A possible model for cell-cell recognition via surface macromolecules. Alternative possibilities for the establishment of the proper cell distribution during embryogenesis are summarized at the beginning, followed by an assessment of the examples known so far where cell-cell recognition is known to be mediated via cell surface components. In the second part the species-specific recognition process which occurs during the sorting-out of dissociated sponge cells is analysed since it may serve as a possible model for cell-cell recognition in higher animals. Three possible mechanisms for the establishment of proper cell distribution are considered. These include, first, chemotaxis: secondly, guidance of cell or cell sheet  movement by extracellular matrix or by surrounding cells and thirdly, random movement followed by recognition at the final point of destination. Recognition is necessary for both of the two latter processes, i.e. for cell guidance as well as for locking the cells into their final position after random movement. Two basically different recognition mechanisms should be distinguished from each other. On the one hand cells may recognize each other with the help of macromolecules situated in or just outside of the plasmamembrane which fit to each other like enzymes and substrates or antibodies and antigens. On the other hand, cells may exchange information by exchanging cytoplasmatic components via vesicles or gap junctions. The species-specific aggregation of dissociated sponge cells is considered to be a possible model for cell-cell recognition in higher animals. A proteoglycan-like intercellular macromolecule called aggregation factor seems to mediate recognition of a given species of cells in the reaggregation process of dissociated cells. The data available at the present time suggest that a monovalent surface macromolecule (baseplate) may mediate the recognition process probably by recognizing the carbohydrate side chains of the multivalent proteoglycan aggregation factor. A cell-free system was devised to mimic this aggregation process. Addition of aggregation factor to baseplate-coated sepharose beads of approximately the size of the original sponge cells has essentially the same characteristics as the cellular system. Macromolecule-coded surface information for the recognition between cells has not been established during the embryogenesis of higher animals and remains an interesting challenge.


Role of the cell surface in selection during transport of proteins from mother to foetus and newly born. The transport of immunoglobulins from mother to foetus and newly born mammal involves selective events which are independent of molecular size, related to immunoglobulin class, structure, and species of origin, and involve considerable protein degradation. Such events are briefly described as background information to a discussion of how selection of proteins might take place during transport across the cellular barriers concerned, namely the yolk sac splanchnopleur, chorio-allantoic placenta, and small intesting. Until recently the Brambell hypothesis has been the most favoured explanation. This implies that selection occurs intracellularly, within endodermal cells of the yolk sac splanchnopleur and small intestine, and within the syncytiotrophoblast of the chorio-allantoic placenta, of certain species. It also suggests that specific receptors are present which give attached proteins protection from degradation when the vesicles containing them fuse with lysosomes; such protected proteins are then liberated from the vesicle by exocytosis. This hypothesis is examined in the light of what is now known about the mechanism of uptake and transport of proteins by the endodermal cells and syncytiotrophoblast. It is suggested that rather than being an intracellular event, involving protection from proteolytic degradation, selection takes place at the cell surface. Evidence is presented, some direct and some circumstantial, that proteins may be selectively endocytosed by coated micropinocytotic vesicles, and non-selectively endocytosed through a complex apical canalicular system leading to macropinocytotic vesicle formation. In the small intesting of the suckling rat these two processes appear to be segregated, selective uptake occurring in the proximal half and non-selective uptake occurring in the distal half. In the endodermal cells of the rabbit yolk sac splanchnopleur, and by implication in the syncytiotrophoblast of man and monkey, it is suggested that both selective, and non-selective, uptake of protein occurs. Non-selective uptake into macropinocytotic vesicles is regarded as an event leading to complete degradation of all contained protein and functioning so as to supply the foetus and newly born mammal with essential amino acids. Selective uptake into coated micropinocytotic vesicles is regarded as an event leading to the transport of immunoglobulins across the cell without any contact with lysosomes, and functioning so as to supply the newly born mammal with protection against invasive organism. Specific receptors are still required but only for the initial uptake and segregation of proteins into coated micropinocytotic vesicles. The role which the glycocalyx might have in such selective binding of proteins is considered and possible difficulties in characterization of specific receptors brought to light in view of the likely overwhelming need for non-specific binding to effect non-selective uptake.


Inhibition of adenosine 3',5'-monophosphate phosphodiesterase by nicotinamide and its homologues in vitro. Inhibition of cAMP phosphodiesterase from rat liver by nicotinamide and its homologues was studied. Among the several compounds tested, such agents as nicotinamide, N2-ethylnicotinamide, N2-methylnicotinamide, N,N-diethylnicotinamide, 3-acetylpyridine, methylnicotinate, and ethylnicotinate showed potent inhibition of cAMP phosphodiesterase, with an over 90% inhibition by 5 mM ethylnicotinate when cAMP was used as substrate at a 0.48 x 10(-7) M concentration. A comparison of the inhibitory curves of theophylline, papaverine, and ethylnicotinate on enzyme activity showed them to be approximately coincident. Furthermore, the plots with abscissa of equal increments per concentration of ethylnicotinate in the presence of theophylline or papaverine coincided with that in the absence of these agents.


Fractures of the mandible. A review of 909 cases. During a 5-year period, 909 fractures of the mandible were treated by the Department of Plastic Surgery at Groote Schuur Hospital in Cape Town. A simple technique based on Gilmer wiring was evolved to deal with those fractures where sufficient teeth were present. This technique is extremely simple and proved efficacious.


Neurotransmitter content of mouse brain after inactivation by microwave heating. Mice were sacrificed by either microwave inactivation of brain enzymes or by cervical dislocation. The norepinephrine, serotonin, 5-hydroxyindoleacetic acid and homovanillic acid content of the brains of the irradiated animals was significantly greater than that of the conventionally sacrificed controls.


Benzodiazepine-induced suppression of estrous cycles in C57BL/6J mice. Female C57BL/6J mice fed diets containing diazepam, chlordiazepoxide, oxazepam, prazepam, flurazepam, or nitrazepam exhibited significant decreases in the frequency of vaginal estrus.


A comparison of the inductive effects of phenobarbital, methaqualone, and methyprylon on hepatic mixed function oxidase enzymes in the rat. The effects of equal doses of three sedative-hypnotics, phenobarbital, methaqualone, and methyprylon, on the hepatic mixed function oxidase enzymes of the rat were investigated and compared. After 5 days of pretreatment, phenobarbital and methyprylon significantly increased aminopyrine demethylation, aniline hydroxylation, and cytochrome P-450 content in hepatic microsomes. Methaqualone pretreatment only increased hepatic aminopyrine demethylase activity and wet liver weights. After 29 days of pretreatment, phenobarbital significantly increases aminopyrine demethylase, aniline hydroxylase activity, liver weight and cytochrome P-450 content. Methaqualone only produced a significant increase in wet liver weight.


Binding of tritiated dehydroretronecine to macromolecules. In vivo and in vitro experiments have shown that the pyrrolizidine alkaloid metabolite dehydroretronecine binds readily to macromolecules. In the in vivo experiment there was a preferential binding of dehydroretronecine to the gastric mucosa. Further extraction of the mucosa revealed a large percentage of the 3H was bound to the protein fraction and to a much lesser extent to DNA and RNA. The influence of pH on the binding of dehydroretronecine was substantiated in the in vitro experiment. Dehydroretronecine bound to calf thymus DNA and bovine serum albumin most readily under acidic conditions. These data suggest a direct correlation of the levels of dehydroretronecine binding to cellular macromolecules with the lesions that develop in affected organs.


Effect of cromolyn sodium and of daily exposure to compound 48/80 on experimental asthma. Guinea pigs were exposed to aerosols of histamine or the mast cell degranulating and histamine releasing substance 48/80. Cromolyn sodium aerosols prevented 48/80 induced asthma but did not influence histamine induced broncho-constriction. 48/80 given daily for 5 days resulted in marked decrease in susceptibility to 48/80, possibly because of depletion of histamine and other active substances. There was no change in histamine sensitivity. After 3 days of rest, normal susceptibility to 48/80 returned.


Some factors influencing colicin activity between pathogenic and commensal Escherichia coli from the pig. Commensal strains of Escherichia coli derived from pigs had a broad spectrum of in vitro colicin acitivity against pathogenic serotypes. Of eight pathogenic serotypes tested, only three were colicinogenic and were active against relatively few commensal strains. Colicin activity was influenced by temperature, pH and oxygen tension as well as by the availability of certain nutrients and the presence of trypsin. Lack of colicin activity in intestinal supernatant fluid was ascribed to the concentration of trypsin present. It was concluded that colicins are unlikely to influence the dominance of pathogenic Escherichia coli serotypes in the pig's intestine, except possibly in the colon and rectum where the concentration of trypsin is low.


Effect of carbacholine and urecholine on pentagastrin-stimulated gastric secretion in healthy subjects. Dose-response studies of pentagastrin-stimulated gastric secretion were performed in 6 healthy volunteers. On different days pentagastrin was given in doses of 0.15, 1.5, and 15 mug/kg/hr either alone or in combination with carbacholine, 2 mug/kg/hr, or urecholine, 60 mug/kg/hr. Carbacholine and urecholine increased acid and pepsin secretion evoked by the lowest dose of pentagastrin while there was no augmentation at the highest dose. The dose of pentagastrin required to elicit half maximal acid output (Km) tended to decrease by simultaneous infusion of carbacholine or urecholine, suggesting that the cholinomimetics increased the sensitivity of the parietal cells to pentagastrin stimulation. Km for pentagastrin alone was higher than previously found in unoperated duodenal ulcer patients.


Gastro-oesophageal acid reflux. Method for 12-hour continuous recording of oesophageal pH with analysis of records. Description of method for continuous recording of oesophageal pH and automatic, electronic analysis of records. Stability of apparatus has been tested over 12 hours in the laboratory with different buffer solutions and gastric secretions with and without bile, and on 36 patients with oesophageal disorders. Drift of apparatus max. 0.2 pH units. The analyser, dividing the pH range into arbitrary intervals, performs a rapid and reliable analysis of single examinations from variable criteria. The temporal distribution of pH values into the intervals is expressed in percentages of the total time of recording. Reliability of the analyser was tested by analysis of a simulated pH curve on which changes in amplitude corresponded to the most rapid changes seen in practice. Deviation below one per cent. Repeated analyses of the same records showed a coefficient of variation for the various analytical interval to be 10-0.01 per cent. The method allows recording of the number of reflux episodes, duration of each episode and total duration of pH within chosen limits.


The stability of human pepsins in stored gastric juice. The effect of storage at -20 degrees C and +2 degrees C on the pepsins of human gastric juice was assessed by quantitative assay and agar gel electrophoresis. Quantitative assays showed no significant loss of peptic activity at either temperature over periods of storage up to 3 weeks, and no correlation between change in peptic activity and the acidity of the gastric juice, although raising the pH of the gastric juice above pH 2 with buffers gave improved stability over gastric juice mixed with buffer of lower pH. At low pH, the addition of bovine serum albumin gave improved stability. Agar gel electrophoresis showed that the pepsins were all stable at -20 degrees C for several days or several weeks, but alterations occurred in the pepsin molecules after several months.


Hydrogen ion, pepsin and bile acid binding properties of hydrotalcite. The hydrogen ion, pepsin and bile acid binding properties of hydrotalcite (Altacite) were investigated in human gastric juice both in vitro and in vivo. Hydrotalcite is an efficient antacid in vivo and between pH 2,5 and 3,5 it binds and inactivates pepsin in vitro. The results in one case suggest that this also occurs in vivo. Hydrotalcite is capable of removing substantial amounts of bile acids from bile-containing gastric juice in vivo. The findings indicate that hydrotalcite could be a useful agent in the treatment of both gastric and duodenal ulcers.


Alfathesin for anaesthetic induction at caesarean section. Anaesthesia was induced with Alfathesin (60 - 70 mul/kg) in 50 healthy mothers undergoing elective Caesarean section. Anaesthesia was maintained with nitrous oxide, oxygen, muscle relaxants and controlled ventilation. The mothers were tilted laterally throughout the operation. Blood gas studies done on the mothers before induction and at delivery, revealed a mild respiratory alkalosis associated with a moderate degree of metabolic acidosis, which appeared to increase during anaesthesia. Umbilical cord blood gas analyses indicated a mild degree of fetal respiratory acidosis (mean pCO2 Uv 45,3


Indications for intubation in laryngotracheobronchitis in black children. Fifty-eight children with laryngotracheobronchitis (LTB) were studied. Many were malnourished, or in the post-measles state, or had some lower respiratory tract or cardiac involvement. In these patients, factors helpful in assessing the need for relief of airway obstruction by tracheal intubation have been evaluated. A set of indications for intubation, which includes clinical and arterial blood gas features of LTB is suggested. Cognisance of complications of disease, where present, has also been taken. If cyanosis or muscular hypotonia or unawareness is present, intubation is urgently needed. When two of the following occur together, the need for intubation has arisen: (a) pulse rate greater than 170/min and respiratory rate greater than 55/min; (b) paCO2 greater than 37 mmHg; (c) paO2 less than 50 mmHg; and (d) a complication (cardiac failure or severe lower respiratory tract infection).


Fanconi's syndrome in an adult.  A case history. A case of the Fanconi syndrome in an adult presenting with life-threatening hypokalaemia is described.  Amino-aciduria, glycosuria, high phosphate and uric acid clearances with reduced plasma levels of these two substances, reduced urinary concentrating power and abnormal urinary potassium loss with a systemic acidosis, were the essential biochemical abnormalities.  Osteomalacia was shown by a radiological bone survey.


The respiratory effects of tilidine hydrochloride. A standard 50-mg dose of tilidine hydrochloride (Valoron) was administered to 20 patients in the postoperative phase of open-heart surgery. No significant change in respiratory status, as measured by arterial blood gas analysis, was demonstrated with this dosage.


Propanidid for anaesthetic induction at Caesarean section. In 50 healthy mothers scheduled for elective Caesarean section, anaesthesia was induced with propanidid (7 mg/kg body weight). Thereafter, ventilation was controlled with nitrous oxide, oxygen and muscle relaxants. A further dose of propanidid (1 mg/kg body weight) was administered 3 minutes after the initial injection of this drug, as a means of preventing maternal awareness during equilibration with the anaesthetic gas mixture. The acid-base status of the mothers before the induction of anaesthesia, and at delivery, revealed a mild degree of respiratory alkalosis with a compensatory metabolic acidosis. Umbilical cord blood gas results indicated the presence of significant fetal acidosis, both respiratory (mean pCO2 Uv 46,3 torr (SD 11,3) and Ua 54,3 torr (SD 12,0)), and metabolic (mean base excess Uv-9 mEq/l (SD 4,2) and Ua-11,8 mEq/l, (SD 5,0)) in origin. The average umbilical cord blood oxygen tensions were Uv 25,9 torr (SD 10), and Ua 15,4 torr (SD 8,5); mean maternal to fetal base-excess gradients were Ma-Uv 4,1 mEq/l (SD 2,8) and Ma-Ua 6,5 mEq/l (SD 3,5). Five mothers (10%) offered convincing evidence of factual recall during surgery, and 3 of these were aware of pain. Nausea and vomiting occurred in 5 patients and in 4 there were clinical signs of postoperative chest infection. The degree of fetal biochemical asphyxia, and the incidence of maternal awareness during surgery, were significantly greater than previously reported when thiopentone was used for the induction of anaesthesia for Caesarean section. The results obtained are discussed, and the conclusion is drawn that propanidid for anaesthesia appears to offer no advantage over thiopentone in obstetric practice.


Hemodynamic effects of intra-arterial infusions of catecholamines on the canine gastric circulation. The effects of intra-arterial infusions of epinephrine, norepinephrine, and isoproterenol upon right and left gastric blood flow were studied in anesthetized dogs. Blood flow was measured electromagnetically before and after adrenergic blockade. Infusion of epinephrine (0.05 mug Kg.(-1) min.(-1)) resulted in vasoconstriction (-50 +/- 6 ml. min.(-1)) with autoregulatory escape in the right gastric artery (RGA) and constriction (-41 +/- 8 ml. min.(-1)) followed by significant dilation (+ 56 +/- 10 ml. min.(-1)) in the left gastric artery (LGA). Alpha adrenergic blockade with phenoxybenzamine produced only a dilator response in both RGA and LGA to epinephrine infusion and beta adrenergic blockade resulted in a constrictor response with no autoregulatory escape. Norepinephrine infusions produced a constrictor response of similar magnitude as epinephrine but with little evidence of autoregulatory escape in either RGA and LGA. Alpha adrenergic blockade significantly attenuated this response in both circulations. Isoproterenol is a dilator in both circulations and its response is attenuated only by beta adrenergic blockade. This study demonstrates that the canine stomach has two regionally distinct circulations with the fundus and body exhibiting a greater dilator response than the antrum and pylorus.


Light scattering changes of edetic acid-treated, washed rat platelets: effect of adenosine diphosphate and calcium ion in relation to chilling. The effe-ts of adenosine diphosphate (ADP) and Ca2+ on platelet morphology were studied in edetic acid (EDTA)-treated, washed platelets of rat by means of light scattering measurement at right angles. ADP caused a transient change of light scattering, which was independent of concentration above 5 muM and of pH between 4 and 10. When the platelets were pretreated with ADP or chilled persistently, Ca2+ also caused another transient change of light scattering. The effect of Ca2+ was abolsihed at pH below 6.5. The velocity of both ADP- and Ca2+-induced response was dependnet on temperature, and the magnitude was reduced by metabolic inhibitors and SH-blocking agents. Adenosine triphosphate (ATP) competitively inhibited the ADP-induced response, but not the Ca2+-induced one. The response to Ca2+ of the control and chilled platelets was compared by scanning electron microgram. Apparent similarity of the effect of chilling on membrane structure to that of ADP was suggested.


The effect of human platelets on lipolytic activity in plasma. A method for assay of lipolytic activity in platelet-poor plasma (PPP), platelet-rich plasma (PRP) and washed human platelets is presented. The lipolytic activity in PRP was about twice the activity in PPP. A significant correlation between lipolysis in platelets and platelet number was established. Molar sodium chloride strongly inhibited lipolytic activity in platelets and a washing procedure did not significantly change the lipolysis. The results indicate that a lipoprotein lipase is bound to human platelets and may play a role in metabolism of triglyceride in platelets.


The activation of human factor IX.

The activation of human factor IX. The activation of factor IX purified from human plasma has been studied. Factor XIa and kallikrein separately activated factor IX to factor IXa. In both cases factor IXa had an apparent molecular wight of about 42-45000 in sodium dodecyl sulphate-polyacrylamide disc gel electrophoresis compared with a molecular weight of about 70000 for the native factor IX. The activation by XIa required Ca2+-ions, wherease Ca2+-in and factor VII or Russell's-viper venom alone did not activate factor IX. Trypsin activated and plasmin inactivated factor IX.


Studies on stereospecific reduction of acetoacetyl CoA and crotonyl CoA in lactating rabbit mammary glands. Stereospecificity as well as the dependency on reduced pyridine nucleotides of the enzymatic reduction of acetoacetyl CoA and crotonyl CoA in lactating rabbit mammary glands are reported. In the reduction of both acetoacetyl CoA and crotonyl CoA as substrates the tritium from NADP3H was stereospecifically incorporated into the beta-position of n-butyric acid. The reaction of acetoacetyl CoA reduction was much more dependent on NADH while the reduction of crotonyl CoA was rather more dependent on NADPH. There was no difference between the dependencies on NADH and NADPH in the reduction of 2-hexenyl CoA.


Radiation effects on rat testes. VIII. Kinetic properties of hydrolases following partial body gamma irradiation of rats. Kinetic properties such as Michaelis constant (Km), maximum velocity (Vmax), temperature coefficient (Q10) and energy of activation (Ea) for hydrolysis of adenosine-5'-phosphate at pH 9.5 and sodium pyrophosphate at pH 8.35 by normal and radiated testes supernatants have been described. Kinetic parameters are related to respective phosphohydrolases (phosphatases). (1) Km values for 5'nucleotidase and inorganic pyrophosphatase of normal testis were determined as 1.25 X 10(-3)M and 0.81 X 10(-3)M respectively; (II) Vmax correspond to 318 mug P/15 min and 430 mug P/15 min for 100 mg tissue respectively; (III) Q10 for 5 nucleotidase is 1.7 and for inorganic pyrophosphatase is 4.2 at a temperature 10-30degreesC; (IV) Ea for hydrolysis of AMP and sodium pyrophosphate were calculated by Arrhenius plots as 17000 and 9000 cal/mol. (V) Km values for irradiated enzymes are similar to the control values suggesting that the binding capacities of these enzymes with their substrates remain unaffected after radiation; (VI) Vmax for radiated enzymes correspond to a value of 500 mug P/100 mg tissue/15 min for 5'nucleotidase and 118 mug P/100 mug tissue/15 min for inorganic pyrophosphatase; (VII) 110 for 5'nucleotidase is 2.2 and inorganic pyrophosphatase 1.16 at 10-30degreesC; (VIII) Ea for irradiated 5'nucleotidase is comparable to those of normal rats whereas for inorganic pyrophosphatase Ea is moderately declined. The observed changes have been related to the different types of metabolic activity in germinal and nongerminal cells of testes.


Long-term survival of cardiac allografts in lethally irradiated rats repopulated with host-type hemopoietic cells. To test the hypothesis that hemopoietic cells within a tissue graft are responsible for its immunogenicity, two experimental protocols were followed. LEW hearts were grafted into (LEW X BN)F-1 host rats and LEW or F-1 lymphocytes were injected into the apex of the grafted heart. The LEW but not the F-1 cells induced a local reaction, apparently because the circulating F-1 cells were the necessary immunogens. The second protocol took advantage of the knowledge that lethally irradiated LEW rats were able to reject WF Ag-B-incompatible hemopoietic cells (but not tissue allografts) within a few days. LEW rats were lethally irradiated and grafted with WF hearts on day 0. A mixture of LEW marrow, thymus, spleen and lymph node cells, or marrow cells only were infused either on day 0 or day 2. Cardiac allografts in hosts repopulated with the mixture of lymphoid cells survived a mean of 11.3 days in hosts infused on day 0, but survived indefinitely if the lymphoid cells were infused on day 2. The 2-day interval also prolonged the survival of allografts in rats infused with only marrow cells. The long-term recipients, without any further treatment, rejected WF skin grafts as first-set reactions 1 year later but did not reject second WF cardiac allografts. Lymphoid cells from long-term recipients imparied the rejection of WF cardiac allografts by LEW host rats. The lack of rejection of the original cardiac allograft supported the hypothesis tested. Certain hemopoietic cells responsible for the immunogenicity of cardiac allografts were probably eliminated in the 2-day interval at least in part by host effector cells capable of rejecting allogeneic hemopoietic cells. However, the mechanism of long-term "unresponsiveness" to WF hearts could have been caused by loss of accessory cells during the 2-day interval followed by infusion of immunocompetent cells. Skin rejections in these recipients may have been attributable to reactions against skin differentiation-specific antigens.


The role of the peripheral sympathetic nervous system in cerebral blood flow autoregulation. The effect of chronic, unilateral superior cervical ganglionectomy on cerebral blood flow and blood flow autoregulaiton to changes in perfusion pressure was examined in seven phencyclidine anesthetized monkeys. Ten to 14 days prior to the experiments Doppler ultrasonic flow transducers were placed on both carotid arteries after ligation of the external carotid branches and removal of one superior cervical ganglion. Autoregulation was tested by exsanguination and metaraminol infusion with the monkeys inspiring from air, 9% and 12% carbon dioxide in air. Immediately following experimentation the cerebral vessels were examined for the presence of noradrenergic fibers. The results of the study demonstrate that: (1) superior cervical ganglionectomy produces a significant reduction in the noradrenergic innervation of ipsilateral extraparenchymal arteries: (2) the peripheral sympathetic nervous system contrivutes to overall cerebral vascular resistance primarily by affecting resistance in extraparenchymal arteries; and (3) as a result, it determines the contribution of the extraparenchymal arteries to overall cerebral blood flow autoregulation.


Purification and characterization of guinea pig lymphotoxin produced by lymph node cells stimulated by phytohemagglutinin. Guinea pig lymphotoxin (LT) produced by stimulation of lymph node cells with Phaseolus vulgaris phytohemagglutinin was purified approximately 1,000-fold in specific activity by ammonium sulfate fractionation, DEAE-Sephadex column chromatography, and gel filtration on Sephadex G-100. The properties of LT tested at the various stages of purification revealed that the LT was a heat-sensitive, protease-sensitive proteinaceous substance, and its approximate molecular weight was estimated to be 50,000 by means of gel filtration. The most purified fraction (Fr. D) was found to be devoid of the activities of migration inhibitory and mitogenic factors, clearly indicating that these activities are mediated by different substances.


Depression of cellular immune response during syngeneic pregnancy as measured by the graft-versus-host reaction. The cellular immune response during the course of intrastrain pregnancies was evaluated in primiparous and multiparous BALB/c females by the graft-versus-host (GVH) activity of the splenic and lymph nodal lymphocytes. Splenic lymphocytes of both primiparous and multiparous pregnant mice had a GVH activity lower than that of the virgin mice. This immune deficit was more evident in multiparous than in primiparous females and occurred at the 6th day and during the last week of pregnancy. A similar although less pronounced phenomenon was found when lymph nodes instead of spleen of multiparous females were used at the source of cells in GVH reaction. The immune deficit was less marked when spleen and lymph node cells of pregnant mice were compared with similar cells of pseudopregnant females. Incubation of virgin BALB/c spleen lymphocytes in the serum of multiparous pregnant mice as compared to normal virgin serum caused a reduction of the GVH activity; incubation of the cells in pseudopregnant serum also inhibited the immune response, but to a lesser extent than the incubation in multiparous sera.


Pial microcirculation in subarachnoid hemorrhage. Microsurgical and microscopic methods were employed in guinea pigs to expose, observe, and measure response characteristics of cerebral cortical pial microvessels and microcirculation to traumatic and nontraumatic experimental subarachnoid hemorrhage. Bleeding produced by vascular micropuncture was associated with a 44.3% arteriolar constriction. Topical application of homologous blood alone produced a 33.2% vasoconstriction. Observed microcirculatory flow characteristics subsequent to such microvascular changes were consistent with those known to be associated with cerebral cortical infarction. These changes could be prevented or reversed by topical application of the alpha adrenergic blocker, phenoxybenzamine. Topical pretreatment with the beta adrenergic blocker, propranolol, prevented blood-induced spasm, but did not reverse such spasm once it had been established. A chemo-mechanical mechanism is suggested as underlying the vasoconstriction association with rupture of pial microvessels. It is thought that consideration of such microvascular characteristics, in conjunction with those known to be associated with larger intracranial vessels, adds to current knowledge of the pathophysiology of subarachnoid hemorrhage and may be extrapolated to bear future clinical import.


The case against a viral aetiology in feline urolithiasis. Efforts have been made to endorse the thory of a viral aetiology for the feline urolithiasis syndrome (FUS) in the male cat. The presence of virus in the urine of three cases could not be confirned nor could the condition be transmitted by intra vesicular inoculation of urine from four affected cats into the bladders of normal cats. Following these negative findings work was done on SPF cats and a survey made of the incidence of urethral obstruction in six SPF cat units in England. No cases were reported from four of these units over a two-year period. Five cases occurred in the other two SPF units, three of which were associated with feeding an expanded dry cat food. There is, thus, no direct evidence that viral agents are involved in the aetiology of FUS.


The effects of sulphydryl reagents on the binding and mixed function oxidation of hexobarbital in rat hepatic microsomes. 1. The effects of the sulphydryl reagents p-chloromercuribenzoate, N-ethylmaleimide and iodoacetamide on the binding spectrum, oxygen consumption and formation of a suspected substrate-cytochrome P-450-oxygen complex for hexobarbital in rat liver microsomes were investigated. 2. The oxygen consumption caused by hexobarbital oxidation was inhibited non-competitively by all three agents, with 50% inhibition at 4 times 10(-5) M for p-chloromercuribenzoate, 3-7 times 10(-4) M for N-ethylmaleimide and 1-9 times 10(-3) M for iodoacetamide. Cysteamine protected and at least partially reversed this inhibition. 3. p-chloromercuribenzoate inhibited the formation of the cytochrome P-450-substrate-oxygen complex, while N-ethylmaleimide and iodoacetamide also inhibited the formation of this complex but to a lesser extent. The p-chloromercuribenzoate inhibition was protected against and reversed by cysteamine. 4. p-Chloromercuribenzoate and N-ethylmaleimide caused a 50% reduction in the magnitude of the hexobarbital-induced binding spectrum, and this was paralleled by the conversion of cytochrome P-450 to cytochrome P-420. Cysteamine protected against this effect but could not reverse it. Iodoacetamide had no effect on the binding spectrum of hexobarbital and failed to convert cytochrome P-450 to cytochrome P-420. 5. Points of attack within the reaction sequence of drug oxidation are tentatively ascribed to the sulphydryl reagents used in this study.


Enzymic reduction of aromatic hydrocarbon epoxides by the microsomal fraction of rat liver. 1. The oxidation of aromatic hydrocarbons to arene oxides and the reduction of these oxides to the parent hydrocarbons are both catalysed by enzymes in the microsomal fraction of rat liver. A suggested name for the enzyme concerned in the reduction of these epoxides is 'epoxide reductase'. 2. 'Epoxide reductase' is NADPH-dependent and is inhibited by oxygen. 3. Preliminary investigations suggest that the enzyme is specific for both 'K-region' and 'non-K-region' arene oxides.


Electron transport systems of lung microsomes and their physiological functions. Enzymic hydroxylation of aniline and steroids. 1. The hydroxylation of aniline by rabbit lung microsomes to rho-aminophenol required oxygen and NADPH, and was inhibited by menadione, ferricytochrome c and carbon monoxide. 2. NADH was a less effective electron donor than NADPH in this reaction, but its addition significantly increased the yield of rho-aminophenol formed in the presence of NADPH. 3. 4,16-Androstadien-3-one and 3beta-hydroxy-5-androsten-17-one were also metabolized by lung microsomes to the same products as are formed by hepatic microsomes.


[Effect of the characteristics of various alimentary stimuli and changes in the pH of the duodenal content on the acid-forming function of the stomach]. In chronic experiments on dogs the effect of introduction of a non-buffered acid gastric juice into the duodenum or/and of drawing outside the bile via a fistula in the common bile duct on theproduction of hydrochloric acid in innervated ventricles (pouches) was studied. The induced pH shifts in the duodenum were checked every 15 minutes in the duodenal chyme samples. In tests involving meat feeding the reduced pH of the duodenal contents acted not only as an inhibitor of the gastric secretion, but much more as its stimulator. Infusion of the juice into the duodenum, or/and elimination of the bile tended to drastically reduce the inhibitory effect of the fat on the acid-formation in the stomach, this being due to inactivation of the pancreated lipase with HCl and to a defective micellar stage of the intraintestinal transformation of fat. This gives ground to assume that the entrance of acid non-buffered gastric juice into the intestine during transtubal feeding of patients with peptic ulcer is counter-productive in this method of treatment. The application of hyposecretory agents concurrent with the tubal feeding is warranted.


The partial purification and properties of a human erythrocyte 4-nitroacetophenone reductase. 1. A soluble enzyme which catalyses the NADPH-dependent reduction of 4-nitroacetophenone to 4-nitrophenylmethylcarbinol has been partially purified from human erythrocytes. 2.inter-individual or intra-individual differences in the enzymic activity were small except for very low activity observed in one subject with glucose 6-phosphate dehydrogenase deficiency resulting in decreased levels of NADPH. 3. The enzyme was inactivated above 50 degrees or on storage at 4 degrees for longer than 24 h. The pH optimum was between 7-0-8-0. 4. the enzyme has been differentiated from NADPH-methaemoglobin reductase, NADPH-cytochrome c reductase, glutathione reductase, alpha,beta-unsaturated ketone reductase and aromatic alpha-keto acid reductase activities, but similarities exist between this enzyme and a rabbit kidney cortex aromatic aldehyde/ketone reductase.


The metabolism of foreign compounds in the cestode, Moniezia expansa, and the nematode, Ascaris lumbricoides var suum. 1. The ability of the cestode Moniezia expansa and the nematode Ascaris lumbricoides var suum to metabolize foreign compounds has been assessed. 2. Both species were unable to oxidase aldrin, aniline, biphenyl, butylbenzene and nitrobenzene or to demethylate aminopyrine, and 4-nitroanisole. 3. M. expansa and A. lumbricoides var suum readily induced 4-nitroanisole, nitrobenzene, 4-nitrobenzoic acid and 4-nitrophenol to the corresponding amines. Azobenzene, dimethylaminoazobenzene, and 1,2-dimethyl-4-(4-carboxyphenylazo)-5-hydroxybenzene were also reduced. 4. Hydrolysis of esters, acetanilide, acetylsalicylic acid, aryl sulphates and aryl phosphates took place readily. However, beta-glucuronides were not hydrolysed. 5. The following reactions were not detected in either species: phosphate, sulphate, beta-glucuronide or beta-glycoside conjugation of phenolic compounds; acetylation of amino compounds, or the formation of glycine conjugates with 4-aminobenzoic acid or benzoic acid. 6. Male nematodes showed a higher rate of drug metabolism than female nematodes.


4-Nitrobenzoic acid reductase of the nematode Ascaris lumbricoides var suum. Localization of the enzyme and optimum assay conditions. 1. Reduction of 4-nitrobenzoic acid by the nematode Ascaris lumbricoides var suum occurred in the intestinal brush border cells. Cuticle, mesenchyme fluid and reproductive tissue showed no activity. 2. 4-Nitrobenzoic acid reductase was in the supernatant fraction after centrifugation of intestinal homogenates at 75 000 g for 2 h. 3. The enzyme required as cofactors NADH2, a low molecular weight thiol such as glutathione or cysteine and a divalent metal ion such as Mn++ or Cu++. The reduction  was not catalysed by NADPH2. 4. Reduction was not inhibited by O2 or CO. Optimum pH of the reaction was about 6.5.


Glutathione conjugates as metabolites of benz[a]anthracene. 1. [3H]Benz[a]anthracene is converted into water-soluble metabolites by microsomal plus soluble fractions of rat-liver in the presence of NADPH and glutathione. Chromatography on Sephadex G25 gave four radioactive peaks; the first contained hydrocarbon or hydrocarbon derivatives bound to soluble protein while the other three peaks contained glutathione conjugates of hydrocarbon metabolites. 2. Conjugates formed when either of the benz[a]anthracene metabolites, 5,6-dihydro-5,6-dihydroxybenz[a]anthracene or 8,9-dihydro-8,9-dihydroxybenz[a]anthracene, were similarly incubated were probably S-(5,6,8,9-tetrahydro-5,6,9-trihydroxybenz[a]anthracen-8-yl)glutathione and S-(5,6,8,9-tetrahydro-6,8,9-trihydroxybena[a]anthracen-5-yl)glutathione respectively. The corresponding peak obtained in the metabolism of benz[a]anthracene probably contains a mixture of these two isomers. 3. The third peak contained the conjugate, S-(5,6-dihydro-l-hydroxybenz-[a]anthracen-k-yl)glutathione, also formed by the conjugation of the "K-region" epoxide of benz[a]anthracene with glutathione. This was not formed in the metabolism of the dihydrodiols. 4. The fourth peak contained a new type of conjugate that is probably S-(8,9,10,11-tetrahydro-8,9,10-trihydroxybenz[a]anthracen-11-yl)glutathione. This conjugate is chromatographically similar to a product obtained from incubation of the 8,9-dihydrodiol, and is probably formed by microsomal oxidation of the 10,11-bond of the dihydrodiol, followed by conjugation of the resulting diol-epoxide with glutathione.


Biochemically aberrant Salmonella enteritidis ser. newington from human sources in Connecticut. Three isolates of a lactose-fermenting, xylose-negative variety of Salmonella enteritidis ser. newington, identical in biochemical and serological reactions and in the antibiogram, were recovered from three patients in different areas of Connecticut in January 1974. Hydrogen sulfide production was not visible in Salmonella-Shigella agar, in triple sugar iron agar, and in Kligler iron agar but was noticed in lysine iron agar and on XLD agar, among others. The amount of fermentable carbohydrates present was found to correlate with failure to show hydrogen sulfide production (pH effect). In contrast to lactose-fermenting Salmonella strains reported by other authors, we could not elicit a direct transfer of the lac(+) character at frequencies above 10(-6). An epidemiological follow-up remained unsuccessful. Recommendations for the recognition of similar strains are presented.


[Danger of fetal acidosis in vaginal delivery from breech presentation (author's transl)]. On the basis of our own experience and the literature the risk of acidosis, which corresponds to the risk of asphyxia, during vaginal delivery of breech presentations is examined. Compared with delivery of vertex presentation it is 3-10 times greater. The risk of acidosis does not depend on the duration of pregnancy and only a little on the parity of the mother. Even with carefully selective indication for primary Caesarean section it cannot be reduced below the high level. Typically there is acute compression of the cord at the end of the first or later stages of labor. Even with intensive intra-partum care it cannot safely be predicted with sufficient certainty. As a criterion of the efficiency of modern obstetrics the impact of acidosis in umbilical blood and its increase in breech presentation is discussed. Only systematic Caesaren section before or early in labor will lower the risk to that of vertex  presentation. This is done and recommended by the authors. Links between acdosis in umbilical blood and permanent cerebral damage are probable but by no means certain and their importance in unknown. Selective  indication for Caesarean section which has to be made generously, represents an acceptable alternative.


[Hypoxia and acidosis--morbidity (author's transl)]. A critical study based on 3291 analyses of blood from the umbilical cord, and the consequences. In routine analyses acidosis (pH 7.10) was found in 2.03%. This percentage is considered too high. Protracted labor carries special risks. Conventional and intensive care supervision, when risks are foreseen, are often ineffective, since 48.8% of acidoses were recognized only post partum. More general intensive observation with internal cardiotocography and microinvestigations of blood appears to be the only way in which the frequency of acidosis could be decisively reduced.


[The action of prostaglandin E2-induced pains on the metabolic and clotting systems of mother and child during birth (author's transl)]. 11 pregnancies at term were terminated by dilatation of the uterine cervix, low amniotomy, and by intravenous administration of PGE2. The average infusion time was 4 hours 20 minutes, and the average total dose of PGE2 amounted to 0.2 mg. Parameters of acid-base changes, carbohydrate and energic state changes, gas metabolism, and changes in coagulation and fibrinolysis in mother and in fetus were analysed during labor and after birth. Labor activity and fetal cardiac action were monitored cardiotocographically. Checked against 50 uncomplicated spontaneous deliveries, we have found no disadvantageous changes in the parameters investigated.


[Addition of gentamicine to polymethyl methacrylate for therapy of infectious bone diseases. Expeimental in vivo tests]. In experimental  in vivo tests the therapeutic effect of Gentamicin added to Palacos is detected in a standardized osteomyelitic infection caused by Staph. aureus haemolyticus. The number of germ populations is reduced significantly by the antibiotic which is released in a microbiologically active concentration. On the other hand the number of germ populations in control trials (without adding an antibiotic) is remaining on a high level. During the time of the tests the appearance of other bacterial is registrated.


[Tissue compatiblity of stomatologic materials in cell cultures]. The paper contains results of investigations on the suitability of cell cultures for biological testing of soluble substances from dental cements and their components. The preparation of the specimens as well as the elution process are described. Furthermore the culture system of cell populations in vitro and representative biological parameters are reported.


[Biolgical problems of a manmade small lake (author's transl)]. In connection with an intensive hygienic supervision of public bathing facilities and the attempts to draw up a law on bathing hygiene, the lack of contamination standards for small lakes available to the public for bathing was particularly conspicuous. In addition to the current chemical and bacteriological routine examinations, a small bathing lake with a surface area of 3000 square metres containing about 7000 m3 of water and which has been in existence for over 10 years was objected to extensive biological investigations during the months July to September. The results of the analysis show a relatively constant hydrogen ion concentration (pH 8.2), small variations in electrolytic conductivity and 8 German degrees of hardness. The mean phosphourus level was 60 mug/L. the orthophosphate levels ranged between 5.2 and 18.5 mug/L. Oxygen saturation during the entire summer months was more than 100%. Altogether, 49 different kinds of phytoplankton- and 12 kinds of zooplancton-organisms were identified. The total biological mass of phytoplancton organisms was between 8.5 and 16 mg fresh weight per litre, bacterial biomass varied between 4.5 and 9.5 mg/l. Particularly striking for summer conditions was the high proportion of Cyclotella- and Synedra species among the phytoplancton mass, while the algal-bloom producing Microcystis flos aquae attained a maximum proportion of 15%. In spite of the great variety of its forms, green algae were present in small numbers only. The comparison of the described findings with the trophic-systems of HUTCHINSON, NYGAARD and RAWSON indicated an eutrophic type of lake; classification according to the types of the various saprobic systems of NAUMANN, KOLKWITZ and LIEBMANN yielded a predominantly beta-mesosaprobic type of lake. The investigations described show, however, that this is probably a lake rich in nutrients, but that there is a relatively stable biological equilibrium and that, consequently, there is no further threat of progressive eutrophism. Special hygienic parameters, contamination due to visitors and phytoplancton production will be reported in a second communication.


[Randomised investigations of some Tyrolean swimming pools for the presence of Trichomonas vaginalis and pathogenic fungi (author's transl)]. Trichomonads were not demonstrated either microscopically or by culture in water samples from 15 public open air swimming pools in the Tyrol. On the other hand, a small number of pathogenic fungi (Candida albicans, Trichophyton mentagrophytes, Tr. verrucosum) were cultured from pool water with a free chlorine content of less than 0.35 mg/l (DPD 1). Dermatophytes could also be demonstrated by cultures of the coatings on the surfaces of tiles in the showers, and of pool surrounding-and cabin tract floors. (Candida albicans, Trichophyton mentagrophytes, Epidermophyton floccosum, Scopulariopsis brevicaulis).


[Isolation of the causative agent of tularemia from Siberian lemmings in Eastern Taymyr]. The authors present the results of bacteriological and serological study for tularemia of 498 lemmings caught in Taimyr. Positive results were revealed in 4 out of 98 sera examined in the indirect hemagglutination test. In carrying out 67 biological tests on albino mice there were isolated for the first time in the Soviet Union 6 cultures of the causative agent of tularemia from the spleen of lemmings. By morphological, cultural and virulent properties the cultures obtained failed to differ from those isolated in other regions of the Soviet Union, and, consequently, we referred to the holoarctic race. Thus, it was established by the authors (both serologically and bacteriologically) that there existed tundra foci of tularemia.


[Development of new types of chemical vaccine for prevention of intestinal infections and problems of their qualitative evaluation. III. Comparative study of immunologic characteristics of experimental antigenic preparations]. As a result of comparative study of the immunological properties and antigenic activity of thyphoid, paratyphoid A and B and Sonne and Flexner dysentery antigenic preparations obtained by various methods there was revealed a decreased toxicity of the experimental preparations of the Tashkent and Leningrad Institute of Vaccines and Sera (in comparison with the tryptic antigens) and a considerable advantages of the experimental thyphoid and paratyphoid antigenic preparations by their capacity to induce the H-antibody formation in the presence of the O-antigenic activity. The preparations of the Moscow Institute of Vaccines and Sera had increased toxicity. The detected advantages of the experimental antigenic preparations of the Tashkent and Leningrad Institute of Vaccines and Sera permitted to recommend the method used in their making to obtain new chemical vaccines against the intestinal infections.


[Pathomorphological and immunofluorescent studies of smallpox vaccine neurotropism]. Experiments were conducted on guinea pigs sensitized with the AK C-vaccine components. In intracardiac injection with smallpox vaccine there was shown a possibility of development of marked hemodynamic disturbances, of the inflammatory-dystrophic processes of irreversibel character, with a subsequent neuronophagia and demyelinization. Injection of smallpox vaccine into the circulation of intact guinea pigs was accompanied by development in the nervous system of insignificant circulatory disturbances and of the inflammatory dystrophic phenomena of reversible character. A method of immunofluorescence was used and the antigen of the vaccine virus was revealed in the neurons of the brain and the spinal cord of the sensitized and intact animals. Marked hemodynamic and insignificant inflammatory-dystrophic processes were revealed in the nervous system of a child which died of the post-vaccinal encephalitis; an antigen of the smallpox virus was found by the immunofluorescent method in the nerve cells and the vessels in various portions of the nervous system.


Complexes of derivatives of 1-nitro-9-aminoacridine with DNA. Substituted 1-nitro-9-aminoacridine derivatives were shown to inhibit RNA and to a lesser extent protein synthesis in cultured human cells. Complex formation between the compounds studied and DNA were considered to be responsible for their cytostatic action. Two types of complexes differing in their binding forces were found. The biological activity of the studied compounds seems not to be dependent on the existence of a positive charge on the acridine ring.


Synthesis and properties on some 2-, 4- and 5-aminopyrimidines. Effect of -NH2 and ortho-C methylation on protolytic equilibria and electronic absorption spectra. A number of new 2-, 4- and 5-aminopyrimidines with sterical hindrance to the amino group rotation were synthesized. The pKa values and u.v. absorption spectra of the aminopyrimidines were measured in order to elucidate the conformation of the amino group depending on the place of substitution.


Tyrosine aminotransferase in frog liver. 1. In frog liver, tyrosine aminotransferase is located mainly in cytoplasm. The enzyme is an anionic protein of mol. wt. 115 000 daltons, specific toward 2-oxoglutarate. The enzyme separates on ion-exchange chromatography into two active forms. 2. Administration of triiodotyronine in vivo induces the activity of the enzyme. Epinephrine and glucagon have no effect, and cAMP and insulin repress this activity by about 70%. 3. Triiodotyronine stimulates incorporation of [14C]leucine into protein, and the amount of the enzyme in the nacent polysome-bound protein is considerably increased.


Radiation-induced aldehydes in collagen. Aldehydes present in native and irradiated (30 krad) collagen were separated from its enzymic degradation products using molecular-sieve chromatography on Bio-Gel P-2 column. From the absorption spectra of N-methylbenzothiazolone hydrazone derivatives it was concluded that two out of five of the aldehydes separated from the irradiated collagen, were identical with those present in control collagen. The possible origin of three other aldehydes is discussed.


Isolation and properties of poly(A)-containing RNA from the cytoplasm of rat liver cells. 1. The described technique for preparative isolation of poly(A)-containing RNA by complexing with poly(U)-cellulose and elution at 45 degrees C, is simple, reproducible, and the poly(U)-cellulose preparation is suitable for repeated use. 2. The poly(A)-containing RNA isolated from the poly(U)-cellulose complex is still contaminated by ribosomal RNA which can be removed by recomplexing. The purified preparation is heterogeneous on polyacrylamide-gel electrophoresis and is located over the region from 5-6S to 28S. Poly(A) segments of cytoplasmic RNA show heterogeneous size distribution. 3. In acidic medium, the poly(A) segments form ordered double-stranded structure. 4. The rapid labelling of rat liver and fibroblast cytoplasmic poly(A)-containing RNAs and their base composition resemble closely the corresponding properties of mRNA.


Changes of the oxidative phosphorylation in mitchondria of rat skeletal muscle following strenous exercise. The rate of oxygen consumption of mitochondria from rat muscles at pH 7.4 is elevated by 1-lactate. The respiratory control ratio and the ADP/O-ratio are decreased under these conditions. Acidification to pH 6.5 in the absence of 1-lactate does not change the interpreted mitochondrial functions. The experimental data are discussed as a partial uncoupling effect of 1-lactate on the oxidative phosphorylation. Similar changes in those mitochondrial functions are found after short-time intensive swimming exercise of rats. These variations might be a reason for the sometimes described reduced aerobic performance after intensive work.


Redox metabolism of glutathione in the red blood cell. A theoretical study of the biochemical and clinical aspects of the reduction-oxidation metabolism of glutathione in the mature red blood cell is presented. A summarizing survey of the literature data has led to the formulation of a mathematical model which comprises the kinetic properties of the enzymes partaking in the hexose monophosphate pathway (HMP) and the oxidation of NADPH and GSH. The model takes the form of a system of differential equations describing the motion of metabolites in one cell. The interactions between metabolites ane enzymes, in particular between glutathione and the SH-dependent enzymes of glucose phosphorylation and HMP have been included into the model...


[Phosphatidylcholine transfer between liposomes and mitochondria. Testing system for the study of specific phospholipid exchange]. A system consisting of liposomes and mitochondria for studying the exchange of specific phospholipids is described. The liposomes were prepared from phosphatidylcholine labelled with 14C-palmitic acid. The transfer of liposomes to the mitochondria is specifically stimulated 2- to 3fold by the pH 5.1 supernatant, and proceeds in a linear fashion for 90 min.


Specific receptors for prolactin in the ovary. A role of prolactin (PRL) in ovarian function has been suggested in several species, but not unequivocally established except in the rat. We, therefore, examined the presence of specific receptor for PRL in ovaries of rat, cow, and human. Human PRL (hPRL) labelled with 125I by the lactoperoxidase method was shown to be capable of specific binding to rat mammary tissue homogenate. Human, cow, and rat ovarian homogenates and/or partially purified plasma membranes were also shown to specifically bind 125I-hPRL. Binding was a saturable phenomenon and was dependent on receptor protein concentration. Optimal binding was observed at pH 7.0 and 37 degrees C. Binding was reversibly inhibited by exposure of membranes to pH 10.0 and irreversibly destroyed by exposure to pH 3.0. Bound 125I-hPRL was displaceable by unlabelled human, ovine, and bovine PRL but not by FSH or LH. However, human chorionic somatomammotrophin (hCS) and hGH showed some competition with 125I-hPRL. Number of binding sites/mg protein was lowest (0.8 x 10(-12)M) during metoestrus and increased during dioestrus (11 x 10(-12)M) reaching the maximum number at  pro-oestrus (24.6 x 10(-12)M). These results demonstrate that presence of specific PRL receptor in the ovaries and are consistent with a role of PRL at the ovarian level.


A new antihistamine hc20-511 compared with dimetinden (fenistil retard) in the treatment of chronic urticaria and other pruritic dermatoses. The new antihistamine, HC20-511 (Sandoz), was compared with Dimetinden (Fenistil retard) in a single-blind comparative study in 42 patients with dermatoses, 28 of whom suffered from chronic urticaria. HC20-511 had a better effect, especially in chronic urticaria, where pruritus, erythema and papules quickly disappeared. The effect appeared somewhat faster than and lasted as long as that of Dimetinden, although HC20-511 is not a retard-preparation unlike the Dimetinden preparation used for comparison. HC20-511 also caused less side effects.


Effect of pH, temperature and incubation period on the study of intrinsic innervation of various organs by thiocholine technique in certain vertebrates. The adrenergic fibres can be demonstrated if the optimal time and temperature are maintained. The cholinergic fibres were demonstrated at pH 5.2, incubation period l4h and temperature 37 degrees C. To ascertain the cholinesterase activity in tongue, heart, lung(birds, mammals and reptiles), gizzard and proventriculus (birds) and penis (rat and embryo of squirrel), pH 4.9, 16h incubation and temperature 37 degrees C, were quite suitable. It was possible to demonstrate the intrinsic innervation in tongue, lung, heart (mammals, birds and reptiles), gizzard and proventriculus (birds) and penis (rat and embryo of squirrel), at pH 5.2, 20h incubation and temperature 40 degrees C.


Ventricular fluid lactate, pyruvate, bicarbonate and ph in unconscious brain-injured patients subjected to controlled ventilation. In 35 unconscious patients suffering from traumatic brain injury and subjected to controlled hyperventilation within 12 hours of the acute trauma, we measured continuously the intraventricular pressure (IVP), ventricular fluid lactate, pyruvate, L/P ratio, pH and bicarbonate during the first 4 posttraumatic days, and related the findings to the clinical course. In patients in whom the follow-up study did not reveal severe mental impairment or dementia, the ventricular fluid lactate never exceeded 4 mmol/litre. In this group, the IVP level never exceeded 40 mmHg/hour, and the mean pressure/day never exceeded 20 mmHg. In half of the patients in whom dementia, vegetative survival or death occurred, ventricular fluid lactate exceeded 4 mmol/litre; the IVP level/hour exceeded 40 mmHg in two of 23 patients, and the mean pressure/day exceeded 20 mmHg in nine of 23 patients. The prognostic value of pyruvate was of less importance, although a significant increase in patients with a poor outcome was seen. No prognostic conclusions could be drawn from ventricular fluid, L/P ratio, pH and bicarbonate.


The influence of changes in the environment on twitching motility. By examining medium composition and cultural conditions quantitatively it was possible to define conditions suitable to bring twitching motility about. Such conditions include the use of freshly poured, relatively thick and only slightly dried plates of a dilute medium in which the agar concentration is not too high. Incubation should take place in a humid atmosphere. In fact, everything points to the humidity as a factor of the utmost importance. Using strains of Acinetobacter calcoaceticus, it was found that a pH value of the medium adjusted to 9.0 enhances twitching motility. Spreading growth was not produced if agarose was applied instead of ordinary agars. Twitching motility was inhibited by a number of different chlorides, potassium nitrate, Tween 80 and sodium taurocholate. Possible interpretations of these observations are discussed.


Boric acid tolerant Vibrio cholerae: biological and physical properties. Two boric acid tolerant variants were developed by passage in boric acid containing media from a mouse virulent parent Vibrio cholerae strain. The variants were stable, exhibited increased acid resistance, were less virulent to mice, but protected against challenge with the mouse virulent parent strain.


Airway effects of slow reacting substance, prostaglandin F2alpha and histamine in the guinea-pig. SRS, PGF2chi, and histamine were administered intravenously or as aerosols to artifically ventilated guinea-pigs in order to asses their capacity to affect tracheal insufflation pressure measured by means of Konzett-Rössler technique, Independently of route of administration all three compounds increased tracheal insufflation pressure, SRS being the most potent one. Bilateral cervical vagotomy did not alter the effect. Relative to histamine SRS and PGF2chi were considerably more active by aerosol administration than by intranvenous injection. The aerosols had little or no effect on systemic blood pressure. On intravenous injection, histamine decreased and SRS and PGF2chi increased arterial blood pressure in a dose-dependent fashion. The airway effects of histamine were correlated to those on blood pressure whereas with SRS and PGF2chi this was not seen when the blood pressure effects were marked. Preadministration of adrenaline or isoprenaline as aerosols antagonized the increase in insufflation pressure, but not the effects on blood pressure, produced by intravenously injected histamine or PGF2chi. It is concluded that SRS, PGF2chi and histamine on intravenous or aerosol adminstration increase tracheal insufflation pressure in the guinea-pig mainly by an action on airway tone. The data emphasize that SRS is a potent bronchoconstricting agent, possibly of pathophysiological significance in guinea-pig anaphylaxis.


Transcapillary fluid movements in sympathectomized intestine and skin during hemorrhagic hypotension. Net transcapilary fluid exchange in skin tissue (paw) and small intestine was observed during a 90 min period of hemorrhagic hypotension at 50 mm Hg in the cat. Reflex fluid transfer was prevented by regional sympathectomy and chi-adrenergic blockade. Early in hemorrhage, fluid absorption from the extravascular space occurred in both tissues, apparently caused by osmosis. The process was thus co-ordinated in time with a positive arterio-venous osmolar difference, in turn caused by a marked arterial hypersomolality. Experimetnal arterial hyperosomolality of similar magnitude, created by i.v. infusion of hypertonic glucose in non-bled animals, led to transcapillary fluid absorption in both intestine and skin and at rates similar to those in bleeding. Regional hypotsionen per se caused no fluid absorption. Later in hemorrhage (greater than 30 min), plasma fluid moved into the extravascular space both in skin and intestine, apparently due to a gradual increase of cappilary hydrostatic pressure. It is concluded that the arterial hypersomolality during bleeding can cause transcapillary fluid absorption in intestinal and skin tissues, as previously shown for skeletal muscle (Järhult 1973). The hemodynamic significance of this process for plasma volume regulation in hemorrhage is, however, much greater in skeletal muscle than in intestine and skin, mainly due to the much larger total mass of the muscle tissue.


Membrane potential and conductance during pinocytosis induced in Amoeba proteus with alkali metal ions. An investigation of the relationship between the polarized state of the membrane and the onset and the intensity of pinocytosis was made in Amoeba proteus. Membrane potential and input resistance was in all instances found to decrease in approximate proportion to the number of channels when pinocytosis was induced by a variety of alkali metal ions at varying pH. Channels began to appear when the membrane was depolarized to -30 mV by the inducer of pinocytosis. With all inducers the maximum pinocytosis was encountered at membrane potentials close to zero. No positive potentials were recorded when the chloride salts of the inducing cations were used. At high concentrations of alkali ions a transient increase of the chloride permeability caused short-lasting hyperolarizations of the membrane. Inhibition of pinocytosis by Ca++ was accompanied by an increase of input resistance and membrane potential. The selectivity of the membrane to different alkali metal ions observed as changes in pinocytosis intensity, membrane potential and input resistance was found to vary with the concentration of the inducer and with the Ca++ concentration of the extracellular solution. Displacement of membrane bound Ca++ appeared to decrease the field strength of charged groups in the membrane altering its selectivity among alkali cations. The formation of pinocytotic channels is suggested to require translocation of Ca++ from the membrane into the cell and would therefore be closely related to the electrical properties of the amoeba.


The blood oxygen transport system. A numerical simulation of capillary-tissue respiratory gas exchange. A theoretical model of the process of respiratory gas exchange between capillary and tissue is described, with special reference to the importance of variations in blood properties. The volume of tissue supplied with oxygen from a single vessel, as a function of the blood flowrate (u), hematocrit (h), and 2,3 diphosphoglycerate concentration (DPG), is calculated from a solution to the set of equations governing species distributions in the blood and tissue. The results, which are presented in the form of crossplots of the three blood parameters (u, h, and DPG) at a constant oxygen supply rate, show the possible significance of in vivo variations in the oxygen affinity of hemoglobin as a compensatory mechanism. Of further physiological interest is the sharp increase in venous erythrocyte pH in response to decreases in hematocrit, once the hematocrit is below a certain level. These results, and those relating DPG to hematocrit at constant O2 supply, are consistent with experimental observations of elevated DPG and pH levels in anemic individualts, and the dependence of erythrocyte DPG concentration upon pH.


The problem of tissue oxygenation in diabetes mellitus. I. Its relation to the early functional changes in the microcirculation of diabetic subjects. The underlying cause leading to the reversible functional changes in the microcirculation of insulin-dependent diabetic subjects early during the disease prior to any clinical signs of retinopathy and nephropathy (functional microangiopathy) is discussed. It is suggested that the initial microvascular dilation observed in diabetics is due to an autoregulatory response to relative tissue hypoxia providing an increased tissue perfusion in order to improve tissue oxygen delivery. Supporting evidence for this suggestion is derived from the findings that diabetics simultaneously may show increased tissue oxygen consumption and decreased ability of the circulating blood to release oxygen to the tissues. The latter defect is likely to be caused by two interrelated factors: 1. an increased proportion of haemoglobin A1c with high oxygen affinity, and 2. difficulties of maintaining a sufficiently high concentration of plasma inorganic phosphate in order to provide an optimal 2,3-diphosphoglycerate (2,3-DPG) content in the erythrocytes. The basal oxygen demand of diabetics may fluctuate even within a few hours dependent upon the state of metabolic control and is increased at times of poor regulation. Hence, diabetics may suffer from innumerable cellular hypoxic injuries, which during the first years of the disease are counteracted in the microcirculation by an autoregulatory response. These microvascular reactions associated with increased plasma permeation may over the years be of major importance for the development of the degenerative microangiopathy in diabetes.


The problem of tissue oxygenation in diabetes mellitus. In order to study the determining factors for oxygen transport the oxyhaemoglobin dissociation curve (ODC), red cell 2,3-diphosphoglycerate (2,3-DPG), and plasma inorganic phosphate were estimated in insulin-requiring juvenile and adult diabetics in various conditions of metabolic control. 2,3-DPG has been shown to vary much more in diabetics than in normals, depending upon the state of metabolic control. These fluctuations of 2,3-DPG are mediated by variations in plasma inorganic phosphate as indicated by a close correlation. While 2,3-DPG was markedly decreased in diabetic ketoacidosis, it tended to be increased in ambulatory, non-acidotic patients. Since in the non-acidotic patients the oxygen-carrying capacity, i.e. the haemoglobin concentration was simultaneously elevated, these findings suggest the presence of relative tissue hypoxia in diabetes. Both in non-acidotic and in ketoacidotic patients there was a strong correlation between the amount of 2,3-DPG and the P50 at actual pH as an experssion of the oxygen affinity of haemoglobin. In order to guarantee an optimal erythrocyte oxygen release in diabetics the content of red cell 2,3-DPG and plasma inorganic phosphate should be higher than normal.


In vivo and in vitro phosphorylation of DNA-binding proteins from Escherichia coli. A deoxyribonucleoprotein (DNP) complex has been isolated from Escherichia coli cells by chromatography on Sephadex G-200. The DNP complex contains phosphoproteins and the content of phosphorus bound to the DNP protein is 3 times higher than in cytoplasmic proteins not bound to DNA. These results have been confirmed by in vivo (32-P-KH2PO4) and in vitro (32-P-ATP) phosphorylation of E. coli DNA-binding proteins isolated by chromatography on DNA--cellulose.


Self-association, conformation and binding equilibria of concanavalin A. The discovery of distinct intact and fragmented forms of Con A, together with the observation that Con A self-associates near neutrality raises questions that may be important when interpreting experiments concerned with the biological actions of the protein. Do intact and fragmented units have the same affinity for carbohydrate? Do intact and fragmented units differ in conformation? Are all dimeric units of a homologous type or do hybrid dimers consisting of one intact and one fragmented unit also exist? Can all dimeric types self-associate to the tetramer form? Do dimer and tetramer species differ in their affinity for carbohydrate? These questions have been made amenable to investigation by the development of a method which separates intact and fragmented species under conditions which do not cause time-dependent or irreversible changes in protein conformation. It is found that intact dimeric units preferentially associate to the tetramer form. Under appropriate conditions of pH and ionic strength, dimer and tetramer species, and therefore fragmented and intact forms, can be separated by chromatography on Bio Gel P-100. Hybrid dimers are not present in appreciable amounts. Both types of homologous dimers (intact and fragmented) have similar affinity for carbohydrate, but dimer and tetramer species show significant differences. The results of near UV circular dichroism studies indicate that fragmented units possess slightly different conformation than intact units. An ionization-linked conformational transition in Con A does not appear to be linked directly with the self-association of the protein between pH 5 and 7. Ligand-induced changes in the conformation of Con A are now being examined in detail. Pflumm et al. (1971) have shown that occupation of the sugar binding site of Con A results in a perturbation of conformation as revealed by near UV circular dichroism measurements. The perturbation is relatively small and does not result in more than 1-2% increase in the rotational relaxation time (Shinitzky et al., 1973). On the other hand, removal of metal ions causes a hydrodynamic change sufficient to increase the frictional coefficient and to decrease the sedimentation coefficient (S20, w) from 3.98 S to 3.78 S. Differences between the native and the apoprotein conformation are now being examined using fluorescence polarization and the hydrophobic fluorescent probe 1-anilinonaphthalene-8-sulfonate.


Studies on the interaction of concanavalin A with glycoproteins. Lectins (phytohaemagglutinin) are known to have the unique property of binding with certain specific sugars, polysaccharides and glycoproteins. Although the kinetics of interaction between lectins and sugar have been extensively studied, the binding characteristics of the lectins with various glycoproteins are not well understood. In this laboratory a systematic study has been initiated in relation to the interaction of lectins with glycoproteins. Concanavalin A is known to bind alpha-glucosides, mannosides and biopolymers having these sugar configurations. A galactose binding protein from caster bean has been purified to homogeneity and was found to contain mannose. This lectin was used as the source of glycoprotein for studying its interaction with concanavalin A. This study showed that the interaction is temperature dependent and the dissociation is time and alpha-methyl glucoside concentration dependent. This has led to speculate a model for cell-lectin interaction. Using concanavalin A it has been shown that all the lysosomal enzymes from brain studied were glycoprotein in nature. Moreover, using Sepharose-bound concanavalin A it has been possible to devise a method by which these lysosomal enzymes could be purified considerably. With the knowledge that the interaction between lectin and glycoprotein is not only dependent on the specific sugar present in the glycoprotein, but also on the nature of the glycoprotein it was possible to develop a novel method for immobilizing various glycoprotein enzymes, such as arylsulphatase A, hyaluronidase and glucose oxidase.


An anlysis of the optical titrations of the 430 and 455 NM chromophores of ethyl isocyanide complexes of mammalian hepatic cytochrome P-450. A computational method is presented from which one may calculate the pK for the spectral changes of the 430 and 455 mm chromophores of ethyl isocyanide complexes of rabbit and rat liver microsomal cytochrome P-450. For the rat liver protein from control or phenobarital-treated animals, the pK for the loss of the 430 nm absorption is approximately equal to the pK for the gain of the 455 nm absorption, confirming that the two chromophores are in pH equilibrium with one another. For a soluble preparation in which the chromophores exhibit the pH equilbrium property, this equilbrium is also maintained even after the addition of ethyl isocyanide. It is concluded that the appearance of a 430 nm absorption after the addition ethyl isocyanide to reduced cytochrome P-450 does not necessarily represent conversion of the protein to cytochrome P-420.


Mixed function oxidation and intermediary metabolism: metabolic interdependencies in the liver. Both hepatic lipogenesis from glucose and gluconeogenesis from lactate are inhibited by substrates for mixed function oxidation such as aminopyrine. These effects are inducible with phenobarbital pretreatment and absent with a demethylated product of aminopyrine metabolism, aminophenylpyrazolone. The data indicate that an interaction between biosynthesis and drug metabolism occurs in the intact liver cell. In the case of lipogenesis, direct competition for cytosolic NADPH occurs, whereas in the case of gluconeogenesisi, enhanced drug metabolism diverts key intermediates, e.g., malate from glucose synthesis toward NADPH generation. Furthermore, rates of mixed function oxidation were modified following perturbations of intermediary metabolism, e.g., fasting, addition of substrates for glucose synthesis, inhibition of energy metabolism, etc. The data support the hypothesis that in the presence of sufficiently high concentrations of substrate and oxygen, the rate-limiting step for mixed function oxidation in the intact cell is supply of NADPH.


Comparison of methods to study enzyme induction in man. A combination of several in vivo parameters has been applied in male healthy volunteers to test the suitability of these parameters to indicate enzyme induction in man: Urinary excretion of D-glucaric acid and 6 beta-hydroxycortisol, activity of serum gamma-glut amyltranspeptidase, and pharmacokinetics of aminopyrine respond significantly to phenobarbital treatment. Glucaric acid excretion is enhanced about 7-fold. Its response to induction overcomes the large individual and inter-individual variations which exist in the untreated state for glucaric acid excretion and the other parameters applied, as well. Total body clearance of aminopyrine as obtained after an oral test dose increases more than twofold from 251 to 551 ml/min upon phenobarbital treatment. This arises from increases in both the elimination constant and the apparent volume of distribution, as well. Urinary excretion of aminoantipyrine during 24 hr is about doubled, whereas the elimination of acetyl-aminotipyrine is not much affected. 6 beta-hydroxycortisol excretion in urine and activity of serum gamma-glutamyltranspeptidase are significantly augmented to about 150% of control values. Half life times of phenobarbital measured after termination of treatment are in normal range, suggesting no self-induction of phenobarbital metabolism. Because of the complexity of drug metabolizing enzymes only a combination of different parameters reliably indicates alterations in this enzyme system by inducing agents.


[Study of Brazilian agricultural workers. A medico-socio-psychological study]. The general state of health of native Brazilian agricultural workers - a total of 750 people from 3 different plantations in the States of Paraná and Saso Paulo - was examined. The main interest of this study war centred on infectious and parasitic diseases, nutritional conditions as well as social, intellectual factors. Apart from the high number of cases of helminthiasis, amounting to 80%, the general state of health of the examined subjects was found to be good - indeed better than that of a hypothetical, comparable group of Europeans. Except for the Chagas' disease, by which 5% of the test subjects were afflicted, infectious diseases posed no serious problems. There was no case of malnutrition. The relatively lower intellectual level can not be attributed to any heriditary factor, and could definitely be improved by proper schooling. Corrective measures: Chagas: Since brick houses have replaced the wooden ones for several years, new infections are unlikely. Helminthiasis: In addition to the anthelminthic treatment, sanitary prophylactic measures should be taken. Social-intellectual factors: The following points should be emphasized: elementary schooling on the plantations; teaching at intermediary level; sewing and cooking courses; general hygience.


[Simple method for obtaining Toxocara canis antigen for the indirect immunofluorescence technic]. A simple method for the preparation of Toxocara canis antigen for the indirect immunofluorescent test is described: Embryonated Toxocara eggs are treated for 12 hours at room temperature with a 1:1 mixture of 2% NaHO and sodium hypochlorite (NaClO) solution with a concentration of 2% free chlorine in order to remove the outer layers of the egg shells. The larvae which are still enclosed in the lipid membrane are freed by mild ultrasonic treatment. Thereafter, the suspension of larvae is washed and purified in a modified Baermann apparatus. In this way large numbers of larvae in pure suspension were gained and used for the production of frozen sections for the indirect immunofluorescent test. Rabbits and mice experimentally infected with embryonated Toxocara canis eggs showed a positive serological reaction (titers between 1/10 to 1/320) in this test with Toxocara larvae as antigen, while in uninfected control animals no antibodies could be detected. The larval antigen exhibited only a weak cross reaction with sera of animals infected with Ascaris suum eggs.


The esterase patterns in the ovaries and the embryonated eggs of Aedes aegypti L. The esterases of Aedes aegypti were studied in the ovary before and during a gonotrophic cycle and also in fully embryonated eggs by means of disc electrophoresis. During oogenesis no significacant changes can be observed besides a marked increase in the total esterase activity. A different esterase pattern is found in eggs after embryogenesis. The electrophoretic mobility of some esterase bands is highly increased in the ovary compared to other organs. One esterase fraction in the ovary and two in the fertilized egg were identified as acetylcholinesterases. All other enzymes are carboxylesterases.  The results are compared to those of previous authors and are discussed in view of possible functions of esterases during reproduction.


Trypanosomes and experimental trypanosomaisis in East African bats. Using the haematocrit centrifuge technique, four hundred and twenty-seven bats from East Africa were examined for trypanosome infections. Approximately 21% of the bats were found to be infected. The infection rate varied from zero to 73.3%. No trypanosome was found in fruit-eating bats (Megachiroptera). Three species of trypanosomes were found in insect-eating bats (Microchiroptera), none of the trypanosome was infective to mice or rats. The trypanosomes encountered in the survey were Trypanosoma (Schizotrypanum) vespertilionis, T. (Megatrypanum) heybergi, and T. (M). mpapuense. New descriptions based on abundant materials are given for each of the species. Trypanosoma rhodesiense and T. brucei produced a much more chronic infection in insect-eating bats (Tadarida condylura) than in mice. Since it is known that some species of Glossina feed on bats, we raised the possibility of insect-eating bats as potential reservoirs of these trypanosomes. In experiments fruit-eating bats seem to be much more susceptible to T. brucei than the insect-eating bats. T. vivax is not infective to bats.


The effect of purine and pyrimidine analogues and virazole on adenovirus replication. The multiplication of adenovirus 19 in HeLa cells was inhibited by various purine and pyrimidine analogues and by virazole.  The formation of infectious virus and of capsid proteins (haemagglutin, group-specific complement-fixing antigen) was inhibited to the same degree, while the viral cytopathic effect (CPE) was not inhibited.  The reversibility of the inhibition after removal of the substances was more complete for purine than for pyrimidine analogues.  The inhibition was counteracted by simulataneous addition of the corresponding nucleosides.  Adenosine was more effected than guanosine against purine analogues; both were partially effective against virazole, but none of them against arabinofuranosyladenine.  The time-dependence of inhibition, the ensuing eclipse period after removal of the inhibitors, and the successive application of two inhibitors led to the conclusion that most of them affect the viral multiplication mainly by inhibition of DNA synthesis.  Azacytidine inhibits the synthesis of structural proteins as well.


Two mechanisms of interferon production by leukocytes from immunized animals. Leukocytes from animals immunized against measles were treated with different doses of interferon and 5-fluorouracil. Under the influence of virus-inducer, two mechanisms of interferon production were realized in the leukocytes: early after immunization (1 day), pre-existing interferon was released; subsequently (4 days) de novo interferon production was started. Interferons induced in cultures of leukocytes from animals at 1 and 4 days after immunization were shown to differ in some physico-chemical properties.


Double-stranded complex of polyguanylic and polycytidylic acids and its antiviral activity in tissue culture. The antiviral activity and conditions of formation of the most active double-stranded complexes of synthetic homopolynucleotides, polyriboguanylic and polyribocytidylic acids, were studied on the model of primary trypsinized chick embryo cells and RNA-containing viruses. The (poly G).(poly C) complex was very active against the viruses tested; their replication in cell cultures was inhibited completely. The antiviral activity of the (poly G).(poly C) complex increased markedly in the presence of diethylaminoethyl- (DEAE-) dextran. After treatment with 1 mug/ml of (poly G). (poly C) for 1 hour in the presence of 100 mug/ml DEAE-dextran, the cell sheet remained protected for 5-7 days. Preparations of (poly G).(poly C) obtained under optimal conditions were as active as (poly I).(poly C) complexes and exceeded them markedly in the level of the therapeutic index which under the present experimental conditions was 5-10 times 10(3) for (poly G).(poly C). Highly purified homopolymers with sufficiently high molecular weight must be used for production of active and stable (poly G).(poly C) complexes.


Neuraminidase activity of influenza virus-infected cells: localization and properties. Infection of chick embryo cell (CEC) cultures with influenza virus results in the appearance of neuraminidase activity lost by the cells as a result of cultivation.  Neuraminidase activity is associated mainly with the lysomal cell fraction.  Different distribution of neuraminidase activity in lysosomes with various densities, different reaction to sodium ethylene diamine tetracetate (EDTA) and differenct optimal pH suggest that at early stages of viral infection the cell enzyme is activated and by the 7th hour of infection viral neuraminidase is synthesized.


Biophysical properties of coronavirus strain OC 43. Coronavirus OC 43 from suckling mouse brains was subjected to sucrose density gradient centrifugation.  Maxima of complement fixing (CF) activities occurred in the ribosomal (density 1.14) and mitochondrial (density 1.19) fraction which contained incomplete, damaged virus particles.  The haemagglutinating (HA) activity was associated with the microsomal soluble protein fraction displayed no antigenic activity.  The isolated purified virus proved to be very unstable.


Further virological and clinical investigations on the attenuated E5"14" virus from the tick-borne excephalitis complex. A total of 22 persons, given a single intramuscular dose of 3.1 times 10(6) newborn mouse ic LD50/ml of the E5"14" virus (stored for 26 months at 4 degrees C in lyophilized state), were subjected to rigorous clinical, clinical-laboratory and virological investigations. Clinical observations and laboratory tests (blood and cerebrospinal fluid cytology and biochemistry) revealed no ill-effects or deviations attributable to the immunization procedure. After administration of the immunizing dose, no viraemia was detected and specific seroconversions from negativity to positivity were found after 9 weeks in 73-82% of the subjects, using different tick-borne encephalitis (western subtype) viruses in the virus neutralization experiments. The results obtained seem to give increased confidence in the further use of this attenuated virus clone with strictly defined characteristics.


A modified plaque method for arboviruses on plastic panels. Autoclavable culture media containing an increased (10---15-fold)concentration of succinate buffer permit a comparitively long-term cultivation of cells in free gas exchange with the atmosphere.  Based on them, an economical technique of plaque titration of arboviruses on plastic panels with methylcellulose overlay was developed.  With seven arboviruses of three different groups and three cell lines (CV-1, PS and PK), the method proved sufficiently sensitive as compared with titrations in mice or tube cell cultures and suitable for plaque reduction tests.


Morphology of A/WSN influenza virus-infected chick embryo cells. A/WSN (H0N1) influenza virus-infected chick embryo cells (CEC) attained a swollen central region 4 hours post infection (p.i.), different from the elongated uninfected cell. From 6 hours p.i. of CEC an increasing number of approximately 100-120 nm, round protrusions were observed by scanning and transmission electron microscopy on the cell membrane, which was smooth on the uninfected cell. At 48 hours p.i. a great number of protrusions were observed on the cell; in addition, there were a number of holes or invaginations in the membranes of the infected cells indicating cell destruction.


Interferon-hyporesponsiveness in mice in connection with dose interval and site of administration of Newcastle disease virus. Reflexion in serum complement levels. Intraperitoneal administration of Newcastle disease virus (NDV) resulted in enhanced serum levels of complement not accompanied by an increase of interferon levels, when measured at 24 hours' intervals. On the other hand, intravenous injection of NDV caused a drop of complement levels of short duration with an accompanying increase of interferon levels. Hyporeactivity to induction of serum interferon could not be achieved by intraperitoneal administration of NDV, but an incomplete hyporeactivity could be achieved by intravenous administration of NDV. It might be assumed that production of interferon in mice occurs in different separated compartments depending on the route of inoculation of the inducer.


Isolation and identification of group B arboviruses from the blood of birds captured in Czechoslovakia. Three West Nile (WN) virus strains were isolated from the blood of birds (Tringa ochropus, Vanellus vanellus and Streptopenia turtur) captured in south Slovakia.  One strain of tick-borne encephalitis (TE) virus was isolated from the blood of Tringa ochropus captured in south Moravia.


Antibody to lymphocytic choriomeningitis virus in children with congenital hydrocephalus. Antibody to lymphocytic choriomeningitis (LCM) virus was found by the indirect immunofluorescence (IF) technique in high titres in 9 out of 28 children with congenital hydrocephalus and in moderate titres in 6 mothers of these seropositive children.  It is suggested that LCM virus or a closely related virus infecting the foetus in utero may play a role in the aetiopathogenesis of some cases of congenital hydrocephalus.


Interaction of plasma membranes with influenza virus. V. Changes in creatine phosphokinase activity. A decrease in creatine phosphokinase (CPK) activity was observed in plasma membranes (PM) treated with native A/WSN influenza virus but not in PM treated with heat-inactivated virus. The decrease in CPK activity depended on the amount of virus added to PM, on the pH of the medium and on the quality of the isolated PM. It was evident already after 10 minutes of incubation at 37 degrees C. The possible mechanism of the inhibition of the CPK activity and the relation to changes in adenosine triphosphatase (ATPase) activity and to fusion are discussed.


Excision of bromodeoxyuridine from T4-DNA by an antimutator polymerase of T4 phage. With gene-43 (DNA polymerase)-ts-mutants of T4 phage, L98 (mutator) and CB121 (antimutator), and the T4 wild type, double labelling of DNA was carried out with (H3)-bromodeoxyuridine (BUdR) and (C14)-thymidine (TdR). Experiments on (C14) TdR for DNA synthesis measurement in the presence of BUdR offered evidence of the ability of the CB121 mutant to excise BUdR from the DNA. This effect took place only at increased temperature. As distinct from DNA synthesis of the host, all T4 phages used preferred TdR rather than BUdR for net synthesis.


Adenosine triphosphate phosphoydrolase activity associated with purified parainfluenza type 3 virions. An adenosine triphosphate phosphohydrolase associated with purified parainfluenza type 3 virions has been characterized. It hydrolyzed ATP to ADP and AMP when activated with Mg-2+ ions. Using Ca-2+ the production of ADP was inhibited but not that of AMP. Neither K+ NOR Na+ ions were required for the expression of maximal activity. Ouabain had no inhibitory effect on enzyme activity even at 10-3M. After exposure of virus preparations to Tween 20, enzyme activity was not affected. A linear relationship between enzyme activity and concentration of virus was observed.


Purification of influenza viruses on wide-pore glass columns. Influenza viruses of different serotypes were purified by gel filtration on wide-pore glass. The virus thus purfied retained completely its haemagglutinating activity and infectivity, and by one purification cycle at least 99% of ballast proteins were removed.


Properties of the cholera phage PL 163/10. Vibrio cholerae phage PL 163/10, belonging to Mukherjee's group I, gave clear plaques with surrounding halos of overall diameters varying between 1 to 4 mm when plated on a lawn of host V. cholerae OGAWA 154. It was fairly stable in the PH range 6-11. Its thermal inactivation was characterised by half lives of 39, 12, 4.5 and 1.0 minutes at 55, 60, 65 and 70 degree C respectively. The thermodynamic parameters deltaH, deltaF and deltaS were determined at these temperatures. The phange was resistant in vitro to sodium deoxycholate, trytrypsin, chloroform, robonuclease, deoxyribonuclease, Tris, Tris + EDTA, Tris + lysozyme and phosphate buffer but rapidly inactivated by sodium lauryl sulfate. Adsorption of this phage was biphasic. Intracelllular growth of the PL 163/10 phage was characterised by an eclipse period of 13 minutes, latent period of 31 minutes, rise period of 29 minutes and an average burst size of about 10 PFU/cell. This phage possessed a hexagonal head 106 plus or minus 18 x x 740 plus or minus 27 A without any tail structure.


Cytogenetic study of the continuous RH cell line and its clones with a different susceptibility to measles virus. A comparative karyological study was carried out on the RH continuous line of human embryo kidney cells and its 3 clones differing in their sensitivity to the cytopathic effect (CPE) of vaccine measles virus (L-16 strain). The clonal variants studied were shown to have different karyotypes and their sensitivities to be genetically determined. There was a correlation between the rate of appearance and the pattern of the CPE and the percentage of cells in which a set of 33 chromosomes and a high level of polyploidy were combined.


A live vaccine against tick-borne encephalitis: integrated studies. I. Basic properties and behaviour of the E5 "14" Clone (Langat virus). The course of plaque segregation from the Langat E5 strain of the E5 "14" clone is described. The virus, displaying an ic+ sc s plus or minus t e u s character, reproduced at 35 and 37, but not at 39 degree C. In subcutaneously (sc) inoculated monkeys, viraemia lasted for 2--4 days without detectable central nervous system (CNS) involvement. In sc inoculated 8--10 g mice, the infection was manifested only by a transient trace viraemia, but caused a marked resistance against challenge with virulent tick-borne encephalitis (TE) viruses, protection indices being 5.7--6.7. Pathogenetic investigations in challenged, live virus-immunized mice showed no signs of a marked productive infection except of a booster effect.


A live vaccine against tick-borne encephalitis; integrated studies, H. Histopathology of mice peripherally immunized with E5 "14" virus and challenged with virulent virus. Histopathological changes in the central nervous and lymphoid systems were semiquantitatively analyzed in subadult mice (M) peripherally immunized with the live, highly attenuated E5 "14" virus from the tick-born encephalitis (TE) complex and in immunized M subjected to virulent challenge. The E5 "14" clone possesses an exceedingly restricted neuroinvasivity. A study of the sc marker, refined and extended by microscopic examinations seems to present a more relevant approach for comparative investigations on TE virus virulence. Morphological findings in immunized M given immunosuppressive doses of cyclophosphamide (CPA) suggest that the high efficiency of live vaccines may be related to the prolonged antigenic stimulation.


A live vaccine against tick-born encephalitis: integrated studies. III. Response of man to a single dose of the E5 "14" clone (Langat virus). The clinical and immunological events in volunteers following administration of 5 and 6 dex newborn mouse intracerebral LD5O (NmicLD5O) of the E5 "14" virus clone, segregated from the Langat TP21 E5 strain (tick-borne encephalitis-TE-complex) and propagated in 7 days old SPF chick embryos, are reported. The experimental vaccine, containing the cloned virus, carrying a set of genetic markers of an ic(+) sc s(+/-) t e u (s) N character, caused in volunteers no clinically recognizable effects, but elicited an immune response in them, as determined in PS-cell culture neutralization tests with various TE (western and eastern subtype) strains. In all vaccines, given the virus intramuscularly, seroconversions from negativity to positivity were observed, with antibody titres ranging from 4-128. The parenteral virus administration was superior to the oral route.


Passive immunity against Junín virus in mice. Passive immunity, naturally acquired from immune mothers or artificially induced by the administration of homologous hyperimmune serum, conferred on suckling mice a high degree of resistance against infection with Junín virus. Maternal antibodies in the circulating blood of the young were not detectable in the first days after birth, but rised rapidly from the 8th to the 20th day of lactation. By cross-foster nursing experiments it was shown that the greater part of the transmission of passive immunity occurred after birth, although there was transmission of a significant, though small part, before birth. The virus passage from mothers to offspring was excluded, since Junín virus was not recovered from brains, livers, spleens and kidneys of uninfected young, born from infected mothers.