Character of the tumors developing in young hamsters by the mixed cell transplantation of para-adeno-virus 12 and SV40 tumors-the similarity to the tumors induced by para-adenovirus 7. Two mixtures of either allogeneic PARA-adenovirus 12 tumor cells or adenovirus (Adeno) 12 tumor cells and SV40 tumor cells in an equal number of 1 multiplied by 10(7) cells were transplanted subcutaneously at two sites of 10 young hamsters. The former two tumor cells were transplantable in nearly half of the transplants and the latter tumor cells in all of the transplants. By the mixture of PARA-Adeno 12 and SV40 tumor cells, 10 tumors were grown, 7 of which were mixed with Adeno and SV40 types and 3 were solely SV40 type. The other 10 tumors produced by the mixture of Adeno 12 and SV40 tumor cells revealed SV40 type only. Morphologically, Adeno type resembled the tumor induced by Adeno, and SV40 type by SV40. The results suggest that the existence of SV40 genetic information in Adeno type tumor cells is indispensable to the formation of mixed tumor in allogeneic tumor cell system. The similarity between the tumors developed by the mixed cell transplantation and those induced by PARA-adeno 7 was discussed both viro-immunologically and morphologically.


Bronchial carcinoid accompanied by thyroid adenomas and adrenal adenomas. A large tumor massively occupying the left pleural cavity had the findings of both typical carcinoid and onococytoma which were thought to be of bronchogenic origin. The ultrastructural observation of the tumor revealed a mixture of rod-shaped granules in addition to usual round neurosecretory ones. In the nuclei of dark cells of the oncocytoma, a latticed or hatched structure was detected. Besides two adenomas and hyperplastic foci of large acidophilic cells in the thyroid, a black adenoma and cortical adenoma in the adrenal gland, were detected. Moreover, there was an ectopic adrenal gland in the retroperitoneum. Briefly it was suggested that the bronchial carcinoid presented may be related to multiple endocrine adenomatosis.


Labyrinthine input to the vestibular nuclei of the awake cat. The labyrinthine input to the vestibular nuclei was investigated in 24 awake cats. Stimulus consisted of electrical shocks given through bipolar silver wire electrodes, implanted in the utricular and lateral ampullar nerves. Throughout the vestibular nuclei, single units were recorded extracellularly with glass micropipettes filled with Fast Green. The tracts of the penetrating electrodes were identified histologically. In all four nuclei units responding to both labyrinths outnumbered unilaterally responding neurones with certain differences between the individual nuclei. Excitatory as well as inhibitory responses were observed, polysynaptic being more common than mono- or disynaptic ones. No monosynaptic contralateral responses were seen. The latency distribution of contralateral responses closely mirrored that of ipsilateral responses within each nucleus. Both excitatory and inhibitory responses fell into relatively segregated populations, based upon latency distribution. This implies separate pathways for labyrinthine input to the vestibular nuclei.


Histological observation of the brain of Tay-Sachs disease with seizure and chronic DPH intoxication--report of an autopsy case. The patients was a 3-year-old boy with psychomotor retardation and attacked by seizures since 8 months of age. On funduscopy, the maculla presented a cherry-red spot. Serum hexosaminidase A activities were as low as 8.2 percent. Both parents were carriers. The patient was diagnosed as classical Tay-Sachs disease by neurological examination. Diphenylhydantoin was continuously given for 2 years and 2 months till his death. Autopsy revealed swelling of the cerebrum, atrophy and sclerosis of the cerebellum, hepatomegaly and mild enlargement of the lymph nodes. Histologically, the cerebrum showed ballooned swelling of nerve cells, slight gliosis and demyelination, while cerebellar Purkinje's cells and granular cells were degenerated and disappeared. The cerebellar cortex showed small focal spongy degeneration. By electron microscopy, membranous cytoplasmic bodies were found in the nerve cells. The change of brain observed in this case were interpreted as a combined result of (i) essential change to classical Tay-Sachs disease, (ii) ischemic change due to frequently repeated seizures, (iii) chronic toxicity by long-term anticonvulsant administration.


A new type of mucolipidosis associated with hereditary thrombocytopathy and color blindness. Autopsy findings of a 22-year-old Japanese male who showed the symptoms of both mucopolysaccharidosis and sphingolipidosis are reported. The patient had a gargoyle-like face, bone change with cherry-red spot and absence of mucopolysacchariduria, and moreover accompanied by hereditary thrombocytopathy and color blindness. Autopsy findings were almost the same as those of mucopolysaccharidosis, histochemically and electron microscopically. Unique findings were, however, present in the hepatocytes, another inclusion containing dense fine granuloreticular structures was found electron microscopically. Some foamy cells in the lymph nodes, liver including sinusiodal cells, bone marrow and spleen contained intracytoplasmic sudanophilic substance in the form of moderate electron dense globules by electron microscopy. The outstanding finding of the enzymatic activity was the decrease of beta-galactosidase in the liver and brain.


Mechanical and metabolic effects of diazoxide in rat uterus. Diazoxide relaxed both polarized and depolarized rat uterus. The drug also conteracted the contractions elicited by Ca2+ in a competitive manner. The relaxing effect was associated with an increase in the tissue level of cyclic AMP. This metabolic effect of diazoxide was inhibited by propranolol-treatment and in preparations from reserpinized animals, while the mechanical effects were only partially reduced. Diazoxide was also found to increase the release of tritium from preparations preloaded with [3H]-noradrenaline. It is suggested that diazoxide  may induce some of its mechanical and metabolic effects by releasing the adrenergic transmittor substance noradrenaline. An effect of diazoxide on the Ca2+-metabolism is also probable.


Effect of adenosine compounds (ATP, cAMP) on renin release in vitro. Renin release by surviving canine renal cortical slices incubated media with ATP or cAMP at concentrations of 5 X 10(-5)--5 X 10(-3) M has been studied. Both adenosine compounds were significantly increasing renin release. A linear correlation was observed between their dose and the renin activity of the medium. The difference between the effects of ATP and cAMP appeared to be caused by phosphodiesterase, since the difference was eliminated if to the medium containing cAMP 5 X 10(-2) M theophylline, a phosphodiesterase inhibitor was added.


Circadian rhythm of hypothalamo-hypophyseal-adrenal activity in the chicken. Corticosteroid production, ACTH content in the adenohypophysis and CRF content in the median eminence were studied in vitro and in vivo in the 3-weeks-old chicken according to SAFFRAN and SCHALLY in four different parts of the day: at 6 and 12 a.m., and 6 and 12 p.m. The chicken adrenals in vitro produced only corticosterone in fluorimetrically measurable amounts, with the maximum during the day and the minimum in the evening and at night. Sensitivity to ACTH of the adrenals in vitro was higher during the day than in the evening or at night. After paper-chromatographic separation, steroid hormones equivalent to corticosterone and cortisol could be detected fluorimetrically in the plasma. The "cortisol" level was about 2-2.5 times higher than the corticosterone level. Plasma "cortisol" level displayed the peak at 6 a.m. and the minimum at 12 p.m., while corticosterone at 12 a.m. and 12 p.m., respectively. ACTH content in the adenohypophysis was lowest at 6 a.m. then increased gradually until 12 p.m. CRF level in the median eminence fell during the forenoon, was lowest at noon, to rise up to 6 p.m. and persist at a high level between 6 p.m. and 6 a.m. Thus, in the 3-week-old chicken the functioning of the hypothalamo-hypophyseal-adrenal system displays a circadian rhythm characteristic of animals showing diurnal activity.


Seasonal rhythm in the hypothalamo-hypophyseal-adrenal function of the chicken. The seasonal rhythm in hypothalamo-hypophyseal-adrenal function was studied in 3-week-old, meat-hybrid chickens, bred under standard conditions, CRF content in the median eminence and ACTH content in the adenohypophysis showed the maximum in February, the minimum in August, to return practically to the February level by November. Adrenal corticosterone production was maximum in February decreased by May and stayed at the low level for the further months of the year. The increase in autumn of the CRF content in the median eminence and of the ACTH content in the adenohypophysis, with unaltered corticosterone production suggests a reduced functioning of the mechanisms regulating hypothalamic release. It is assumed that the winter maximum in the activity of the corticotropic system is due to the endogenous annual rhythm.


Suppression of the excitatory adrenergic neurotransmission; a possible role of cholinergic nerves in the retractor penis muscle. The excitatory adrenergic response to transmural nerve stimulation of the isolated retractor penis (rp) of dog, cat, horse, boar, elk, bull, ram and goat, as well as the evoked release of 3H from dog rp preloaded with 3H-noradrenaline were studied. In untreated preparations a low concentration of scopolamine could markedly increase the excitatory adrenergic response. On the contrary, physostigmine per se suppressed this response and so did acetylcholine. Scopolamine or atropine totally prevented these suppressions. They also immediately restored the suppressed responses in spite of continuous presence of physostigmine and/or acetylcholine. In the rat rp there was no suppression of the response by physostigmine. Physostigmine decreased the evoked release of 3H and this effect was counteracted by scopolamine. The rp of dog, gelding, boar, hog, bull and ram contained considerable amounts of noradrenaline and also of acetylcholine. It is suggested that one action of cholinergic nerves in the rp is muscarine suppression of the excitatory adrenergic neurotransmission.


Frequency-dependence of 3H-noradrenaline secretion from human vasoconstrictor nerves: modification by factors interfering with alpha-or beta-adrenoceptor or prostaglandin E2 mediated control. Isolated superfused field  stimulated human omental arteries and veins, preincubated with 3H (-)-noradrenaline (NA) were used to study the frequency dependence of NA secretion and of the mechanisms  for its local feedback control. 3H-NA secretion per shock was found to be basically a simple hyperbolic function of the stimulation frequency from 1 to 30 Hz, as long as secretion was restricted by prostaglandin E2 (PGE2). In the absence of restriction, or during facilitation, 3H-NA secretion per shock reached its maximum at 10 Hz and then declined at 30 Hz, indicating 'overload' in some link in the secretory mechanism. 3H-NA secretion was depressed by exogenous NA and by PGE2, and enchanced by isoprenaline, phentolamine and by blockade of PGE2 formation. Most of these effects were inversely related to the stimulation frequency. Attempts were made to study interactions between the different control mechanisms and to evaluate possible in vivo consequences of disturbance of adrenergic neuroeffector transmission by interference with the local control of the secretory mechanisms.


Comparison of two muscle relaxant drugs on human sleep: diazepam and parachlorophenylgaba. The effects of two muscle relaxant drugs of different chemical structures, parachlorophenylgaba and diazepam, were investigated in the sleep of normal young adults. Parachlorophenylgaba did not show any significant and dose-related effect. In contrast, diazepam induced clear modifications, including a decrease of stage 4 and of REM-sleep, an increase of spindles and a decrease of K potentials in stage 2, a decrease of rapid eye movements in REM-sleep, and finally a decrease of body movements. In addition, the recovery of REM-sleep upon withdrawal of the drug was much faster than the recovery of stage 4. These results add a new contribution to known differences in the pharmacological profiles of these two compounds ; for diazepam, the possible role of GABA mediated monoaminergic mechanisms is discussed.


The characteristics of human plasma lipoprotein metabolism: influence of low caloric diet on hyperlipidemia. We have classified hyperlipidemia into three groups according to different levels of VLDL-TG, and postulated the effect of low calorie-diets on plasma TCh are as follows: 1) Low calorie-diets are effective on VLDL-TG in every type of hyperlipidemia, except hyperlipidemia with VLDL-TG under 180 mg/100ml; 2) There was a reciprocal relation between the decreased amount of plasma TCh and the change VLDL-TG induced by the diet; 3) Catabolism of VLDL was accelerated by an increased esterification of VLDL-FC; 4) In the the group with VLDL-TG less than or equal to 180 mg/100ml, the LDC showed a high level of TCh: protein ration, and in the group with VLDL-TG greater than or equal to 260 mg/100ml, there was to be low level of TG: protein ratio in VLDL: 5) In hyperlipidemia with plasma TCh unchanged by the diet LDL-TCh increased significantly without any increase in LDL protein and LDL-TG.


The origin and fate of HDL.

The origin and fate of HDL. High density lipoproteins are synthesized both in the liver and intestine. All apoproteins can be synthesized in the liver and intestine except the "C" apoproteins which could not be synthesized by the intestine. After injection of 125I-HDL its different apoproteins are removed from the circulation at a similar rate. Most of the HDL is removed by the liver and the degradation of the protein moiety of HDL was localized to the lysozomes of the parenchymal cells. Cholesterol feeding alters the distribution of lipoprotein fractions and results in the appearance of a new intermediate density class, in which all HDL apoproteins are present in addition to apo-B. Cholesterol feeding also results in an alteration of apoprotein composition of HDL.


Regulatory function of delta6 desaturate -- key enzyme of polyunsaturated fatty acid synthesis. The delta6 desaturation of unsaturated acyl-CoA is the first reaction involved in the normal biosynthesis of all polyunsaturated fatty acids families in animal microsomes. Due to this key position it can regulate the biosynthesis of the fatty acids of the series. The reaction is modified by competition with substrates and products, ATP, and acyl-CoA acceptors. Dietary glucose and fructose inhibit the enzyme whereas protein diets and essential fatty acid deficient diets enhance the reaction independently of hormonal effects. The enzyme is sensitive to hormones concentration. Insulin enhance the reaction but the effect is eliminated by protein synthesis inhibition. Hyperglucemic hormones as glucagon, and epinephrine depress the activity of the delta6 desaturase by reactions triggers by an increase of cAMP concentration. The lateral relation of linoleic or alpha-linolenic microsomal elongation is insensitive to insulin, glucagon, epinephrine and protein. All these effects have been proved by either in vivo experiments or cell culture using linoleic or alpha-linolenic acids as substrates.


Electron paramagnetic resonance studies of ethanol on membrane fluidity. An optimum level of fluidity in the membrane appears to be important for some physiological functions. The present studies examined the effects of ethanol in erythrocyte and brain membrane preparations from Swiss Webster mice using electron paramagnetic resonance (EPR) techniques with a fatty acid spin label. The spectral parameter measured was the order parmeter, S, an index of membrane fluidity. Synaptosomal membranes were more fluid than myelin but less fluid than mitochondrial membranes. In low concentrations of 1 and 2 mg/ml of ethanol membrane fluidity was increased in mitochondrial, synaptosomal, and erythrocyte membranes. Dose-related increases in membrane fluidity were also observed at higher concentrations of 4, 8 and 16 mg/ml of ethanol for all of the membranes except myelin. These data indicate that non-lethal concentrations of ethanol may increase membrane fluidity in vivo.


Regulatory factors of acetaldehyde metabolism in isolated rat liver mitochondria. The factors affecting acetaldehyde oxidation were studied by direct measurement of redox changes of NADH and cytochrome c in isolated rat liver mitochondria which contain the natural complement of mitochondrial substrates and co-factors, etc. Thus, this system affords a quantitative model for mitochondrial acetaldehyde metabolism simulating in vivo conditions. The activity of acetaldehyde dehydrogenase, as measured by the turnover number of cytochrome c, k3, depends upon the substrate concentration in a complex way. It reaches a maximum below 0.033 mM acetaldehyde and decreases abruptly at higher acetaldehyde concentration, interpreted here to be due to substrate inhibition. When mitochondria undergo hypotonic swelling, the maximal value of k3 is lowered by a factor of 15 and the substrate inhibition largely disappears. When mitochondria are stripped of the outer membrane and are suspended in pyrophosphate buffer, similar effects are obtained. It is concluded that acetaldehyde oxidation in mitochondria is dependent upon the state of mitochondria and the substrate concentration, and that the mitochondrial metabolism of acetaldehyde cannot be accurately predicted from in vitro data of solubilized enzyme.


Changes in receptor sensitivity of the cerebral cortex and liver during chronic ethanol ingestion and withdrawal. The evidence was reviewed supporting the noradrenergic sub- and supersensitivity hypothesis of ethanol withdrawal pathogenesis. New data indicates that the cAMP generating system linked to noradrenergic receptor sensitivity is in a steady state in the brain during ethanol withdrawal since the cAMP levels were not different from controls in vivo. Propranolol blocking experiments indicated the beta-adrenergic receptor is a necessary link in the increased cAMP response induced by ethanol withdrawal. The changes in sensitivity observed in the ethanol withdrawn rats were reproduced by acute and chronic reserpinization.


The metabolism of biogenic amines in experimental animals and in human subjects during acute and chronic administration of ethanol. Ethanol has been shown to induce a shift in catecholamine metabolism peripherally from normally oxidative pathways to reductive pathways. The mechanisms of this effect may result from competitive inhibition of aldehyde dehydrogenase by acetaldehyde. The shift in metabolism cannot be found in brain and alterations in catecholamine function may reflect changes in the turnover of these amines.


Alcohol, sleep and cerebrospinal fluid changes in alcoholics: cyclic AMP and biogenic amine metabolites in CSF. Relative to baseline, one day of alcohol ingestion by chronic alcoholics demonstrated significant increases in SWS, decreases in REM sleep and decreases in CSF levels of cyclic AMP and 5-HIAA.


Asymptomatic gonorrhea.

Asymptomatic gonorrhea. Asymptomatic gonorrhea is significantly common in both males and females. A patient with one sexually transmitted disease (STD) is more likely to have an asymptomatic gonorrheal infection. A physician seeing a patient with an STD should obtain a culture for Neisseria gonorrhoeae to check for a possible asymptomatic infection. Routine culturing of this sexually active population will help to control the increasing incidence of gonorrhea. Appropriate treatment should be initiated by the physician who makes the diagnosis of gonorrhea.


LDH isoenzyme electrophoretic pattern differences between colloid and other adenocarcinomas of the colon. LDH isoenzyme electrophoretic patterns have been studied in 27 adenocarcinomas of the colon. The different content of M monomer in poorly differentiated tumors is statistically significant (P less than 0.001) when compared to well-differentiated tumors of the colon. Such a difference has not been found for two Broders groups of well-differentiated tumors. A commentary is made on the possibilities of histochemistry and cytochemistry providing better classification of these tumors.


Effects of blood glucose levels on aspirin-induced gastric mucosal damage. In female rats aspirin-induced gastrin mucosal damage was increased and glycoprotein synthesis decreased by fasting and by insulin administration. Glucose added to the drinking water during the fasting period reduced mucosal damage and increased glycoprotein synthesis to control levels. Alloxan diabetes did not affect mucosal damage or glycoprotein synthesis. Alloxan diabetes plus insulin restored blood glucose levels to normal, and susceptibility to aspirin damage and glycoprotein synthesis were also normal. Alloxan diabetes plus fasting restored blood glucose levels to normal but increased aspirin-induced mucosal damage and reduced glycoprotein synthesis. In vitro incubation of gastric mucosal homogenates showed that diburyryl cyclic AMP and theophylline inhibited glycoprotein synthesis but dibutyryl cyclic GMP had no significant effects. The importance of an adequate supply of glucose to the gastric mucosa and the effects of cyclic nucleotides on glycoprotein synthesis are discussed.


Epidemic keratoconjunctivitis at a Vietnamese refugee camp in Florida. During the summer of 1975 an ongoing outbreak of conjunctivitis occurred among Vietnamese refugees temporarily housed at a U.S. mainland camp. Twenty-two per cent of surveyed refugees gave a history of the disease and 10% were documented as having clinical conjunctivitis at the time of the survey. Fifty-six per cent of documented cases were in children less than 10 years of age. The attack rate among American camp personnel was 4%. Comprehensive microbiologic analysis revealed multiple potential pathogens in most cases, but the recovery of adenovirus 8 (AV8) in 81% of cases cultured within two weeks of onset implicated AV8 as the principal cause of the epidemic.


Effect of maternal isoxsuprine on fetal rabbit lung biochemical maturation. Isoxsuprine was given intramuscularly for 24 hours to pregnant rabbits at 26 to 27 days' gestation. There was no effect on fetal weight within one day of the last injection. Fetuses of isoxsuprine-treated mothers had higher lung lecithin (6.67 micron per gram +/- 2.81) and higher tracheal wash lecithin/sphingomyelin (L/S) ratios (5.85 +/- 4.5) than did untreated fetuses--lung lecithin (5.35 micron per gram 1.23) and tracheal wash L/S (4.08 +/- 2.3). In contrast, phosphorylcholine glyceride transferase activity of the fetal lung was less in isoxsuprine-treated fetal lung tissue.


Differences in growth of transplants of liver, liver hyperplastic nodules, and hepatocellular carcinomas in the mammary fat pad. Transplantation of fragments of normal rat liver autologously and isologously into the inguinal mammary fat pad permitted survival for up to 75% of grafts for 38 weeks, the longest interval studied. Similarly transplanted hepatocarcinomas grew rapidly and progressively in this site. Neither autologous or isologous transplants of liver hyperplastic nodules displayed obvious growth, although like normal liver, they also persisted for up to 38 weeks. Some persisting hyperplastic cells retained certain characteristic features, but others appeared to revert to a normal morphology. Thus, there is a stage in which hyperplastic cells do not possess the progressive growth ability of neoplastic cells and appear to be capable of reversion to a normal phenotype.


Effect of oxygen tension on cellular energetics. Simultaneous measurements of the mitochondrial [NAD+]/[NADH], the cytoplasmic [ATP]/[ADP] x [Pi], and the respiratory rate were carried out in suspensions of cultured kidney cells in a range of defined oxygen tensions. The results show that as the extracellular oxygen concentration falls there is a decrease in the respiratory rate, which is accompanied by a decrease in the [ATP]/[ADP] and a progressive reduction of cytochrome c. Even at low O2 tensions the mitochondrial respiratory chain between the NAD couple and cytochrome c remains at near equilibrium with the ATP synthesizing reactions. It is concluded that limited oxygen supply affects cellular metabolism at much higher concentrations than the P50 value for the oxygen dependence of respiration, but the respiratory rate remains relatively unchanged due to compensatory changes in the [ATP]/[ADP] X [Pi] and progressive reduction of cytochrome c. These metabolic changes may form a basis for the phenomenon of tissue oxygen sensing at near physiological oxygen tensions.


Considerations on the preneoplastic lesions of the mammary gland. The general characteristics of the preneoplastic lesions of the human mammary gland, as they are known through histologic description, are outlined, and data obtained from the experimental analysis of mammary gland preneoplasia in five areas of endeavor are discussed. Results obtained with transplantation procedures and aimed at defining the growth potential of hyperplastic outgrowths are reported. Information derived from the study of events able to induce benign hyperplastic outgrowths or their malignant transformation in the murine mammary gland is summarized. Attempts to predict neoplastic transformation in morphologically hyperplastic epithelium of human and rodent glands are discussed. The present status of efforts toward the prophylaxis of preneoplastic lesions of the mammary gland is described. Considerations of the relationship between preneoplasia and tumor dormancy conclude the presentation.


Estimates of quantal-release and binomial statistical-release parameters at rat neuromuscular junction. Quantal-release and binomial statistical-release parameters were examined in the isolated rat diaphragm phrenic nerve preparation. The muscle resting potentials were reduced by cutting the muscle fibers to prevent muscle action potentials and contractions. The cutting technique was modified to allow persistent observation of miniature end-plate potentials (MEPPs) and end-plate potentials (EPPs) with intracellular recording techniques. Direct estimates of quantal release were examined and compared with predicted binomial and Poisson distributions. The results indicate that release is binomial when the nerve is stimulated with low- or high-frequency stimuli. The indirect method of estimating quantal release (variance method), which assumes release is described by a Poisson distribution, seriously overestimates quantal release. The binomial analysis indicates that the statistical store is small (less than 90 quanta) and that most of these quanta are released with each nerve impulse.


Rat adrenal cell sensitivity to angiotensin II, alpha-1-24-ACTH, and potassium: a comparative study. Rat adrenal glomerulosa and fasciculata-reticularis cell sensitivity to comparable molar doses of angiotensin II (AII) (2.4 X 10(-12) to 2.4 X 10(-4) M) and ACTH (alpha-1-24-adrenocorticotropin) (3.5 X 10(-13) to 3.5 X 10(-8) M) as well as small increments in potassium (K+) (3.7 to 13 meq/liter) was investigated. Glomerulose cells responded to physiological levels of AII (2.4 X 10(-10) M) and alpha-1-24-ACTH (3.5 X 10(-12) M), whereas an increment of as little 0.3 meq/liter in medium K+ also significantly increased aldosterone production. Of the three stimuli, alpha-1-24-ACTH caused the greatest aldosterone rise (11 times control). Fasciculata-reticularis cells responded only to alpha-1-24-ACTH. Whereas the threshold sensitivity was no lower than with glomerulosa cells, the maximum response was significantly greater (63 times control). These findings are consistent with findings documented in vivo in man, suggesting that the control of aldosterone secretion is similar in these two species.


Cardiopulmonary sympathetic afferent influences on renal nerve activity. Cardiopulmonary sympathetic afferent nerves may affect renal control of intravascular volume by influencing renal sympathetic nerve activity. This influence was evaluated in alpha-chloralose anesthetized, vagotomized, sino-aortic denervated cats. When the afferent nerves were activated with a single electrical stimulus, the renal nerve responded with an excitatory burst of activity followed by a long period of inhibition. This response had characteristics of a supraspinal reflex. Repetitive stimulation of the sympathetic afferent nerve either inhibited or excited renal nerves and increased or decreased systemic blood pressure. The direction of these changes depended on stimulus parameters. No obligatory correlation in the direction of change of nerve activity and blood pressure was observed. Activation of cardiopulmonary sympathetic afferent nerves by intravascular volume expansion inhibited renal nerve discharge. Inhibition was elminated by sectioning the sympathetic afferent nerves. Volume expansion had no effect on lumbar sympathetic discharge monitored simultaneously with renal nerve activity. This observation suggests specificity of reflex influences of these afferent nerves on the kidney. In conclusion, cardiopulmonary sympathetic afferent nerves can reflexly alter renal nerve activity, and therefore may affect renal control of intravascular volume.


Sleep and hibernation in ground squirrels (Citellus spp): electrophysiological observations. Electroencephalogram (EEG), electrooculogram, electromyogram, and electrocardiogram were recorded from ground squirrels (Citellus beldingi and C. lateralis) during the summer and also during the hibernation season. Summer recordings revealed that the animals spent an average of 66% of the 24-h period asleep (49% of the 12-h light period and 84% of the 12-h dark period); 19% of the total sleep time (TST) consisted of rapid-eye-movement (REM) sleep, and 81% of TST consisted of slow-wave sleep (SWS). Recordings obtained during the hibernation season showed that hibernation was entered through sleep, but the distribution of sleep states was different than in euthermic sleep. During the early entrance when brain temperature (Tbr) was between 35 and 25 degrees C, the animals were asleep 88% of the time, but only 10% of the TST was spent in REM sleep. The EEG amplitude declined with decreased Tbr so that classical sleep stages could not be identified below a Tbr of 25 degrees C. The frequency of the EEG increased as Tbr decreased; but activity in the 0-4 cycles/s band occupied the majority of the record even at a Tbr of 10 degrees C. Below a Tbr of 10 degrees C the EEG was isoelectric except for intermittent bursts of spindles. It was concluded from these and other results that the entrance into hibernation represents an extension of the thermoregulatory adjustments that occur during SWS.


Paget's disease of the vulva. A histogenetic study of five cases including ultrastructural observations and review of the literature. Five cases of extramammary Paget's disease of the vulva were studied by histologic, ultrastructural, and immunologic methods. In one of these cases, the associated invasive adnexal carcinoma was studied in a similar fashion. In all five cases, carcinoma-in situ of the sweat glands was observed. In two cases, the glands were identified as aporcrine. In the other three it could not be determined whether they were apocrine or eccrine. In one case tumor cells were seen extending from a focus of underlying carcinoma-in-situ of sweat glands through the excretory duct into the surface epithelium. The ultrastructural findings indicated that the Paget's cells as well as the cells of the invasive carcinoma were adenocarcinoma cells, but the findings were not conclusive as to whether the cells were apocrine or eccrine. There was no evidence of origin of the Paget's cells from keratinocytes. We believe that the bulk of evidence favors an extraepidermal origin for extramammary Paget's disease. The tumor is most commonly derived from the secretory portion of sweat glands, probably either of apocrine or eccrine types, or the dermal or poral portion of the sweat duct. Some cases of Paget's disease are derived from other types of glands.


Complete devascularization of hepatic lobe with nonresectable hepatoma. Complete dearterialization of the entire liver was carried out in three patients with nonresectable liver cell carcinoma in whom the right or left portal vein had already obstructed due to tumor thrombosis or invasion. Although remittent fever ensued for two to three weeks postoperatively in all patients, hepatic infaction never occurred in the devascularized hepatic lobes. The effect of the treatment on tumorous lesions appeared satisfactory. The results indicated that the tumor-bearing liver can tolerate nearly complete deprivation of hepatic arterial and portal blood in at least one lobe of the liver if it is not simultaneous.


Response of plasma corticosteroids and circulating leukocytes in cattle following intravenous injection of different doses of adrenocorticotropin. The relationships among exogenous adrenocorticotropin (ACTH), plasma corticosteroids, and circulating leukocytes were studied in 7 lactating cows. Blood samples were obtained from jugular cannulas at -2, -1, and 0 hours before ACTH was injected (base line) and 0.25, 0.50, 1, 2, 3, 6, and 24 hours after injection. Plasma corticosteroids were increased progressively by injecting doses of ACTH between 1 and 200 IU. Plasma corticosteroids reached peak concentrations between 15 and 30 minutes and returned to base line within 1 to 3 hours after 1, 5, and 10 IU doses of ACTH were injected, but required as long as 6 hours after injection of 100 and 200 IU. Base line counts of circulating leukocytes averaged 7.3 X 10(3) cells/mm3 and remained unchanged after injecting 0 and 1 IU of ACTH (P less than 0.05). Significant dose-dependent increases in circulating leukocytes were detected within 2 hours after administering 5, 10, and 100 IU of ACTH. Responses to 100 and 200 IU were similar. The average concentration of leukocytes increased up to 6 hours after ACTH administration and returned to base line values within 12 to 24 hours in cows injected with 5 and 10 IU, but not until 48 hours in cows injected with 100 and 200 IU of ACTH. In contrast to the delayed and sustained responses observed for leukocytes, corticosteroid responses were rapid and transient. Moreover, the administration of 200 IU of ACTH was considered to increase circulating corticosteroids and leukocytes beyond that found in dairy cattle exposed to stress associated with overmilking, acute coliform mastitis, or parturition.


Detection of turkey coronaviral enteritis (bluecomb) in field epiornithics, using the direct and indirect fluorescent antibody tests. In Minnesota, efforts have been made over the past 10 years to eliminate turkey coronaviral enteritis (TCE, bluecomb) by controlled depopulation and decontamination with a rest period before restocking. In 1973, clinical observations indicated that bluecomb was restricted to one limited area in Minnesota. Five epiornithics occurred during late 1973 and 1974, involving 5 different farms in this limited geographic area. During 1975, 3 epiornithics of TCE were investigated, involving 185,000 turkeys in 17 flocks, of which approximately 17,000 died. Naturally infected turkeys representing 7 operations between 1973 and 1976 were examined by both the direct fluorescent antibody test and indirect fluorescent antibody test (IFAT). The direct fluorescent antibody test detected coronaviral antigen in intestinal tissues during the acute phase of the disease, and the IFAT was highly useful in detecting TCE serum antibodies of turkey flocks that had recovered and were potential carriers. Therefore, an IFAT surveillance program was instituted for replacement flocks on farms where clinical epiornithics of TCE had occurred in 1974 through 1976. Operation 5 involved TCE epiornithics over a 2-year period and illustrate the importance of complete depopulation with an intensive decontamination program.


The prognosis of lung cancer originating as a round lesion. Data from the Philadelphia pulmonary neoplasm research project. A population of 6,027 men 45 or more years of age was screened every 6 months for 10 years with chest photofluorograms and questionnaires regarding symptoms. Although volunteers, they were similar to older men in the general population with respect to age, race, and smoking habits. Of 121 men who developed lung cancer after the beginning of observation, 48 had neoplasms appearing as round lesions at the time of radiographic detection. Only 8 per cent of the 48 men survived 5 years or more, a rate identical to that of men in whom cancer first appeared in some other form. There was an inverse relationship between initial size of the cancer and survival. Two thirds of the tumors were squamous cell carcinomas. Comparison with the literature suggests that selection accounts for the favorable prognosis of round lung cancers in hospital-based series.


Type III hyperlipoproteinemia: diagnosis in whole plasma by apolipoprotein-E immunoassay. Because the cholesterol-rich very low density (VLD) lipoproteins of subjects with type III hyperlipoproteinemia are distinctively enriched in apolipoprotein E, a radial immunodiffusion assay for apolipoprotein E in whole plasma was developed. Its diagnostic usefulness was tested in randomly selected (n = 174) and hyperlipidemic (n = 61) subsets of an adult employee population and a hyperlipidemia clinic referral group (n = 63), which included 18 patients with well-documented type III hyperlipoproteinemia. Apolipoprotein-E levels were normally distributed among the random population subset, were equal between the two sexes, and increased little with age. The mean and 99th percentile values were 24.6 and 40.1 mg/dl, respectively. All subjects with type III patterns as assigned by standard criteria from both population (n = 4) and referral sources exceeded this 99th percentile (chi +/- SD = 54.7 +/- 9.7 mg/dl). Hence a plasma apolipoprotein-E concentration exceeding 40 mg/dl appears diagnostic of type III hyperlipoproteinemia, representing the first application of an apolipoprotein immunoassay to improved diagnosis of the hyperlipoproteinemias.


Type III hyperlipoproteinemia: paradoxical hypolipidemic response to estrogen. Exogenous estrogens (ethinyl estradiol, 1 microgram/kg body weight per day), which stimulate triglyceride production in normal women and those with endogenous hypertriglyceridemia, were found to exert a paradoxical, hypolipidemic effect in six subjects (five women, one man) with type III hyperlipoproteinemia on diets both of normal and of fat-free, high-carbohydrate composition. Moreover, very low-density (VLD) lipoprotein lipid and apolipoprotein composition and electrophoretic mobility became normal during estrogen administration in these subjects. Levels of normal VLD lipoproteins remained mildly to moderately elevated in a type IV lipoprotein pattern. Estrogen withdrawal promptly restored the type III pattern with its abnormal enrichment of VLD lipoproteins with apolipoprotein E (the arginine-rich peptide). These findings suggest that estrogens facilitate the assimilation of chylomicron and VLD lipoprotein remnants, a defect that appears likely to represent the metabolic abnormality underlying type III hyperlipoproteinemia.


Primary type V hyperlipoproteinemia. A descriptive study in 32 families. We have studied 32 kindreds identified by propositi with primary type V hyperlipoproteinemia. The clinical presentation, metabolic associations, and natural history confirm the distinctiveness of primary type V hyperlipoproteinemia from other lipoprotein abnormalities. Although the underlying defect(s) remains unknown, several factors such as obesity, alcohol, drugs, and diet are able to modify the glyceridemia, the major manifestation of this disorder. Abnormalities of postheparin lipolytic activity or its subfractions do not appear to be involved in the pathogenesis of primary type V. The prevalence of hyperuricemia, diabetes, pancreatitis, and xanthomatosis appears high among the 32 propositi; the last two entities are much less prevalent in the relatives, even among those relatives classified as hyperglyceridemic. There is no evidence in these families of excessive coronary artery disease prevalence. Triglyceride levels are positively associated with age in this population, especially among women. Average triglyceride levels were lower for women than for men before age 50.


Evacuation of orbital hematoma by continuous suction. An orbital hematoma, unusual in its delayed onset and association with indirect trauma, is described. Successful therapy consisted of continuous suction through a polyethylene catheter. Various methods of therapy, conservative and surgical, have been described in the ophthalmic literature, but no previous report was found of the employment of continuous suction.


The deficiency of a lysosomal acid hydrolase in two clones derived from the human lymphoblastoid line F137 after mutagen treatment. Two clones (out of a total of 181 clones tested) derived from the human lymphoblastoid (lymphoid) line F137 after mutagen treatment were found to be deficient in a lysosomal acid hydrolase. The clone N32 derived from EMS-treated F137 is deficient in N-acetyl hexosaminidase A and B but contains normal levels of N-acetyl hexosaminidase C and low levels of an enzyme resembling N-acetyl hexosaminidase S. Thus the enzyme deficiency in this clone appears to resemble the so-called Sandhoff variant of Tay-Sachs disease, a disease inherited as an autosomal recessive condition. The clone G3 derived from MNNG treated F137 is deficient in alpha-galactosidase A. This clone resembles the situation in X-linked Fabry's disease. Karyotype analysis of the clones failed to reveal any chromosome rearrangement or losses of chromosomal material that might have accounted for the mutations and it is suggested that a single point mutation might in each case account for the loss of enzyme activity. No storage of the natural substrates of the two enzymes could be demonstrated in the clones.


Family studies with the chromosome 9 markers ABO, AK1, ACONs and 9qh. We have failed to measure the recombination fraction between 9qh and the ABO:AK1 linkage group. Since the total male map distance along chromosome 9 is likely to be at least 120 cM, calculated from male chiasmata counts (Hultén, 1974) they may well not be within measurable distance of each other. ACONs does not lie between ABO and AK1, but there is so little information on this marker from family studies that it could lie almost anywhere else on chromosome 9 and might be within measurable distance of 9qh. The map of chromosome 9 based on studies on families with normal karyotypes is summarized in Fig. 1. We hope to improve this map in a subsequent paper based on observations on families with abnormal karyotypes.


Lysosomal enzymes in human lymphoblastoid lines: unusual characteristics of RAJI and DAUDI. The lysosomal enzymes N-acetyl hexosaminidase, alpha-galactosidase and alpha-mannosidase vary in electrophoretic mobility in different human lymphoblastoid (lymphoid) lines. The relative mobilities of these three enzymes and three out of four other lysosomal enzymes tested correlate well with each other. The patterns appear to be relatively stable characteristics of each line. The lines RAJI and DAUDI show a strikingly fast electrophoretic mobility for all of these enzymes. N-acetyl hexosaminidase is also markedly deficient in DAUDI.


Antiviral activity of ribavirin in rotavirus gastroenteritis of mice. Ribavirin was inactive against the rotavirus of murine gastroenteritis; this may be due to the presence of guanosine inhibitors in the gut.


Growth properties of mecillinam-resistant bacterial variants in urine. Resistant variants are often formed when susceptible Enterobacteriaceae are grown in media containing mecillinam. These variants have not yet been detected in patients or identified as a cause of treatment failure in limited clinical trials. Mecillinam-resistant organisms were formed by 5 out of 13 urinary isolates of Enterobacteriaceae incubated in urine containing mecillinam. The mean generation time of these five variants in urine containing therapeutic concentrations of mecillinam was 3.2 times that of normal organisms in antibiotic-free urine. When three of these five resistant variants were subcultured in antibiotic-free urine, the generation time, morphology, and antibiotic susceptibility returned to normal. On antibiotic-free agar medium, all five mecillinam-resistant variants readily reverted to the "susceptible" form and were therefore more likely to be phenotypic rather than genotypic mutants. In a second series of experiments, the flushing effect of the bladder on the clearance of organisms was partly simulated by frequent subculture in urine. Under these conditions, the cell density of cultures of all bacteria remained high during 10 subcultures over 52 h. However, bacterial populations progressively decreased in urine containing mecillinam until none could be detected at 28 h or thereafter. The slow growth rate of mecillinam-resistant variants may explain why detectable numbers of these organisms fail to colonize the urinary tract during treatment. Other factors may reinforce the postulated effect of the reduced growth rate in vivo.


Latent herpes simplex virus infections in sensory ganglia of mice after topical treatment with adenine arabinoside and adenine arabinoside monophosphate. Adenine arabinoside (Ara-A) and Ara-A monosphosphate (Ara-AMP) ointments were able to prevent the fatal outcome of herpes simplex virus (HSV)-induced skin infection of the lumbosacral area in hairless mice. Ara-A and Ara-AMP had no irritating effect on the skin, but in a number of animals a protracted healing time of the skin lesions after the treatment was observed. The compounds conferred only a partial protection against the establishment of latent HSV infection in the spinal root ganglia of the treated animals. The immune response, as judged from levels of HSV-specific neutralizing serum antibody titers, was not impaired by the antiviral treatment.


Production of uricase by Candida tropicalis using n-alkane as a substrate. Production of uricase (urate oxidase, EC 1.7.3.3) by n-alkane-utilizing Candida tropicalis pK233 was studied. Although the yeast showed very low enzyme productivity under growing conditions on glucose or an n-alkane mixture (C10 to C13) (less than 2 U/g of dry cells), enzyme formation was enhanced markedly in an induction medium consisting of potassium phosphate buffer, MgSO4, uric acid, and an n-alkane mixture (47 U/g of dry cells) or glucose (21 U/g of dry cells). Of the carbon sources tested, the n-alkane mixture was the most suitable for enzyme production. Appropriate aeration also stimulated uricase formation. In addition to uric acid, xanthine, guanine, adenine, and hypoxanthine were also effective for inducing uricase. Under optimum conditions, the maximum yield of the enzyme was 91 U/g of dry cells. Uricase thus induced was localized in the microbodies of the yeast.


Granular cell myoblastoma in Nigerian Igbos. Three cases of granular cell myoblastoma occurring in the Igbos of Nigeria are reported. In this ethnic group the tumor has shown several typical features as follows: it is rarely diagnosed preoperatively; it is found more frequently in the oral cavity and breast than in most other sites; and it is in the maxilla of girls that it usually presents congenitally. A unique presentation in this ethnic group occurred in the perineum. The lesion found in this site appears to be the first to be reported in the literature.


Three cases of rare tumors of the mammary gland. The cases are reported of three patients with rare tumors of the mammary gland: one osteogenic sarcoma, one benign mixed tumor and one reticulum-cell sarcoma of the breast. The pathological and clinical aspects of these tumors are discussed with reference to the literature.


Chemical composition and swelling of normal and osteoarthrotic femoral head cartilage. II. Swelling. Studies of the equilibrium, partition, and diffusion of tritiated water were carried out on normal and osteoarthrotic cartilage as well as on cartilage from which proteoglycans had been extracted chemically. All the water was found to be freely exchangeable in both normal and degenerate specimens. The diffusiviity of the water was equal to about 40% of the value in free solution, with an activation energy equal to that in free solution. It was concluded that the swelling of degenerate tissue is not due to any changes in the state of the water, but to a failure of the damaged collagen network to oppose the swelling pressure of the proteoglycans. Swelling similar to that observed in fibrillated cartilage was also found in initially normal specimens treated with collagenase.


The influence of various methods of sensitization on delayed hypersensitivity to Bordetella pertussis in the Sprague Dawley Rat. In the rat, the injection of Bordetella pertussis produces, after prior sensitization, a delayed hypersensitivity inflammatory reaction within the pleural cavity. The influence of various methods of sensitization on this hypersensitivity was studied in the Sprague Dawley rat. The reaction was very variable according to the experimental conditions used. Optimal sensitization was obtained following the injection of antigen mixed with complete Freund's adjuvant into the dorsal surface of the paws.


Influence of cyclic nucleotides and a phosphodiesterase inhibitor on in vitro human blood neutrophil chemotaxis. Cyclic AMP and dibutyryl cyclic AMP as well as several 8-substituted analogs of cyclic AMP inhibited in vitro human blood neutrophil chemotaxis. Cyclic GMP enhanced chemotaxis. 8-substitution on the cyclic GMP nucleus markedly dampened the cyclic GMP effect. SQ 20,009, a potent phosphodiesterase inhibitor when tested alone inhibited chemotaxis; it potentiated the inhibition or enhancement of chemotaxis when combined with cyclic AMP or cyclic GMP, respectively.


Bilateral diaphragmatic paralysis in association with neurogenic disease. A patient had complete diaphragmatic paralysis and his disease progressed to include systemic neurological deficit. The cause is unknown. Appropriate diagnostic tests are emphasized, and a discussion of potential etiologies with a review of the literature of idiopathic diaphragmatic paralysis is presented.


Spontaneous cytomegalovirus mononucleosis with conjunctivitis. Cytomegalovirus (CMV) was isolated from the conjunctiva of a patient with mild unilateral catarrhal conjunctivitis associated with the typical clinical features of CMV mononucleosis syndrome in a previously healthy adult. The diagnosis was confirmed by a marked rise in the complement fixation antibody titers. Prospective studies are needed to determine the frequency of conjunctival involvement in CMV mononucleosis and the relation, if any, to sporadic cases of conjunctivitis.


Qualitative x-ray spectrometric study of E. histolytica trophozoite nuclei. Qualitative X-ray energy spectroscopical analysis was performed in two strains of Entamoeba histolytica trophozoites: HK-9; NIH strain, and HM-2; IMSS strain, cultured in axenic and monoxenic conditions, respectively. The trophozoites were fixed in buffered-glutaraldehyde, embedded in Epon, thick sectioned )50-200 nm), and mounted on nickel grids. X-ray emissions were analyzed with an Ortec X-ray energy detector and multichannel and analyzer adapted to a transmission electron microscope. The study was concentrated on the refractive intranuclear bodies, characteristically numerous in axenically grown E. histolytica trophozoites. Nuclei of trophozoites with those intranuclear bodies contained phosphorus, calcium, and magnesium in greater amounts than nuclei not containing those structures. This was true for all axenically grown trophozoites and two monoxenically grown trophozoites which contained four intranuclear bodies. It is concluded that the composition of the refractive intranuclear bodies in E. histolytica trophozites does not appear to change with the culture conditions or with the composition of the culture media.


Routine titration of foot-and-mouth disease virus suspensions by analytical ultracentrifugation 2nd communication: sedimentation equilibrium method. A routine method for the determination of the virus concentration in FMD virus cultures and vaccines was developed. This method was based on sedimentation equilibrium in the analytical ultraviolet scanning ultracentrifuge. The virus suspension was first clarified. The virions were then sedimented in a preparative ultracentrifuge. The resuspended virions were diluted in a Cesium chloride solution and brought to equilibrium in the density gradient generated in the analytical ultracentrifuge. The optical density of the virus band was measured by the UV scanning system. A calculation procedure was developed to compute the density at the limits and at the maximum of the virus band. The virus concentration expressed as weight, was calculated for the original virus suspension.


Location of antigens associated with bovine leukemia virus. Antigens associated with bovine leukemia virus (BLV) infection (gp 44/55 and p23) were identified by neutralization and immunofluorescent antibody (FA) tests. Only serums containing antibody to gp 45/55 neutralized the virus; antibody to p23 did not. These results indicate that gp 44/55 may be located in the virus envelope. In indirect FA tests, both a cytoplasmic fluorescence and a membrane fluorscence were detected in BLV-infected cells. The serum used in the FA test was then absorbed with either p23 or gp 45/55. The results of these absorption tests indicate that the membrane fluorescence maybe associated with gp 45/55 and the cytoplasmic fluorescence may be associated with p23.


Inhibition of bovine leukemia virus release by antiviral antibodies. Peripheral blood lymphocytes from bovine leukemia virus (BLV) infected cattle were grown in vitro with serums from BLV-infected and uninfected cattle, sheep and rabbits. Only serums from infected animals which had antibody to gp 45/55, the major glycoprotein antigen, inhibited the release of virus from the cells. Although viral antigens could be detected in the cells themselves, none were detected in the supernatants of cultures grown with these serums. Normal serums and serums with antibody only to p23, the internal BLV antigen, did not inhibit virus release. To identify the factor responsible for virus release inhibition, 2 inhibitory serums were absorbed with either gp 44/55, p23 or tissue culture cells from BLV-infected and uninfected cells lines. The results indicated that antibody to gp 45/55 is the factor responsible for virus release inhibition.


Effect of tunicamyein on the morphogenesis of Semliki Forest virus and Rous sarcoma virus. Semliki Forst Virus (SFV) infected chick embroyo cells (CEC) incubated in the presence of Tunicamyein, a drug which specifically inhibits glycosylation, were studied under the electron microscope. No extracellular virions were produced although intracellular viral nucleocapsids in paracrystalline form were detected. Under these conditions the structures, referred to as type 1 and 2 cytopathic vacuoles (cpv-1 and cpv-2, respectively) as well as membraneous spherules at the plasma membrane could still be observed. CEC rpoductively infected and transformed by Rous sarcoma virus (RSV) incubated for 24 hours in the presence of Tunicamycin (1 microgram/ml) still showed budding and extracellular parties. Intracellur nucleocapsid could could not be detected.


The polypeptide composition of avain infectious bronchitis virus particles. Egg grown avian infectious bronchitis virus (IBV) centrifuged on sucrose density gradients was found to consist of a major virus peak of density 1.17 to 1.18 g/cm(3). Three different IBV strains were examined and no morphological differences were detected between virus particles of different densities or from different strains. The polypeptides of the different density virus particles from the three IBV strains were analysed on polyacrylamide gels. In all cases 7 polypeptides were observed, although there were differences in the proportions of these polypeptides in particles of different densities and those from the different strains. The polypeptides have been called VP1 (molecular weight 130,000), VP2 (105,000), VP3 (97,000), VP4 (81,000), VP5 (74,000), VP6 (51,000) and VP7 (33,000). Additional polypeptides were produced if slightly harsher treatments were used.


Detection of transmissible gastroenteritis virus neutralising antibody in cats. High titres of neutralizing activity to transmissible gastroenteritis virus (TGEV), a porcine coronavirus, were found in sera and peritoneal fluids from cats infected with feline infectious peritonitis (FIP). A small proportion of cats, from a hospital population unaffected by FIP, also had neutralising activity. Procedures to remove non-specific viral inhibitors, including treatment by heat inactivation, trypsin, sulphydryl reagent and kaolin absorption were unsuccessful. The active component was unable to neutralise another porcine coronavirus, haemagglutinating encephalomyelitis virus or the porcine enterovirus, Talfan. Gel filtration of feline sera and peritoneal fluid demonstrated high levels of the neutralising activity in the area corresponding to 7S IgG, which could be removed by absorption with specific anti-IgG serum and these properties are suggested to be consistent with those of antibody. These findings imply that there is a coronavirus in cats which is antigenically related to TGEV and its possible nature is discussed.


Ocular manifestations of focal dermal hypoplasia syndrome. Although 40 percent of patients with the focal dermal hypoplasia syndrome have ocular abnormalities, to our knowledge, only one case of this condition has been previously described in the ophthalmic literature. Two additional cases are presented, and the systemic and ocular findings are reviewed.


Malignant fibrous histiocytoma of the orbit. A 31-year-old woman had a mass in the posterior inferior orbit that progressively increased in size for almost two years. Histopathologic examination of an orbital biopsy specimen showed pleomorphic neoplastic cells arranged in a storiform pattern. The tumor cells were composed of hyperchromatic nuclei with prominent nucleoli and admixtures of fibroblasts. Electron microscopy demonstrated histiocyte-like cells with complex infoldings of plasma membrane, prominent mitochondria and golgi, and free ribosomes. Fibroblast-like cells displayed abundant rough endoplasmic reticulum and adjacent collagen fibrils. The diagnosis was malignant fibrous histiocytoma. An exenteration was performed and postoperative systemic chemotherapy with doxorubicin hydrochloride and methotrexate sodium sulfate was commenced. There was no evidence of recurrence two years later.


Herpesvirus in sensory and autonomic ganglia after eye infection. In herpesvirus hominis (HVH) infections, virus harbored in the sensory ganglia is now thought to be the main source of recurrent infection at peripheral sites. Experimental HVH infection of the external eye in rabbits produces an acute infection and then latent infection of the trigeminal ganglion. In this study, acute infection of the automatic ganglia serving the eye (superior cervical and ciliary) as well as trigeminal ganglia occurred after HVH inoculation of rabbits' corneas with a herpes type 1 strain (RE). Latent virus infection was detected in the trigeminal ganglion of one of five animals tested six months after initial infection. Since the superior cervical and other autonomic ganglia serving the eye become infected during acute herpes simplex virus infection of the external eye in rabbits, it is possible that these ganglia are also sources of reinfection in recurrent herpetic disease of the eye. Following the initial eye disease with this virus strain, HVH shedding could not be demonstrated even after induction attempts by topically applied epinephrine or systemic use of cyclophosphamide. Thus, establishment of latent HVH infection in the ganglia and chronic shedding of virus into the external eye is not a constant feature of this animal model, but may depend on the specific strain of herpesvirus used.


Herpes simplex uveitis in immune rabbits. Priming effect of nonherpetic uveitis. Systemic immunization of rabbits with herpes simplex virus (HSV) had two opposite effects on the outcome of subsequent efforts to produce primary HSV uveitis, the difference depending on whether or not the rabbits had had nonherpetic uveitis before the HSV challenge. In normal eyes, systemic immunization with HSV provided complete protection against the production of primary uveitis by an intraocular injection of HSV; but in eyes that had had a bout of experimentally induced nonherpetic uveitis before the challenge, the same systemic immunization was not protective. In these eyes, an immune-mediated uveal inflammation developed. Nonherpetic uveitis had apparently "primed" the eyes of the HSV-immune rabbits for subsequent immune-mediated HSV uveitis.


Acute peripheral facial palsy. Part of a cranial polyneuropathy? In 14 of 16 consecutive patients with acute peripheral facial palsy, one or more (up to four) other nerves were involved. The nerves affected in addition to the facial nerve were as follows: trigeminal (ten patients), vestibular (eight), cochlear (six), vagus (one), and upper cervical (five). Virus was not isolated from any of the patients. A fourfold increase or decrease in complement-fixing antibody titers was present in eight patients (in four, varicella-zoster; in one, varicella-zoster and mumps; in two, cytomegalovirus; in one, mumps). Further, two of the patients with varicella-zoster antibodies showed clinical signs of herpes zoster oticus. About one fourth of all patients had an increase of ESR and of alpha2-globulins in serum, and two thirds of them had increased gamma-globulins in CSF. Acute peripheral facial palsy seems to be part of a cranial polyneuropathy and may be caused by a viral infection.


Preweaning mortality in the pig. Pathological findings in piglets dying between birth and weaning. During a survey of preweaning mortality 538 piglets which died between birth and weaning were autopsied. The results of laboratory examinations permitted a division of the findings into a number of syndromes which were considered to be associated with the immediate cause of death. The non-infectious conditions such as trauma, starvation and suffocation were the most common. Seventy-eight piglets were included in the trauma group because of depressed cranial bone fractures or visceral rupture leading to haemorrhage into the serous cavities. An absence of alimentary tract contents was detected in each of the 92 cases of starvation, fatty metamorphosis of the liver was found in 28 while the hepatocyte cytoplasm of the remainder stained uniformally. Cyanosis and visceral congestion and haemorrhage were the main features observed in the 159 deaths ascribed to suffocation. Bacterial septicaemias and infections with enterotoxic strains of E. coli were the most prevalent infectious conditions. E. coli and beta haemolytic streptococci were the most common causes of septicaemia, being isolated from 36 and 25 cases, respectively. The post-mortem findings in these cases were non-specific except for fibrinous polyserositis which was observed in 11 of the carcasses from which E. coli was recovered. Twenty-six piglets yielded E. coli in heavy pure culture from the upper small intestine and the most common serogroup involved was 08. Encephalomyocarditis virus infection was associated with the deaths of 19 piglets. Pathologically, it was characterised by myocardial degeneration and a variable influx of mononuclear inflammatory cells. No significant lesions were found in 41 piglets.


Studies on poxvirus isolated from a magpig in Queensland. An avian poxvirus was isolated from a black-backed magpie in Queensland. The virus produced pocks on the chorio-allantoic membrane of chicken embryos and lesions on the skin of chickens. Comparison by passive haemagglutination test and neutralisation test indicated that the virus was related more closely to pigeonpox virus than to fowlpox virus.


Intracardial gas bubbles of altitude after negative pressure breathing. The influence of negative pressure breathing on the appearance of intracardial gas bubbles at a subsequent decompression to altitude was investigated in five subjects using the precordial Doppler ultrasound technique. Every subject was tested for a suitable exposition that, after 30 min oxygen breathing at surface, caused both intracardial bubbles and decompression sickness. An identical exposition followed a week later, except that oxygen breathing at the surface now included negative pressure breathing. In all cases, negative pressure breathing caused a delay of the onset of both intracardial bubbles and bends and, in some cases, neither bubbles nor bends appeared at all. The total amount of bubbles was always less after negative pressure breathing. Thus, negative pressure breathing in connection with decompression may reduce the amount of intracardial bubbles and the risk of decompression sickness. This may be of importance in diving procedures and in the construction of diving devices.


[The fructose induces "glycogenosis". III. Histochemical and morphometrical studies of glycogen metabolism in mouse liver after fructose overload (author's transl)]. After feeding fructose for 7 days rat liver cells show an accumulation of glycogen, a high activity of glucose-6-phosphatase combined with a SER- and RER-reduction. This result was reviewed by mouse liver cells using histochemical and morphometrical methods. 60% fructose in drinking water was given mice as only nutritional source. Controls had free access to Altromin-R-standard diet and drinking water. Glycogen and glycogen metabolizing enzymes are demonstrated in the course of an 1-14 days fructose diet. After a 7 days diet liver tissue was analysed morphometrically. Feeding of fructose leads to a high glycogen content, combined with a high activity of glycogen-phosphorylase and glucose-6-phosphatase in the liver parenchyma of mouse. Glycogen-synthetase activity falls to a low level. The SER and RER and the peroxisomes are reduced. The single volume of the hepatic nucleus is decreased and the hepatocellular chondrioma is transformed in a smaller number of larger mitochondria. Compared with the rate the analysed organelles and enzymes of mouse liver show only slight quantitative differences. The increase of glucose-6-phosphatase and simultaneous reduction of endoplasmic reticulum-membranes is illustrated by the dynamic structure of endoplasmic reticulum-membranes, which adapt to metabolic changes. The variable turnover of different parts of endoplasmic reticulum-membranes seems to be very important.


Cytochrome c oxidase activity in T/t6 (balanced lethal) mutant mice. R-1 (1450g) and R-2 (25,000g) liver fractions from T/t6 and B6CBAF1 hybrid mice were analyzed for their protein content, mitochondria concentrations, and activities of three respiratory-chain enzymes of the mitochondrial inner membrane: cytochrome c oxidase (ferrocytochrome c: oxygen oxidoreductase, E.C. 1.9.3.1.), alpha-glycerophosphate dehydrogenase [L-glycerol-3-phosphate: (acceptor) oxidoreductase, E.C. 1.1.99.5], and succinate-cytochrome c reductase. Only cytochrome c oxidase activity, calculated as units per 10(10) mitochondria, was significantly lower in both R-1 and R-2 fractions of T/t6 mice. Cytochrome c oxidase activity varied greatly among T/t6 mice, as did their liver mitochondria concentrations and body weights. Cytochrome c oxidase activity in the R-1 fraction of T/t6 mice, averaged about 40% lower than in B6CBAF1 mice. alpha-Glycerophosphate dehydrogenase activity was often elevated in T/t6 mice, particularly in the R-2 fraction. The T/t locus, a complex genetic locus on chromosome 17, may contain genes important to the function and biogenesis of mitochondria.


The gross architecture of an antibody-combining site as determined by spin-label mapping. 1. A series of Dnp (dinitrophenyl) nitroxide spin labels was used to map the dimensions of the combining site of the Dnp-binding immunoglobulin A myeloma protein MOPC 315. The method compares the observed e.s.r. (electron-spin-resonance) hyperfine splittings with those calculated on the basis of different postulated motions for the spin label. The analysis is complicated by the sensitivity of the e.s.r. hyperfine splitting to the overall ;tumbling' time of the antibody-hapten complex and the polarity of the spin-label's environment. When these effects are considered quantitatively, it is then possible to determine the degree of mobility of each hapten which is allowed by the shape of the combining site. 2. The dinitrophenyl ring is rigidly held, and the depth of the site is 1.1-1.2nm and has lateral dimensions at the entrance to the site >/=0.6nmx0.9nm. The analysis of the results for spin-labelled haptens with chiral centres allows these lateral dimensions to be refined to 0.8nm and 1.1nm, and it is shown that the site is asymmetric with respect to the plane of the dinitrophenyl ring. 3. A polarity profile of the combining site was also obtained and a positively charged amino acid residue, possibly arginine-95(L) (light chain), was located at the entrance to the site. 4. The binding of Gd(III) to the antibody-hapten complexes results in quenching of the e.s.r. signal of the nitroxide. By using La(III) as a control, the paramagnetic contribution to the quenching is measured. 5. Analysis of the differential quenchings of the enantiomers of two five-membered nitroxide ring spin labels gives two possible locations of the metal-binding site. One of these is equidistant (0.7nm) from each of the three dinitrophenyl aromatic protons, and nuclear-magnetic-resonance relaxation studies, at 270MHz, on solutions of dinitrobenzene, Gd(III) and the Fv fragment (variable region of heavy and light chain) from protein MOPC 315 support this location for the metal site. 6. The e.s.r. and metal-binding data were then compared with the results of a model of the combining site constructed on the basis of framework invariance in immunoglobulins [Padlan, Davies, Pecht, Givol & Wright (1976) Cold Spring Harbor Symp. Quant. Biol.41, in the press]. The overall agreement is very good. Assignments of possible chelating groups for the metal can be made.


Comparison of the dimensions of the combining sites of the dinitrophenyl-binding immunoglobulin A myeloma proteins MOPC 315, MOPC 460 and XRPC 25 by spin-label mapping. The mouse immunoglobulin A myeloma proteins MOPC 315, MOPC 460 and XRPC 25 all possess dinitrophenyl (Dnp)-binding activity. Differences in specificities were shown by measuring the affinities of a variety of haptens. By using a series of Dnp-spin-labelled haptens, the dimensions of the binding sites of the three myeloma proteins were compared by the method described for protein MOPC 315 [Sutton, Gettins, Givol, Marsh, Wain-Hobson, Willan & Dwek (1977) Biochem. J.165, 177-197]. The dinitrophenyl ring is rigidly held in all three sites. The depths of the sites are all 1.1-1.2nm, but there are differences in the lateral dimensions at the entrance to the sites. For protein XRPC 25 these dimensions are 0.75nmx0.8nm, which may be compared with 0.85nmx1.1nm for protein MOPC 315 and >/=1.0nmx1.1nm for protein MOPC 460. The site in protein MOPC 460 is more symmetrical with respect to the plane of the dinitrophenyl ring than in either of the other two myeloma proteins and also allows greater penetration of solvent. In protein XRPC 25 a positively charged residue was located at the entrance to the site, similarly positioned to that reported for protein MOPC 315 [Sutton, Gettins, Givol, Marsh, Wain-Hobson, Willan & Dwek (1977) Biochem.J.165, 177-197]. All three proteins possess lanthanide-binding sites, but only in protein MOPC 315 is there antagonism between lanthanide and hapten binding. However, the effects of the diamagnetic La(III) on the electron-spin-resonance spectra of bound Dnp spin labels in both proteins MOPC 460 and XRPC 25 suggest an interaction between the two sites. Comparison of this effect with that caused by the addition of the paramagnetic Gd(III) enables the distance between the lanthanide- and hapten-binding sites to be calculated. In both proteins MOPC 460 and MOPC 315 the metal site is approx. 1.0nm from the nitroxide moiety of the spin-labelled hapten, but in protein XRPC 25 this distance is at least 2.0nm.


Variable-temperature magnetic-circular-dichroism spectra of cytochrome c oxidase and its derivatives. A detailed study of the effect of temperature on the m.c.d. (magnetic circular dichroism) spectra of cytochrome c oxidase and some of its derivatives was undertaken to characterize the spin states of haem a and a(3). The fully reduced enzyme contains haem a(3) (2+) in its high-spin form and haem a(2+) in the low-spin state. This conclusion is reached by comparing the spectrum with that of the mixed-valence CO derivatives and its photolysis product. The cyanide derivative of the fully reduced enzyme contains both haem a and a(3) in the low-spin ferrous form. The m.c.d. spectra of the fully oxidized derivatives are consistent with the presence of one low-spin ferric haem group, assigned to a, which remains unaltered in the presence of ligands. Haem a(3) is high spin in the resting enzyme and the fluoride derivatives, and low spin in the cyanide form. The partially reduced formate and cyanide derivatives have temperature-dependent m.c.d. spectra due to the presence of high- and low-spin haem a(3) (3+) respectively. Haem a is low-spin ferrous in both. A comparison of the magnitude of the temperature-dependence of haem a(3) (3+) in the fully oxidized and partially reduced forms shows a marked difference which is tentatively ascribed to the presence of anti-ferromagnetic coupling in the fully oxidized form of the enzyme, and to its absence from the partially reduced derivatives, owing to the reduction of both Cu(2+) ions.


Aldolase-like imine formation in the mechanism of action of phosphonoacetaldehyde hydrolase. Phosphonoacetaldehyde hydrolase (EC 3.11.1.1), the bacterial enzyme that catalyses the reaction HCO-CH2-PO(OH)2+H2O leads to HCO-CH3+Pi, is inactivated by borohydride if either phosphonoacetaldehyde or acetaldehyde is present. This supports the suggestion that the substrate forms an imine with an amino group of the enzyme. Such imine formation would labilize the C-P bond in the same way that aldolase and related enzymes labilize C-C and C-H bonds (Scheme 1a).


Chromous ion reduction of mammalian cytochrome oxidase and some of its derivatives. The reduction of cytochrome c oxidase by Cr2+, followed by means of stopped-flow spectrophotometry, exhibits two phases: the faster Cr2+-concentration-dependent reaction has an initial rate constant of 1.1 X 10(4)M-1-S-1, but reaches a rate limit at high concentration of reductant; the slower phase is concentration-independent with a rate of 0.3S-1. The activation energies of the fast and the slow processes are 35 and 71 kJ/mol respectively. The reduction kinetics of the mixed-valence CO complex and the cyanide-inhibited enzyme were compared with those of the fully oxidized forms: both the liganded species have a fast phase identical with that found in the oxidized oxidase. A comparison of the kinetic difference spectra obtained for the fast phase of reduction of oxidized oxidase with those obtained on reduction of the liganded species suggests that the rapid phase arises from the reduction ofhaem a, and the slow phase from the reduction of haem a3.


The effects of amphiphilic cationic drugs and inorganic cations on the activity of phosphatidate phosphohydrolase. 1. Phosphatidate phosphohydrolase from the particle-free supernatant of rat liver was assayed by using emulsions of phosphatidate as substrate. 2. The inhibition of the phosphohydrolase by chlorpromazine was of a competitive type with respect to phosphatidate. The potency of various amphiphilic cationic drugs as inhibitors of this reaction was related to their partition coefficients into a phosphatidate emulsion. 3. The effect of chlorpromazine on the phosphohydrolase activity was complementary rather than antagonistic towards Mg2+. Chlorpromazine stimulated the phosphohydrolase activity in the absence of added Mg2+ and was able to replace the requirement for Mg2+. However, at optimum concentrations of Mg2+, chlorpromazine inhibited the reaction, as did Ca2+. The phosphohydrolase activity was also stimulated by Co2+ and to a lesser extent by Mn2+, Fe2+, Fe3+, Ca2+, spermine and spermidine when Mg2+ was not added to the assays. 4. It is concluded that the inhibition of phosphatidate phosphohydrolase by amphiphilic cations can largely be explained by the interaction of these compounds with phosphatidate, which changes the physical properties of the lipid, making it less available for conversion into diacylglycerol. 5. The implications of these results to the effects of amphiphilic cations in redirecting glycerolipid synthesis at the level of phosphatidate are discussed.


The isolation and characterization of corticotropin from the pituitary gland of the ostrich Struthio camelus. 1. Avian corticotropin (ACTH) was purified from both fresh and aged pituitary glands of the ostrich Struthio camelus. 2. The isolation of corticotropin in pure form involved acid/acetone extraction, NaCl fractionation, CM-cellulose chromatography and Sephadex G-50 chromatography. 3. The hormone preparations from fresh and aged glands behaved as single substances on polyacrylamide-gel electrophoresis, and both preparations were found to consist of 39 amino acid residues, in identical molar proportions for the different amino acids. 4. The isoelectric points of the two hormone preparations were estimated to be in the range pH 8.3-8.7, indicating possible differences in amide content, and the N-terminal amino acid of both preparations appeared to be serine. 5. The hormone preparations from fresh and aged glands exhibited similar biological potencies (73 and 77 i.u./mg respectively), as measured by steroidogenesis in vitro. 6. Apart from possible differences in amide content, the corticotropin preparations obtained from fresh and aged glands appear to be indistinguishable.


Binding proteins for adenosine 3':5'-cyclic monophosphate in bovine adrenal cortex. Five peaks of cyclic AMP-binding activity could be resolved by DEAE-cellulose chromatography of bovine adrenal-cortex cytosol. Two of the binding peaks co-chromatographed with the catalytic activities of cyclic AMP-dependent protein kinases (ATP-protein phosphotransferase, EC 2.7.1.37) of type I or type II respectively. A third binding protein was eluted between the two kinases, and appeared to be the free regulatory moiety of protein kinase I. Two of the binding proteins for cyclic AMP, sedimenting at 9S in sucrose gradients, could also bind adenosine. They bound cyclic AMP with an apparent equilibrium dissociation constant (K(d)) of about 0.1mum, and showed an increased binding capacity for cyclic AMP after preincubation in the presence of K(+), Mg(2+) and ATP. The two binding proteins differed in their apparent affinities for adenosine. The isolated regulatory moiety of protein kinase I had a very high affinity for cyclic AMP (K(d)<0.1nm). At low ionic strength or in the presence of MgATP, the high-affinity binding of cyclic AMP to the regulatory subunit of protein kinase I was decreased by the catalytic subunit. At high ionic strength and in the absence of MgATP the high-affinity binding to the regulatory subunit was not affected by the presence of catalytic subunit. Under all experimental conditions tested, dissociation of protein kinase I was accompanied by an increased affinity for cyclic AMP. To gain some insight into the mechanism by which cyclic AMP activates protein kinase, the interaction between basic proteins, salt and the cyclic nucleotide in activating the kinase was studied.


Biological properties and clinical application of propolis. V. The action of ethanol extract of propolis (EEP) on laboratory animals. Histochemical investigations. In the experiment on rats the ethanol extract of propolis (EEP) injections caused an activation of all experimental enzymes. The greatest effect EEP exerted was on NADPH2 tetrazolium reductase and glucose-6-phosphatase.


Effect of lithium on cyclic 3',5'-adenosine monophosphate formation in brain slices and its influence on phosphodiesterase. The effect of lithium on cyclic 3',5'-adenosine monophosphate formation from precursor [8-14C]-adenine in guinea pig and rat brain slices and its influence on cAMP phosphodieesterase activity were studied. It was demonstrated that lithium stimulated cAMP formation in brain slices in correlation to its concentration. In guinea pig brain slices it stimulated cAMP formation in the presence of neurotransmitters and a phosphodiesterase inhibitor (histamine less than norepinephrine less than 5-OH-tryptamine). In rat brain slices, norepinephrine was also more effective than histamine. Phosphodiesterase inhibition was demonstrated only in preparations in which Triton X 100 was used.


[Receptor occupation and pharmacological activity as demonstrated on opiates (author's transl)]. The availability of highly radioactively labelled opiates with high receptor affinity has made it possible to identify and characterize opiate receptors in vitro as well as in vivo. These investigations revealed that the affinity of the compounds to the receptor decides about the pharmacological potency of the compounds. The differing action of opiate agonists and antagonists are explained by an allosteric receptor model. In vivo experiments showed that the dose range at which opiates induce specific pharmacological actions is identical with those doses at which increasing and finally complete receptor occupancy is obtained. Toxicological aspects of receptor occupation are discussed.


Adjuvant polyarthritis. III. Evidence in support of viral etiology. Intraperitoneal administration of a relatively small volume (0.2 ml) of plasma obtained from Sindbis virus--infected animals containing approximately 480 units of interferon/ml effectively suppressed the development of adjuvant arthritis. Similarly, administration of interferon (624 units/rat/day, ip) prepared in vitro by exposing rat embryo fibroblast culture to poly I:C in the presence of cycloheximide also suppressed the development of adjuvant arthritis without affecting the humoral or the cell-mediated immune response. The findings provide circumstantial evidence to the speculation that a virus or a virus-like organism plays a role in the pathogenesis of this disease.


Reproducibility of measurements of oestrogen-receptor concentration in breast cancer. The reproducibility of measurements of oestrogen-receptor activity has been examined in multiple specimens from a rabbit uterus, a rat mammary tumour and human breast tumours. The relationship between receptor concentration and tumour histology has also been investigated in 11 large primary tumours. In the animal tissues, receptor measurements were relatively reproducible (coefficient of variance: wet wt. basis 16-17%, protein basis 16-21%) but in human breast tumours receptor activity varied considerably (c.v.: wet wt. basis, 22-125%; protein basis, 28-72%). In addition to these variations in receptor activity within tumours, there was a difference between tumours, as demonstrated by an analysis of variance (P less than 0.01). In the 11 primary breast cancers selected for study, the level of receptor activity was related to menopausal status and the tumour content of the specimen. We conclude that the receptor activity detected varies within a tumour and depends upon the tumour content of the biopsy specimen. Predictions based on precise quantitation of receptor concentrations may therefore necessitate replicate tumour sampling and correction for the fraction of non-tumour tissue in each sample.


Electron spin resonance of caeruloplasmin and iron transferrin in blood of patients with various malignant diseases. Electron spin resonance studies have been made of caeruloplasmin and iron transferrin levels in whole blood of healthy controls and patients with a variety of malignant conditions receiving various forms of treatment. A small difference was found in caeruloplasmin level between normal males and females, although normal females receiving contraceptive steroids had an elevated level. No difference was found in iron transferrin level. Various conditions increased the caeruloplasmin and some also decreased the iron transferrin level in patients with malignant disease. These included surgery and the approach of a terminal phase of disease. Once allowance for these factors was made, the remaining small differences in Cu and Fe between patients with either squamous cell carcinoma or breast cancer and controls appeared to have no clinical significance.


Effect of hormone replacement therapy on glucose tolerance in postmenopausal women. Oral glucose tolerance tests were performed on 50 symptomatic postmenopausal women before and after three months of hormone replacement therapy. All patients were randomly allocated to one of five groups treated with various synthetic or so-called naturally occurring oestrogens. Therapy produced a significant deterioration of carbohydrate tolerance with sequential preparations containing 100 microgram of ethinyl oestradiol or graduated doses of mestranol up to 50 microgram. The conjugated equine oestrogen (1.25 mg daily) and oestrogen valerate (2 mg daily) treated groups did not show abnormal glucose tolerance. The decreased glucose tolerance may be due as much to dosage levels as to any metabolic characteristics of the various oestrogens prescribed.


Lipid binding by fragments of apolipoprotein C-III-1 obtained by thrombin cleavage. We have used thrombin to cleave apolipoprotein C-III-1 into two fragments constituting residues 1-40 (apoLP-C-III-A) and 41-79 (apoLP-C-III-B). The lipid binding properties of these fragments with dimyristoyl- and 1-palmitoyl-2-oleoylphosphatidylcholines have been determined using circular dichroic and intrinsic tryptophan fluorescence spectroscopy. The peptide-phospholipid mixtures were fractionated by density gradients of cesium chloride. ApoLP-C-III-A showed disordered structure in the absence and presence of DMPC and no significant amount of peptide-phospholipid complex was isolated. ApoLP-C-III-B showed conformational changes in the circular dichroic spectrum and a shift in the intrinsic tryptophan fluorescence spectrum. Ultracentrifugation in cesium chloride gradients yielded peptide-phospholipid complexes isolated between density 1.10 and 1.18. The molar ratio of lipid to protein was 12:1. The results of these studies and the examination of space filling models of apoLP-C-III provide evidence that an amphipathic alpha helix which contains a nonpolar face and a polar face is the basic structural unit for binding of phospholipid by the plasma apolipoproteins. These results also provide direct evidence that the hydrophobicity of the nonpolar face is important in lipid binding since the nonpolar face of residues 1-40 is considerably less hydrophobic than the nonpolar face of residues 41-79.


Angiotensin I converting enzyme from human plasma. The angiotensin I converting enzyme was purified 101 000-fold to homogeneity from human plasma by a combination of chromatographic and electrophoretic techniques. The enzyme is similar to other angiotensin I converting enzymes. It is an acidic glycoprotein consisting of a single polypeptide chain of molecular weight 140 000 with an isoelectric point of 4.6. The enzyme requires chloride ion for activity and is inhibited by ethylenediaminetetraacetic acid, angiotensin II, bradykinin, bradykinin potentiating factor nonapeptide, and 3-mercapto-2-D-methylpropanoyl-L-proline (SQ-14,225). The purified preparation cleaves bradykinin as well as angiotensin II and hippuryl-L-histidyl-L-leucine. Its specific activity with angiotensin I is 2.4 units per mg and with hippuryl-L-histidyl-L-leucine is 31.4 units per mg.


Effect of coenzymes and temperature on the process of in vitro refolding and reassociation of lactic dehydrogenase isoenzymes. Dissociation and deactivation of the H4 and M4 isoenzymes of lactic dehydrogenase in strong denaturants may be reversed with a yield of reactivation up to 100%. The products of reconstitution are indistinguishable from the native enzymes as far as the Michaelis constants and the dissociation constants for substrate and coenzyme as well as spectral and hydrodynamic properties are concerned. The presence of NAD+ and NADH does not affect either the conformational state of the product of reconstitution, or the kinetics of reactivation, using the pure apoenzymes as a reference. At 20 degrees C the kinetics of reactivation for LDH-M4 in the presence and absence of coenzyme may be quantitatively described by a second-order rate equation (k2 = 23.4 +/- 2.6 mM-1S-1) while LDH-H4 is characterized by a uni-bimolecular reaction sequence (k1 = 1.45 +/- 0.45 X 10(-3)-S-1, k2 = 5 +/- 1 mM-1S-1), in agreement with earlier observations (Rudolph, R., et al. (1977), Biochemistry 16, 3384-3390). Regarding the influence of temperature on the rate of reactivation no significant anomalies are detectable within the range of 0-25 degrees C. The (apparent) activation energies, taken from the linear Arrhenius plots, are 58 kcal/mol for the association reaction of LDH-M4, and 41 kcal/mol for the transconformation reaction of LDH-H4.


Mutations in Escherichia coli altering an apurinic endonuclease, endonuclease II, and exonuclease III and their effect on in vivo sensitivity to methylmethanesulfonate. The levels of endonuclease II, an apurinic endonuclease, and exonuclease III in the parent strains (AB 1157) of Escherichia coli and in various mutants were determined by chromatography on DEAE-cellulose. AB 3027 and NH 5016 lacked endonuclease II and exonuclease III. BW 2001 lacked the apurinic endonuclease and exonuclease III while BW 2007, BW 9093, and BW 9059 lacked only exonuclease III. Deletion mutants BW 9101 and BW 9109 lacked all three enzymes. The latter mutants locate the genes for the two endonucleases in the region of exonuclease III (chith) of 38.2 min (White et al., 1976). All of the mutants which were sensitive to methylmethanesulfonate in vivo lacked exonuclease III, but not all mutants lacking exonuclease III were MMS sensitive. The deletion mutants and NH 5016 were the exceptions.


Purification and characterization of baboon endogenous virus DNA polymerase. An RNA-directed DNA polymerase was purified from baboon endogenous type-C virus by successive column chromatography on DEAE cellulose, phosphocellulose and hydroxyapatite. The purified DNA polymerase has a molecular weight of 68 000, a pH optimum of 8.0, a Mn2+ optimum of 1 mM, and a KCl optimum of 40 mM. The purified enzyme transcribes heteropolymeric regions of viral 60--70 S RNA isolated from different type-C viruses. The purified enzyme is immunologically related to a similarly purified polymerase from the cat endogenous type-C virus RD114.


Conformational changes in yeast tRNATyr revealed by EPR spectra of spin-labelled N6-(delta2-isopentenyl)-adenosine residue. Temperature-induced conformational changes in the anticodon region of yeast tRNATyr were studied by EPR spectroscopy. The spin label 4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl was attached to the N6-(delta2-isopentenyl)-adenosine residue in tRNATyr, previously made reactive by iodination. The labelled tRNATyr gave an asymmetrical triplet spectrum typical of rapidly tumbling nitroxide, with a rotational correlation time (tauc) of 0.65 ns. Spin-labelled tRNATyr was exposed to heating and cooling in three different buffers each with or without MgCl2. In each case the Arrhenius plot of --log tauc vs. inverse absolute temperature gave two straight lines, intersecting at a critical temperature (tcr). Above tcr, the anisotropy of the spectrum was not reduced and the activation energy of motion increased, indicating that the transition is associated with a conformational change of the macromolecule. Transitions in 0.05 M potassium phosphate (pH 8.0) and 0.02 M Tris - HC1 (pH 7.0) were observed at potassium phosphate (pH 8.0) and 0.02 M Tris - Hc1 (pH 7.0) were observed at approx. 37 degrees C. When 0.01 M mgCl2 was present in these buffers, transitions were shifted to 46 degrees and 53 degrees C, respectively. Transitions in 0.01 M sodium cacodylate were observed at temperatures which are significantly lower. Since all these transitions occur at temperatures considerably below those required to melt the helical regions of tRNA, and at least approximately 10 degrees C below those reported to break tertiary interactions, it is supposed that they reflect some reorientation of the anticodon region, e.g. a change in tilt of the bases.


Cytoplasmic and nuclear protein kinases during the cell cycle. Nuclear and cytoplasmic protein kinases were measured during the traverse of synchronous CHO cultures through G1 into S phase. Cells were synchronized by selective detachment of cells blocked in metaphase using colcemid. Nuclei were isolated and the protein kinases extracted from the nuclear preparation with 0.6 M NaCl. This procedure solubilized greater than 90% of the total protein kinase activity present in the nuclear preparation. DEAE chromatography of this extract showed 5 apparently different ionic forms of nuclear protein kinases. The nuclear protein kinases preferred casein and phosvitin to histone as substrates and were cyclic AMP-independent. Nuclear protein kinase activities increased greater than two-fold, when expressed as units of activity per cell nucleus, during G1 phase traverse, concomitant with a 70% increase in nuclear non-histone proteins (those soluble in 0.6 M NaCl). This resulted in only a 40% increase in the specific activities (units/microgram protein in 0.6 M NaCl extractable nuclear fraction) of these enzymes as cells progressed through G1 into S phase. This was in contrast to cytoplasmic cyclic AMP-dependent protein kinase activities which also increased two-fold during progression through G1 phase while total cellular protein increased less than 20%. Activation of, as well as synthesis of, cyclic AMP-dependent cytoplasmic protein kinases during G1 phase suggests a regulatory mechanism for precise temporal phosphorylation, whereas the constant specific activity in nuclear kinases during cell cycle is more compatible with the maintenance of bulk phosphorylation processes in the nucleus.


Myeloperoxidase inactivation in the course of catalysis of chlorination of taurine. Myeloperoxidase (donor: hydrogen-peroxide oxidoreductase, EC 1.11.1.7) was isolated from leukocytes of patients with chronic granulocyte leukemia. In the presence of H2O2 and Cl- at pH 4.0-6.6 the myeloperoxidase catalyses chlorination of taurine to monochloramine taurine and simultaneously undergoes inactivation. The myeloperoxidase inactivation rate depends on the concentration of H2O2 and Cl-: both the initial rate of chlorination and myeloperoxidase inactivation rate increase with increasing concentration of H2O2. However, an increase in concentration of Cl- results in a decrease in enzyme inactivation. At a given H2O2 concentration, myeloperoxidase inactivation is a first order reaction, which implied that the enzyme may react with a substrate a limited number of times.


Some properties of purified phosphoprotein phosphatases from rabbit liver. The activity of two purified homogeneous phosphoprotein phosphatases types P I and P II) (phosphoprotein phosphohydrolase, EC 3.1.3.16) from rabbit liver (Khandelwal, R.L., Vandenheede, J.R., and Krebs, E.G. (1976) J. Biol. Chem. 251, 4850-4858) were examined in the presence of divalent cations, Pi, PPi, nucleotides, glycolytic intermediates and a number of other compounds using phosphorylase a, glycogen synthase D and phosphorylated histone as substrates. Enzyme activities were usually inhibited by divalent cations with all substrates; the inhibition being more pronounced with phosphorylase a. Zn2+ was the most potent inhibitor among the divalent cations tested. The enzyme was competitively inhibited by PPi (Ki = 0.1 mM for P I and 0.3 mM for PII), Pi (Ki = 15 mM for P I and 19.8 mM for P II) and p-nitrophenyl phosphate (Ki = 1 mM and 1.4 mM for P I and P II, respectively) employing phosphorylase a as the substrate. The compounds along with a number of others (Na2SO4, citrate, NaF and EDTA) also inhibited the enzyme activity with the other two substrates. Severe inhibition of the enzyme was also observed in the presence of the adenine and uridine nucleotides; monophosphate nucleotides being more inhibitory with phosphorylase a, whereas the di- and triphosphate nucleotides showed more inhibition with glycogen synthase D and phosphorylated histone. Cyclic AMP had no significant effect on enzyme activity with all the substrates tested. Phosphorylated metabolites did not show any marked effect on the enzyme activity with phosphorylase a as the substrate.