsomal enzymes and lipogenesis by isolated hepatocytes. Modification in the enzymatic complement and lipogenic functions of rat liver endoplasmic reticulum (ER) were shown to occur during pneumococcal sepsis. Glucose-6-phosphatase, 5'nucleotidase, esterase, and NADPH cytochrome C reductase decreased in activity by as much as 50% with respect to controls. Hydroxymethylglutaryl-CoA and NADH cytochrome C reductases were increased 6-and 2-fold, respectively. Alkaline phosphatase and inosine-5'-diphosphatase did not differ with respect to fasted controls. The lipogenic capacity of the ER was shown to be enhanced. In vitro [14C]acetate incorporation into cholesterol and other lipids by hepatocytes isolated from infected rats was increased 2-to 10-fold. It is concluded that the flow of acetyl-CoA in liver cell of Streptococcus pneumoniae-infected rats is toward lipogenesis rather than ketogenesis.


Prostaglandin E2-hemoglobin AA and SS erythrocyte interaction (prostaglandin-erythrocyte interaction). Prostaglandin E2 (PGE2), at concentration larger than or equal to 5 x 10(4) ng/ml, induced discocyte leads to echinocyte transformation of saline-suspended hemoglobin (Hb) AA and SS erythrocytes. This erythrocyte transformation is concentration-dependent and is reversible at room temperature after 90-120 min. The Hb SS erythrocytes treated with PGE2 did not exhibit accelerated sickling or increased formation of sickled echinocytes. Erythrocytes suspended in autologous plasma treated with PGE2, 2 X 10(6) ng/ml, did not exhibit echinocytic transformation probably because of drug binding to the plasma proteins. Other in vitro studies showed that PGE2 of concentrations of 10-500 ng/ml had no adverse effects on intact, plasma-suspended Hb SS erythrocytes. These Hb SS erythrocytes were examined for changes in morphology, potassium and calcium flux, and blood viscosity under oxygenated and hypoxic conditions.


Biological activity of dilute isoproterenol solution stored for long periods in plastic bags. The effect of long-term storage in polyvinyl chloride bags on the biological activity and the chemical stability of isoproterenol was studied. A freshly prepared solution of isoproterenol, isoproterenol solution stored for six months and isoproterenol stored for 30 months were infused into the coronary circulation of 14 isolated rabbit hearts; heart rates were determined. Spectrophotometric studies to determine stability at various pH levels were also done. Aging of the solutions at pH 4 and at a concentration of 4 microgram/ml caused no significant impairment of their potency. Solutions above pH 6 were unstable.


Continuous subcutaneous pH measurement in human fetuses: correlations with scalp and umbilical blood pH. The subcutaneous pH values of 42 fetuses has been measured continuously during labor with the use of the electrode designed by Stamm and associates. We have obtained 60 per cent good tracings. The results have confirmed the reliability of the subcutaneous pH measurement, if the electrode is correctly inserted, when compared with both capillary and umbilical artery blood pH measurements (r = 0.91, p less than 0.001 and 0.83, p less than 0.001, respectively). Technical improvements appear to be necessary prior to widespread clinical use of continuous pH measurement.


Rapid emergence of carcinogen-induced hyperplastic lesions in a new model for the sequential analysis of liver carcinogenesis. Hyperplastic liver lesions which develop following administration of hepatocarcinogens have been implicated as probable precursors for the cancers which ultimately develop. Some, and possibly all, of these putative precursor lesions are resistant to the necrogenic and growth-inhibitory action of hepatocarcinogens and other hepatotoxins. An in vivo assay system based on this resistance phenomenon has been developed which encourages the rapid selective growth of carcinogen-altered hepatocytes, facilitating their early identification. The system consists of a) single carcinogenic dose of diethylnitrosamine (DEN), b) short-term dietary exposure to 2-acetylaminofluorene (2-AAF) sufficient to suppress growth of virtually all normal hepatocytes, and c) partial hepatectomy (PH) to actuate rapid growth of DEN-altered hepatocytes not suppressed by 2-AAF. Following PH, the DEN-altered hepatocytes grow out as basophilic foci which are distributed randomly throughout the 2-AAF-suppressed parenchyma. Within 1 week they can be seen as tiny, discrete, translucent nodules on the capsular and cut surface of the remaining lobes. The lesions continue to proliferate and become histologically indistinguishable from typical carcinogen-induced hyperplastic liver nodules frequently described in the literature. These in turn appear to be precursor lesions for at least some hepatocellular carcinomas. Future experimentation based on this phenomenon of "selective resistance to cytotoxicity" should prove valuable in answering specific questions about the carcinogenic process in liver and possibly in other tissues.


Vitamin K2 colonic and ileal in vivo absorption: bile, fatty acids, and pH effects on transport. Colonic and ileal absorption of vitamin K2 ([2-methyl-3H]menaquinone-9) was investigated in the conscious rat. When the absorption rate was plotted against the perfusate concentration, a linear relationship was found between these two parameters in the ileum and colon. The absorption rate of menaquinone by the ileum was increased as the bile salt concentration, degree of unsaturation of the added long-chain fatty acids, hydrogen ion concentration, and perfusate flow rates were increased. Colonic menaquinone absorption decreased as the bile salt concentration was increased. Menaquinone colonic absoprtion increased as the pH decreased, but no change was noted as the perfusion rate was increased. The present experimental observations in vivo, coupled with prior observations in vitro, indicate that absorption of menaquinone by the ileum and colon occurs by a passive diffusion process that is modified by variations in the perfusate bile salt concentration, the presence of unsaturated fatty acids, and the perfusate pH. The present observations indicate that the mammalian colon and terminal ileum can provide a constant source of vitamin K to aid hemostasis despite episodic lack of dietary vitamin K.


Glucose utilization and role of blood in endotoxin shock. The present study was conducted to explore influences modifying glucose uptake in canine blood administered LD100 E. coli endotoxin. Particular emphasis was given to assay the role of the white blood cell (WBC) in glucose utilization. Significant increases in glucose uptake and lactic acid production, attributed to increased activity of the WBC, were observed 1-3 h after endotoxin was added to blood in vitro. Although a net increase in glucose utilization was noted, endotoxin simultaneously exerted adverse effects by depressing glucose uptake below predicted values (Q10 = 2.12 with LD100 endotoxin vs. 2.78 in saline controls) and increasing WBC mortality rate. Blood from dogs pretreated with sublethal doses of endotoxin in vivo utilized glucose at an accelerated rate when subjected to endotoxin in vitro. Excess glucose was consumed because of elevated numbers of white blood cells although additional glucose requirements after endotoxin were independent of temperature between the ranges of 34-41 degrees C. All animals pretreated with daily sublethal injections of endotoxin  for 3 days survived superlethal doses of endotoxin.