In vivo effects of some tertiary alkylaminoethyl esters with choline acetyltransferase inhibitory properties. The tertiary nitrogen derivatives of two choline esters, chloroacetylcholine and acrylcholine, known to inhibit choline acetyltransferase (ChAc) in vitro, were tested for their effects in the whole animal, including peripheral and central cholinergic systems. These esters are N,N-dimethylaminoethyl chloroacetate (Cl-DMA) and N,N-dimethylaminoethyl acrylate (acryl-DMA). The peripheral preparations studied included a neuromuscular junction, a sympathetic ganglion and a postganglionic parasympathetic exocrine preparation. Both Cl-DMA and acryl-DMA blocked responses in these preparations when injected intravenously. The LD50 values for Cl-DMA and acryl-DMA were 640 mg/kg and 183 mg/kg, respectively. Cl-DMA and acryl-DMA were also able to inhibit brain ChAc when injected intravenously by 32% and 18.5%, respectively. The brain levels of acetylcholine (ACh) were significantly reduced by about 25% with Cl-DMA but not significantly with acryl-DMA when the animals were forced to exercise after injection. It is obvious that ChAc inhibition is not effective in decreasing ACh levels significantly under normal conditions.


Possible role of prostaglandin E1 on adrenergic neurotransmission in the guinea-pig taenia coli. The effects of prostaglandin (PG) E1 were investigated on the responses to adrenergic and non-adrenergic inhibitory nerve stimulation using the perivascular nerve-taenia preparation of the guinea pig. PGE1 caused a rapid and sustained contraction and markedly inhibited the response to adrenergic but not to non-adrenergic inhibitory nerve stimulation. It was also observed that PGE1 had some desensitizing action to exogenous noradrenaline on the postjunctional site. Although indomethacin decreased the tone of the preparation, it potentiated the response to adrenergic nerve stimulation without any effects on the response to non-adrenergic inhibitory nerve stimulation. From these observations, it was concluded that endogenous PGE1 may also play a regulatory role in adrenergic inhibitory neurotransmission, mainly by inhibitory action on noradrenaline release and partly by a similar action on the postjunctional site.


Effects of hydralazines on canine muscarinic ganglion transmission. The effects of hydralazine, dihydralazine and 4-propyl-1-hydrazinophthalazine (4-propylhydralazine) were investigated on reflex pressor responses to increased intracranial fluid pressure (IIP) during occlusion of the abdominal aorta and the vena cava (MVO). It has been shown that application of MVO, caudal to the renal arteries, produces two independent vascular zones in the animal (Steinberg and Hilton, 1966a). Increasing intracranial fluid pressure during MVO elicits a reflex pressor response which consists of two components. One component is blocked by the nicotinic ganglionic blocking agent, chlorisondamine, and the other component is blocked by small doses of atropine. It was found that hydralazine and dihydralazine were effective in blocking residual pressor responses following partial blockade of reflex pressor responses to IIP with chlorisondamine. 4-Propylhydralazine, which is chemically similar to hydralazine and dihydralazine, was less active in inhibiting the residual pressor responses. It is suggested that hydralazine may act in part by interfering with muscarinic ganglionic transmission.


A method for site selectivity analysis applied to opiate receptors. A method has been developed to determine whether or not a receptor system consists of several sites with different binding specificity. Several radioactive indicator drugs are used in solutions of identical chemical composition. Homogeneity of the receptor system is tested graphically. We have applied the method to a study of opiate receptors. It appeared that there were at least two different binding sites. Several different competitors with various characteristics have been tested against three indicator solutions with radioactive dihydromorphine, naloxone and naltrexone respectively. Three different classes of opiates were observed in the system, morphine-like, naltrexone-like and an intermediate group. A morphine-like substance shows high selectivity for one of the sites while naltrexone-like compounds show practically no discrimination between the sites and a group of partial agonists such as nalorphine is intermediate.


Stimulated formation of cyclic AMP in different areas of chick brain. Marked accumulation of cerebral cyclic AMP was induced in the chick by the i.v. administration of isoprenaline or histamine of by decapitation. These responses were examined in various cerebral regions and compared to the responses produced in vitro when maximal concentrations of isoprenaline, histamine or adenosine were added to appropriate brain slices. Histamine only markedly stimulated cyclic AMP formation in the cerebral hemispheres. Isoprenaline was equally potent in the cerebellum and the cerebral hemispheres though the medulla-diencephalon and optic lobes were less responsive. There was a lack of correlation between the effects of decapitation in vivo and the responses to adenosine in vitro. Similar accumulations of cyclic AMP were produced by decapitation in the cerebral hemispheres and cerebellum though adenosine was much more effective on cerebellar slices in vitro. The topographical distribution of catecholamine and histamine sensitive cyclic AMP forming sites do not correlate with endogenous levels of noradrenaline or histamine in chick brain and possible interpretations are discussed.


Calcium uptake by myometrial membranes: effect of A 23187, a calcium ionophore. A subcellular fraction, relatively enriched with plasma membranes, showed an ATP-dependent Ca2+ uptake. The addition of a Ca2+. Other drugs (angiotensin, prostaglandin F2alpha, 5-hydroxytryptamine and cyclic nucleotides) had negligible effects on Ca2+ transport. The preparation appears to transport Ca2+, although binding is negligible.


Weakening of genetic resistance. II. The effect of injection of the macrophage toxic agents silica and carragheenan and the cytostatic drugs cyclophosphamide, busulphan and vinblastin. Silica (4 mg/i.v.) and Carragheenan (20 mg/i.p.) were shown to be capable of weakening hybrid resistance in the spleen whereas no improved growth was observed in the femoral bone marrow. Histological data did not indicate that the agents in the doses used caused significant macrophage mortality. It was concluded that the weakening effect of the agents was not due to death of the effector cells but possibly to a stimulation of the mononuclear phagocytic system in the spleen. Cyclophosphamide (250 mg/kg) induced a weakening of hybrid resistance which was observed both in the spleen and the femoral bone marrow. Experiments with the alkylating agent Busulphan and with Vinblastin showed that both agents were only capable of inducing a moderate stimulation of growth of the transplant in the spleen. A direct specific influence of cyclophosphamide on the microenvironment was proposed. Xenogeneic resistance was studied by transplantation of rat bone marrow to mice. Silica and cyclophosphamide were shown to be capable of inducing a weakening of the resistance in the spleen. The development of the graft in the femoral bone marrow did not appear to be affected by the administration of silica or cyclophosphamide.


Localization of evoked potentials in the digastric, masseteric, supra- and intertrigeminal subnuclei of the cat. Extracellular focal potentials were evoked and mapped in the trigeminal motor nucleus and its surrounding borderzone in the cat. Graded electrical stimulation was used for orthodromic and antidromic excitation of the masseteric and digastric motoneurones and for orthodromic stimulation of the lingual and inferior alveolar nerves. The method of referring Horsley Clarke coordinates of microelectrode recording positions to their location of the actual histological section was studied and the total error affecting the method was calculated for the H, AP and L axes. The characteristics and the distribution of the evoked focal potentials were described and related to the histological section from the actual experiment. A phase reversal of the negative focal potential evoked by the lingual and inferior alveolar nerves in the main sensory nucleus and in the intertrigeminal nucleus was observed to indicate the dorso-lateral border of the motor nucleus. Other borders were given by the antidromic potentials evoked in the nucleus. Digastric motoneurones were found medially in the caudal third and ventro-medially in the middle third of the motor nucleus. The masseteric motoneurones were located laterally in the middle and rostral thirds of the nucleus. Potentials evoked in the supratrigeminal and intertrigeminal subnuclei, adjacent to the motor nucleus, were considered and discussed in relation to the available evidence of interneurones subserving trigeminal reflex arcs.


Further evidence for common antigens in intracytoplasmic A particles of mouse mammary tumors and B type virions of murine milk. Using the indirect immunofluorescence technique and radioimmunoprecipitation test (RIP test) according to ABELEV and ELGORT (1970) immunological investigations intracytoplasmic A particles isolated from mouse mammary tumors were performed. In order to purify the virus particles sucrose gradients were employed according to TANAKA et al. (1972) and in some details according to SMITH and WIVEL (1973). In immunofluorescence studies on mammary tumor slices a complete cross-reaction between antisera against A particles from mammary tumors and MTV-B particles from mouse milk was observed. Both kinds of antisera reacted in identical fluorescence pattern types with intracytoplasmic A particle clusters and with extracellular accumulations of B type virions, respectively, in tumors of different histological structures. RIP test studies suggest that these cross-reactions are attributed to three common antigens in A and B type viruses. Thus, there does not remain any doubt that intracytoplasmic A particles in murine mammary tumors are MTV-related. All these A particles antigens were identically detected using an antiserum against A particles isolated from mouse leukemia cells (serum kindly provided by Dr. TANAKA). The biological role of intracytoplasmic A particles in mouse tissues is discussed. In contrast to the precursor theory, it is believed that they could represent something like a nonsense way of particle maturation in MTV-infected cells.


The mannophosphoinositides of Nocardia asteroides. The phospholipid of N. asteroides has been investigated. It was found to contain phosphatidyl ethanolamine, cardiolipin, phosphatidyl inositol and a family of mannophosphoinositides. Dimannophosphoinositides with 3 and 4 moles of fatty acid per phosphate residue represented the major glycophospholipids besides small amounts of other more glycosylated mannophosphoinositides.


Adenosine promoted accumulation of adenosine 3',5'-monophosphate in rabbit vagus nerve. Desheathed rabbit vagus nerve has been found to form cyclic AMP when incubated with adenosine. This accumulation of cyclic AMP is inhibited by theophylline but not by antiadrenergic agents, anticholinergic agents or local anaesthetics. Depolarizing media are not able to promote cyclic AMP accumulation in this preparation.


Effects of stress and adrenocorticotrophin administration on plasma corticosterone levels at different stages of pregnancy in the mouse. Stress or administration of ACTH to pregnant mice gave rise to much higher plasma corticosterone levels in the second half of pregnancy than in the first half, suggesting that there may be increased adrenal sensitivity to ACTH or decreased metabolism of corticosterone during the second half of pregnancy.


Some aspects of neurophysiological basis of insecticide action. When the insecticide parathion was administered to awake, unrestrained rats with chronically implanted brain electrodes, it was observed that the latency of the averaged flash-evoked potential in the visual cortex and superior colliculus was increased and the amplitude was decreased 2 to 4 hours later with responses returning to pretreatment levels about 8 hours after administration. Similarly, after administration of several dose levels of parathion in the rat, durations of phases of the maximal electroshock seizure (MES) pattern were altered to the greatest extent 4 hours later, but effects disappeared at 24 hours. These effects of parathion on the MES and evoked potentials coincided with a fall in blood and brain acetylcholinesterase (AChe) activities but disappeared after AChe inhibition had reached its peak and stabilized. Brain AChe activities required 2 to 4 weeks for recovery whereas blood AChe activity recovered in 1 week following inhibition by parathion (at least 2 mg/kg body weight). Studies in the monkey demonstrated similar results. Because these measurements of central nervous system function returned to normal despite continued inhibition of AChe activity, the results are interpreted to mean either that adaptation of evoked potentials or MES responses to prolonged AChe inhibition can occur in the rat and monkey after parathion administration or that some of the effects of parathion do not depend on AChe inhibition. Administration of DDT (100 mg/kg by mouth) to awake, unrestrained rats markedly increased the amplitude of spontaneous electrical activity in the cerebellum, whereas there was much less effect on electrical activity recorded simultaneously in the occipital cortex, reticular formation, and medial geniculate body. Similarly, DDT administration had marked effects on the averaged, sound evoked potential recorded in the cerebellum; DDT caused the appearance and increased the amplitude of an early component of this response not usually present during control recordings. Sound-evoked potentials recorded simultaneously from the frontal and occipital cortex and reticular formation were affected less or were decreased in amplitude by administration of DDT.


[Effect of acoustic stimulation on lipid metabolism, indices of the blood coagulation system and development of experimental atherosclerosis in rabbits]. The 14-day sound stimulation of healthy rabbits increases the level of unesterified fatty acids and the blood coagulability. Morphologically the hypertrophy of the heart coronary arteries and necrotic foci in the myocardium are revealed. In rabbits with experimental atherosclerosis subjected to sound stimulation during 14 and 28 days, in spite of a high level of lipids in the blood and hypercoagulatory shifts, the developing conditions aid to resorption of lipids from aortal plagues and coronary arteries. Apart from that, morphological studies reveal hypertrophy and oedema of the vascular wall, necrotic foci, local and diffuse fatty infiltration of the myocardium.


[Effect of catecholamines on sodium resorption and oxygen tension in rat kidneys]. Functional significance of alpha- and beta-adrenoreceptors for the mechanism of catecholamines effect on sodium reabsorption and oxygen tension in the rat kidney, was studied. Adrenaline inhibited sodium excretion decreasing its filtration in glomeruli and stimulating its reabsorption in tubules. The oxygen tension in these conditions did not change in the renal cortex while oxygenation of the external cortical layer was significantly increased. The blocking agent for alpha-adrenoreceptors phentolamin abolished the inhibiting effect of adrenaline on the glomerular filtration and somewhat decreased the degree of oxygen tension growth in the external cortical layer, leaving the tubular effect unaltered. The blocking agent for beta-adrenoreceptors inderal did not affect the inhibitory action of adrenaline on the glomerular filtration but completely prevented its activating effect on the tubular reabsorption of sodium and on oxygenation of the external cortical layer. A conclusion was drawn that catecholamines stimulation of sodium reabsorption in the rat kidney follows excitation of beta-adrenoreceptors. The increase in oxygen tension in the external cortical layer under effect of catecholamines is supposed to improve energetic supply of the sodium active transport in the ascending portion of the Henle loops.


Transformation of Chinese hamster embryo cells with an avian sarcoma virus ts mutant. Chinese hamster embryo cells were transformed with temperature-sensitive mutant of the Schmidt-Ruppin strain of Rous sarcoma virus-subgroup A (ts NY68S Ra-A). The developed cell line elicited the morphological transformation at a permissive temperature of 35 degrees C, whereas after a shift to a non-permissive temperature of 40 degrees C the cells reverted to a normal phenotype. Tests for the presence of avian sarcoma group-specific antigen showed that the antigen was not expressed both at permissive and non-permissive temperature. However, the transient expression of gs antigen could be achieved by treatment with 5-IUdR only at the permissive temperature. There was a difference in the rate of 14C-2-deoxy-D-glucose uptake between cells cultured at the permissive and non-permissive temperature.


Regulation by glucose and cyclic nucleotides of the glucagon and insulin release induced by amino acids. The glucagon and insulin release induced by amino acids was studied in the presence of glucose, dibutyryl cyclic AMP (dbcAMP) or theophylline on the splenic part of the pancreas of new born rats (48 to 64 hours). The results were compared to the literature data. Arginine or a mixture of the three amino acids (A.A.), arginine, lysine or alanine, stimulate glucagon secretion at 1.6 mM glucose. This stimulation is suppressed by 16.7 mM glucose. On the other hand, 16.7 mM glucose potentiates the effect of arginine or of the 3 A.A. on insulin release. At 1.6 mM glucose, theophylline potentiates the effect of 3 A.A. (10 mM each) on glucagon and insulin release : this effect reaches a maximum at 5 mM of theophylline; dbcAMP also potentiates the effect of 3 A.A. on glucagon and insulin release, and the effect of arginine, alanine or lysine on glucagon release. On the beta cell, the lack of potentiation observed between dbcAMP and arginine, lysine or alanine indicates that these A.A. interact positively when mixed together. In the presence of arginine or of the three A.A., the percentage stimulation of glucagon and insulin release depends on the dbcAMP dose and does not vary with the glucose concentration. The increase of glucagon and insulin release observed when the NaCl concentration in the incubation medium decreases cannot account for our results. Cyclic GMP (4 mM) does not modify the glucagon or insulin secretion induced by different concentrations of glucose or by the mixture of A.A. (10 mM each). The stimulating effect of acetylcholine on insulin release would not be related to the cyclic GMP molecule. In conclusion, instead of modifying the specificity of substrate, theophylline or dbcAMP accentuate it: glucose stimulates specifically the beta cell whereas 3 A.A. are more effective on the alpha2 cell than the beta cell. Cyclic AMP suppresses the glucose effect on glucagon release induced by the amino acids. Because of its interaction with glucose and amino acids, cyclic AMP seems to be a very important element in the regulation of the release of these pancreatic hormones.


[Energy metabolism in the perinatal period (author's transl)]. Neonatal hypoglycemia is of frequent occurrence in fasted newborn babies or animals but the origin of this hypoglycemia is not fully understood. Studies performed in newborn rats have shown that liver glycogenolysis and gluconeogenesis occur immediately after birth and that the increase in the activities of key regulatory enzymes (phosphorylase, glycogen synthetase and phosphoenolpyruvate carboxykinase) results probably from the rise of plasma glucagon and the fall of plasma insulin induced by the "stress" of birth. When the liver glycogen stores have been exhausted, i.e. between 6 and 16 hours after birth, a profound hypoglycemia develops in fasting newborn rats. The inability of hepatic gluconeogenesis to produce sufficient glucose to meet the energy requirement of the newborn tissues results from a lack of fat-derived (free fatty acids and ketone bodies) and gluconeogenic (lactate, amino acids) substrates. The stage of appearance and the mechanisms regulating gluconeogenesis in other species including human are discussed.


Measurement and partial characterization of immunoreactive glucagon in gastrointestinal tissues of dogs. We have reported previously that increasing amounts of immunoreactive glucagon (IRG), measured by four specific antisera, appeared in plasma of depancreatized insulin-deficient dogs. It was therefore concluded that pancreatectomy was not accompanied by glucagon deficiency in the dog, but instead excessive amounts of extrapancreatic IRG could contribute to the diabetic syndrome. In order to locate the source of extrapancreatic glucagon, tissue extracts were assayed with anti-glucagon sera 30-K and K-44, which cross-react minimally with crude gut extracts. IRG was detected in all gastrointestinal tissues and in the salivary glands, but not in extracts of liver, kidney, brain, heart atrium, and adenohypophysis. Immunologic dilution curves of extracts from all gastrointestinal tissues were parallel to those of the pure pancreatic glucagon standard, and both antisera (30-K and K-44) measured the same concentrations. The highest concentration of gastrointestinal IRG was found in the fundus and corpus of the stomach. Presence of IRG in gastrointestinal tissues of depancreatized dogs indicates that gastrointestinal cells can not only secrete but also store large amounts of IRG. Extracts of mucosa of stomach fundus were further purified by gel filtration on Biogel P-30 columns. The immunoreactivity in the eluate was assayed by 30-K and a strongly crossreacting antibody, K-4023. One pooled fraction corresponding to marker pancreatic glucagon in its elution volume was found to contain the largest amount of IRG and the highest specific immunoreactivity (IRG/protein concentration). This fraction showed also the highest activity in a glucagon-receptor assay system. Disc gel electrophoresis in the presence of urea resolved this fraction into three immunoreactive components, one of which was identical to pancreatic glucagon in its electrophoretic mobility. It appears, therefore, that mucosa of the upper stomach in the dog contains a polypeptide similar to pancreatic glucagon. We conclude that (a) hyperglucagonemia in the dog can result from excessive secretion of IRG not only by the pancreatic alpha cells but also by cells of the gastrointestinal tract; (b) the highest IRG concentration was found in fundus and corpus of the stomach and lower concentrations throughout the gastrointestinal tract; (c) the IRG component in the stomach displayed immunologic and physical properties similar to pancreatic glucagon.


Intracellular enzymes of collagen biosynthesis in rat kidney in streptozotocin diabetes. The activities of the four enzymes catalyzing intracellular post-translational modifications of the collagen polypeptide chains were assayed in the kidneys of rats with streptozotocin diabetes. When the changes in the four enzyme activities were expressed per milligram of protein in the 15,000 X g supernatant of the kidney homogenates, there were no changes in any of the enzyme activities at four weeks and only slight increases in the prolyl and lysyl hydroxylase activities at 12 weeks after the induction of diabetes. When the changes were expressed as total enzyme activities per two kidneys, again no changes were found in any enzyme activity at four weeks, but at 12 weeks significant increases were found in all four enzyme activities, namely prolyl hydroxylase, lysyl hydroxylase, collagen galactosyltransferase, and collagen glucosyltransferase. The data would be consistent with an increased collagen synthesis in diabetic kidneys, but they do not support the hypothesis that there might be specific changes in some of these enzyme activities or in the level of certain posttranslational modifications of the collagen polypeptide chains in this disease.


Infant human pancreas. A potential source of islet tissue for transplantation. Twelve pancreases from human infants one year old or less were analyzed for tissue insulin and amylase content before and after dispersal of pancreatic fragments by mincing and collagenase digestion. Tissue insulin and amylase content provide an index of pancreatic islet mass and exocrine digestive enzyme content, respectively. The results were compared with similar anaylses performed on juvenile and adult human pancreases before and after islet isolation and on intact and dispersed neonatal rat and adult rat pancreas. Infant human pancreas has an average tissue insulin concentration of 1,128 mug./gm. of tissue and a total insulin content of 1,718 mug/pancreas, as against values of 140 mug./gm. of tissue and 7,209 mug./pancreas for adult human pancreas. Average tissue amylase concentration is 0.24 mg./gm. of tissue in infant human pancreas and 3.0 mg./gm. of tissue in adult human pancreas. The insulin/amylase ratio in infant pancreas is 4,800, as against 46 in the adult pancreas. Neonatal rat pancreas, which can be dissociated and transplanted without separation of islet and exocrine components, has a similarly high tissue insulin and low tissue amylase content when compared with adult rat pancreases. Infant human pancreas has a total islet mass 24 per cent that of an adult human pancreas, and neonatal rat pancreas has a total islet mass 11 per cent of that of an adult rat pancreas. One neonatal rat pancreas prepared by minimal collagenase digestion can cure diabetes when transplanted via the portal vein to a rat. Following dispersal of infant human pancreas by collagenase digestion, the islet content and the insulin/amylase ratio of the recovered tissue equals or exceeds that which usually can be isolated from adult cadaver pancreases. Infant human pancreas is a rich source of islet tissue that is relatively uncontaminated by exocrine digestive enzymes. After dispersal, infant human pancreas may be ideal for transplantation to selected diabetic patients.


Peritoneoscopy and guided biopsy in the diagnosis of intraabdominal disease. In 145 cases of intraabdominal disease, a laparotomy was considered the next diagnostic step, but peritoneoscopy was performed instead. In 37 cases with a suspicion of metatastic carcinoma, peritoneoscopy with guided biopsy demonstrated carcinoma in 29. In 32 cases, with biopsy-proven cirrhosis of the liver with high suspicion of a hepatoma, peritonescopy demonstrated the presence of hepatoma in 12. In 28 cases, protracted unexplained jaundice was present; nonsurgical causes for jaundice were found in 15. In 48 cases an exudative (protein greater than 2.5 per 100 ml) ascites was present. In 19 cases, either tuberculosis or carcinomatous implants of the peritoneum were found, and ovarian carcinoma was found in 9. Peritoneoscopy with guided biopsy obviated the need for laparotomy in 90% of these cases.


Interaction of peptides related to secretin with hormone receptors on pancreatic acinar cells. Three analogues of the carboxyl-terminal tricosapeptide of secretin (S5-27), one with glutamine replacing glutamic acid in position 9 (9-Gln-S5-27), a second with asparagine substituted for aspartic acid in position 15 (15-Asn-S5-27), and a third with both replacements (9-Gln-15-Asn-S5-27) were tested for their ability to interact with hormone receptors on dispersed pancreatic acinar cells. Each of these analogues inhibited binding of 125I-labeled vasoactive intestinal peptide (VIP). None of the analogues increased cellular cyclic AMP but each inhibited the increase in cellular cyclic AMP produced by secretin or VIP. At the high affinity VIP receptor (the low affinity secretin receptor) each analogue had an apparent affinity which was greater than that for S5-27, whereas at he low affinity VIP receptor (the high affinity secretin receptor), each of the analogues had an apparent affinity which was the same as that for S5-27. Thus, in S5-27, substituting glutamine in position 9 or asparagine in position 15 makes the fragment more VIP-like but not less secretin-like. These results also provide additional evidence that the receptor having a low affinity for secretin and a high affinity for VIP is functionally distinct from the receptor having a high affinity for secretin and a low affinity for VIP.


Effect of propranolol on bile acid- and cholera enterotoxin-stimulated cAMP and secretion in rabbit intestine. Stimulation of net secretion by deoxycholic acid (DCA) in the colon and by cholera enterotoxin (CE) in the jejunum is mediated by cAMP. Propranolol (Pr) inhibits adenylate cyclase (AC) activity and net secretion induced by bile acid in the colon. The aim of this study was to assess the organ specificity of DCA and CE as well as the selectivity of Pr inhibition. Three colonic and three jejunal loops were prepared in each of 8 rabbits treated intravenously with Pr, 4 mg per kg, 1/2 hr before loop construction and in each of 10 untreated control rabbits. One milliliter of DCA, 6 mM, CE, 10 mug per ml, or heat-inactivated CE or 0.9% NaCl, as basal controls were injected in random order into each of the loops. The volume of luminal fluid and mucosal AC were measured in each intestinal loop 5 hr later. DCA in the colon stimulated AC 2-fold (P less than 0.01) and luminal fluid 15-fold (P less than 0.01). CE in the jejunum stimulated AC 2.3-fold (P less than 0.01) and luminal fluid 9-fold (P less that 0.01). No significant effects on volume or AC occurred in response to CE in the colon or to DCA in the jejunum. Pr pretreatment completely prevented the stimulation of AC and luminal fluid by DCA in the colon but did not affect the action of CE in the jejunum of the same animals. Thus, DCA and CE are organ-specific stimulants of cAMP systems, and Pr is a selective inhibitor of certain inducers of cAMP and net secretion.


Effect of indomethacin on cholera-induced fluid movement, unidirectional sodium fluxes, and intestinal cAMP. Cholera enterotoxin (CT) produces intestinal secretion associated with an elevation of intestinal cyclic AMP (cAMP). Indomethacin, a potent inhibitor of prostaglandin (PG) synthesis, decreases CT-induced secretion, although a role for PG in this process has not been demonstrated. The purpose of this study was to measure the effects of indomethacin on net fluid movement and unidirectional Na fluxes in rabbit jejunal loops exposed to CT and to correlate these findings with intestinal cAMP levels. In untreated animals (no indomethacin), CT loops secreted 0.37 ml per cm per 4 hr compared to absorption in control loops of 0.23 ml per cm per 4 hr (P less than 0.001). In indomethacin-treated animals, there was a striking reduction of secretion in CT loops (0.07 ml per cm per 4 hr, P less than 0.001). Absorption in control loops in indomethacin animals was greater than in untreated animals (0.42 ml per cm per 4 hr, P less than 0.02). Unidirectional Na fluxes were greatly depressed in indomethacin animals at 1 h in both CT and control loops. This effect disappeared by 4 hr. Intestinal cAMP levels in CT loops, although not elevated at 1 hr despite the onset of secretion, were significantly elevated at 4 hr. Indomethacin did not alter cAMP levels at 1 and 4 hr in either cholera or control loops. These studies support the view that PG synthesis is not involved in CT-induced elevation of intestinal cAMP. Indomethacin may depress intestinal secretion by inhibiting a PG-mediated step beyond the generation of cAMP or by acting on some other, as yet unidentified, biological mechanism involved in intestinal secretion.


[Attempted protection against experimental endotoxic shock]. Protection against endotoxin challenge. The AA. have summarized shortly some experimental data dealing with trials of protection against endotoxin challenge pointing out, mostly, the effects of phosphate-containing compounds, as well as dextrane, anticoagulants, platelets, polymyxin B, and serum antiendotoxin and Lipid A antiserum. They have reported some experiments carried out by means of Limulus amebocyte lysate.


Stimulation of colonic glycoprotein synthesis by dibutyryl cyclic AMP and theophylline. The effects of dibutyryl cyclic AMP (B2cAMP) and theophylline on glyoprotein synthesis in rabbit colon were studied in mucosal organ cultures using [3H]glucosamine as a precursor. Addition of B2cAMP (1 mm) to culture medium caused a significant increase in glycoprotein synthesis after 12 and 24 hr compared with biopsies cultured in control medium. The increase in glycoprotein synthesis was observed only if the cyclic nucleotide was present continuously in the incubation medium for at least 12 hr. The stimulatory effect of B2cAMP on [3H]glucosamine incorporation was blocked by cycloheximide. B2cAMP also stimulated mucosal uptake of glucosamine into the intracellular pool and markedly enhanced specific activity of mucosa galactosyltransferase, an enzyme involved in glycoprotein synthesis. Addition of 5mM theophylline caused a greater than 2-fold increase in cAMP levels, which was also accompanied by an increase in glucosamine uptake and incorporation into mucosal glycoproteins. This study demonstrates that elevation of intracellular cAMP concentration in colon epithelium in vitro is associated with an increase in glycoprotein synthesis. These effects may be mediated in part by (1) increased uptake of glycoprotein precursors such as glucosamine, and (2) increased activity of glycoprotein synthetic enzymes.


[Pathogenesis, morphology, prognosis, clinical course and therapy of cystosarcoma phyllodes mammae (author's transl)]. Our investigation of 20 cases of C. ph. of the breast in women aged between 12 and 70 years yielded the following results: 1. In all cases a fibroadenoma or remnants of such a tumor were seen alongside with C. ph. A C. ph. therefore always develops on the base of a benign fibroadenoma of the breast. 2. The mitotic proliferation of the epithelium induces a proliferation of the myoepithelial cells which form the stromal portion of the tumor and can differentiate in to fibroblasts, lipoblasts, smooth muscle cells, reticulum cells, cartilage cells, osteoid cells or bone cells. The C. ph. of the female breast should thus be seen as an adenomyothelioma. Malignant degeneration of stromal cells possibly with haematogenous metastasis is observed in some cases. Concurrent malignant change of the epithelium (carcinosarcoma) is extremely rare and leads to lymph node metastases. 3. At the time being, there is no standardized and/or schematic treatment available. The therapy should be performed according to the size and growth of the tumor into the surrounding tissue, from simple excision to mastectomy. Radiation therapy ist not effective.


Serum immunoreactive corticotrophin and response to metyrapone in old age in man. Attempts have been made to establish the level of immunoreactive corticotrophin (IR-ACTH) in serum in healthy subjects with partts were investigated and the clinical and biochemical condition s of selection specified. Serum IR-ACTH was determined by a radioimmunoassay procedure using an N-terminal antibody produced against the 1--24 sequence of the conrticotrophin molecule. The morning level of serum IR-ACTH was determined in 115 fasting healthy subjects, range less than 16--210 pg/ml, median 94 pg/ml (3rd International Standard preparation). The level was found to be unchanged within the age interval 20--94 years, irrespective of sex. Thy nycterohemeral variation of the IR-ACTH concentration in serum was studied in 11 sibjects. A rhythmic variation of the IR-ACTH level was not constantly found and age-related difference in the temporal pattern was not demonstrated. In 10 young and 10 elderly subjects a metyrapone test was carried out and the IR-ACTH response was determined. The rise in serum IR-ACTH concentration varied between 20 and 739 pg/ml. Any significant age-related difference in response was not found. The interpretation of the present results with respect to the decreased secretory activity of the glucocorticoid producing system in the aged is discussed. It was concluded that the findings in this study did not indicate a primary deficiency of the axis of the glucocorticoid producing system, neither at the pituitary level nor at the adrenocortical level, as a causal factor to the decreased glucocorticoid production in old age.


[Ulnar nerve neuropathy in the elbow region: surgical findings and conclusions about the etiologic mechanism and indications for surgery]. The operative findings of 191 cases of so-called tardy ulnar neuritis are demonstrated. They may be divided into 2 groups, neuropathies associated with (67 cases) and without dislocation or subluxation of the nerve (124 cases). Cases of ulnar neuritis associated with dislocation of the nerve showing no other pathologic changes indicate that dislocation of the nerve alone may account for clinical signs. The next step is the formation of adhesions followed at a later date by formation of a pseudoneuroma. As this condition is mostly of congenital origin an additional factor is needed for including clinical manifestations such as direct or chronic professional trauma. Not enough importance has been attached to the strong triceps, with large muscle mass reaching far down to the olecranon, which might cause irritation of the nerve by pressing it against the wall of the sulcus or dislocating it over the epicondyle. In cases of neuropathy without dislocation/subluxation there is always a pathological finding even when there is no pseudoneuroma. The m. epitrochleoanconaeus is found in this category (14 cases, 11%). The cubital tunnel syndrom of OSBORNE in a very large sense (all possible causes of compression distal to the sulcus) has been found 28 times. Therapeutic measures aim at taking the nerve away from the causative irritation (anterior transposition) or at erradicating the cause (resection of the epicondyle, section of the arcus tendineus, excision of a tumor etc.). OSBORNE's operation has been performed only thrice. Subluxation or dislocation of the nerve may follow this procedure or the nerve is left more exposed to pressure than before. Deep submuscular transposition is preferred in patients less than 50 years old but other factors may determine the choice between deep and subcutaneous transposition such as the cause of the neuropathy, arthritis of the elbow joint, strength of the flexor muscle mass etc.


[Ulnar nerve neuropathy in the wrist region: surgical findings]. The author describes the operative findings in 20 cases of compression of the ulnar nerve at the wrist. Adhesions with or without compression of the nerve, neuroma formation and compression by a ganglion are the most frequent findings. In some cases an enlarged artery is compressing the nerve. In one case a loose pisiform bone was causing the irritation. Despite clinical and electromyographical symptoms there was no pathological finding in 2 cases.


[Decompression of individual nerves in anatomical strictures in polyneuropathies]. During clinical and electromyographical examination of one patient with diabetic and one with alcoholic polyneuropathy we found an especially severe lesion of the ulnar nerve at the elbow on one side. As the polyneuropathy generally affects the nerves symmetrically we presumed in addition some other most probably ulnar nerve compression syndrome at the elbow. Both patients were operated and the nerves were transposed (Prof. NIGST). The local finding during the operation, the improvement of the paresis of ulnar nerve and the worsening of the polyneuropathy--found simultaneously at control after 2 and 4 years--confirmed our supposition. Should in cases of polyneuropathy an especially severely damaged nerve be found, one ought to think of the possibility of an additional mechanical lesion and treat the patient accordingly.


[The fibrolipomatous hypertrophy of the main nerves of the upper extremity]. 10 cases of lipofibromatous hypertrophy of the main nerves in the upper limb in 7 female and 3 male patients are described. In 1 case at the level of the middle third of the upper arm and in 6 cases at the level of the wrist and neighboring areas. In 2 patients these pathological changes were found in the ulnar nerve, in 1 case the hypertrophy was limited to the upper arm, in the other it extended all along the extremity from below the axilla to the wrist. Finally, in 1 case a 20 cm long hypertrophy was found at the level of the deep branch of the radial nerve (posterior interosseus nerve) in the lower arm below the elbow. The fibrous and lipomatous tissue snugly surrounds the fascicles and cannot be separated from them without damaging them, even if the finest microsurgical techniques are used. As the disease has been diagnosed in children as young as five years a congenital etiology has to be considered. The condition is distinct though from the disease of DEJERINE-SOTTAS or from that of von RECKLINGHAUSEN. The hypertrophic nerve is progressively submitted to a chronic compression in the naturally narrow passages (carpal tunnel, ulnar grove at the elbow, arcade of FROHSE) or it becomes painful and paretic after even minor trauma which leads probably to local oedema, intraneural bleeding and epineural compression. An external and internal decompression has to be carried out. Resection or defatting of the hypertrophic fascicles, however, should be avoided. In 2 of our cases the disease was anamnestically present for 45-50 years and clinically followed in another case for 18 years. The disease does not seem progressive. These cases show, that this disorder, which first appeared in the medical literature in 1964 and have been described by several authors in the median nerve, exists as well in the two other main nerves of the upper extremity.


Comparison of protein components of the plasma membrane fraction from rat liver and Morris hepatomas 5123D and 7777. Isolated plasma membrane fractions from rat liver and Morris hepatoma 5123D and 7777 were labelled with radioiodine 125I by a chemical or enzymatic procedure and then were solubilized in 2 per cent solution of sodium dodecyl sulphate containing 1 per cent 2-mercaptoethanol. Solubilized proteins were separated into 20--22 zones staining with Coomassie Brilliant Blue R-250 after disc gel electrophoresis (7.5 per cent polyacrylamide gel). The high similarity of electrophoretic patterns of polypeptide components of all three preparations of cellular membranes was found in distinction to apparent differnces in the amount and disposition of substances stained with Schiff's reagent. Some tentative conclusions were drawn on the disposition of proteins within membrane structure studied by the method of labelling by chemical and enzymatic procedures (distinguishing between extrinsic and integral proteins).


Histoenzymatic and ultrastructural changes in the hepatocytes of gamma-irradiated rabbits. Ultrastructural changes and intracellular enzyme activities in the hepatocytes were studied in rabbits irradiated with 550 rads of gamma rays at 1,3,6,9,15 and 30 days after irradiation. Swelling and marked rarefaction of the mitochondrial matrix observed on the first day were followed by gradual condensation of the matrix between the 6th and 9th day. This state was accompanied by marked reduction in the succinate dehydrogenase activity, ehich gradually returned to the normal by the 30th day of observation. In the hyaloplasm, the most intense changes developed between the third and sixth day and were manifested by clearing of the cytoplasm and marked fragmentation of the endoplasmic membranes, with concurrent negligible decline of the lactate dehydrogenase activity and unchanged glucose-6-phosphatase activity. In the Golgi apparatus, vacuolization of the cytoplasm and fragmentation of smooth membranes were most pronounced on the 6th day and were correlated with a weakened and diffuse reaction for thiamine pyrophosphatase. The alkaline phosphatase activity was irregularly distributed in the lobule. The activities of lysosomal hydrolases, i.e. acid phosphatase, beta-glucuronidase and non-specific esterase, had various localizations within the lobules. The strongest deviations from the normal and of longest duration. (up to 9 days) were seen in the Browicz-Kupffer cells. Complex studies on the same material conducted concurrently with the use of different methods showed that radiation damages structure and function in unequal degrees. Moreover, within the same organ the cellular response to ionizing radiation varies according to the character, localization and functional state of the cells. Deviations from the normal state occur between the first and ninth days, most of the structural and functional elements showing sings of return to the normal about the 15th day after irradiation.


Histochemistry of oxidative enzymes in experimental allergic encephalomyelitis. The histoenzymic pattern of oxidative enzymes (G3PD, IDH, SD, G6PD,HBD, NADPH: dehydrogenase) was investigated in experimental allergic encephalomyelitis (EAE) produced in rats according to PATERSON [13]. The results obtained lead to following conclusions: (1) The neuroglia, including the white matter oligodendroglia of immunized rats, exhibits increased oxidoreductase activities; (2) The neuroallergic reaction induces some stimulation of the oxidoreductive metabolism of oligoden-droglia; (3) The enzymatic hyperactivity in EAE does not show any relation to the morphological signs of alterations of the myelin sheath.


The long-term prognosis infantile spasms--the present condition of cases of infantile spasms followed in school age. A follow-up study has been made of 25 cases with infantile spasms, all of whom were six years old or more at review. Only four (16%) out of 25 cases made a full recovery and attended normal school. Spasms ceased in 96% of all cases, but fits other than spasms (grand mal, tonic seizure, atonic seizure, myoclonic seizure, atypical absence and psychomotor seizure) occurred subsequently in 11 cases (44%). The EEG became normal in two cases (8%), but still showed modified hypsarhythmia in three cases (12%), "epileptic non-hypsarhythmic" discharges in 17 cases (68%) and non-specific abnormalities in three cases (12%). The important factors associated with good prognosis were normal development before the onset of spasms, late onset (seven months old or over) and short duration of spasms, the absence of other types of fit following spasms and lack of neurological abnormality. A bad prognosis was associated with abnormal development prior to the onset of spasms, early onset and long duration of spasms, the presence of other types of fit following spasms and evidence of any neyrological abnormality This follow-up may confirm that the therapy with ACTH-A has no significant effect on final mental state.


[Improved demonstration of normal lymph nodes in lymphangiograms (author's transl)]. Lymphography with Lipiodol ultra fluid was carried out in eight domestic pigs. The purpose of the experiment was to obtain even distribution of the oil in normal lymph nodes in order to improve its diagnostic value. This was attempted by changes in pressure, temperature and by the addition of benzopyrone. By increasing injection pressure from 0.4 to 0.8 atmospheres, using a pump, it was possible to show more of the lymphatic tissue. Whether this would be possible in man is still not certain. The use of a benzopyrone preparation also improved contrast distribution in lymph nodes. Additional pelvic lymphatics were also seen. The improved distribution of oil is indicated by the altered pattern, strikingly even filling of the vacuoles and 10% incfreased filling rate.


[Cytomegalic virus infection in normal and disturbed immunity. Clinical and animal experiment studies]. Virological and serological studies of cytomegalovirus infections revealed following results: 1. The distribution of cytomegalovirus infections in the area of Munich can be compared with other European areas. 57% of adults have antibodies against the cytomegalovirus. 2. Cytomegalovirus is an important agent initiating infectious liver diseases in childhood. 3. The virus is also an important factor for severe cerebral retardation. 4. Studies in adult mice showed that a non-pathogenic strain of mouse cytomegalovirus can produce a fatal infection, when the immunological resistance was depressed by cytotoxic agents (i.e. cyclophosphamide, azathioprine, antilymphocyte serum). 5. Cytosine arabinoside inhibiting in vitro the replication of DNA viruses, increases in mice significantly the cytomegalovirus infection. Therefore cytosine arabinoside should be used very critically in the therapy of human cytomegalovirus infections.


[Pseudohypoparathyroidism. Disease of the second messenger]. Pseudo-hypoparathyroidism (PHP) is a hereditary disorder with typical dysmorphic signs and symptoms of hypoparathyroidism, which is resistant to parathyroid hormone (PTH). The knowledge of the role of cyclic adenosine monophosphate (cAMP) as a mediator of the action of PTH and the finding, that parathyroid extract in PHP could not stimulate the urinary excretion of cAMP, were necessary to explain the hitherto unknown pathogenesis of Pseudo-hypoparathyroidism. The PTH-sensitive adenylcyclase-system as second messenger is here unable to mediate the action of PTH on its target cells causing disturbances in calcium and phosphorus metabolism.


[Changes of the renin-angiotensin-aldosterone system under contraceptive steroids. Contribution to the etiology of hypertension under hormonal contraceptives]. Treatment with hormonal contraceptives leads to a considerable stimulation of the renin-angiotensin-aldosterone-system. First of all, there is a 2,5- to 4fold increase of renin substrate synthesis in the liver. As a consequence, more angiotensin I and angiotensin II are released. Angiotensin II stimulates the secretion of aldosterone in the adrenals, thus producing a higher aldosterone concentration in plasma. The urinary excretion of aldosterone is elevated to a lesser degree, probably because of the simultaneously increased binding of aldosterone to plasma proteins. The release of renin is suppressed to 50% by negative feedback mechanisms. Some possible factors in the etiology of hypertension induced by oral contraceptives are discussed.


13C-NMR studies of the membrane structure of enveloped virions (vesicular stomatitis virus). The mobility of the lipids in the bilayer of the envelope of vesicular stomatitis virus has been probed over its complete space by the biosynthetic incorporation of [N-13CH3]- choline as a probe for the polar head groups and [3-13C]- and [11-13C] oleic acid and [16-13C]- palmitic acid for the hydrophobic region of the bilayer. These precursors were effectively incorporated as established by the concomitant administration of the same precursors in radioactive form. Spin lattice relaxation time measurements (T1) of the 13C enriched segments in complete virus envelope allowed estimation of their mobility. The mobility of the polar head groups is restricted, probably due to ionic interactions with neighbouring acidic phospholipids (phosphatidylserine) and/or acidic side chains of the glycoprotein (G-protein). The rigidity of the hydrophobic part of the bilayer is due to the high cholesterol content and interaction with the immersing polypeptide chains of the G- and possibly M-protein. The rigidity is limited to a depth of about 15 A ranging from the inner and outer surface, whereas the inner core of the bilayer is fluid. Tryptic cleavage of the hydrophilic part of the G-protein allows the lipophilic immersing polypeptide fragment to enter further the bilayer which then reduces the fluidity of the hydrocarbon chains in the core region by lipid-protein interactions.


In vitro activation of a soluble cholesterol esterase from bovine adrenals by a cAMP-dependent protein kinase. Properties and partial purification of the bovine adrenal cholesterol esterase from the 100000 X g supernatant fraction were investigated. Variations of the enzyme activity with time-dependent (enzymatic) and time-dependent (non enzymatic) effects have been demonstrated. Mg2 has been proved to inhibit the enzyme activity by a non-enzymatic effect in 50mM Tris/HCl buffer, pH 7.4. A time-dependent inactivation of the cholesterol esterase has been observed in the same buffer. The enzyme could be protected from this enzymatic inactivation by its substrate, cholesterol oleate. cAMP, ATP and Mg2 cuase a time-dependent stimulation of the enzyme in 50mM Tris/HCl buffer, pH 7.4. This result suggests that corticotropin activates the soluble cholesterol esterase from bovine adrenals via cAMP-dependent protein kinase. This view is strengthened by the incorporation of 32P radioactivity from [gamma-32P] ATP into the protein fraction of the 100,000 X g supernatant. The protein-bound 32P radioactivity could be co-purified with the enzyme activity during the partial purification of the soluble cholesterol esterase.


3': 5'-Nucleotide phosphodiesterase in the superior cervical ganglion of the rat. Evidence for multiple forms and influence of the beta-adrenergic receptor system in the electrophoretic pattern of the enzyme. Two forms of a cAMP phosphodiesterase were found in the superior cervical ganglia of the rat. The influence of various detergents on solubility and activity of the enzyme was studied. Two Km values were determined, one of 0.9mum and the other 270mum cAMP. Separation by polyacrylamide disc electrophoresis revealed four distinct peaks of enzyme activity. One of these activities was increased when ganglia were incubated with adrenaline for at least 4 h. The increase caused by adrenaline was unaffected when ganglia were pre-incubated with cycloheximide, but was blocked in the presence of propranolol, a beta-adrenergic antagonist. Incubation with dibutyryl cAMP for 4 h mimicked the effect of adrenaline. Brief incubation with adrenaline was without effect. Electrophoresis in the presence of 0.1% Lubrol W, a non-ionic detergent, showed that one band of activity disappeared, while the main peak of enzyme activity increased in size. Two-dimensional starch gel electrophoresis confirmed the results of the disc electrophoresis.


The role of the certified social worker in England's social services. With the reorganization of England's social services into a single unified system, the professional called the certified qualified social worker is playing a key role in the administration and delivery of community services. The author describes some of the worker's activities and also presents an overview of hospital, day treatment, and outpatient services for adult psychiatric patients in the county of Warwickshire. One of the most effective programs is a system of voluntary group homes, operated by an area council of churches, for former psychiatric inpatients.


An educational model for teaching living skills to long-term patients. Establishing rehabilitation programs for long-term mental patients outside mental health settings can save mental health funds and further the patients' integration into the community, the author declares. He describes an approach that uses an educational model to teach patients the basic skills of everyday living. As "students," they enroll in a course taught by a credentialed teacher and sponsored by the adult education department of the local high school. Mental health professionals are used only for consultation, and the cost to the local mental health program is minimal. A key element in the course is having the students participate in planning the curriculum.


Fetal rhabdomyomatous nephroblastoma-a variant of Wilms' tumor. The fetal rhabdomyomatous nephroblastoma is considered to be a predominantly monophasic mesenchymal variant of Wilms' tumor, which has not been seen in patients older than four years. It acts less aggressively than a Wilms tumor despite its much larger size. Its better prognosis appears to be related to either the absence of or the insignificant amounts of neoplastic epithelium. The bilaterality of this tumor in one-third of the cases, however, may negatively affect the overall prognosis, because complete resection may be impossible or because renal failure ensues.


Intracytoplasmic inclusion bodies in the chondrocytes of type I lethal achondrogenesis. Lethal achondrogenesis in the past has been frequently confused with achondroplasia. Clinical and radiologic advances in the last decade have led to clear differentiation of this condition from other types of bone dysplasia. It is further separated into two types on the basis of radiographic and pathologic findings. Re-evaluation of the histologic features has led to the recognition of heretofore unrecognized intracytoplasmic inclusion bodies in the chondrocytes of type 1 achondrogenesis. The finding of inclusions strengthens the differentiation of the two types and may have implications for the pathogenesis of the form of chondrodystrophy.


Propagation of mouse mammary tumor cell lines and production of mouse mammary tumor virus in a serum-free medium. Five different mouse mammary tumor cell lines were propagated in a serum free medium. Evaluation of growth characteristics, including logarithmic growth, cell population increase, protein production and days to confluency, showed serum-free medium comparable to serum-containing medium. Mouse mammary tumor virus expression and production, in C3H and GR tumor cell lines, as determined by virus particle counting and RNA dependent DNA polymerase assays, subsequent to dexamethasone stimulation revealed equivalent to higher levels of virus in serum-free medium as compared to serum-containing medium.


The co-carcinogenic activity of 4-nitropyridine-1-oxide (4-NPO) and prevention of transformation by type-specific anti-viral antibodies. Fischer rat embryo cells chronically infected with Rauscher murine leukemia virus, and known to be sensitive to transformation by potent chemical carcinogens, were transformed by the weak carcinogen 4-nitropyridine-1-oxide. Transformed cells grew in semi-solid agar and produced tumors in newborn Fischer rats. Transformation was inhibited by antisera specific for the ecotropic Rauscher murine leukemia virus, but not by antisera of equal toxicity specific for xenotropic Swiss mouse AT-124 virus.


[Atypical pneumonia, etiology and possibilities for the diagnosis (author's transl)]. Beginning with the antimicrobial chemotherapy a decrease in the incidence of bacterial pneumonias is accompanied by a relative increase in the incidence of the so-called atypical pneumonia. This disease syndrome is predominantly caused by Mycoplasma pneumoniae, Chlamydia psittaci, Coxiella burneti and various viruses. In addition, bacteria which are usually involved in lobar pneumonia may occasionally cause atypical pneumonias. The present publication is concerned with the most frequently occurring causative agents of atypical pneumonia, the epidemiology of the disease and the possibilities for the diagnosis.


Immunoglobulin content and antibody activity in an artificial body cavity. Artificial body cavities (ABC) were created by the insertion of hollow polyethylene balls in the subcutaneous tissue of rabbits. After two months no inflammatory reaction could be detected, the ABC was enveloped by a membranous structure, and the cavity contained about 20 ml of fluid. The protein concentration was about 3 times, and the IgG about 8 times, lower in the ABC fluids than in the corresponding serum. At the same time the antibody titers against sheep red blood cells (SRBC), human IgG and herpes simplex virus type 1 (HSV) were about 20 200 times lower than in the corresponding serum samples. The IgG molecules appeared to be undamaged as shown by the presence of various allotypes and by the elution pattern from G-200 column. Testing the protective activity of ABC fluids and of serum against HSV infection showed that ABC fluids had no protective activity. The experiments suggest that the membranous structure selected among classes of Ig and probably among other serum proteins. In addition, the IgG molecules with antibody activity against various inoculated antigens were selectively excluded to penetrate in the ABC. The ABC might be considered analogous to virtual cavities of the body surrounded by membranes and possibly to the extravascular compartment.


Lymphocyte activation by cell separation procedures. Cell separation techniques normally used to obtain subpopulations of lymphocytes were shown, under certain conditions, to render the cells cytotoxic towards a number of target-cells including autologous lymphocytes. To cause cytotoxicity, it was necessary to pass cells through glass wool and/or nylon wool columns equilibrated with media containing fresh plasma or serum. Cells lost activation upon overnight in vitro culture or treatment with trypsin. In addition to direct cell cytotoxicity, cells released heat labile cytotoxic factors and antibody during the separation procedures. The implications of the results for the interpretation of cell separation studies designed to attribute immunological effects to one or another cell type were discussed.


Mode of interaction of different polyanions with the first (C1,C1) the second (C2) and the fourth (C4) component of complement. IV. Activation of C1 in serum by polyanions. Treatment of serum with dextransulphate polyvinylsulphate or polyanetholsulphonate resulted in a dose-dependent activation of C1 and C3; this was found for normal serum as well as for C4-deficient guinea-pig serum. Activation of C1 and C3 occurred at the same concentration of polyanions. The consumption of C3 in C4 deficient serum and the requirement of factor D of the alternative pathway indicate that C3 is activated via the alternative pathway.


Comparison of serological tests for antibody to hepatitis A antigen, using coded specimens from individuals infected with the MS-1 strain of hepatitis A virus. To compare serological tests for antibody to hepatitis A antigen (anti-HA), we tested 15 paired serum specimens, submitted under code, from individuals infected with the MS-1 strain of hepatitis A virus. Immune electron microscopy (IEM), immune adherence hemagglutination (IAHA), and solid-phase radioimmunoassay (RIA) tests for anti-HA were performed with hepatitis A antigen (HA Ag) derived from human stool; results were also compared with previously reported titers determined by IAHA with HA Ag derived from marmoset liver. Antibody titers (IAHA and RIA) and ratings (IEM) determined with stool-derived HA Ag compared favorably, and a seroresponse to HA Ag was detected by all three methods for every serum pair tested. Differences in titers were noted between IAHA tests with liver-derived and with stool-derived HA Ag, but the discrepancies could be accounted for by differences in test technique. The agreement found in this study among the three techniques was quite good and confirms the specificity and sensitivity of tests for anti-HA that are done with stool-derived HA-Ag.


Viral infections in renal transplant recipients. Cytomegalovirus (CMV) infections are common in renal transplant recipients. We studied 23 recipients prospectively to determine whether infections by other herpes-group and non-herpes-group viruses were also present. Sera, obtained at the time of surgery and periodically thereafter, were tested for antibody to CMV, herpes simplex virus (HSV), Epstein-Barr virus (EBV), parainfluenza viruses types 1, 2, and 3, and the viruses of measles and rubella. We found no evidence of an unusual incidence of primary or secondary infection by the non-herpesviruses tested. Rises to CMV, HSV, and EBV antibody titers occurred in 43, 38, and 32% of patients, respectively. All  serological rises to herpes-group viruses occurred in patients seropositive at the time of transplantation, with the exception of three patients who experienced primary CMV infections. We conclude that reactivation of all herpes-group viruses tested may occur in transplant recipients. Morbidity was associated only with primarly CMV infection.


Synergistic effect on mortality in mice with murine cytomegalovirus and Pseudomonas aeruginosa, Staphylococcus aureus, or Candida albicans infections. A synergistic effect on mortality was demonstrated in a combined infection of mice with murine cytomegalovirus (MCMV) and Pseudomonas aeruginosa, Staphylococcus aureus, or Candida albicans. Mice infected intraperitoneally with a 0 to 20% lethal dose inoculum of MCMV 3 days prior to the intravenous injection of a 0 to 20% lethal dose inoculum of either the bacteria or fungus demonstrated a striking enhancement of mortality. MCMV-infected mice given Pseudomonas or Staphylococcus exhibited a 90 to 100% mortality within 24 to 48 h, whereas 80% of viral-infected animals injected with Candida died in 5 days. Injection of the bacteria or fungus at various times during the MCMV infection resulted in enhanced mortality on days 0,1,2, and 3 of the viral infection. Greatest synergism was observed on day 3, with a progressive decline in death rates thereafter. Immunization with MCMV abrogated the synergistic effect on mortality in all three combined infections. Immunization with Pseudomonas reduced mortality in the combined MCMV-Pseudomonas infection. These results indicate that mice exhibit a markedly enhanced susceptibility to bacterial and fungal infections during the course of the MCMV infection and suggest that the enhancement may be related to viral-induced alterations in host resistance.


Clinical aspects of dental anomalies.

Clinical aspects of dental anomalies. Many inherited disorders have oral manifestations which can be detected on dental radiographs as alterations in the morphology or chemical composition of the teeth. Thus the dentist may be the first to detect disorders of development and metabolism of importance to the general health of the patient and his family. In one group of conditions the pulp chamber is larger than normal. This may be associated with taurodontism in such conditions as polyploidy of the -X chromosome and trisomy-21 or Down's syndrome. Taurodontism also occurs in a variety of other syndromes including the tricho-dento-osseous syndrome described by Robinson, Miller and Worth (1966) and Mohr's syndrome. It may also be associated with scanty hair endoligodentia. In certain metabolic conditions the pulp chamber may be enlarged but the teeth are of relatively normal from (cynodont). Such cases include hypophosphatemic vitamin D-resistant and dependent rickets, pseudo-hypoparthyroidism otodental syndrome and hypophosphatasia. Small pulp chambers and associated anomalies of root morphology also occur in hereditary disorders either alone or as part of various syndromes such as dentino-osseous dysplasia and brackioskeletogenital syndrome. Dentinogenesis imperfecta may occur alone or as one manifestation of osteogenesis imperfecta. Other developmental defects including pulpal dysplasia, labodontia and dens invaginatus are also associated with small pulp chambers.


Indomethacin enhancement of spleen-cell responsiveness to mitogen stimulation in tumorous mice. Spleen cells removed from C57Bl/6J mice bearing a methylcholanthrene-induced fibrosarcoma (MC-16) demonstrate suppressed responsiveness of phytohemagglutinin (PHA) and bacterial lipopolysaccharide (LPS) induced mitogenesis as compared to non-tumorous mice. A similar depression of PHA-induced mitogenesis was observed with spleen cells from C3H/HeJ mice bearing syngeneic mammary adenocarcinomas (C3HBA). The administration of indomethacin, a non-competitive irreversible prostaglandin (Pg) synthesis inhibitor, (75 or 100 mug/mouse, IP) on an alternate day basis to groups of tumor-bearing mice of both strains, significantly enhanced immune cell responsiveness to mitogenic stimulation. The addition of indomethacin (10 mug/ml) to cultures of spleen cells from these tumor-bearing mice, as well as to DBA/1J mice bearing the Cloudman S-91 melanoma, enhanced spleen-cell responsiveness to mitogen-induced DNA synthesis by as much as 156%. Indomethacin administration in vivo or in vitro had no significant effect on mitogen-induced DNA synthesis of spleen cells from non-tumor-bearing animals. It is hypothesized that tumors, or tumor-cell antigens, increase Pg production of a population of spleen cells, and that the increased Pg content of the spleen may be important in controlling immune responsiveness in mice.


Expression of feline type-C virus in normal and tumor tissues of the domestic cat. Tumor and normal tissues from domestic cats were examined for FeLV and RD-114 specific nucleic acids and proteins. Virus-positive tissues showed a 100- to 200-fold higher level of FeLV and a 200- to 400-fold higher level of FeLV p30 than virus-negative tumors and normal cat tissues. In contrast, the levels of RD-114 viral RNA and p30 antigen were comparable in the majority of tumorous and normal tissues examined. When the DNA of these tissues was examined for FeLV-related sequences, the virus-positive tumors were found to contain extra copies of FeLV DNA. Virus-negative tumors contained DNA sequences homologous with FeLV but the copy number of these sequences could not be distinguished from normal tissues. The results indicate that in certain FeLV-induced type-C lymphoproliferative diseases, extra type-C sequences not present in normal tissues can be identified.


Antibody responses to membrane antigens in monkeys infected with Herpesvirus saimiri. The antibody response to Herpesvirus saimiri (HSV)-induced membrane antigens (MA) was followed in HSV-infected owl monkeys using the membrane immunofluorescence (MF) and antibody-dependent lymphocyte cytotoxicity (ADLC) assays. These responses were correlated with the loss of T-cell responsiveness to general mitogens. Antibody titers to MA as determined by ADLC but not MF increased to remarkably high titers in those monkeys that developed malignant disease following virus infection, while remaining at relatively low and constant levels in those monkeys that developed a chronic virus infection in the absence of malignant disease. This disease-related antibody response pattern was paralleled by the loss of T-cell function in diseased animals. The development of leukemia in HVS-infected owl monkeys did not suppress the ability of peripheral blood lymphocytes to act as effector cells in the ADLC assay. The relationship of these immune response patterns to the development of malignant disease in this system is discussed.


Undulating membraneous structures associated with the endoplasmic reticulum in tumour cells. The morphology and occurrence of tubuloreticular and undulating membraneous structures (TRS and UMS) associated with the endoplasmic reticulum were examined under normal and pathologic conditions. It was concluded that TRS and UMS are cytoplasmic inclusions of similar type but of different organization. The occurrence of UMS in a human cell line derived from a carcinoma of the lung and in a transplantable chicken hepatoma derived from an MC-29 virus-induced liver tumour is described.


Immunological monitoring and immunotherapy in carcinoma of the lung. One hundred and seven patients with carcinoma of the lung underwent immunologic testing, and 62 of these patients were randomized to an immunotherapy protocol comparing the effects of Pasteur strain BCG, either alone or combined with allogeneic tumor cells, to the effects of no immunotherapy. Patients with residual disease left at the time of surgery or with metastatic disease at the time of diagnosis showed no increase in survival as a result of this form of immunotherapy. An insufficient number of patients with less advanced disease, in whom we would expect the most beneficial effect, have been entered in this study. In general, we were unable to document substantial effects of immunotherapy on the immunologic parameters tested. Only in recall antigen skin testing was there a statistically significant increase in reactivity in the immunotherapy groups. Tests of general immune status appeared to have a predictive value in monitoring lung cancer patients. Anergic patients had a poorer prognosis than did patients who demonstrated skin test reactivity. Patients with normal percentages of lymphocytes (T cells) forming rosettes with sheep erythrocytes at 29 degrees C were generally normal in other tests of immune competence. In serial studies of rosette formation, all patients who developed recurrent disease had a pattern of depressed or falling rosette values, and these abnormalities occurred an average of 3.1 months prior to clinical detection of recurrence. Patients with large-cell anaplastic carcinoma were found to have a significantly higher incidence of depressed rosette levels than the other histologic types. Both large and small-cell anaplastic patients had significantly depressed lymphocyte proliferation by mitogens and allogeneic cells. Although lung cancer patients have been described as immunologically depressed, they are capable of recognizing tumor-associated antigens. When tested in leukocyte migration inhibition assays with tumor-associated antigens, the majority of the patients in our study were found to be reactive. The use of a 3 M KCl extract of pleural effusion cells from a patient with pulmonary adenocarcinoma has given good reactivity and specificity in lung cancer patients of all histologic types. In addition, these patients have been shown to respond in a mixed lymphocyte/tumor interaction to tumor-associated antigens (Dean, 1976b).


Studies on the microcytotoxicity test. I. Evidence that the effects of normal lymphoid cells on tumour cell growth in microtest plates may be caused by non-immunological modifications of the culture medium. Lymph node cells from normal Wistar rats were found to affect the growth/survival of syngeneic chemically-induced solid tumour cells in the microcytotoxicity test. Whether inhibition or enhancement of tumour cell numbers occurred depended on the particular tumour cell type and, in some cases, on the particular in vitro subline used. Fractionation of LNC on nylon wool columns revealed that the two effects were mediated by distinct subpopulations of lymphoid cells: column-retained cells showed predominantly an inhibitory effect and column-eluted cells predominantly an enhancing effect. When column-retained and column-eluted cells were cultured under the conditions of the microcytotoxicity test but in the absence of tumour cell growth. The inhibitory activity was maximal within one hour of lymphocyte culture, while the enhancing effect developed slowly during incubation. Furthermore, the tumour cells themselves were found to produce growth-enhancing activity. It is proposed that interaction between these various supernatant activities accounts at least in part for the non-specific effects of normal lymphoid cells in the microcytotoxicity test.


Expression of viral proteins in mammalian cells transformed by avian sarcoma viruses. The expression of viral proteins in nine lines of hamster and rat cells transformed by avian sarcoma viruses (ASV) was studied by indirect immunofluorescence with monospecific antisera to purified gp85 and p27 of AMV-B and a polyvalent antiserum to all the p proteins of this same virus. The lines of ASV-transformed cells were either low virus producers (VP) or inducible or non-inducible non producers (NP). Cytoplasmic expression of p proteins was observed in all the cell lines except the least inducible NP cell line, and cytoplasmic expression of gp85 in all the cell lines. The degree of expression varied widely with the lines and was not related to the class of permissiveness or inducibility. However, in the inducible NP class, the expression of p proteins and gp85 was higher in the most inducible cell lines. The data also suggest that the expression of the p proteins must be uncoordinate in at least some cell lines and must also be uncoordinate with the expression of gp85. In the VP cell lines and the most inducible NP lines, g85 and some p proteins other than p27 were also expressed on the cell membrane. The membrane expression of gp85 and the p proteins which were expressed appeared to be coordinate and to parallel the degree of cytoplasmic expression. In contrast, no, or a negligible expression of viral proteins was observed on the membrane of the least inducible and the non-inducible cell lines. These results suggest that there may exist translational and/or post-translational controls of the expression of viral proteins in the ASV-transformed mammalian cells and that the permissiveness and the inducibility of the cells may depend on the insertion of viral proteins in the cell membrane. The failure of p27 to insert in the cell membrane could account for the low permissiveness or the non-permissiveness of the cells.


Properties of a baboon lymphotropic herpesvirus related to Epstein-Barr virus. Three lymphoblastoid cell lines were established from splenic lymphocytes of a lymphomatous baboon (Papio hamadryas) by co-cultivation of the lymphocytes with X-irradiated cells of marmoset or baboon lymphoblastoid cell cultures; the baboon splenic lymphocytes failed to grow when cultured alone. A herpesvirus, associated with each cell line, was identified by immunofluorescence, molecular hybridization and electron microscopy. Antigenic comparison with Epstein-Barr virus (EBV) showed that the baboon herpesvirus and EBV shared cross-reacting viral capsid antigens (VCA): 20 of 20 (100%) anti-VCA (EBV)-positive human sera and 55 of 62 (89%) baboon sera reacted with the baboon lymphoblastoid cells and baboon sera stained EBV VCA in P3HR-1 and EB-3 cells. No nuclear antigen, as assayed by anti-complement immunofluorescence tests, was detected in baboon lymphoblastoid cells when human or baboon anti-VCA positive sera were used. Baboon anti-VCA-positive sera also failed to stain EBV nuclear antigens (EBNA) in Raji or P3HR-1 cells. Preliminary molecular hybridization studies showed only approximately 40% homology between viral DNA of baboon cell lines and DNA of EBV derived from P3HR-1 cells.


Leukemogenic activity of murine type C viruses after long-term passage in vitro. Cloned stocks of several murine leukemia viruses (MuLVs) were shown to be leukemogenic for susceptible mice after more than nine years of in vitro passaging in mouse embryo fibroblasts. Tissue culture-grown Rauscher (R-) MuLVs injected into newborn or young adult BALB/c mice induced lymphocytic leukemias in 100% of the animals beginning 80 days post-inoculation. No erythroblastic leukemia was observed even after passaging the tissue-culture-grown R-MuLVs twice through mice, indicating that the component responsible for that disease had been lost or attenuated during growth in fibroblasts. The tissue-culture-grown stock of Moloney (M-) MuLVs likewise induced lymphocytic leukemias in 94% of injected newborn BALB/c mice, and the tissue culture-grown Gross (G-) MuLVs induced lymphocytic leukemias in 42% of injected newborn C3Hf mice. The host range and neutralization characteristics of viruses recovered from animals that became leukemic after injection with the tissue-culture-maintained MuLVs were found to be identical with those of the injected viruses. These data implicate the injected MuLVs in the induction of the leukemias and suggest that the capacity to induce the disease is stably inherited as part of the viral genome even in the absence of expression.


Adrenocorticotropin. 48. Synthesis and biological activity of (15, 16-D-lysine, 17, 18-D-arginine)-adrenocorticotropin-(1-19) and an all-D-retropeptide related to the amino terminal octadecapeptide of adrenocorticotropin. The peptide [15, 16-D-lysine, 17, 18-D-arginine]-adrenocorticotropin-(1-19) and an all-D-retropeptide related to the amino terminal octadecapeptide of adrenocorticotropin have been synthesized by the solid-phase method. The nonadecapeptide was shown to possess 10-15% of the steroidogenic activity and 3% of the lipolytic activity of adrenocorticotropin-(1-19). The all-D-retropeptide showed no activity and exhibited no inhibitory activity in steroidogenesis and lipolysis.


X-ray-induced luminescence and optical absorption in DNA films. For DNA films, X-irradiated at 77 K, a comparative study was performed on (a) light emission (fluorescence and phosphorescence) during irradiation, (b) afterglow and thermoluminescence, (c) optical absorption after irradiation, and (d) free-radical reactions in similar gamma-irradiatied DNA samples. Spectra are reported for the various kinds of light emission and for the optical absorption. From temperature-dependence studies, it was concluded that the light-emission phenomena are unrelated to the reactions of the free radicals as studied by electron paramagnetic resonance. The optical absorption on the other hand, is strongly correlated to the presence of free radicals in the DNA. Some mechanisms possibly involved in the observed luminescence reactions are discussed.


Radical formation in salts of pyrimidines III. Cytosine . H2O cyrstals. The radicals produced by X-irradiation at 77 K and at 300 K in cytosine monohydrate crystals have been analysed by electron-spin-resonance (e.s.r.) spectroscopy. Three radicals have been identified at 77 K: the anion radical and the cation radical of the cytosine molecule, together with the radical resutling from H-abstraction from the nitrogen N1. Irradiation at 300 K produces radicals resulting from H-adition at three different positions of the cytosine molecule. These are the C5-addition radical, the C6-addition radical, and the O2-addition radical. The results are compared with those found previously by other authors.


Experimental disciform edema and necrotizing keratitis in the rabbit. The development of experimental disciform edema and necrotizing keratitis in the corneas or rabbits following intrastromal inoculation with the RE strain of herpes simplex virus is described. Following an initial episode of conjunctivitis and epithelial keratitis, a mild, centrally localized, stromal edema developed on the fifth day. Stromal edema, opcification, and neovascularization of the cornea reached maximum severity on the seventh to twenty-second day, and began to fade in most eyes thereafter. On the twenty-ninth day most corneas have attained a resolved state characterized by subepithelial granular opacities. Several eyes were observed which developed central necrotizing keratitis. Marked similarities between the animal model and human herpetic stromal keratitis were apparent. Histological observations show that early necrotizing keratitis in the rabbit is characterized by an infiltration of plasma cells and lymphocytes in the limbus, with polymorphonuclear leukocytes, lymphocytes, and macrophages in the central cornea.


Differences in adenylate cyclase activities in murine normal cells and bladder tumor cells in tissue culture. Cyclic AMP may be involved in the modulation of cell growth. The present work sought to further define differences between normal cells and tumor cells in their cyclic AMP system. Mouse embryo fibroblasts and murine bladder transitional epithelium tumor cells were grown in vitro; at various times, adenyl cyclase activity was assayed by measuring the conversion of [alpha32P]ATP to cyclic AM32P; stimulation by prostaglandin E1 or sodium fluoride was also determined. Base line and fluoride-stimulated enzyme activity were significantly greater in normal cells than tumor cells (P less than 0.01), and reached a peak at day 2; at confluency, levels in both systems decreased. Prostaglandin E1-stimulated levels, in contrast, were greater in tumor cells, there being a 10 fold greater relative stimulation in these cells compared to normal cells (P less than 0.01). Findings of a possibly greater sensitivity in these tumor cells may be important in a therapeutic modulation of tumor growth.


In vitro stimulation of human lymphocytes by purified cytomegalovirus. Lymphocytes from 19 healthy donors were tested against purified cytomegalovirus (CMV) strain Ad 169, in a lymphocyte-transformation test. The test was performed in microcultures using various preparations and concentrations of antigen. These included, besides purified CMV, CMV-infected cells, herpes simplex virus type 1 antigen, and control antigens. Lymphocytes from CMV-seropositive donors were stimulated by purified virus and infected cells, whereas lymphocytes from seronegative donors did not respond. Similarly, only lymphocytes from herpes-seropositive donors did respond to herpes antigen. With these preparations of antigen the test was found to be sensitive and specific.


Plaque dissociation of herpes simplex viruses: biochemical and biological characters of the viral variants. Polycaryocytogenic (P) and non-polycaryocytogenic, or aggregating (A), stable variants were selected from a herpes simplex virus type 1 (HSV-1) and from a herpes simplex virus type 2 (HSV 2) which had not been deliberately exposed to known mutagenic agents. The P variant of HSV-1 (FP) differed from the A variant (FA) in polypeptides and glycoprotein patterns, but no gross differences were evident between the two variants of HSV-2 (GP and GA). Each P variant proved more 'specific' than each A variant in immune neutralization tests. At high multiplicity, GP produced polycaryocytes but FP did not. Virulence tests in mice showed FP to be much more virulent than FA but GA to be more virulent than GP. Finally, A and P variants of each type could not be differentiated with respect to thermal resistance of virons, capacity to grow at high temperature, and buoyant density of DNA.


Genetic factors influencing endogenous type-C RNA viruses of mouse cells: control of viral polypeptide expression in the C57BL/10 strain. Type-C viral polypeptides are expressed at very low levels in C57BL/10 liver and embryo cells as compared to cells of other mouse strains. The 12,000-mol. wt. polypeptide in C57BL/10 cells was purified and shown to possess type-specific antigenic determinants indistinguishable from those of a previously described class of endogenous virus. The very low levels of endogenous virus-specific RNA in cells of C57BL/10 as compared to other strains suggests control of this endogenous virus at the level of transcription. Analysis of viral antigen expression F1 hybrids of C57BL/10 with strains characterized by intermediate and high levels of viral antigen expression indicates that the restriction of this endogenous virus by C57BL/10 cells is not due to simple dominance by a repressor of virus expression.


Inhibition of herpes simplex virus multiplication by 2,3-bis(acetyl mercaptomethyl)-quinoxalin. The drug 2,3-bis(acetyl mercaptomethyl)-quinoxalin (BAMMQ) was 99.9% inhibitory for herpes simplex virus (HSV) multiplication in cell cultures at concentrations of 1.6 X 10(-5) M or less. The drug was not inactivating for the virus, did not interfere with adsorption and penetration of the virus, and was still active when added as late as 12 h after infection with HSV. BAMMQ did not prevent HSV DNA replication, but interfered with a late stage of virus assembly and/or maturation.


An unusual defective genotype derived from herpes simplex virus strain ANG. A defective genotype of herpes simplex virus strain Angelotti (HSV ANG) accumulates in the course of controlled serial high MOI virus passages representing 50-60% of the total number of mature virions in the seventh of these passages. Defective HSV ANG significantly differs from other defective HSV genotypes described so far: the DNA of the defective particles has the same buoyant density as nondefective DNA. In contrast to non-defective HSV ANG DNA, it is not attacked by the restriction endonucleases Eco R I, Hpa I and Hind III. Defective virions strongly suppress the formation of progeny virus. They do not interfere, however, with the synthesis of HSV-specified thymidine (TdR) kinase.


[Scintigraphic findings of liver and spleen in dermatomyositis]. In 3 patients with dermatomyositis scintigraphic examination of liver and spleen revealed in every case diseases of the liver such as hepatoma, hepatomegaly and damage of the liver parenchyma. Therefore scintigraphic examination of liver and spleen in patients with dermatomyositis is suggested to exclude primary or secondary affections of these organs.


Immune inhibition of virus release from herpes simplex virus-infected cells. By treatment of herpes simplex virus-infected cells with virus antiserum with or without complement, the yield of infectious extracellular virus was significantly reduced. This was shown to be due to an immune alteration of the cell membrane which inhibited release of virus particles from the infected cells and not due to neutralization; both type-common and type-specific antigens of herpes simplex virus were involved. The phenomenon was also evident with antisera directed against cell determinants. The experimental findings are presented and their significance in the immunological defense mechanisms of the body and in viral immunotherapy is discussed.


Some histochemical studies on the prostate, urethral and bulbourethral glands of the one-humped camel (Camelus dromedarius). The histochemical localization of carbohydrates, ribonucleoproteins (RNA), lipids, some hydrolytic enzymes, succinate and lactate dehydrogenase and acetylcholinesterase were investigated in the prostate, urethral and bulbourethral glands of the camel. These glands probably secrete carbohydrate-protein complexes. In the bulbourethral glands, they are sulphated mucopolysaccharides. RNA was seen in the cytoplasm of the prostate and urethral glands. Neutral lipids were cytoplasmic and present in moderate amounts in the prostate and urethral glands and in traces, in the bulbourethral gland. Acid phosphatase-containing granules were abundant in the prostate, moderate in the urethral glands and in traces in the bulbourethral glands. Alkaline phosphatase was observed in the apical cytoplasm of the prostate and bulbourethral glands and in the ducts of the urethral glands. ATPase and adenosine 5-monophosphatase were seen in the basal laminae and interstitial tissue. In the urethral glands, adenosine 5-monophosphatase was distributed diffusely in the cytoplasm. Succinate dehydrogenase was seen in the urethral and bulbourethral glands. Varying degrees of lactate dehydrogenase activity was observed in all the glands. Acetylcholinesterase was confined to neural elements. The pars disseminata and the urethral glands were considered as two distinct glandular zones along the pelvic urethra. The significance of these histochemical results is discussed.


Agarose gel filtration of concentrated fecal extracts containing virus-like particles associated with hepatitis A. Virus-like particles shown to be associated with hepatitis A infection were recently visualized by immune electron microscopy in human and chimpanzee acute-phase fecal extracts. Agarose gel filtration of concentrated chimpanzee fecal extracts containing those 27-nm diameter particles separated more than 99% of the high molecular weight fecal impurities into two major peaks as determined by absorbance at 280 and 260 nm. The hepatitis A-associated particles were found between these two peaks by both immune electron microscopy and a new immunoradiometric assay.


Contact-mediated changes in ATPase activity at the surface of primary cultured hepatoma cells. Hepatoma cells grown in monolayer culture display certain alterations in their Mg-ATPase activity present on the cell surface as a function of time during a exponential growth. Levels of enzyme estimated biochemically and expressed as activity per cell increase as the cell population density increases. Histochemical investigation shows that Mg-ATPase activity is located intensively on the surface of cell contact and the activity is not encountered on the cell surface facing the free space. No enzyme activity is detected histochemically on the cell surface of sparse culture. Deposits of acidic polysaccharide are also seen on the surface of cell contact.


Serologic comparison of hepatitis A antigen to echovirus 25 isolate CR69(076) recovered from a patient with viral hepatitis. A serologic comparison was performed by immune electron microscopy between hepatitis A antigen (related to the MS-1 strain of hepatitis A virus) and a viral isolate, CR69(076), from the stool of a patient in Costa Rica with viral hepatitis. CR69(076) was not antigenically related to hepatitis A antigen but was related to echovirus 25. Thus, it is unrelated to the MS-1 strain of hepatitis A virus.


Rapid eye movement (REM) activity in normal and autistic children during REM sleep. Thirty normal children (aged 3--68 months) and 16 autistic children (aged 36--62 months) were recorded during nonmedicated sleep and data pertaining to rapid eye movements (REM) were measured during the first three REM periods of the night. When time of night from which data were gathered was held constant, normal children showed a significant relationship between age and the organization of eye movements into discrete bursts. When autistic children were compared to age-matched normal controls, they showed an immaturity in this phenomena, their results being similar to those found in children less than 18 months of age. Such an immaturity could result from dysfunction at a number of diverse levels and sites in the central nervous system.


Fosfomycin resistance: selection method for internal and extended deletions of the phosphoenolpyruvate:sugar phosphotransferase genes of Salmonella typhimurium. Selection for resistance to the antibiotic fosfomycin (FOS; L-cis 1,2-epoxypropylphosphonic acid, a structural analogue of phosphoenolpyruvate) was used to isolate mutants carrying internal and extended deletions of varying lengths within the ptsHI operon of Salmonella typhimurium. Strains carrying "tight" ptsI point mutations and all mutants in which some or all of the ptsI gene was deleted were FOS resistant. In contrast, strains carrying ptsH point mutations were sensitive to FOS. Resistance to FOS appeared to result indirectly from catabolite repression of an FOS transport system, probably the sn-glycerol-3-phosphate transport system. Resistant ptsI mutants became sensitive to FOS when grown on D-glucose-6-phosphate, which induces an alternate transport system for FOS, or when grown in the presence of cyclic adenosine 3',5'-monophosphate. A detailed fine-structure map of the pts gene region is presented.


Properties of phenylalanine hydroxylase of cultured hepatoma cells. The kinetic and immunologic properties of phenylalanine hydroxylase of adult rat liver were compared to the properties of the similar enzyme present in cultured H4-II-E-C3 hepatoma cells. The enzymes from the two sources could not be distinguished by the Km values for either phenylalanine or 6,7-dimethyltetrahydropterin. Analysis by double immunodiffusion showed that phenylalanine hydroxylase from the two sources had identical immunologic determinants, but immunotitrations revealed a small but significant difference between the enzyme of the normal adult rat liver and the enzyme of cultured hepatoma cells. The results of double immunodiffusion and immunotitration experiments indicated also that the increased levels of phenylalanine hydroxylase seen in the hepatoma cells grown in the presence of hydrocortisone resulted from the accumulation of enzyme protein, but it could not be decided whether this accumulation resulted from an increased rate of synthesis or decreased rate of degradation.


31P NMR studies of the arginine kinase reaction. Equilibrium constants and exchange rates at stoichiometric enzyme concentration. The arginine kinase reaction, the reversible transfer of the terminal phosphoryl group of ATP to L-arginine, has been investigated by the technique of 31P NMR at catalytic and stoichiometric concentrations of the enzyme. Three of the four substrates, ATP, ADP, and P-arginine produce easily distinguishable resonances in the 31P NMR spectrum, thus permitting a determination of equilibrium constants from the integrated areas of the resonances. From the linewidths, the exchange rates between reactants and products may be evaluated. At pH 7.25 and a temperature of 12 degrees, the equilibrium constant at catalytic enzyme concentration: Keq = [MgADP] [P-arginine]/[MgATP] [L-arginine], is found to be 0.10 +/- 0.02 and that at stoichiometric enzyme concentration: K'eq = [E-MgADP] [E-P-arginine]/[E-MgATP] [E-arginine] to be 1.56 +/- 0.5. Thus, as the enzyme concentration increased, the production of P-arginine is increasingly favored. From the NMR line shapes in the presence of excess enzyme, the rate of the single step, the transfer of the phosphoryl group on the surface of the enzyme is found to be 192 +/- 15 s-1 in the forward direction, i.e. from E-MgATP, and 154 +/- 15 s-1 in the reverse direction from E-P-argine. At 12 degrees and pH 7.25, the rate of the overall reaction in the forward direction was determined from kinetic measurements to be 19 s-1, an order of magnitude slower than the rate measured by NMR. It can, therefore, be concluded that the interconversion of substrates on the surface of the enzyme is not the rate-determining step in the overal reaction. From the equilibrium constants and other known data the dissociation constant of P-arginine from its enzyme complex can be determined and is found to be 100 muM.