Effect of ethanol on cyclic nucleotides in vivo: consequences of controlling motor and respiratory changes. Many psychotropic drugs alter cerebellar cyclic guanosine-3',5'-monophosphate (cGMP) content. Whereas apomorphine increased levels, central depressants such as ethanol, chlordiazepoxide or barbiturates, reduce the content of cerebellar cGMP without altering levels of cyclic adenosine-3',5'-monophosphate (cAMP). Additional data indicate that tolerance develops to this reduction of cerebellar cGMP by ethanol. In paralyzed animals, the increase in cerebellar cGMP content induced by apomorphine and the decrease caused by ethanol were dramatically attenuated. Since relatively high doses of ethanol were needed to decrease blood CO2 tension in spontaneously moving rats, changes in respiratory function appear to be  of only minor importance in the ethanol-induced decrease in cerebellar cGMP. It is concluded that ethanol-induced changes in content of cerebellar cGMP in vivo may be secondary to alterations in motor and, to a lesser extent, in respiratory function.


Cyclic nucleotides in the development of alcohol tolerance and dependence: a commentary. This paper aims to outline some of the caveats inherent in studies on first and second messengers in the central nervous system as related to tolerance and dependence. Analysis of systems based on measures of first and second messengers may be an over-simplification, since the actual molecular consequences of neurotransmission might result in a multilevel cascade of many sequential messages including protein kinase activation and protein phosphorylation and/or dephosphorylation.


Effects of electric stress on glucose metabolism, glucose-stimulated cyclic adenosine 3',5'-monophosphate accumulation and 45 Ca++ efflux in isolated pancreatic islets from rats fed with a high fat diet. The effects of the electric stress on glucose oxidation, cyclic adenosine 3', 5'-monophosphate (AMP) accumulation and 45Ca++ efflux in response to glucose were studied in pancreatic islets isolated from rats fed on a control (C) or a high fat diet (F) for 12 weeks. The half of rats on each diet were subjected to electrical shocks in the random time schedule for 1 hr per day for the last 3 weeks of the feeding period (group C-S and F-S). The remaining rats were not given any shocks (group C-NS and F-NS). The rats in F-S group had the high levels of plasma epinephrine, dopamine and blood glucose. The basal content of cyclic AMP after 20 min of incubation with 2.8 mM glucose was decreased in islets from F-S group without affecting insulin release. After 20 min of incubation with 25 mM glucose, the cyclic AMP content in islets from F-S group, which was identical with that in F-NS group, was only 50% of that in C-S group. Insulin release in response to high glucose was significantly inhibited in islets from F-S group. In spite of a remarkable increase of cyclic AMP content in islets from C-S group, insulin release did not differ from that in C-NS group. Glucose (16.7 mM)-stimulated 45Ca++ efflux from the perfused islets was greatly inhibited by the high fat diet rather than by stress. The rate of glucose oxidation with 16.7 mM glucose was decreased in islets from F-S group. It is suggested that the decreased insulin release in response to glucose provoked by the combined effects of the feeding of a high fat diet and electric stress may be mediated by changes of the adenylate cyclase-cyclic AMP system on the plasma membrane of the B-cell or be related to changes in glucose metabolism in islets.


A case of the syndrome of inappropriate secretion of TSH. A girl aged 4 years with goiter and accelerated physical and skeletal growth was found to be hyperthyroid on the basis of elevated serum thyroid hormone level, nevertheless both the basal TSH and TSH responsiveness to TRH were maintained within the normal range. Serum TSH was suppressed by exogenous T3 and dexamethasone administration, but not significantly changed after propylthiouracil (PTU) treatment. The diurnal rhythmicity of anterior pituitary hormones was preserved with the high nocturnal peak of TSH and prolactin. Clinically, neither thyrotoxic signs nor evidences of pituitary tumor were observed. Her accelerated growth and elevated thyroid hormone level appeared to be induced by inappropriate secretion of TSH. In view of the literature, this is the first case of the syndrome of inappropriate secretion of TSH excluding the neoplastic origin in Japan.


Lack of inhibition of ACTH-induced aldosterone stimulation by des-asp1-,ileu8-angiotensin II in man. In 5 normal men an intravenous injection of 0.5 mg of synthetic 1-24 ACTH caused a significant increase in plasma aldosterone and a simultaneous intravenous infusion of 600 ng/kg/min of des-asp1-, ileu8-angiotensin II (AIIIA) did not inhibit this increase. Since this dose of AIIIA is known to inhibit an angiotensin II-induced increase in plasma aldosterone in normal men, the present results suggest that the ACTH-induced aldosterone stimulation is mediated by an adrenocortical receptor which is different from angiotensin II receptors.


Physical training under the influence of beta-blockade in rats. II. Effects on vascular reactivity. Rats were treated by daily swimming or running exercises for 7 weeks. One group of rats was also trained under the influence of propranolol, while another group received daily propranolol injections only. The rat groups trained without beta blockade maintained a higher tail skin temperature when exposed to 5 degrees C after the 7-week training period. This phenomenon was not observable in the animals having received their training under the influence of beta-blockade. Both rat groups trained without beta-blockade showed increased vasodilatatory response to isoprenaline, as judged from a higher elevation of the tail skin temperature in response to the drug. This response was absent in the animal group having performed its training periods under the influence of propranolol. After the injection of phenylephrine the trained rats had a higher tail skin temperature than did the controls or propranolol-treated rats. The present results suggest an elevated sensitivity of beta 2-adrenoceptors and/or decreased sensitivity of alpha-adrenoceptors in trained rats. It is suggested that for the development of these changes repeated activation of the sympathetic nervous system by exercise periods is needed. That is why they are preventable if the training is performed under the influence of beta-blockade.


Purification and properties of a phosphorylase (phosphoprotein) phosphatase associated with an alkaline phosphatase of Mr 35000 from bovine adrenal cortex. A metal-ion-independent, nonspecific phosphoprotein phosphatase (Mr = 35000) which represents the major phosphorylase phosphatase activity in bovine adrenal cortex has been purified to apparent homogeneity. An alkaline phosphatase activity (p-nitrophenyl phosphate as a substrate) of the same molecular weight, which requires both a metal ion (Mg2+ greater than Mn2+ greater than Co2+) and a sulfhydryl compound for activity, has been found to co-purify with the phosphoprotein phosphatase throughout the purification procedures. Characterization of the phosphoprotein and the alkaline phosphatase activities with respect to their catalytic properties, substrate and metal ion specificities, relationship with large molecular forms of the enzymes and responses to various effectors has been carried out. The results indicate that the phosphoprotein phosphatase can be converted by pyrophosphoryl compounds (e.g. PPi and ATP) to a metal-ion-dependent form which, subsequently, can be reactivated by Co2+ greater than Mn2+ but not by Mg2+ or Zn2+. The results also indicate that, although the phosphoprotein and the alkaline phosphatase activities are closely associated, they exhibit distinct physical and catalytic properties. Discussions concerning whether these two activities represent two different forms of the same protein or two different yet very similar polypeptide chains have been presented.


Premature appearance of hepatic phosphoenolpyruvate carboxykinase in fetal rats, not mediated by adenosine 3':5'-monophosphate. Injection of streptozotocin in utero to fetuses elicited a premature appearance of cytosolic hepatic activity of phosphoenol pyruvate carboxykinase. This was due to a precocious initiation of the synthesis of the enzyme. The streptozotocin-induced appearance of enzyme activity was not mediated by adenosine 3':5'-monophosphate since the concentration of the cyclic nucleotide in the liver was unaffected by the antibiotic, the administration of dibutyryladenosine 3':5'-monophosphate to streptozotocin-treated fetuses elicited an additive increase in enzyme activity, and insulin administration in utero repressed the streptozotocin effect while the effect due to dibutyryladenosine 3':5'-monophosphate was not inhibited by simultaneous insulin injection. Streptozotocin treatment also caused a small but consistent retardation of fetal growth and a marked reduction of liver wet weight. Histological analysis of the liver demonstrated a premature loss of some hematopoietic elements, while hepatocytes appeared normal. Hepatic protein synthesis was unaffected by the streptozotocin treatment. Streptozotocin treatment had no effect on fetal renal phosphoenol pyruvate carboxykinase activity or kidney wet weight.


Mechanism of carbamoyl-phosphate synthetase. Properties of the two binding sites for ATP. Carbamoyl phosphate synthethase I synthesizes carbamoyl phosphate from ammonia, HCO3- and two molecules of ATP, one of which, ATPA, yields Pi while the other, ATPB, yields the phosphoryl group of carbamoyl phosphate. Pulse-chase experiments with [gamma-32P]ATP without added HCO3- demonstrate separate binding sites for ATPA and ATPB. Bound ATPA dissociates readily from its site (t1/2 approximately 1--2 s) and the Kd is 0.2--0.7 mM. For the ATPB binding site the t1/2 for dissociation is 5--12 s and the Kd approximately 10 mM. Kd for ATPA seems to increase with enzyme concentration whereas Kd for ATPB does not change. HClO4 releases the ATP unchanged from the enzyme . ATPB and enzyme . ATPB . ATPA complexes. In the presence of HCO3-, ATP and N-acetylglutamate, an enzyme . ATPB . HCO3- . ATPA complex is formed. Its formation by the addition of HCO3- to the enzyme . ATPB . ATPA complex appears to involve an initial bimolecular addition reaction followed by an isomerization. Treatment with HClO4 releases Pi from ATPA but ATPB is released unchanged. Spontaneous hydrolysis of ATPA is responsible for the ATPase activity of the enzyme. Thus, a covalent bond may form between HCO3- and ATPA. However, ATPA can dissociate rapidly (t1/2 less than 10 s). The Kd for ATPA is approximately 0.2 mM. ATPB appears unable to dissociate from the enzyme . ATPB . HCO3- . ATPA complex since the t1/2 for dissociation of ATPB from the enzyme is lengthened about five times in the presence of 19 mM HCO3- and at 1 mM ATP. ATPA may also hydrolyse in this complex and be replaced by another molecule of ATP in the absence of exchange of ATPB. However, the ATPA binding site must be occupied to prevent ATPB release. ATPB may be bound in a pocket which becomes inaccessible to the solution when HCO3- and ATPA also bind. In contrast, HCO3- does not inhibit the binding of ATPB to the enzyme. Various intermediate steps in the formation of the enzyme . ATPb . HCO3- . ATPA complex are discussed. Additional evidence is presented that the ATPB binding site is only periodically accessible to ATP in solution and that ATPB in the steady-state reaction binds when the products leave. Since greater than 1.3 mol ATPB and greater than 1.8 mol ATPA bind/mol enzyme dimer, the enzyme monomer may be an active species.


Aerobic lactate synthesis by cardiac muscle. Cardiac lactate production under aerobic conditions is absolutely dependent upon the availability of extracellular pyruvate. In the steady state, aerobic lactate output is largely independent of cardiac work load, but increases slightly when octanoate is included in addition to pyruvate in the perfusion fluid. Transient episodes of supra-normal lactate production are seen after sudden increases in cardiac work output, and also after transitions from octanoate to pyruvate in the perfusion media. These pulses of lactate production are invariably associated with the slow activation of pyruvate dehydrogenase in response to a sudden change in cardiac metabolic state, and they are abolished by pre-perfusion with dichloracetate, which converts pyruvate dehydrogenase into the fully active form. A second, additional component of the lactate pulses is sensitive to pre-perfusion with the transaminase inhibitor aminooxyacetate. The size of the second component is markedly dependent upon the precise protocol adopted for the experiment, and these variations suggest that the second component is associated with a major redistribution of cardiac Krebs' cycle intermediates and amino acids following the initial exposure to pyruvate-containing media. Steadystate aerobic lactate production is insensitive to both dichloroacetate and aminooxyacetate, and is thought to result from a direct exchange of malate for oxaloacetate across the heart mitochondrial membranes.


Polygraphic study during whole night sleep in infantile spasms. The whole night EEG were polygraphically recorded and analyzed in 9 patients with infantile spasms prior to ACTH therapy. The subjects were divided into two groups, favorable and unfavorable, depending upon the response to the ACTH therapy. (1) Among the unfavorable group, the deep sleep stage was not observed; while the light sleep stage tended to dominate. (2) REM sleep period was noted less among the unfavorable than among the favorable group. REM density also tended to be lower among the unfavorable group. (3) Of the 4 unfavorable cases, 2 did not manifest REM sleep at all. Of the remaining 2, 1 had a remarkably long REM interval period. Even among the favorable cases, REM sleep tended to be short and appear frequently. (4) The period of muscle atonia during NREM sleep was markedly prolonged in all cases. (5) Body movements of both types (gross and twitch) were less frequent comparing to those of normal younger children, more remarkably in unfavorable cases. From the above findings, a disorder of the pontine reticular formation would be suggested in cases of infantile spasms. Reduction of body movements at each sleep stage might indicate abnormalities of monoamine metabolism in the brain stem of patients with this condition.


Computerized automated syndromic and topographical neurologic diagnosis of lesions of the cranial nerves. A system is described which, by means of a digital computer, computes from a previously automatically recorded objective neurologic finding a syndromic and topographical conclusion. For the present the system is designed and checked for lesions of the cranial nerves, but its principle is appropriate for the whole syndromic and topographical neurologic diagnosis.


Cyclic neucleotides in progressive muscular dystrophy. The authors radioimmunoassayed cyclic nucleotide concentrations in plasma and biopsied muscles of muscular dystrophy and muscles of chicken embryo. c-AMP concentrations in plasma were significantly lowered in Duchenne-type muscular dystrophy and this lowered degree was correlated with the stage of progression. Plasma c-GMP levels were also depressed in Duchenne-type dystrophy. In biopsied muscles, c-AMP concentrations per milligram of non-collagen protein were within normal limits. Therefore, the decrease of plasma c-AMP concentrations might be an expression of total metabolic changes rather than a pathologic process of the muscle itself. As for the dystrophic chicken embryo, both c-AMP and GMP concentrations were decreasing in the pectoral muscles in parallel with the advancement of hatching stages.


The action of neurotensin on single myenteric neurones. The action of neurotensin was studied on single myenteric neurones within ganglia of the myenteric plexus isolated from the guinea-pig ileum. Drugs were applied by adding them to the perfusing Krebs solution. Extracellular recording with glass suction electrodes indicated that neurotensin (100 pM-300 nM) caused a dose-dependent excitation of about 50% of myenteric neurones; the remaining neurones were unaffected. This effect persisted in calcium-free solutions. Intracellular recording showed that a similar proportion of Type 1 myenteric neurones were depolarized by neurotensin: this was associated with an increase in membrane resistance. Type 2 cells were either depolarized or hyperpolarized by neurotensin. The depolarization persisted in calcium-free solutions. The hyperpolarization disappeared in calcium-free solutions, suggesting either that the potential change itself is calcium-dependent or that it was due to release by neurotensin of a hyperpolarizing substance.


Indirect intestinal stimulatory effects of heroin: direct action on opiate receptors. The effects of bolus intra-arterial doses of heroin and other stimulant drugs were studied in vascularly perfused isolated segments of dog small intestine. Heroin caused dose-related increases in intraluminal pressure similar in appearance to those caused by morphine. Perfusion with Krebs bicarbonate solution containing naloxone selectively abolished intestinal responses to heroin. Perfusion with cinanserin, a 5-hydroxytryptamine (5-HT) antagonist, decreased intestinal responses to 5-HT and heroin without affecting responses to dimethylphenylpiperazinium (DMPP) or bethanechol (BeCh). Tetrodotoxin reduced responses to heroin, 5-HT and DMPP but not responses to BeCh. Atropine antagonized contractile responses to all 4 stimulatory agents. These data indicate that heroin interacts with a conventional opiate receptor in the intestine and that the intestinal stimulatory effect of heroin is mediated by the release of endogenous 5-HT which activates intramural cholinergic neurons.


Pretreatment with delta 1-tetrahydrocannabinol and psychoactive drugs: effects on uptake of biogenic amines and on behavior. Injection of delta 1-tetrahydrocannabinol (THC) into mice increased the uptake into brain synaptosomes of radioactive DA, NE, 5-HT and GABA, with the effect on GABA being the greatest; uptake of leucine was not stimulated, indicating that THC does not facilitate the transport of amino acids in general. The effect of THC was stereospecific because pretreatment with the non-psychoactive isomer, (+) delta 6-THC had no effect on uptake of DA into cortical synaptosomes. The effect on DA uptake was correlated with psychoactive potency. THC enhanced uptake more than did SP-111 (a water soluble ester of THC but less potent than THC) and much more than cannabidiol (a non-psychoactive ingredient of marijuana). THC increased the uptake of DA into striatal synaptosomes but much less than into cortical synaptosomes. The enhancing effect of THC on uptake in cortex showed tolerance after chronic (1 week) treatment with THC. Catecholaminergic receptor antagonists (chlorpromazine, propranolol), like THC, stimulated the uptake of DA or NE into cortical synaptosomer. In contrast, pretreatment with MAO (pargyline) or uptake (tricyclic antidepressants) inhibitors, or with amphetamine, decreased uptake. Thus THC does not inhibit MAO uptake, or stimulate the release of catecholamines but may interact with a receptor. The notion of a THC--receptor interaction is supported by behavioral experiments.


The role of calcium and cyclic AMP in the contractile action of angiotensin II upon rat descending colon. The effect of procedures which affect calcium availability and tissue levels of cyclic AMP were investigated upon the contractions of rat colon to angiotensin II, PGE2 and KCl. Calcium free Tyrode containing EDTA (25 microM) reduced the response to angiotensin more than to PGE2 and KCl. An increase in calcium concentration to 3.6 mM potentiated responses to angiotensin and KCl. A further increase to either 7.2 mM or 10.8 mM still potentiated responses to angiotensin but those to KCl wre reduced. SKF 525A (2.6 x 10(-5) M) and verapamil (3.5 x 10(-6) M) inhibited the responses to KCl but the responses to PGE2 and angiotensin were less affected. Isoprenaline, Theophylline and dibutyryl cyclic AMP preferentially reduced the responses to angiotensin and PGE2. It is suggested that part of the response to angiotensin involves an influx of calcium, independent of depolarisation.


Urotensin I effects on intracellular content of cyclic AMP in the rat tail artery. Rat tail artery strips were incubated in the presence of 4 x 10(-3) M theophylline and urotensin I (UI). At the concentrations of 1.50, 7.50 mU/ml but not of 0.75 mU/ml UI, the content of cAMP increased significantly. Cyclic AMP content and muscle tension were measured at 5--30 sec intervals during noradrenaline (NA)- and potassium chloride (KCl)-induced contractions and during subsequent urotensin I-induced relaxation. Cyclic AMP content declined initially after addition of NA but returned to baseline levels during the contraction. Significant dose-related increases in cAMP were noted upon addition of UI, prior to the onset of the relaxation response followed by a marked decrease during the relaxation response. An increase in cAMP was observed after initiation of the contractile response to KCl-contracted tissues, but no further increase in cAMP after addition of UI was detected. These findings suggest an involvement of cyclic AMP in the initial events leading to the relaxation response to urotensin I in the NA-contracted rat tail artery. Urotensin I-induced relaxation in KCl-contracted tissues appears to proceed by an alternative mechanism which does not require increased levels of cAMP.


Norepinephrine stimulated cyclic AMP accumulation in rat limbic forebrain slices: partial mediation by a subpopulation of receptors with neither alpha nor beta characteristics. Both isoproterenol and norepinephrine (NE) increase cyclic AMP in slices of the rat limbic forebrain and the responses are enhanced in the presence of the phosphodiesterase inhibitor RO 20-1724. However, even in the presence of RO 20-1724, no accumulation of cyclic AMP was observed after the addition of dopamine, serotonin or the alpha-agonists methoxamine and phenylephrine. This suggests that these agents do not activate adenylate cyclase in this preparation or that their respective receptors--unlike the beta-receptor--are not coupled to adenylate cyclase. Isoproterenol, which has a high affinity for this adenylate cyclase system but only 20-30% of the maximal activity of NE, does not interfere with the agonist activity of NE. Moreover, the effect of isoproterenol is not additive with that of NE thus suggesting that isoproterenol is acting on a subpopulation of NE receptors. The results indicate that two populations of NE receptors coupled to adenylate cyclase are present in slices of rat limbic forebrain: one which has beta-characteristics and the other with neither alpha- nor beta-characteristics based on agonist studies.


Loss of beta-adrenoceptor binding sites in rat striatum following kainic acid lesions. Intrastriatal injection of kainic acid (5 nmoles) to rats led to severe destruction of nerve cell bodies throughout the caudate-putamen complex and an extensive proliferation of glial cells. Lesioned striata displayed a significant 23% loss of beta-adrenoceptor binding sites 21--24 days after injection of kainic acid. Further analysis of these changes revealed that this loss of sites was selectively within the beta 1 receptor population. Although these results do not rule out a partial glial cell localisation for beta-adrenoceptors, they do indicate that at least a proportion of beta 1 receptors are present on striatal perikarya.


Inhibition by digoxin and SC4453 of (Na+ + K+)-ATPase prepared from human heart, guinea-pig heart and guinea-pig brain. SC4453 is a digoxin analogue with a pyridazine instead of a lactone ring on C17 beta. SC4453 was compared with digoxin with respect to inhibition of (Na+ + K+)-ATPase prepared from human heart, guinea-pig heart and guinea-pig brain. SC4453 was slightly less potent than digoxin but showed a similar sensitivity to K+. As for cardenolides, species differences in sensitivity to SC4453 were accounted for by differences in the rate of dissociation from the receptors. These observations confirm that the human heart is one of the tissues most sensitive to cardiac glycosides.


Effect of naloxone, haloperidol and propranolol on cyclic 3',5'-adenosine monophosphate content of rat amygdala. Naloxone (0.4 mg/kg, i.p.) causes an increase of cyclic adenosine monophosphate levels in the amygdala, but not in the hippocampus, caudate, or hypothalamus, of rats. The effect is antagonized by haloperidol (0.5 mg/kg, i.p.) and by propranolol (0.5 mg/kg, i.p.). This is consistent with the hypothesis of a tonic inhibitory influence of endogenous opiates on central dopaminergic and beta-noradrenergic systems. Haloperidol had an effect of its own on amygdala cyclic adenosine monophosphate levels which was blocked by propranolol. This suggests the possibility of an antagonistic interaction between dopaminergic and beta-noradrenergic innervation on this structure.


Inhibition of cardiac sympathetic neurotransmission by adenosine. Experiments were performed to study the effect of adenosine on sympathetic neurotransmission to the myocardium. Adenosine administration to pentobarbital-anesthetized dogs resulted in decrease in blood pressure and significant impairment of cardioacceleration produced by stimulation of cardiac sympathetic nerves. The positive chronotropic effect of intravenous norepinephrine was not affected by adenosine. The blood pressure lowering as well as the inhibitory effect of adenosine on cardiac sympathetic nerve function could be significantly antagonized by theophylline. These results provide in vivo evidence in support of the hypothesis that adenosine caused inhibition of sympathetic neurotransmission via an action on purinergic receptors located on sympathetic nerve terminals.


Sanguinarine: a positive inotropic alkaloid which inhibits cardiac Na+,K+-ATPase. In isolated, isometrically contracting left guinea pig atria, sanguinarine, a benzophenanthridine alkaloid from the papaveracea Sanguinaria canadensis, produced a concentration-dependent positive inotropic effect. Between 2.3 x 10(-6) M and 6.5 x 10(-5) M, sanguinarine increased contractility by 108% which was comparable to the maximal inotropic effect of ouabain. Within the same concentration range, sanguinarine caused inhibition of Na+,K+-ATPase isolated from guinea pig myocardium. 100% inhibition of Na+,K+,ATPase activity occurred at 1 x 10(-4) M sanguinarine. The I50 for enzyme inhibition and the ED50 for the inotropic action of sanguinarine were the same (6-6.5 x 10(-6) M) indicating that both effects may be causally related.


Fatal herpesvirus tamarinus infection in cotton-topped marmosets (Saguinus oedipus). Fatal herpesvirus tamarinus infection was observed in cotton-topped marmosets (Saguinus oedipus) imported from South America via the United States on August 26, 1976. In addition to the lesions hitherto reported in herpesvirus tamarinus infection, severe degenerative and necrotic changes of ganglion cells were recognized with intranuclear inclusion bodies in the plexus of the digestive tract and the sympathetic nerves and their ganglions in the abdominal cavity. Inflammatory or regressive changes were also noted in the central nervous system. A large number of basophilic or eosinophilic intranuclear inclusion bodies frequently recognized in multinucleated giant cells were observed in various organs and tissues, and they showed different shapes at the electron microscopic level. Morphological findings indicated that herpesvirus tamarinus infection seemed to be similar to herpes simplex virus infection in man. The findings of the susceptibility of a variety of cell cultures to the virus isolate serologically identified as herpesvirus tamarinus and physicochemical characteristics of the virus isolate were in general agreement with the findings of herpesvirus tamarinus already reported by previous workers.


[Synthesis and pharmacological activity of 1,2,3-triazole derivatives of naphthalene, quinoline and pyridine]. The preparation of three series of 1,2,3-triazol derivatives, with a naphthalene, quinoline or pyridine ring in 1 position, is described. Preliminary pharmacological screening of some of these compounds showed no appreciable activity.


[Variations of B- and T-lymphocytes in the peripheral blood and variations of the stimulability of cyclic nucleotides in the T-lymphocytes of patients suffering from malignant melanoma (author's transl)]. The portion of B- and T-lymphocytes at the whole white blood count have been estimated in patients with malignant melanoma. An elevation of the B-lymphocytes and a decrease of the T-lymphocytes could be observed in comparison to controls. The CAMP and CGMP level in lymphocytes of patients suffering from melanoma shows a correlation to the degree of metastasing changes. The cyclic nucleotides by mealanoma patients are influenced differently than by normal persons, after addition of physiologic stimulators (isoproterenole, PGE, PGF, acetylcholine). The beta-adrenergic system shows a decreased stimulability and the cholinergic system an increased one depending upon the disease stage in comparison to controls.


Tissue-specific expression of onco-fetal antigens during embryogenesis. It has become evident from recent literature that especially in tumor virus systems, cell transformation leads to an arrest of differentiation or to a retrodifferentiation. This may be reflected by the expression of embryonic antigens and it is therefore particularly important to characterize such antigens according to their specificity as well as to their specificity during embryogenesis. We have demonstrated the expression of embryonic antigens which are cross-reactive in avian fibroblasts transformed either by Rous sarcoma virus or by methylcholanthrene. This paper is intended to demonstrate that these embryonic antigens are detected only at a certain period of embryogenesis and particularly in muscle cells. They are detected only occasionally or not at all in cells of other tissues such as brain, liver, lung, and the digestive organs. These antigens are absent from the target cells before transformation and are consequently induced by the transforming agent, either viral or chemical. Therefore, these results suggest that by transformation mechanism, cells become specifically reverted to an earlier stage of differentiation (retrodifferentiation).


Increase in levels of cyclic AMP during avian limb chondrogenesis in vitro. In the present study the level of cAMP was measured during in vitro chondrogenesis of wing mesenchyme of stage 24 chick embryos and was found to increase significantly from 6.3 pmol/mg protein at the end of the first day of culture to 9.7 pmol/mg protein on the second day, when chondrogenic expression is first detected by the appearance of an Alcian blue staining extracellular matrix. Nonchondrogenic cultures derived from wings of stage 19 embryos had a lower level of cAMP (4.4 +/- 0.07 pmol/mg protein). The level of cAMP in intact wings was 4.5 +/- 0.4 pmol/mg protein and did not change between stages 19 through 25. The correlatin between increased levels of cAMP and the onset of chondrogenesis is consistent with a role of cAMP in the expression of differentiated functions in chondrocytes, as well as in some other cell types.


True divergent differentiation in a cellular slime mold, Polysphondylium pallidum. Three abstract models of divergent cell differentiation to multiple cell types are presented. These differ primarily in the proportion of developmental events common to the pathways leading to each cell type. Two experimental approaches are outlined to determine which type best describes divergent differentiation occurring in a particular organism. The first technique is to describe and compare changes in labeling of specific polypeptides which characterize development to the several cell types. The second is to observe the ability of mutants which are blocked in one pathway to develop along alternate pathways. These approaches are applied to the case of Polysphondylium pallidum, where amebae develop into stalk cells, spores, or microcysts. It is concluded that cell differentiation in P. pallidum is of the truly divergent type in which developing cells show identical sequences of events until a branch point, and thereafter very different sequences of events.


Plasma lecithin: cholesterol acyltransferase activity in liver disease. The plasma lecithin: cholesterol acyltransferase was determined in patients with various liver diseases and the relationship between this enzyme activity and the other liver function tests were studied including long term observations. Lecithin: cholesterol acyltransferase activity in fulminant hepatitis and liver cirrhosis showed a significant decrease in comparison with normal volunteers. Although the enzyme activity of hepatoma showed significant decrease, they were ascribed to the influence of concomitant liver cirrhosis. The enzyme activity showed insignificant changes in the acute and chronic hepatitis and alcoholic liver disease. Lecithin: cholesterol acyltransferase activity was correlated with the concentration of cholesterolester rather than with the ratio of esters to cholesterol. In addition, it was well correlated with pseudocholine esterase and serum albumin. The lecithin: cholesterol acyltransferase activity in the cases during follow-up period varied in good parallel with cholesterol-esters concentration and pseudocholine esterase in the cases with acute hepatitis; with serum albumin in the cases with liver cirrhosis. Furthermore, it varied inversely with SGPT in the cases with acute hepatitis. In a case with hepatoma, lecithin: cholesterol acyltransferase activity decreased more sharply than the cholesterolesters concentration and serum albumin immediately before death.


Incidence and clinical significance of HBe antigen and antibody in HBsAg-positive various liver diseases. The HBeAg was detected in 5 of 24 patients with acute type B hepatitis (20.8%), 33 of 95 with chronic hepatitis (34.7%), 6 of 33 with liver cirrhosis (18.2%), and 3 of 39 with hepatocellular carcinoma (7.7%). On the other hand, anti-HBe was found in 4.2% of acute hepatitis, 18.9% of chronic hepatitis, 9.1% of liver cirrhosis, and 12.8% of hepatocellular carcinoma. We found that an early detection of HBeAg in patients with acute hepatitis is of no prognostic value, but its persistence may provide the earliest evidence of potential chronicity. In chronic liver diseases, HBeAg-positive cases showed remarkable fluctuations of serum transaminase levels, severe histological changes and poor responses to treatment. Many of the HBeAg-positive patients lost their initial positivity of HBeAg within six months or one year and in some cases serocoverted to anti-HBe after acute exacerbation. Follow-up study more than several years revealed that the presence of anti-HBe reflect an inactive stage and a more favorable outcome, whereas persistence of HBeAg may provide an active and continuing hepatocellular damage. From these results, we believed that serial measurements of HBeAg/anti-HBe system are useful prognostic marker in patients with HBsAg-positive liver disease.


Isolation and purification of human large bowel mucosal lymphoid cells: effect of separation technique on functional characteristics. Human large bowel lamina propria lymphoid cells have been isolated using both mechanical and enzymatic techniques. Their separation from other cell types after isolation was effected with greater efficiency by sedimentation on isokinetic gradients than by filtration through glass bead columns. After being purified, the capacity of the lamina propria lymphocytes to function in vitro as effector cells in antibody-dependent cellular cytotoxicity was determined. Mechanical distruption of the mucosa gave low yields of lymphoid cells, which lacked the capacity for cytotoxicity. Enzymatic digestion of mucosal tissue, by comparison, yielded large numbers of viable lymphoid cells which retained a significant level of cytotoxic activity. Investigation revealed that mechanical homogenisation stimulated the synthesis of prostaglandin E2, and inhibitor studies showed that this mediator was responsible for the lack of cytotoxic activity in mechanically-liberated lymphocytes.


[Epstein-Barr virus and human chromosomes: Close association of the resident viral genome and the expression of the virus-determined nuclear antigen (EBNA) with the presence of chromosome 14 in human/mouse hybrid cells (author's transl)]. Fourteen hybrid clones derived from the fused cultures of human lymphoblastoid FV5 cells and 5-bromodeoxyuridine-resistant mouse fibroblastic MCB2 cells grown in HAT selective medium were examined for the presence of Epstein-Barr virus (EBV) DNA, the expression of the virus-determined nuclear antigen (EBNA), and the presence of human chromosomes, in the course of serial passage in vitro. Among the hybrid clones tested, three were positive for EBV DNA and EBNA, whereas the remaining 11 clones were totally negative. The chromosome investigations showed that human chromosome 14 was consistently involved in all three EBV genome-positive and EBNA-positive hybrid clones, but not in any negative clones. In 10 subclones isolated from one of the three positive clones, all of which contained chromosome 14 alone as human chromosomes, a concordant segregation of EBNA, EBV DNA and No. 14 chromosome was evident. These findings suggest that the resident EBV genome is closely associated with chromosome 14 and the presence of this particular chromosome alone is sufficient for the maintenance and the expression of EBV genetic information in human lymphoblastoid cells.


[Study on glycolipids in human lung carcinoma of histologically different types (author's transl)]. Human lung carcinoma tissues with histological types of adenocarcinoma, squamous cell and small cell undifferentiated carcinomas were investigated for glycolipids. Carcinoma tissues, as well as normal adult and embryonic lungs contained ceramide mono-, di- and trihexosides, globoside and hematoside as major glycolipids. In addition to them, sulfatide which was identified as ceramide 3-sulfate-galactoside, was isolated in much lesser amount. The content of sulfatide was markedly increased in adenocarcinoma than that in other carcinomas and normal lung. Adenocarcinoma was also characterized by significantly lower level of total glycolipids which was largely due to the diminished contents of ceramide mono- and dihexosides, and hematoside, as compared to those in other two carcinomas. Squamous cell carcinoma had a characteristic pattern with an increment of hematoside. In small cell undifferentiated carcinoma, glycolipid contents were similar with those in squamous cell carcinoma but the relative composition of major glycolipids was markedly differed from that in other types. All the types of carcinoma examined showed marked increase of ceramide mono- and dihexosides (except for ceramide dihexoside in adenocarcinoma) compared to those in normal adult lung. Overall feature of glycolipids in embryonic lung appeared to be an intermediate between carcinomas and normal adult lung.


[Clinical study on aldolase isoenzyme--the development of the method of cancer diagnosis with muscle type aldolase (author's transl)]. A specific radioimmunoassay was developed for the quantitation of human muscle type aldolase (M-ALD) in human serum. The amount of M-ALD antigen present in 135 sera from normal healthy subjects, noncancer patients and cancer patients was determined. The serum M-ALD value for the 41 normal healthy subjects averaged 171 +/- 39 ng/ml and they had a range from 130 ng/ml to 210 ng/ml. In 33 noncancer hospital patients, excluding patients with muscle diseases, the serum M-ALD values averaged 164 ng/ml and had a range of 125 to 220 ng/ml. In contrast the 61 cancer patients had serum M-ALD values which averaged 586 ng/ml and had a range of 85 to 5800 ng/ml. Eighty two percent of the cancer patients had serum concentrations of M-ALD which were outside of the normal range. The CEA value was determined in the serum of the cancer patients and thirty five percent of the patients had elevated serum concentrations. The measurement of M-ALD values in human serum may be an additional laboratory test in the diagnosis and follow up of the cancer patients.


[Experimental and clinical studies on blood coagulation and lysis system after hepatic resection (author's transl)]. Blood coagulation and lysis system was studied with other biochemical serum analysis in three groups of mongrel dogs; laparotomy without hepatic resection (group I), with 50% hepatic section (group II), and 70% hepatic resection (group III). In clinical studies, six cases of hepatic tumors and one case of hepatoma with cirrhosis were selected for examinations. All the data examined in group I were restored toward normal within 48 hours. Total serum protein level was significantly decreased after hepatic resection. The protein level in group III was lower than in group II on the 4th postoperative day (P less than 0.01). In clinical studies, low serum protein levels did not reach a preoperative value even 3 weeks after extended right hepatic lobectomy. Time course of Al-P and transaminase changes in clinical studies was similar to that in experimental study with dogs. Al-P and transaminase showed an abnormally high level in the patient with hepatic cirrhosis. Serum bilirubin levels were not increased after hepatic resection. In blood coagulation and lysis system, serum fibrinogen levels were markedly decreased: 50% reduction in group III and 30% reduction in group II on the first postoperative day. The degree of decrease in the fibrinogen level was proportional to the size of resected volume of the liver. On the other hand, in clinical studies fibrinogen levels were slightly decreased. In the case of hepatic resection with cirrhosis, however, the values were markedly decreased: 40% reduction on the 4th and 25% on the 21st postoperative days. Fibrinolysis system was accelerated group II and III. The acceleration continued until 3 weeks after hepatic resection. From these results it may be concluded that analysis of blood coagulation and lysis system after hepatic resection is useful in evaluating a residual hepatic function after partial resection and in selecting a treatment suitable for hepatic insufficiency.


Intrinsic ADP-ribose transferase activity versus levels of mono(adp-ribose)protein conjugates in proliferating Ehrlich ascites tumor cells. Transition of proliferating Ehrlich ascites tumor cells (3 days after transplantation) to the non-proliferating status (8--14 days after transplantation) was associated with an increase in total mono (ADP-ribose) protein conjugates. This increase was largely confined to the NH2OH-resistant subfraction. When the amounts of mono-(ADP-ribose) conjugates from 20% trichloroacetic acid precipitates were compared with those from 5% perchloric acid precipitates, no significant differences were seen. This fact excludes histone H1 as a major mono (ADP-ribose) acceptor in vivo in these cells. Transition to the resting state was also associated with a small decrease in NAD levels, and with no significant changes of total ADP-ribose transferase activity. However intrinsic ADP-ribose transferase activity as expressed in permeabilized cells was increased, being correlated with the changes in the level of the NH2OH-resistant mono (ADP-ribose) protein conjugates. This shows that alterations in intrinsic transferase activity may, in general, indicate similar alterations in major subfractions of ADP-ribose conjugates. Intrinsic ADP-ribose transferase activity exhibited an inverse relationship to ornithine decarboxylase activity.


Cerebral microvessels and derived cells in tissue culture: isolation and preliminary characterization. Microvessels isolated from mouse forebrain were used as the source material for the derivation of cerebral vascular endothelium and smooth-muscle cells in culture. The microvessels were isolated by a mechanical dispersion and filtration technique, and were maintained in vitro as organoid cultures. A microvessel classification system was developed and proved to be useful as a tool in monitoring culture progress and in predicting the type(s) of microvessel(s) that would give rise to migrating and/or proliferating cells. The isolated cerebral microvessels were heterogeneous in diameter, size of individual vascular isolate, and proliferative potential. The isolated microvessels ranged in diameter from 4 micron to 25 micron and in size from a single microvascular segment to a large multibranched plexus with mural cells. The initial viability, determined by erythrosin B exclusion, was approximately 50% on a per cell basis. All microvessel classes had proliferative potential although the rate and extent of proliferation were both microvessel class- and density-dependent. The smaller microvessels gave rise to endothelial cells, whereas the large microvessels gave rise to endothelial and smooth-muscle cells. The viability and progress of a microvessel toward derived cell proliferation seemed to be directly proportional to the number of mural cells present.


Effects of culture conditions on the synthesis of human chorionic gonadotropin by placental organ cultures. Culture conditions for maintaining first trimester human placenta in organ culture, which enhance the secretion of human chorionic gonadotropin (hCG), are described. Nutrient medium, oxygen tension and Gelfoam support matrix influence the synthesis of hCG by these cultures. Placental tissue remained viable for the duration of experiments (12 days) as judged by the incorporation of tritiated thymidine into DNA and the lack of release of incorporated [125I]iododeoxyuridine. Optimal conditions for hCG synthesis in placental organ culture included an atmosphere of 95% air and 5% Co2 (approximately 20% O2), CMRL 1066 medium containing fetal human or bovine serum, insulin, hydrocortisone and retinal acetate. Multiple pieces of placenta could be cultured in the same dish with an additive effect on hCG secretion. The functional responsiveness of these placental cultures was demonstrated by modulation of hCG synthesis with theophylline and 3'5' dibutyryl cyclic AMP.


Biology of mouse thymic virus, a herpesvirus of mice, and the antigenic relationship to mouse cytomegalovirus. Mouse thymic virus (TA) is a herpesvirus which produces extensive necrosis of the thymus of newborn mice 7 to 14 days after infection. Infectious virus can be recovered from the thymus for only 10 days after infection, with highest titers occurring between days 5 and 7. In mice 5 days old or less, TA infects thymus cells and produces massive necrosis. TA also infects the salivary glands and persists as a chronic infection. Newborn mice infected with TA have no detectable humoral immune response. Infected adult mice respond, and humoral antibody is detected 7 days after infection. Titers are maintained for months thereafter. Regardless of the age of the mice inoculated with TA, persistent infection was established in the salivary glands, but no evidence for thymus involvement was observed when adults were infected. TA does not cross-react serologically by immunofluorescent, complement fixation, or virus neutralization tests with mouse cytomegalovirus; however, interestingly, the epidemiology of the two herpesviruses are similar. Both mouse cytomegalovirus and TA were isolated from the same animals in populations of laboratory and wild mice. Evidence of infection with mouse cytomegalovirus and TA were most apparent by virus isolations, since humoral antibody responses are rarely observed. All strains of mice tested were susceptible to TA infection. However, in some strains maximum necrosis occurred at 7 days, compared with 10 to 14 days for other strains. The difference in age susceptibility and the target tissue of thymus in newborn mice suggests that TA is a model herpesvirus for studying the effects of viral infections on humoral and cell-mediated immunological functions.


Difference in capacity of Sendai virus envelope components to induce cytotoxic T lymphocytes in primary and secondary immune responses. Studies were made on the abilities of Sendai virus envelope components to induce primary and secondary generations of virus-specific cytotoxic mouse T lymphocytes (CTL). The primary CTL response in BALB/c mice was induced by reassembled envelope particles that had fusion activity but not by envelope glycoproteins without fusion activity, although both preparations induced a humoral immune response. Reconstitution of membrane-bound envelope proteins from envelope glycoproteins with lipids restored the fusion activity and the capacity to induce CTL. Target cells susceptible to virus-specific CTL could be induced by reassembled envelope particles, but not by envelope glycoproteins or LLC-MK2 cell-grown Sendai virus, neither of which had fusion activity. On the other hand, all the viruses and envelope components tested were found to stimulate a virus-specific CTL response in the in vitro secondary generation of CTL from virus-primed spleen cells. These results suggest that Senaei virus fusion activity is involved in primary induction of the CTL response as well as in target cell formation, but that it is not essential for secondary stimulation of the CTL response.


Interaction of murine cytomegalovirus with separated populations of spleen cells. Mouse spleen cultures were infected with murine cytomegalovirus, either before or after separating the cells into various classes, and the growth of virus and production of infectious centers were measured in each cell class. The separation techniques, which were used in various combinations, comprised: adherence to plastic surfaces; adherence to nylon wool columns; gamma irradiation; and treatment with anti-immunoglobulin serum plus complement. The "macrophage population" took up most of the input virus, as shown by autoradiography of spleen cells infected with radioactive virus. These cells formed infectious centers but replicated the virus only transiently. In contrast, the "B-lymphocyte population" contained a minor cell fraction which was permissive for replication. The "T-lymphocyte population" appeared to be nonpermissive for replication and did not produce infectious centers.


Effect of intracisternal injection of ACTH on blood glucose and hepatic glycogen in dogs. The intracisternal administration of ACTH in a dose of 0.2 muU in mongrel dogs produced a significant (P less than 0.001) rise in blood glucose (BGL) and a fall (P less than 0.01) in hepatic glycogen concentration (HGC): in contrast its intravenous administration was devoid of this action. These changes were markedly reduced in the hepatic denervated animals and were completely abolished in animals with spinal cord transectomy. The above changes suggest that ACTH on the intracisternal administration causes a rise in the BGL by an action on the liver through the sympathetic fibers.


Phagocytosis-induced injury of normal and activated alveolar macrophages. The present study tested the hypothesis that BCG-activated macrophages become injured when they phagocytose certain particulates. The data indicate that alveolar macrophages obtained from Mycobacterium bovis BCG-sensitized animals were more susceptible to cell death after in vitro incubation with BCG or zymosan than were macrophages from normal animals. Increased susceptibility was dependent on phagocytosis, since incubation with cytochalasin B, a phagocytosis inhibitor, abrogated the effect. Catalase, cytochrome c, and ascorbic acid offered partial protection to the macrophage, suggesting the involvement of free radicals in the generation of cytotoxicity. Not all of the cells from the alveolar populations were equally susceptible to cell death, thus suggesting either heterogeneity in the cell population or a requirement of more than one cell type in the induction of necrosis or both.


Induction of an inhibitor of interferon action in a mouse lymphokine preparation. An inhibitor of interferon action was identified in mouse lymphokine preparations. The inhibito was first detected in the supernatant fluid of mouse spleen cells at 72 h after stimulation by staphylococcal enterotoxin A. Inhibitor was not detected in supernatant fluids from unstimulated cultures. This inhibitor blocks the antiviral activity of both immune and fibroblast interferons. The inhibitor was purified 1,000-fold by two-step column chromatography. The partially purified inhibitor blocked the antiviral activity of up to 400 U of interferon. The immunosuppressive effect of interferon was also blocked by the inhibitor, suggesting that the inhibitor may modulate the immunoregulatory function of interferon.


Epstein-Barr virus specific IgA serum antibodies in nasopharyngeal and other respiratory carcinomas. In order to better understand the relationship between IgA and IgG antibodies to Epstein-Barr virus (EBV) in nasopharyngeal carcinoma (NPC), we analyzed 230 NPC sera but also a series of sera from patients with other carcinomas selected for their high EBV/IgG antibody titres. We were surprised to find that 21 out of 46 sera from bronchopulmonary carcinomas (BPC) and 6 out of 7 carcinomas of nasal fossae were IgA anti-VCA positive, and 9 BPC sera and 5 nasal fossae sera were also positive for IgA and anti-EA. Bronchial undifferentiated small-cell carcinomas may represent a particular group associated with high EBV profile.


Antibody-dependent cell-mediated cytotoxicity against Epstein-Barr virus membrane antigen in African Burkitt's lymphoma. Serial samples of sera from patients with African Burkitt's lymphoma were tested for antibody against Epstein-Barr virus (EBV)-specific membrane antigen (MA) by the antibody-dependent cell-mediated cytotoxicity assay (ADCC). Titers of patients in the long-term survivor group were generally higher than those found in the sera of patients in the short-term survivor group. Although ADCC titers to EBV-MA were not useful in predicting which patients would relapse there was a definite relationship between ADCC titer and prognosis. The individual differences in ADCC titers in patients in remission may explain the variability of responses that have been reported in studies on serotherapy with remission plasma.


Biochemical studies on nickel toxicity in weanling rats -- influence of vitamin C supplementation. The influence of high intake of vitamin C in the young growing rats under administration of nickel sulphate in toxic doses has been studied. Ingestion of nickel sulphate depresses the growth rates of rats, alters the vitamin C status in different tissues, inhibits certain enzymes of vitamin C metabolism and changes the activities of alkaline phosphatase and succinic dehydrogenase in the liver and kidney tissues. The acid phosphatase activity of liver, kidney and brain tissues of rats and glucose-6-phosphatase activity in liver, and serum GOT activity were stimulated, with reduction in the in the liver GOT activity. There is stimulation in the activities of rat brain inorganic pyrophosphatase and cholinesterase. Kidney tissues of rats were found to be more susceptible towards nickel toxicity as compared to the hepatic tissues in respect of morphological alterations. There is almost no alteration in the hepatic lipid composition. Administration of vitamin C in high doses to rats fed nickel salts in toxic doses can restore not only the growth rates but also certain enzyme activities to a significant extent.


Hypercalcemia and elevated parathyroid hormone levels in association with nephroblastoma. A three-month-old girl with nephroblastoma had at the same time a hypercalcemia and elevated parathyroid hormone levels. Both biochemical findings disappeared after surgical removal of the tumor.


Structural investigation of the antibiotic ristomycin A. The amino acid constituents. Hydrolysis of the O-methylated aglycone of ristomycin A by a mixture of KOH and NaBH4 yielded a mixture of aromatic amino acids which, after N-acetylation and O-methylation, were separated by chromatography on silica gel. Compounds III approximately VII were isolated and identified by pmr and mass spectroscopy. Compounds V approximately VII were also oxidatively degraded to the corresponding benzoate esters. Componds III and IV are derived from ristomycinic acid (I) and V from actinoidinic acid (II), both of which had been obtained in earlier acid hydrolyses of the antibiotic. Compounds VI and VII had not been detected previously nor glycine which was also found to be a product of base hydrolysis. It is postulated that the new products arise from bisdechlorovancomycinic acid (X). It is concluded that aglycoristomycin A comprises I, II and X which also constitute the aglycone of ristocetin A.


Comparison of Stomacher and Waring Blendor for homogenizing foods to be examined for Clostridium perfringens. The Colworth Stomacher Model 400 homogenizer was compared with the Waring Blendor for preparing food homogenates to be examined for Clostridium perfringens. Forty-eight samples representing 6 different food types were inoculated with C. perfringens and examined by the AOAC official first action method for enumeration of C. perfringens in foods. Identical paired specimens of each food type were blended with the 2 devices, and plate counts were made as specified in the official first action method. The effects of frozen storage on plate counts were determined by examining 24 food samples that had been stored for 3 days at -68 degrees C and homogenized both devices. Results of a statistical analysis of the experimental data indicated no significant difference overall (P greater than 0.05) in the plate counts of homogenates prepared with the Waring Blendor or the Stomacher 400, either before or after frozen storage of the food samples. However, the overall plate count average of the 48 samples was slightly higher with the Waring Blendor than with the Stomacher 400 homogenizer.


A double-alpha c-type cytochrome, cytochrome c-555, 549, from an extreme thermophile, Thermus thermophilus HB8. A "double-alpha" c-type cytochrome, cytochrome c-555, 549, was isolated from the membrane fraction of an extreme thermophile, Thermus thermophilus HB8, and highly purified by chromatographies on DEAE-cellulose and Sephadex G-75 and by isoelectric focusing. The absorption maxima were at 554.8, 548.6, 522, and 417 nm in the reduced form, and at 528, 409, and 360 nm in the oxidized form. The double alpha-peak of this cytochrome was enhanced at liquid nitrogen temperature. The cytochrome contained one heme c group per protein molecule. The isoelectric point, midpoint redox potential and molecular weight were pH 4.0, +0.206 V and about 10,000, respectively. Cytochrome c-555, 549 is highly thermostable.


Stereospecificity of peptidyl dipeptide hydrolase (angiotensin I-converting enzyme). The stereospecificity of peptidyl dipeptide hydrolase [EC 3.4.15.1] was investigated. Six free and N-blocked alanyl peptides containing D-alanine were synthesized and tested as substrates. Their susceptibilities were determined by measuring Ala-Ala release by cation exchange column chromatography. Their Michaelis constants, Km values, and velocity maxima, Vmax values, were also determined. The enzyme showed high stereospecificity for an amino acyl residue in position 3 from C-terminus: it had an absolute requirement for the alanyl residue of the L-configuration in this position. An alanyl residue of the L-configuration in position 1 or 2 increased, but was not essential for activity. The enzyme showed little stereospecificity for an alanyl residue in position 4 from the C-terminus.


Concanavalin A-induced increase in the membrane fluidity of chicken erythrocytes. To determine the fluidity of the membrane lipid phase, chicken erythrocytes were labeled with a stearic acid derivative spin label. When chicken erythrocytes were treated with concanavalin A (Con A), ESR spectra showed a change in the peaks of the labels in membrane lipids, indicating an increase of membrane fluidity. The degree of the increase in fluidity of the membrane lipid phase depended on the valency of the lectin used. Tetravalent Con A induced an increase of membrane fluidity at a concentration as low as 30 micrograms/ml, while a monovalent derivative of Con A did not affect membrane fluidity. This increase in membrane fluidity was observed within 10 min after the addition of Con A. If bound Con A was removed with methyl alpha-D-mannoside later than 60 min after its addition, a complete return of the fluidity to the normal level could not be observed. However, no change was found in the composition of phospholipids or in the fatty acid compositions of phosphatidylcholine and phosphatidylethanolamine of chicken erythrocytes after the addition of Con A, indicating that this increase in membrane fluidity is not caused by a change of lipid composition. The clustering of membrane receptors of chicken erythrocytes for Con A was demonstrated when the two-dimensional distribution of ferritin-conjugated Con A on the membranes was assayed by transmission electron microscopy. Furthermore, it was shown that major receptors for Con A of chicken erythrocytes were transmembrane glycoproteins having apparent molecular weights of 100K, 45, and 33K.


Regulation of phosphodiesterase and ornithine decarboxylase by cAMP is cell cycle independent. Cyclic AMP (cAMP) causes growth arrest in G1 and induction of cAMP phosphodiesterase and decrease of ornithine decarboxylase in S49 mouse lymphoma cells. Dibutyryl cAMP treatment of partially synchronized cells causes similar changes in activities of both enzymes, regardless of position in the cell cycle. This suggests that cAMP regulation of these enzymes is not mediated by growth perturbation.


A variant vascular endothelial cell line with altered growth characteristics. A variant endothelial cell type was found to arise spontaneously from cultures of bovine aortal endothelial cells. This variant showed no contact inhibition and overgrew confluent cultures of wild-type endothelial cells. Unlike other reported variants of this cell type produced by chemical mutagenesis or by withdrawal of polypeptide growth factor, this variant retained the capacity to synthesis factor VIII antigen, but showed no alteration from wild-type in capacity to adsorb platelets. The variant also had an increased capacity to bind FITC-conjugated con A to its surface.


Growth-promoting actions of extracts from mouse submaxillary glands on human endothelial cells in culture. Extracts of submaxillary glands from two different strains of inbred mice were mitogenic for human endothelial cells in culture. The mitogenic activity of extracts from glands of males of the SWR/J and C57BL/10J strains were equivalent, and the growth stimulating effect was unrelated to renin or esteroproteolytic activity. Mitogenic activity in extracts from SWR/J females was less than that from males, and extracts from C57BL/10J females were inactive. The polypeptide growth factors, epidermal (EGF) and fibroblast (FGF) growth factors, also stimulated replication of endothelial cells. Cells from either umbilical arteries or veins responded to submaxillary extracts, EGF, or FGF with a similar increase in cell number, increase in protein and enhanced uptake of 3H-thymidine. The proliferative response was associated with decreased activity of angiotensin I converting enzyme which is localized on the endothelial surface. Nerve growth factor (NGF) was not mitogenic for endothelial cells. Extracts of submaxillary glands from male mice of either strain contained approximately 20 times more EGF than extracts from females, as determined by immunodiffusion. Mitogenic activity of the extracts was completely inhibited by antiserum to EGF, suggesting that the active component of these preparations is EGF.


Induction of an oligodendroglial enzyme in C-6 glioma cells maintained at high density or in serum-free medium. The relationship between cell density and the activity of 2':3'-cyclic nucleotide 3'-phosphohydrolase (CNP), an enzyme believed to be specific to oligodendroglial cells and myelin in the brain, has been studied in cultured C-6 glioma cells. Over a 12-day period, the specific activity of CNP underwent a 4-fold increase in conjunction with an increase in the cell density (total protein/flask) and a decline in the growth rate of the cultures. In contrast, the specific activity of Na+,K+-ATPase was not influenced by cell density. Experiments with cultures seeded at different initial densities indicated that the increase in CNP activity coincided with the attainment of a specific cell density rather than with the length of time that the cells were maintained in culture. Arrest of cell proliferation in non-confluent C-6 cells by means of thymidine blockade was not sufficient to cause an increase in the activity of CNP; however, removal of serum from the culture medium resulted in a 3-fold induction of the enzyme in the absence of a high degree of cell contact. The induction of CNP in cells maintained in serum-free medium paralleled the development of a series of distinct morphological changes reminiscent of glial differentiation, which occurred within 48 hours after removal of the serum. Inhibition of protein synthesis by cycloheximide prevented the induction of CNP in serum-free cultures. The demonstration that an enhancement of an oligodendroglial characteristic in C-6 glioma cells can be obtained by growing the cells to high density or by removing serum from the medium, provides further support for the suggestion that these cells may be analogous to the glial stem cells present in the developing brain.


Phenotypic characterization of ouabain-resistant Aedes albopictus cells. The phenotype of a ouabain-resistant Aedes albopictus cell line has been partially characterized. Treatment of ouabain-sensitive cells with 0.005-1.0 mM ouabain resulted in an 80% reduction in the uptake of 86rubidium (86Rb+), an ion with an affinity for the K+ pump binding site; ouabain-resistant cells showed only a 40% reduction with 1.0 mM ouabain. When ouabain-sensitive cells were incubated in the presence of ouabain (0.1 mM) for one and one-half to three hours, the molar ratio of intracellular Na+/K+ rose from 0.2 to 4.2. In ouabain-resistant cells, a similar treatment had very little effect. Based on [3H] ouabain-binding studies, ouabain-resistant cells were estimated to have 60% fewer binding sites per cell than ouabain-sensitive cells. The spontaneous mutation rate from ouabain sensitivity to ouabain resistance was calculated to be 1-6 x 10(-8) mutations/cell/generation, a value similar to that reported for mammalian cells at the analogous locus.


Yolk transport in the ovarian follicle of the hen (Gallus domesticus): lipoprotein-like particles at the periphery of the oocyte in the rapid growth phase. Thin sections of the oocyte periphery and surrounding granulosa layer from 1--5 day preovulatory follicles were examined by transmission electron microscopy. With the use of certain procedures in tissue preparation, notably the tannic acid method, numerous particles in the range of 15--40 nm with a mean diameter of 27 nm were observed in both extra- and intracellularly. The particles were abundant in the granulosa basal lamina, in the spaces between the granulosa cells and in the perivitelline space. They appeared to adhere to the oolemma as a continuous double layer which was also observed to line the coated vesicles, 200--350 nm in diameter, invaginating from the oolemma. The layer of particles was not found on the plasma membranes of the granulosa cells, nor were particles present within the cells. In the peripheral cytoplasm of the oocyte the yolk spheres, ranging upwards from 250 nm diameter, were membrane-bound and contained tightly packed particles similar to those on the oolemma. Bodies displaying features intermediate between coated vesicles and yolk spheres suggested that, on entry into the cell, loss of the cytoplasmic coat and obliteration of the vesicular lumen gave rise to nascent yolk spheres which then fused together to form the larger spheres. The extracellular layer, coated vesicles and smaller yolk spheres were absent in oocytes fixed after a 10-min delay. The evidence indicated that 27-nm particles were transferred from the basal lamina to the oocyte surface via the intergranulosa cell channels, incorporated into the cell by adsorptive endocytosis and then transferred to the yolk spheres with little morphological alteration. The identity of the particles with very low density lipoproteins, the major components of the yolk solids, was discussed.


Electron-microscopy of intact nuclear DNA from human cells. Structures retaining many of the morphological features of nuclei may be released by lysing human cells in a non-ionic detergent and 2 M NaCl. Such nucleoids contain all the nuclear DNA packaged within a flexible cage of RNA and protein. HeLa nucleoids have been spread at an air-water interface and viewed in the electron microscope. A tangled network of superhelical fibres surrounds the collapsed cage. Irradiation with gamma-rays abolishes supercoiling and treatment with the untwisting enzyme or a low concentration of ethidium reduces it. A high concentration of ethidium induces supertwisting. The nuclear DNA of higher cells can be isolated naked, supercoiled and intact.


High-performance liquid chromatographic assay for acid and alkaline phosphatase in serum. High-performance liquid chromatography was used to assay serum acid and alkaline phosphatase. Samples were incubated with adenosine-5'-monophosphoric acid (AMP) in a buffer of required pH, 5'-nucleotidase was inhibited with Ni2+ ions, and the phosphatase activity was determined by measuring the concentration of the reaction product, adenosine. The analysis time, after the incubation is terminated, is short (7 min), and the assay is quantitative and reproducible. Complete separation of the reaction product from the substrate and the naturally occurring serum constituents and the high sensitivity of the ultraviolet detection system eliminate some of the problems commonly encountered in spectrophotometric assays.


Stability-indicating assay for pivmecillinam hydrochloride and pivmecillinam hydrochloride capsules. A high-pressure liquid chromatographic assay has been developed for pivmecillinam hydrochloride and pivmecillinam hydrochloride capsules. The method separates five reported degradation products and a number of esterified, related compounds. The accuracy of this method is comparable to the published ultraviolet assay and the relative standard deviation of a series of six replicate assays was better than +/- 0.7%.


Use of protein A-treated sera in unmasking herpes simplex virus type 1 (HSV-1) immunoglobulin A and identifying HSV-1 immunoglobulin G as the predominant neutralizing antibody. Treatment of human sera with protein A reduced the amounts of immunoglobulin G (IgG), IgA, and IgM detected by radial immunodiffusion. This treatment also decreased the amount of herpes-specific IgG and IgM detected by radioimmunoassay, whereas it increased and even unmasked the amount of herpes-specific IgA detected. Comparison of protein A-treated sera with untreated sera indicated that herpes simplex virus type 1 IgG was responsible for more than 92 to 99% of the serum neutralizing activity.


Enzyme-linked protein A: an enzyme-linked immunosorbent assay reagent for detection of human immunoglobulin G and virus-specific antibody. A general-purpose reagent capable of reacting with immunoglobulin G in a modified enzyme-linked immunosorbent assay technique was prepared by using protein A coupled with horseradish peroxidase. The reagent detected low levels (0.003 to 1.0 microgram/ml) of human immunoglobulin G and was also applied in an enzyme-linked immunosorbent assay for titration of antibody to human cytomegalovirus. The antibody titers to human cytomegalovirus determined by enzyme-linked immunosorbent assay and by complement fixation were compared. The correlation coefficient between the two techniques was 0.85, but the enzyme-linked immunosorbent assay was 10 times more sensitive than complement fixation in terms of antibody titers detected.


Comparison of rates of virus isolation from leukocyte populations separated from blood by conventional and Ficoll-Paque/Macrodex methods. One hundred fifty-two blood specimens, largely from immunocompromised patients, were collected in heparinized Vacutainer tubes and divided into paired aliquots of equal volume. Buffy-coat preparations, containing mixed leukocyte and separate mononuclear and polymorphonuclear leukocyte populations were obtained by treatment of blood with conventional and Ficoll-Paque/Macrodex (F-P/M) methods. The development of cytopathic effect in monolayers of WI-38 fibroblasts inoculated with cell suspensions derived from the two methods was used to assess virus infectivity. Twice as many virus isolations were obtained using F-P/M. Of those viruses isolated by both conventional and F-P/M, the development of cytopathic effect was more extensive using the latter method. Moreover, a greater variety of viruses was isolated using F-P/M method, as compared to the conventional method. The F-P/M method is no more time consuming than conventional procedures, is readily adaptable for use in the diagnostic virology laboratory, requires only minimal additional cost, and is a particularly suitable and effective means of monitoring viremia.


Enteric adenoviruses: detection, replication, and significance. Adenoviruses can be demonstrated readily in the stools of pediatric gastroenteritis patients by electron microscopy or counterimmunoelectrophoresis, but in 45% of these cases the virus will not grow in cell culture. Indirect immunofluorescence microscopy can be used to detect nongrowing strains of adenovirus; these strains have a unique single-cell fluorescence pattern. Hematoxylin and eosin staining reveals adenovirus-like inclusion bodies in the same distribution as fluorescent cells. Pretreatment of nongrowing adenovirus with convalescent-phase patient serum neutralized its ability to infect the cell culture and produce fluorescent cells. Indirect immunofluorescence microscopy compared favorably with electron microscopy in demonstrating virus in the stools of patients.


Lipoproteins as substitutes for serum in Mycoplasma culture medium. Crude lipoprotein-containing fractions obtained from sera of three different animal species were tested, in combination with bovine serum in Mycoplasma pneumoniae culture medium. All sera yielded at least one lipoprotein-containing component which was considerably more effective in promoting mycoplasma growth than the unfractionated serum sample from which it was derived. The very low activity of certain whole-serum samples tested in this investigation suggests that toxic substances may be present in whole serum which are not contained in the lipoprotein preparations. The greatest activity appeared in the high-density lipoprotein-containing components of bovine and horse sera and the low-density lipoprotein-containing components of human serum. The high degree of growth-supporting activity of these crude lipoprotein-containing serum components suggests that they may be useful as serum substitutes in mycoplasma culture media.


Enzyme-linked immunosorbent assay for detection of antibodies to murine hepatitis virus. An enzyme-linked immunosorbent assay was developed for the detection of antibodies to murine hepatitis virus. A high prevalence of antibody to murine hepatitis virus was found by the enzyme-linked immunosorbent assay in colonies with a low prevalence of complement-fixing antibodies. Murine hepatitis virus strain A59 was found to be broadly reactive as an enzyme-linked immunosorbent assay antigen.


Effects of somatostatin on insulin and glucagon in patients with insulinoma. Effects of somatostatin on fasting and arginine-or tolbutamide-stimulated insulin release were studied in four patients with insulinoma. Somatostatin (bolus or bolus + infusion) reduced fasting insulin values in all patients; insulin response to tolbutamide was partially reduced in two patients; somatostatin bolus impaired the insulin response to arginine. Fasting glucagon levels and glucagon response to arginine were also reduced by somatostatin. These results indicate the potential usefulness of somatostatin in the diagnosis of insulinoma even if its effect on insulin is only partial.


Insulinoma: clinical manifestations, diagnosis and treatment. The significance of the prolonged fasting test and of the fasting blood glucose-insulin relationship. This paper describes the clinical manifestations and the significance of the fasting blood glucose-insulin relationship and the prolonged fasting test in the diagnosis of insulinoma. Pre-and postoperative results are reported. In addition, a description is given of the angiographic demonstration and the results of the surgical removal of the insulinomas.


Epidemiological, clinical, and pathomorphological characteristics of epidemic poliomyelitis-like disease caused by enterovirus 71. In May-September, 1975, an outbreak of epidemic disease clinically and pathomorphologically simulating nearly all known forms of poliomyelitis occurred predominantly among young infants in Bulgaria. Most cases presented benign aseptic meningitis, sometimes with a short period of general cerebra- symptoms. Paralytic forms, such as bulbar polioencephalitis, anterior poliomyelitis, isolated pareses of the facial nerve, occasional cases of encephalomyocarditis, etc., were observed in about 21% of all cases. Over one-fourth of the paralytic cases with bulbar symptoms ended fatally. In March, 1976 another sporadic fatal case of this disease was examined. No new cases occurred in 1977. Histopathological examinations in all fatal cases regularly revealed lesions in the grey matter of the medulla and spinal cord typical of acute anterior poliomyelitis and bulbar polioencephalitis, with some peculiar features of localization and depth of the involvement of the brain stem. The similarity to poliomyelitis and precariously rapid increase in the incidence led to the decision to urgently vaccinate the entire human population with Sabin's live poliovirus vaccine simultaneously in the whole country in order to produce interference with the circulating agent. This aim appeared to have been achieved partially because soon the number of new cases of the disease began to decrease; however, no sharp and complete break in the curve of the incidence occurred. By the time of mass vaccination, the results of virological examinations started before were not yet available. Later, comprehensive complete evidence was obtained that over 25 fatal and many other typical cases of the disease were associated with an enterovirus proved to be antigenically related to enterovirus 71. The diseases in Bulgaria, 1975, differ considerably in the frequency of paralytic forms and in severity from the epidemics caused by enterovirus 71 in Sweden, 1973, Australia, 1972--1973, USA, 1969--1972, and Japan, 1972--1973.


Influence of magnesium on the secretion and action of parathyroid hormone. The interactions of serum levels of magnesium and parathyroid hormone (PTH) have been studied in a patient with hypomagnesemia and hypocalcemia following intestinal bypass surgery for obesity. When serum magnesium was low serum PTH was not stimulated by hypocalcemia. With correction of magnesium deficiency hypocalcemia was associated with elevation of serum PTH levels. Infusion of exogenous PTH induced a clearly detectable renal response in the presence of hypomagnesemia but the response was diminished when serum magnesium was elevated. In this patient it appears that hypomagnesemia suppressed parathyroid gland activity, leaving the renal action of PTH intact.


Radioimmunoassay for antibodies in varicella-zoster virus serology. The method of RIA for antibodies was employed with success in VZ virus serology. The method is suitable for testing VZ antibodies in the course of varicella or herpes zoster disease as well as for determining anamnestic titres. Its advantages are stability of antigen, objective reading of results and applicability to testing large serum sets.


Haemagglutination-inhibiting and neutralizing antibodies to type 3 and 5 rhinoviruses in human sera and nasal washings. Presence of antibodies to RV 3 and RV 5 was tested by HIT and NT in 60 human sera. Antibodies to RV 3 were detected in 23 sera by HIT in a titre range of 1:4--1:64 and in 19 sera by NT in a titre range of 1:4--1:256. Antibodies to RV 5 were detected in 31 sera by HIT in titres of 1:4--1:268 and 27 sera by NT in the same titre range. In a group of 22 persons with unequivocal serum antibodies nasal secretory antibodies were found in 11 subjects in titres of 1:4--1:32. In a group of 16 persons without detectable serum antibodies, presence of secretory antibodies (titre 1:4) was only found in four cases.


Responses of anterior pituitary hormones to heat exposure. To determine anterior pituitary response to mild hyperthermia, plasma GH, ACTH, PRL and TSH were measured every 10 min in five subjects exposed to hot air (120 min, 50 C, 20 mbar) in a climatic chamber. Mean rectal temperatures increases of 0,8 C and mean sweat losses of 760 g were observed. GH levels increased from 4.4 +/- 0.4 ng/ml to 22.0 +/- 7.0 ng/ml (mean +/- SE) and reached maximum 80 min after the beginning of heat exposure and then declined before the end of the heat stress. These peak levels varied widely with the individual, whereas no significant changes in ACTH, PRL and TSH levels were observed during the exposure period. These results suggest that the mechanisms regulating GH secretion are more sensitive to an acute heat exposure than those of the other pituitary hormones studied.


Enhancement of fluorescent antibody staining of viral antigens in formalin-fixed tissues by trypsin digestion. The staining of viral antigens present in formalin-fixed, paraffin-embedded tissues by fluorescent antibodies is markedly enhanced by trypsin digestion. When the trypsin digestion method was used to detect viral antigens present in hamster brain following experimental infection with measles virus, the results were comparable to those obtained with acetone-fixed, freshly frozen tissues that had been sectioned with a cryostat. Measles antigens were readily identified in brain cells from a patient with subacute sclerosing panencephalitis and in lung and liver tissue from a patient with acute giant cell pneumonia, following preparation of the tissues for routine histologic examination. Viral antigens were detected in brain tissue that had been taken from patients with herpes simplex encephalitis and stored in paraffin for up to 15 years. Cells containing antigen could be precisely identified without loss of histologic detail by restaining the same tissue sections with hematoxylin and eosin.


Lack of susceptibility of marmosets to human non-A, non-B hepatitis. Infectious sera from three humans with chronic non-A, non-B hepatitis, whose blood or serum had transmitted non-A, non-B hepatitis both to other humans and to experimentally inoculated chimpanzees, were inoculated into five marmosets. A sixth uninoculated marmoset served as a control. No elevations in levels of serum alanine aminotransferase or isocitric dehydrogenase occurred in serum samples obtained weekly from any of the marmosets during three months following inoculation. This study indicates that certain species of marmoset, which are susceptible to and provide well-documented animal models for hepatitis A and GB-agent hepatitis, do not appear to be susceptible to the agent(s) of human non-A, non-B hepatitis. In addition, this study suggests that the agent(s) of human non-A, non-B hepatitis and the GB agent are probably different.


Phagocytosis and intracellular fate of Sporothrix schenckii. Human polymorphonuclear leukocytes (PMNLs) phagocytized and killed yeast-phase cells of Sporothrix schenckii in vitro in the presence of 10% unheated serum. The combination of H2O2, KI, and human PMNL myeloperoxidase was lethal to yeast-phase S. schenckii. The results of this investigation suggest that PMNLs may play an important role in host resistance to infection with S. schenckii.


[Arterial patterns in congenital deformities of the hand (author's transl)]. Etiology of congenital deformities of the hand is still unknown in most cases. But anomalous development of arteries has been suspected to play a role in faulty development of the hands. Senior (1926) and Singer (1933) have reported arterial anomalies in the developmental stage of these vessels. Between 1968 and 1978, 367 patients with 453 congenital deformities of the hand visited the hand service of Kyoto University Hospital. These hand anomalies were classified according to Swanson's classification. Angiography was performed on 74 hands of 70 patients in this series, and attempts were made to correlate these with the types of the deformities. From the standpoint of embryological development of arteries, it is concluded that failure of formation of parts (arrest of development), failure of differentiation (separation) of parts and duplication are generated in the same stage from 48 days postovulatory (18 mm embryo), overgrowth (gigantism) of the hand and congenital constriction band syndrome are generated after 52 days postovulatory (23 mm embryo). Generalized skeletal anomalies are considered to be generated in the various developmental stages depending on the each malformed hands. Developmental stage of malformed hands with generalized skeletal abnormalities is discussed with special reference to the embryological development of other anomalies.


Erythrocyte membrane enzyme abnormalities in two hereditary disorders of muscle. Enzymatic properties of erythrocyte membranes in Duchenne muscular dystrophy (DMD) and malignant hyperthermia (MH), two genetically determined abnormalities of skeletal muscle, were examined. Acetylcholinesterase (AChE) and ATPase activities were chosen for investigation since alterations in these enzymes have been demonstrated in animal models of dystrophy. A significant decrease in Na+,K+-ATPase activity was noted in DMD patients and a number of possible DMD carriers, suggesting that this enzyme may provide a useful marker of the carrier state in carriers not exhibiting an elevation in plasma creatine phosphokinase activity. No abnormalities in AChE were demonstrable in any of our DMD patients, indicating that human dystrophy is biochemically distinct from certain animal models of dystrophy (e.g., dystrophic mice) where erythrocyte AChE is decreased. In contrast, evidence was found in two known MH carriers, who had normal erythrocyte ATPase activities, for the presence of an altered membrane AChE characterized by an increase in substrate affinity and a large decrease in maximal hydrolytic rate. While the exact relevance of this membrane defect, if any, to the pathogenesis of MH remains to be seen, the presence of this modified enzyme may serve to identify those individuals in a family where a positive history of MH exists who are at risk of developing a hyperthermic crisis during anesthesia.


Red cell deformability: physiological, clinical and pharmacological aspects. Twenty five thousand billions of red cells circulate through 300 miles of capillary network. The red cell diameter is 7 to 8 microns and the capillary diameter 3 to 5 micron. Red cells deformability is therefore an essential condition for survival. It is impaired in a certain number of clinical conditions. A French task force, covering 40 University Hospitals, is presently investigating the clinical implications of red cell filterability, and its possible therapeutical consequences.


Specificity of attachment of certain enterobacteriaceae to mammalian cells. The specificity of adherence of various Enterobacteriaceae to different mammalian cells was studied in vitro. 3H-Labelled organisms of the same species isolated from various clinical sources differed significantly in their abilities to adhere to the same mammalian cells. Bacteria frequently adhered better to cells derived from sites other than those analogous to their original source. Bacteria did not display consistently 'high' or 'low' attachment to a variety of human and tissue-cultured cells and little selective adherence was demonstrable.


Effects of thyroxine treatment on the hepatic plasma membrane ATPase activity in rats. The effects of T4 and T3 upon ATPase activity of plasma membrane preparations produced by different methods has been studied. Media without Na+ or K+ was used to identify Mg++ activated ATPase, and media containing 100-120 mM Na+ and 20-30 mM K+ in addition to Mg++ was used to identify the augmented activity of Na-K ATPase. We also utilized media containing Na+, K+, and Mg++ in the presence and absence of ouabain to identify the Na+-K+ activation. We found no effect of doses of thyroxine in the physiologic range upon the Na+-K+ ATPase activity. A dose-response study indicated that high doses of T4 may stimulate Mg++ activated ATPase. In the P2 preparation of the Touster method we found minimally enhanced Na+-K+ ATPase activity. The quantity of T4 needed to cause such enhancement is approximately 50 times the physiologic replacement dose needed to restore thyroidectomized animals to a euthyroid state. We conclude that thyroid hormones do not stimulate Na+-K+ ATPase of plasma membranes when administered in a physiologic quantity, but may enhance Mg++ atpase activity when administered in a pharmacologic or toxic quantity.


Inhibition of the replication of parvovirus X14 by 5-iodo-2'-deoxyuridine pre-treatment of cell cultures. Pre-treatment of rat embryo cell cultures with 5-iodo-2'-deoxyuridine (IdUrd) inhibits the replication of parvovirus X14. Reduced yields of haemagglutinating and infectious particles were observed. Adsorption of virus to cells was not affected, but both virus protein and DNA synthesis were inhibited. Fewer cells were capable of supporting protein or antigen synthesis as determined by immunofluorescence. Virus-specific DNA was detected in IdUrd pre-treated cells, but the amount synthesized was considerably less than that from control cultures. Cellular DNA synthesis was also inhibited in IdUrd pre-treated cells. Therefore, the replication of parvoviruses appears dependent upon host cell factors involved in cellular DNA synthesis.


A solid-phase system (SPACE) for the detection and quantification of rotavirus in faeces. This report describes the development of a solid-phase haemadsorption system using chromic chloride-linked, antibody coated erythrocytes. It is proposed to call this technique solid phase aggregation of coupled erythrocytes (SPACE). The system is suitable for the detection of virus antigens, such as from rotavirus infections, which are present in 'dirty' or 'mixed' preparations such as faeces, urine or exudates. The test uses microtitre U-form plates coated with specific antivirus antibody; faecal suspensions are added and virus or antigen allowed to adsorb. The plates are then washed and adsorbed antigens are detected by the addition of virus-specific IgG-coated erythrocytes. The resultant settling pattern is read in the same manner as a conventional haemagglutination test. The system is compared with electron microscopy and fluorescent antibody techniques.


The mitochondrial activation of silicate and its role in silicosis, black lung disease and lung cancer. Silicate substitutes for phosphate in the transitory uncoupling of rat liver mitochondria induced by hydrazine when beta-hydroxy-butyrate is the substrate. Uncoupling is blocked by rutamycin. Just as in the case when phosphate is combined with hydrazine, ATP, ADP, PPi, and Mg++ protect against hydrazine when silicate is combined with hydrazine. A high level of ADP in the absence of added phosphate, but in the presence of silicate, induces a pseudo state three of the mitochondria. Silicate, like sulfate and arsenate which have been reported previously, is activated by the enzymes which mediate oxidative phosphorylation. These results serve to explain a role for silicate in silicosis, black lung disease, and cancer. In addition, since there is suggestive evidence in the literature that lung tissue solubilizes asbestos fibers, these results not only expand the confluence between oxidative phosphorylation and chemical carcinogenesis but are correlated with the synergistic carcinogenicity of asbestos and smoking observed by epidemiologists.


Integration of herpes simplex virus type 1 DNA into the DNA of growth-arrested BHK-21 cells. The ability of HSV-1 DNA to become associated with host cell DNA in an alkaline-stable form has been demonstrated following infection of a ts baby hamster kidney growth mutant (ts BTN-1), at the non-permissive temperature (39.5 depgrees C). After 8 h pre-incubation at 39.5 degrees C, ts BTN-1 cells infected at this temperature using m.o.i. ranging from 0.5 to 200 p.f.u./cell fail to replicate virus DNA even though transport of input virus genomes to the nucleus is the same at both permissive and non-permissive temperatures. Virions containing 3H-labelled DNA were used to infect ts BTN-1 cells at 39.5 degrees C, and the total cellular DNA isolated from these cells was resolved into host and virus material by repeated CsCl equilibrium gradient centrifugation. A significant amount of the input radioactivity was found as a distinct band in the host region in both neutral and alkaline CsCl gradients, strongly suggesting a covalent association between host and virus DNAs. Evidence for this association was strengthened by demonstrating that radioactive material (virus DNA) banding in the host region of CsCl gradients could be driven towards the density expected for virus DNA following degradation of the putative hybrid molecules by shearing.


Analysis of adipose tissue and serum from PBB (polybrominated biphenyl)-exposed workers. In the course of clinical examinations of individuals exposed to PBB (polybrominated biphenyl(s) in Michigan, employees of the chemical company which manufactured PBB also were examined. Analysis of serum and adipose tissue from these workers revealed quantitative and qualitative differences, compared with farm residents exposed to PBB. Serum and fat concentrations of PBB were higher among chemical workers, and the concentrations of PBB homologs were also higher, attributable to different intensity, route (skin, inhalation, direct ingestion vs. ingestion of animal foodstuffs), and composition (unchanged vs. animal-mediated material) of exposure in chemical workers vs. farmers. The unusual nature of occupational exposure was further evident in measurement of serum and fat DDE and identification of serum tetrabromobenzene (TBB). DDE concentrations were higher among employees than among farmers, consistent with the workers' earlier occupational exposure to DDT, which had been produced by the company. Serum TBB was not explicable by direct exposure to the chemical, but has been ascribed to exposure to hexabromobenzene, which was shown to give tetrabromobenzene as a metabolite in the rat.


In vivo and in vitro phosphorylation of murine mammary tumour virus proteins. A comparative study of in vitro and in vivo phosphorylation of murine mammary tumour virus, a type Brna virus, is reported. The protein kinase activity associated with murine mammary tumour virus catalysed the in vitro phosphorylation of endogenous virus polypeptides. This kinase activity required a divalent metal cation, a non-ionic detergent, and was stimulated in the presence of dithiothreitol. Exogenous cyclic AMP was not required. The 32P-labelled products of the in vitro reaction were completely sensitive to pronase digestion and the phosphate was attached mainly by phosphomonoester linkage to serine residues. As determined by SDS-polyacrylamide gel electrophoresis, heterogeneous labelling of major and minor virus polypeptides was observed under in vitro conditions. In contrast, the in vivo labelling of type B virus produced by a continuous cell line (MuMT-73), established from pooled mammary adenocarcinomas of Balb/cfC3H mice, demonstrated specific phosphoproteins associated with murine mammary tumour virus. The major phosphorylated proteins were found to have mol. wt. of 18 000 and 12 000 (p18 and p12) after isolation by molecular sieving chromatography and analysis by gel electrophoresis.


Host-cell response to herpes virus infection in central and peripheral nervous tissue in vitro. In an organotypic nerve cell culture system, all cells in both the central and the peripheral nervous system (CNS, PNS) components supported replication of herpes simplex virus types 1 and 2 (HSV 1, HSV 2). In HSV 1 infection, cellular response was particularly characterized by the formation of small syncytia (which involved neurons) and by the presence of bundles of interwoven fine filaments within the nuclei of infected cells. In HSV 2 infection, groups of parallel tubules characteristically formed in the nuclei of infected cells. All cells in the CNS or PNS succumbed to virus infection, some within 24 h (e.g. oligodendrocytes) and others after 48 h (e.g. neurons), with the exception of astrocytes. Although among the first cells to develop virus nucleocapsids in their nuclei, astrocytes became swollen and filled with increased numbers of bundles of glial filaments within 24 h after infection; by 48 h the actual number of astrocytes was increased by as much as three- to fourfold over the number in controls. The results suggest that astrocytes may have a unique mechanism which modifies virus infection and the cells not only survive, but can also become reactive.


Inactivation by u.v.-irradiation of interfering herpes simplex virus particles: interference requires a functional genome. The sensitivity of interfering herpes simplex virus (HSV) particles to u.v.-irradiation was studied in a virus stock of HSV-1 strain ANG that contained an excess of interfering over infectious particles. Following u.v.-irradiation, samples of this virus stock were assayed for their plaque-forming capacity and their capacity to interfere with the replication of unirradiated standard virus. Depending on the assay conditions, interfering particles appeared to be less, equally, or more sensitive to u.v. light than infectious particles. It is concluded that interference is a gene function of interfering particles rather than being exerted directly by structural constituents of these particles.