Variability of transmitter quanta released during incorporation of a false transmitter into cholinergic nerve terminals. 1. Electrophysiological techniques have been used to investigate the variability of quanta released during various stages of the incorporation of acetylmonoethylcholine (AMECh) into the transmitter store at the neuromuscular junction.2. Stimulation of the rat phrenic nerve-diaphragm preparation at 3 Hz for 15-25 min reduced the time constant of decay tau(m.e.p.c.) of miniature end-plate currents from 1.61 +/- 0.08 (mean +/- S.E. of mean) msec to 1.32 +/- 0.09 msec; this represents 45% replacement of acetylcholine (ACh) by AMECh. Total replacement of ACh was achieved by a further 30-60 min stimulation which reduced tau(m.e.p.c.) to 0.93 +/- 0.06-msec.3. At the intermediate stage of exchange, the coefficient of variation c.v.(tau) of tau(m.e.p.c.) increased from the control value of 0.094 +/- 0.005 to 0.154 +/- 0.012. The latter value was less than the value (0.29) calculated for two distinct populations of quanta (i.e. pure ACh and pure AMECh quanta). Moreover, frequency distribution histograms of tau(m.e.p.c.) did not reveal a bimodal distribution. Total replacement of ACh by AMECh achieved by the second period of stimulation decreased c.v.(tau) towards the control value.4. Pre-incubation of muscles with MECh (to replace any stored choline) or stimulation in the presence of a cholinesterase inhibitor (to prevent ACh destruction and hence re-uptake of choline) did not produce any greater degree of heterogeneity than was seen in the absence of these procedures.5. The increase in c.v.(tau) observed at the intermediate stage of replacement disappeared spontaneously with a half-time of about 1 hr at 20 degrees C.6. These results are discussed in terms of the vesicle hypothesis.


The metabolic and endocrine effects of circulating catecholamines in fetal sheep. 1. Adrenaline and noradrenaline have been infused into the fetal sheep to produce plasma concentrations comparable to those seen during hypoxia. The effects have been compared with those of isoprenaline and methoxamine and the sensitivity to beta- and alpha-adrenergic antagonists has been followed. 2. Adrenaline caused an alpha-mediated increase in blood glucose that is associated with a fall in plasma insulin concentration. It also caused a beta-mediated increase in plasma lactate, free fatty acid and amino acid concentrations. 3. Noradrenaline was much less effective than adrenaline at eliciting metabolic responses. It had not cause a significant change in plasma glucose concentration, although this was associated with a small increase in plasma insulin concentration. It caused a small rise in the concentration of lactate and free fatty acids in fetal plasma but had no effect on plasma amino acids. 4. The beta- and alpha-adrenergic antagonists propranolol and phentolamine alone were without effect on any of the plasma metabolites or hormones assayed. Isoprenaline increased plasma glucose, lactate, free fatty acid, alanine and insulin concentrations, while methoxamine only increased plasma glucose and lactate, and this was associated with a fall in insulin concentration. 5. The concentration of ACTH in fetal plasma was increased by adrenaline and to a lesser extent by noradrenaline and methoxamine; these were blocked by phentolamine. Isoprenaline also caused a small increase in ACTH. There were no corticosteroid changes associated with the increase in ACTH. 6. The results have been discussed in relation to the adrenergic and pancreatic control of metabolism in the fetal sheep.


Contacts between receptors and electrophysiologically identified neurones in the retina of the larval tiger salamander. 1. Following the intracellular recording of bipolar and horizontal cell responses, each unit was injected with horseradish peroxidase and a histochemical staining used to identify it at the level of the light and electron microscopes.2. Centre-depolarizing bipolar cells made contact with rods and cones at basal and ribbon junctions, the latter being fewer. Centre-hyperpolarizing bipolar cells made the same types of contacts with the receptors, but ribbon junctions predominated.3. It appears, therefore, that there is no fixed relationship between the sign of synaptic transmission from receptors to bipolar cells and the junctional features revealed by present methods for electron microscopy of tissue sections. Accordingly, the reason for the existence of more than one type of contact between receptors and bipolar cells remains to be determined.4. Light microscopy of the peroxidase-injected horizontal cells gave further support to the notion that type B responses are recorded from the cell body, and type A responses from the axon terminal, of a single type of horizontal cell. Electron microscopy showed that the processes (dendrites) originating from the cell body make ribbon and distal junctions with rods and cones, just as shown before for the axon terminals.5. As a result of these observations, it is possible to exclude one of two alternative hypotheses previously proposed to account for the properties of the surround responses recorded from the horizontal cell bodies.


Physiological roles of adenosine derivatives which are released during neurotransmission in mammalian brain. 1. Experiments using synaptosome beds suggested that ATP was released from presynaptic sites and degraded to adenosine in the synaptic cleft and that the resulting adenosine was taken up again into nerve endings where it was re-phosphorylated to ATP. 2. Adenosine derivatives in the synaptic cleft inhibited the postsynaptic potentials in olfactory cortex slices in vitro, presumably by the inhibition of Ca2+ influx into nerve endings which resulted in the reduction of transmitter release. 3. The adenosine derivatives also increased the level of cyclic AMP in the slices under the same conditions as above. 4. Although the nature of the "adenosine receptors" for both functions was remarkably similar, the increase of cyclic AMP did not mediate the inhibitory action, but the presynaptic increase of cyclic AMP induced by adenosine derivatives might mediate the facilitation observed in the olfactory cortex. 5. Possible physiological roles of extracellular adenosine derivatives in mammalian brain were classified, at different sites of action around the synapses, with different time courses and modes of action, directly or via the increase of intracellular cyclic AMP.


5'-adenine mononucleotides in preparations from guinea pig cerebral cortex. Accumulation, translocation and release. Isolated nerve terminals (synaptosome beds) were prepared from the neocortex of guinea pig and their ability to accumulate and release adenine nucleotides was studied. Synaptosome beds prelabelled with [14C]adenosine released newly synthesized [14C] adenine derivatives on superfusion. Electrical stimulation and K+ depolarization gave augmented output of both [14C] adenine derivatives and lactate from the preparations. Action of metabolic inhibitors on this output was examined. During incubation and superfusion, the synaptosomes displayed glycolysis and synthesis of ATP. Supply of adenine derivatives to the nerve terminals also occurred by translocation from other parts of the tissue.


The release of ATP triggered by transmitter action and its possible physiological significance: retrograde transmission. 1. When a slice of electric organ of Torpedo is stimulated and superfused with a solution containing a firefly lantern extract, it is possible to measure the release of ATP after each nerve impulse as a light emission. 2. The postsynaptic action of released ACh induces the release of ATP by the postsynaptic cell. Most of the released ATP is of postsynaptic origin. 3. Ion fluxes associated with depolarization, or depolarization itself, trigger the release of ATP from postsynaptic and presynaptic membranes (synaptosomes). 4. ATP is able to block ACh release; a postsynaptic "retrograde transmission" able to control presynaptic transmitter release is possible.


Mechanisms of depression of neuromuscular transmission by ATP and adenosine. 1. The effects of ATP in the presence of theophylline and imidazole were investigated on the twitch tension of a partially magnesium blocked rat phrenic nerve-diaphragm preparation. Both theophylline and imidazole facilitate the neuromuscular transmission and prevent the effects of ATP. 2. The effects of adenosine in solutions with low calcium concentrations in the frog-sartorius and in solutions with very low calcium concentrations in the rat-diaphragm were studied on the miniature end-plate potentials. Adenosine caused a similar reduction of the frequency of the miniature end-plate potentials in both low and very low calcium concentrations. 3. The results are discussed in relation to the cyclic AMP and calcium hypothesis.


Adenine nucleotides in cholinergic transmission: presynaptic aspects. 1. Isolated nerve terminals (T-sacs and synaptosomes) prepared from the purely cholinergic Torpedo electric organ have been studied for their ability to incorporate and metabolise [2-3H] adenosine and to degrade 5'-AMP to adenosine. 2. A temperature-dependent, saturable uptake system for adenosine was found with kinetic properties similar to nucleoside transport systems in other cells. The uptake system in Torpedo nerve terminals was inhibited by 2'-deoxyadenosine, a known inhibitor of adenosine transport. 3. Intraterminal adenosine is rapidly metabolised to a number of products including AMP, ADP and ATP. 4. Isolated nerve terminals contain considerable 5'-nucleotidase activity, most of which resides on the outer face of the external membrane. The Km of the enzyme is congruent to 5 micron and it is inhibited by a phosphonate analogue of ADP, alpha-beta-methylene-ADP. It is suggested that this 5'-nucleotidase plays an important role in the production of adenosine from a nucleotide pool in the synaptic cleft.


[The role of beta and alpha adrenergic receptors in the lipolytic effect of catecholamines on dog adipocytes (author's transl)]. Pharmacological properties of adrenergic receptors have been investigated in fat cells isolated from omental adipose tissue of the Dog. From the results, the following points can be stated. 1. Lipolysis is markedly enhanced by isoproterenol. This effect is competitively inhibited by  propranolol (a beta-adrenoceptor blocking agent). (Fig 1). 2. The beta 2-sympathomimetic salbutamol is found to have only a slight effect on lipolysis rate (Fig. 2). 3. The epinephrine-induced lipolysis is potentiated by phentolamine (an alpha-adrenoceptor blocking agent) only on large sized adipose cells (mean fat cell size 96.7 +/- 5.3 micrometer; Fig. 5). 4. The isoproterenol-induced lipolysis is partially inhibited by phenylephrine (an alpha-adrenomimetic drug) (Table I). These findings show that beta 1 nature of the receptors involved in the adrenergic control of lipolysis in Dog adipose tissue. Moreover an antilipolytic effect of epinephrine, via alpha-adrenergic receptors, is observed, especially in large adipose cells.


Surface smoothness and marginal fit with phosphate-bonded investments. Two phosphate-bonded investments and one calcium sulfate investment were evaluated for the surface smoothness and marginal fit they impart to gold castings. A modified technique was also evaluated for each phosphate-bonded investment, where the silica sol was not diluted and the spatulation time was reduced. The results of this study lead to the following conclusions: 1. The marginal fits obtained with all four phosphate-bonded methods were comparable to each other and superior to that obtained with the calcium sulfate investment. 2. The presence of nodules on the surface of the castings was more prevalent with the phosphate-bonded investments. However, this effect was not statistically significant. 3. Clinical assessment of the roughness of the castings revealed that all the methods tested produced clinically acceptable castings. 4. Visual observation by five dentists revealed that both the recommended and modified techniques for one of the phosphate-bonded investments (Ceramigold) produced a smoother surface than any other investment tested. Rating of scanning electron microscope photographs (X600) revealed no difference in the surface roughness between any of the castings. Consequently, no definitive relation between investment type or technique and surface roughness was established. 5. No correlation was demonstrated between surface roughness, as evaluated by either clinical observation or scanning electron microscope photography, and marginal fit of the castings.


Some infectious diseases of wild vervet monkeys (Cercopithecus aethiops pygerythrus) in South Africa. Our surveys indicate that nearly all batches of vervet monkeys arriving at the National Institute for Virology from various areas of South Africa, are infected with foamy viruses and there is evidence that intra-urterine infection also occurs. Monkeys from certain areas of South Africa are apparently infected with the protozoal parasite Entopolypoides macaci in a sub-clinical state which becomes active when the monkeys are splenectomised. Serological studies indicate that infections with schistosomes, tick-bite fever (Rickettsia conori), chlamydsiae and occasionally by leptospirae, occur in monkeys in the Kruger National Park. Complement fixation tests for antibodies to R. mooseri and R. prowazeki were also sometimes positive; a finding of doubtful importance. There was a high percentage with positive antibody tests to chikungunya virus in a certain age group suggesting that there had been an outbreak of this disease in the Kruger Park in the late summer and autumn of 1976. This coincided with a human outbreak. Large numbers of vervet and baboon sera were tested for antibodies against SA 12 virus, a polyoma virus. A high percentage of baboons but only a small number of monkeys was found to be positive and it is concluded that SA 12 is probably a baboon virus which is occasionally transmitted to monkeys by contact. It is pointed out that these studies only indicate diseases from which monkeys recover and they do not indicate prevalence of severe disease causing grave illness or death.


Nucleosides. 110. Synthesis and antiherpes virus activity of some 2'-fluoro-2'-deoxyarabinofuranosylpyrimidine nucleosides. A series of 5-substituted 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)cytosines 7a-d and their corresponding uracils 9a-d,f were prepared by condensation of 3-O-acetyl-5-O-benzoyl-2-deoxy-2-fluoro-D-arabinosyl bromide (5) with appropriately trimethylsilylated pyrimidines followed by saponification of the protected nucleosides 6 or 8. 1-(2-Deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine (7e) was obtained by iodination of 7a. Iodination of 8a followed by removal of the protecting acyl-protecting groups afforded the 5-iodo nucleoside 9e. Several of these 2'-fluoro-substituted nucleosides completely obviated replication of herpes simplex virus type 1 (HSV-1) in monolayers of Vero cells at concentrations of 10-100 microgram/mL. The 5-iodocytosine analogue 7e was the most effective, showing 99.5% suppression of viral replication even at concentrations of 0.1 microgram/mL. The cytotoxicity of 7e to L5178Y or P815 cells in culture was minimal. A comparison of the efficacy of 7e against HSV-1 with other known nucleoside antiviral agents indicates that further in vitro and in vivo evaluation of 7e is warranted.


Hypobetalipoproteinemic agents. 2. Compounds related to 4-(1-adamantyloxy)aniline. While the previously used displacement reaction of sodim 1-adamantyl oxide on 4-fluoronitrobenzene was applicable to the preparation of 4-(1-adamantyloxy)aniline and several related compounds, certain derivatives were not easily accessible by this route. Thus the recently reported ortho alkylation of anilines and the dicyclohexylcarbodiimide-promoted coupling of 1-adamantanol with phenols were useful in the preparation of aromatic-substituted derivatives. Furthermore, addition of phenylmagnesium bromide to 1-cyanoadamantane provided entry to the 4-(1-adamantylmethyl)aniline series. 4-(1-Adamantyloxy)aniline (3) is herein reported to be a more potent hypobetalipoproteinemic agent than the previously reported bicyclooctyloxy analogue. Replacement of the oxygen atom of 3 with sulfur (74) or methylene (62), but not nitrogen (71), results in active compounds. In the oxygen series derived from 3, the widest scope of substitution on nitrogen resulting in activity is found. The N-ethoxycarbonyl (5), acetyl (6), methyl (12), ethyl (13), N-methyl-N-(2-hydroxyethyl) (19), N-methyl-N-formyl (22), N,N-dimethyl (26), pyrrolidine (14), and piperidine (15) derivatives are active. Aromatic ring substitution also provided the active 3-chloro (44b), 2-fluoro (41b, 42, and 43), and 2-methylthiomethyl (48) compounds. Thus these active compounds are identified for further development as hypobetalipoproteinemic agents.


3-Substituted adenines. In vitro enzyme inhibition and antiviral activity. The direct alkylation of adenine at the 3 position has been extended to produce series of 3-alkyl-, 3-allyl-, and 3-(substituted benzyl)adenines. When these compounds were tested for enzyme inhibition and antiviral activity in vitro, 3-n-pentyladenine was found to be the most active compound in inhibiting the enzyme dopamine-beta-hydroxylase, and 3-(2-bromobenzyl)adenine showed the most striking inhibition of multiplication of Vaccinia virus and of Herpes simplex virus in tissue culture.


Synthesis and pharmacologic characterization of an alkylating analogue (chlornaltrexamine) of naltrexone with ultralong-lasting narcotic antagonist properties. Chlornaltrexamine (CNA) produces ultralong-lasting (3--6 days) narcotic antagonism in mice and persistent stereospecific binding to rat-brain homogenate. Protection studies in mice suggest that CNA mediates its narcotic antagonist effects by interacting with the same receptors that are occupied by naloxone. A single icv dose of CNA also has been found to inhibit the development of physical dependence in mice for at least 3 days. These studies suggest that CNA exerts its sustained effects by selective covalent association with opioid receptors.


Easily hydrolyzable, water-soluble derivatives of (+/-)-alpha-5-[1-(indol-3-yl)ethyl]-2-methylamino-delta2-thiazoline-4-one, a novel antiviral compound. The preparation of a series of indole N-acyl and N-carbamic esters of (+/-)-alpha-5-[1-(indol-3-yl)ethyl]-2-methylamino-delta2-thiazolin-4-one (1) is reported. These derivatives were synthesized as potential water-soluble precursors of the antiviral thiazolinone 1, for evaluation by intranasal administration against influenza and other respiratory infections caused by viruses. Salts of the basic carbamic esters (16--19) possess the required water solubility, undergo rapid hydrolysis and decarboxylation at pH values greater than 6, and have high activity against influenza A2 and Coxsackie B1 viruses in vitro. In influenza A2 infected ferrets a representative ester (16) reduced the severity and duration of disease symptoms and reduced nasal wash virus titres but caused local irritancy.


A model for thyroid hormone--receptor interactions. Theoretical electronic structure calculations on the thyroid hormones and analogues, as well as model hormone--receptor interactions, have been carried out. These studies (a) support the concept that the 4'-OH group is a H-bond donor to the in vivo nuclear receptor and suggest that at the receptor this OH group is trans to the 3' (distal) substituent; (b) indicate that there is an important intramolecular interaction between 3' and 4' substituents, and those 3' substituents that most favor both 4' OH orientation trans to the 3' group and a more acidic OH group substantially increase binding and biological activity; and (c) support the concept that there is a direct correlation between the conformational free energy of the aromatic rings and biological activity.


Deoxymorphines: role of the phenolic hydroxyl in antinociception and opiate receptor interactions. Several 3-deoxy opioids and 3,6-dideoxydihydromorphine was synthesized to ascertain the effect of the phenolic hydroxyl group on antinociceptive potency and receptor binding affinity. Catalytic reduction of the 3-tetrazolyl ether derivatives of dihydromorphine provided the entry into the 3-deoxydihydro series. The prototype, 3-deoxymorphine, was prepared by lithium aluminum hydride reduction of 3-deoxy-N-carbethoxymorphinone, obtained via its 7-(phenylseleno) derivative. 3-Deoxydihydromorphinone and 3,6-dideoxydihydromorphine were found to be about as potent as, or more potent than, morphine in standard antiociceptive assays. Each of them, however, was less potent than the comparable 3-hydroxy analogue, and their binding affinity to the opiate receptor was substantially decreased. The epoxy ring in 3.6-dideoxydihydromorphine was found to increase the antinociceptive potency of the compound.


Prodrugs of 9-(beta-D-arabinofuranosyl)adenine 2. Synthesis and evaluation of a number of 2',3'- and 3',5'-di-O-acyl derivatives. A number of 2',3'- and 3',5'-di-O-acyl derivatives of 9-beta-D-arabinofuranosyladenine (1) were prepared and evaluated as antivirals. These compounds, designed as prodrugs of 1, offer a range of solubilities and lipophilicities, as well as a resistance to adenosine deaminase, that render some as being attractive as possibly useful antiviral agents. Of particular note is 9-(2,3-di-O-acetyl-beta-D-arabinofuranosyl)adenine that was found to be effective as a topical agent in a guinea pig model of genital herpes.


Paradoxical effects of N-cyanoalkyl substituents upon the activities of several classes of opioids. The pharmacological effect of the N-(beta-cyanoethyl) moiety is dependent on the opioid on which it is substituted. It caused a large increase in antinociceptive potency, in (--)-3-hydroxymorphinan and (--)-normetazocine, as compared with the N-methyl opioid. These cyanoethyl compounds do not substitute for morphine in morphine-dependent monkeys. This moiety also appears to greatly increase the ability of the opiate receptor to differentiate enantiomers. An ca. 100,000-fold difference in binding was noted between the epimeric N-(beta-cyanoethyl)-3-hydroxymorphinans and the normetazocines. The levo enantiomers have little acute toxicity and showed excellent therapeutic ratios. In contrast, the N-(beta-cyanoethyl) moiety on normorphine, norcodeine, and noroxymorphone did not appear to improve their pharmacological properties. Homologous N-cyanoalkyl opioids were less potent antinociceptives.


Angiokeratoma corporis diffusum (Anderson-Fabry disease) in a single large family in Nova Scotia. Eighteen males, 17 of whom were members of a single family, affected with angiokeratoma corporis diffusum were examined in detail to determine the extent of clinical variation of the expression of what was almost certainly the same X-linked mutation in each. The commonest symptom was episodic bouts of severe, painful dysaesthesia in hands and feet. This was a major complaint of 12, a minor complaint of 5, and absent in 1. In over half the subjects, the skin rash that is considered a characteristic sign of the disease was absent or inconspicuous. All exhibited mild clubbing of fingers and toes, and 15 showed variable limitation of active and passive extension of the 5th fingers bilaterally. Only 2 (age 36 and 47) had evidence of significant renal disease. Electrocardiograms showed abnormally short PR intervals in 4, and right ventricular conduction disturbances in 5. Echocardiograms on 9 showed no evidence of myocardial dysfunction. The marked variation of the expression of some features of the disease indicates that the clinical expression of the mutation is likely to be subject to considerable genetic or environmental modification in each individual.


The treatment of ACTH Paraneoplastic syndrome with aminoglutethimide. A 60-year-old white female with oat-cell carcinoma of the lung presents with a recurrent pulmonary nodule and Cushing syndrome. The tumor is uncontrollable with MeCCNU and cyclophosphamide (Cytoxan), consequently the symptoms of Cushing disease become severe. Cushing syndrome is documented by laboratory examinations. Aminoglutethimide, an anticonvulsant, is used to reverse the symptomatology and also the laboratory values. When aminoglutethimide is discontinued, the laboratory findings return to pretreatment levels.


Lipid peroxidation activity mediated by NADPH-cytochrome C reductase purified from rabbit liver microsomes. Purified NADPH-cytochrome c reductase of rabbit liver microsomes was examined to determine whether or not the reported low lipid peroxidation activity of rabbit liver microsomes is due to the enzyme, NADPH-cytochrome c reductase. NADPH-cytochrome c reductase was purified from phenobarbital-treated rabbit liver microsomes to a specific activity of 14.9 to 21.4 unit per mg of protein with a yield of 15.2 to 16.4%. The purified sample (21.4 unit/mg of protein) was almost homogenous as determined by sodium dodecylsulfate gel electrophoresis. This sample was used for determining lipid peroxidation activity. EDTA and ferrous ion but not ADP were essential requirements for the activity. FMN enhanced the activity when low concentrations of the NADPH-cytochrome c reductase were used for the assay. NADP and 2'-AMP, which are inhibitors of NADPH-cytochrome c reductase, inhibited the lipid peroxidation activity. a-Tocopherol and p-chloromercuribenzoate (PCMB) also inhibited the activity. From these results, we confirmed the rabbit liver microsomal enzyme NADPH-cytochrome c reductase plays a role in lipid peroxidation activity. The reported low lipid peroxidation activity in rabbit liver microsomes does not appear to be caused by the NADPH-cytochrome c reductase.


[Kidney function and Na+-K+-ATPase and succinate dehydrogenase activity in the cells of the renal tubules of rats with constriction of the thoracic portion of the vena cava inferior]. Circulatory insufficiency in rats was induced by constriction of the thoracic portion of vena cava inferior. Two-three days after the operation renal function was studied and the activity of succinate dehydrogenase and Na+K+-ATPase in the cells of the renal tubules was determined cytochemically. On infusion of 0.9% NaCl solution into the stomach, the filtration in the "caval" rats did not change whereas the excretion of sodium and its concentration in the urine were much lower than in the sham operated rats. At the same time, the activity of Na+K+-ATPase and succinate dehydrogenase in the "caval" rats increased in the cells of the proximal tubule, diminished in Henle's loop, and remained unchanged in the cells of the distal convoluted tubule. Comparison of the results of functional and cytochemical study of the kidneys in caval rats allows the conclusion that intensified proximal reabsorption may be the main cause of antinatriuresis in these animals. The lesser load experienced by the distal segment promotes fuller reabsorption of sodium in this part of the nephron too.


[Sodium-potassium-adenosine triphosphatase-activity in red cell ghosts of patients with essential hypertension (author's transl)]. Increased sodium concentration and high influx of Na22 are reported in erythrocytes of patients with essential hypertension. It was speculated, that these findings are due to a disturbed transport for sodium across red cell membranes. We found a significantly increased activity of the ouabainsensitive Na-K-ATP'ase in red cell ghosts of 27 patients with essential hypertension compaired with 32 normotensive controls. There existed no difference in Mg-ATP'ase-activity between the two groups. These findings suggest an increased activity of the Na-pump in red cell membranes of patients with essential hypertension.


Glomus tumor of the middle ear: origin, symptomatology, and treatment. The glomus tumor is considered to be the most common of all tumors that involve the middle ear. This report defines the glomus tumor and discusses the symptomatology of the tumor with respect to the various developmental courses it may follow. Forty clinical cases of glomus tumor are reviewed, with emphasis upon the incidence of the tumor and its symptoms, including the audiological manifestations noted in this series of cases. Current medical management of the glomus tumor recommends surgical removal of the lesion when possible; however, irradiation and embolization of the tumor may also be considered when its invasion into surrounding structures has been extensive. The pathology and its treatment may have marked temporary or permanent effects upon the hearing of the patient, and the importance of ongoing audiological management is therefore emphasized.


Lymphatic dynamics of liver by hepatic lymphography using lipiodol ultra fluid. Hepatic lymphography by intra-parenchymal injection of four to ten millilitres (ml) of lipiodol ultrafluid, our modification of functional hepatography, performed on sixty one patients helped study lymphatic dynamics of liver. In conditions associated with significant hepatic venous outflow obstructions, such as hepatic cirrhosis, inflammatory diseases of liver and primary or secondary malignant lesions of the liver, this study delineated liver lymphatics and portal and para aortic lymph nodes. In one case mediastinal nodes were also delineated by flow of lipiodol from the bare area of the liver via trans-diaphragmatic and pleural lymphatic. The lymphangio-architecture of the opacified nodes depicted the nature of pathology inflicting them and the liver. Lipiodol in the lymphatic system, staying longer than the freely diffusible aqueous contrast, provided more detail and better information.


Activation of sterol ester hydrolase of bovine corpus luteum by N6,O2'-dibutyryl cyclic adenosine 3':5'-phosphate. The direct activation of sterol ester hydrolase (E.C. 3.1.1.13) in homogenates of bovine corpus luteum by N6O2'-dibutyryl cyclic adenosine 3':5'-phosphate, (dibutyryl cAMP), adenosine triphosphate (ATP), and Mg2+ has been demonstrated. Variability in the extent of activation by the additions was minimized by homogenization of the tissue in 5 mM Mg2+'. Baseline sterol ester hydrolase activity was primarily associated with the 105,000 X g soluble fraction, and significant activation of the enzyme preparation preincubated with dibutyryl cAMP, ATP and Mg2+ occurred within the first 15 min, prior to addition of substrate. A requirement for protein kinase in the system was demonstrated by blocking the cofactor-dependent enzyme activation with commercial protein kinase inhibitor.


Distribution of high density and other lipoproteins in selected LRC prevalence study populations: a brief survey. Selected lipid and lipoprotein data from the Lipid Research Clinics (LRC) Prevalence Study are presented, with particular emphasis given to high-density lipoprotein (HDL) values. Cross-sectional age- and sex-specific mean values are shown for 7007 white participants in the ten North American LRCs. Comparisons are drawn for males and females (including the pediatric group) and for females using or not using sex hormones. The US-USSR Collaborative Program is summarized, and selected comparisons are noted for the Soviet and United States samples.


HDL-cholesterol levels in the Multiple Risk Factor Intervention Trial (MRFIT) by the MRFIT Research Group 1,2. Preliminary data from the Multiple Risk Factor Intervention Trial (MRFIT) have been examined for evidence that the program has an influence on plasma HDL-cholesterol. The overall mean level of this lipoprotein in the initial cohort of 1,084 men was not altered by two years of participation in this risk factor reduction project. However, changes did occur, both upwards and downwards, in some individuals. There were significant negative associations between change in HDL-cholesterol and changes in body mass, VLDL-cholesterol, LDL-cholesterol, and serum thiocyanate (a measure of cigarette smoking exposure); and there was a small positive association with change in reported alcohol intake. Multiple regression analysis revealed each of these associations to be independent of the others. The fat-controlled diet designed to lower total serum cholesterol did not decrease HDL-cholesterol levels. We conclude that conventional risk reduction programs are not likely to lower circulating HDL-cholesterol, and that program components such as weight reduction and smoking cessation may increase the levels.


Serum high density lipoprotein and its relationship to cardiovascular disease risk factor variables in children--the Bogalusa heart study. Serum high density lipoprotein is increasingly recognized as a negative risk for cardiovascular disease. The distribution and interrelationship of serum lipids, lipoproteins, anthropometric measurements and blood pressures were determined in some 5,000 children. Children had mean +/- S.D. alpha-lipoprotein cholesterol levels (mg/100 ml) of 36 +/- 15 at birth, 51 +/- 22 at 6 mo, 53 +/- 18 at 1 yr, 60 +/- 19 at preschool age (2 1/2-5 1/2 yr) and 68 +/- 22 at school age (5-14 yr), reflecting a sharp increase in alpha-lipoprotein between birth and school-age years, when these levels remained relatively stable through age 14. Although white children tended to have higher levels of total cholesterol and alpha-lipoprotein at birth than black children, during childhood this trend was reversed and the differences were pronounced in school-age children (p less than 0.0001). Unlike in adulthood, boys had slightly higher levels of alpha-lipoprotein than girls. The alpha-lipoprotein was negatively correlated with pre-beta-lipoprotein and to a lesser extent with beta-lipoprotein classes. There was an inverse relationship between alpha-lipoprotein and obesity with a consistently significant relationship (p less than 0.01) in older children (10-14 yr). Children with higher levels of alpha-lipoprotein have lower levels of blood pressure, beta-lipoprotein and a lower obesity index.


Membrane-bound phospholipid desaturases. This review covers studies on membrane-bound phospholipid desaturases in yeast and rat liver carried out in this laboratory. In yeast the desaturase system was shown to effect the direct desaturation of dioleoyl-lecithin to dilinoleoyl-lecithin. In rat liver the desaturase was capable of converting 2-eicosatrienoyl-lecithin to 2-arachidonoyl-lecithin. Both systems required reduced pyridine nucleotides, O2 and cytochrome b5. Eicosatrienoyl-lecithin desaturase along with eicosatrienoyl-CoA desaturase of rat liver microsomes was solubilized with detergents and purified 7-8-fold from the microsomal pellets. Both activities were reconstituted in the presence of deoxycholate on addition of the other components of the cytochrome b5-electron transport chain (cytochrome b5 and NADH-cytochrome b5 reductase) to the solubilized desaturase; addition of lecithin further stimulated the activities. The demonstration of desaturation of eicosatrienoyl-lecithin by a solubilized and partially purified desaturase provides strong evidence for the direct desaturation of the lecithin substrate without prior conversion to the acyl-CoA thiolester.


[Effect of phosphate and glucose on the growth kinetics and phsophohydrolase activity of Penicillium brevi-compactum]. The kinetics of growth of Penicillium brevi-compactum and the activity of exocellular phosphohydrolases were studied on media with different concentrations of glucose and phosphate. Exocellular phosphohydrolases were synthesized when the growth was decelerated due to the deficiency of phosphate rather than that of glucose. These enzymes were not synthesized if the growth was not limited by phosphate. Synthesis of exocellular phosphohydrolases was repressed by phosphate when the growth was limited by glucose. Apparently, a complex of exocellular phosphohydrolases is synthesized by the culture if its growth is limited by phosphate.


[Alternative oxidation pathways in the respiratory chain of Brevibacterium flavum]. A scheme is proposed for the branched respiratory chain in Brevibacterium flavum 22 LD. Three branching sites in the main electron transport pathway are suggested: at the initial stage of NADH2 oxidation (before menaquinone) and at the levels of cytochromes b and c. Up to 40% out of the total oxygen is consumed in the NADH2-dependent alternative oxidative pathway in the presence of 5.10(-3) M cyanide or after the near UV treatment. This bypass differs from the main electron flow with a lower P/O ratio. Exogenous NADH2 catalyses the univalent reduction of oxygen through superoxide production in the bacterial membranes. Alternative oxidation pathways are discussed regarding their regulatory role in the bacterial energy metabolism.


Nephroblastoma (Wilms tumor): tubule density and prognosis. Relative differences in tubules density of the primary tumors have been suggested to be prognostically useful in nephroblastoma (Wilms tumor). Forty cases from our institution were retrospectively graded according to tubule density. There were significant differences in disease-free survivals between histologic grades. When compared to other clinical and pathologic staging criteria, tubule density was not more useful prognostically than the staging systems tested. However, when used in conjunction with clinical or pathologic stage, tumor grade improved prognostic sensitivity. Regardless of grade or stage, patients less than 24-months-old at diagnosis had better disease-free survival.


[Localization of hydrogenase in the cells of the thermophilic hydrogen bacterium, Pseudomonas thermophila]. Spheroplasts that were osmotically stable in 0.2M Tris-HCl--0.02M EDTA were prepared from the autotrophically grown cells of Pseudomonas thermophila K-2. The spheroplasts possessed 90--95% of the hydrogenase activity of the whole cells. The half-life time of hydrogenase in the spheroplasts at 80 degrees C was 8.5 min. A spectrophotometric technique was developed for determining the membrane-bound hydrogenase in the presence of sulfhydryl compounds with methylene blue as electron acceptor. The maximal specific activity of hydrogenase in extracts prepared in the anaerobic conditions in the presence of dithiothreitol and Mg2+ and Mn2+ ions was 10 +/- 3 units per 1 mg of protein, which closely corresponded with the activity of hydrogenase in the whole cells. Almost all activity of hydrogenase assayed with methylene blue was localized in the membrane fraction. The activity of soluble NAD-specific hydrogenase was not detected. Large particles located in 60-70% sucrose had the highest hydrogenase activity upon fractionation in a continuous sucrose concentration gradient. The second, lower peak of the hydrogenase activity was detected in fractions of 40--50% sucrose. As was found by electron microscopy, the size of membrane vesicles with the hydrogenase activity varied within the range of 68--156 nm. The membrane preparations possessed the activity of NADH-dehydrogenase, NADH-oxidase and succinate dehydrogenase as well.


[Clinico-statistical study of the period of formation and mineralization of the 3d molar]. Examination of 540 Italians indicated that the third molar may form between the 6th and 13th year of life. The statistical possibilities of forecasting agenesis of this tooth at different periods of life are evaluated in this light of the data obtained.


[Absorption and magnetic circular dichroism spectra of heme proteins in nonequilibrium states. IV. Cytochrome c and its derivatives]. Absorption and magnetic circular dichroism spectra (77 degrees K) of nonequilibrium cytochrome c and its derivatives reduced by thermolysed electrons was studied. The low temperature spectral characteristics of reduced cytochrome c (pH 7.0, 1.6 and 10.0), dimer, carboxymetylated and formylated derivatives at netral and acid pH, also fluoride complexes differ from the characteristics of equilibrium reduced forms. The temperature increase (up to 177 degrees K) induces relaxation of nonequilibrium states. These effects are due to structural differences in the heme vicinity of the reduced and oxidized forms.


The induction of dominant lethal mutations in Tilapia mossambica by alkane sulphonic esters. A serial mating system has been used to enable study of the effect of chemical mutagens in Tilapia mossambica. Embryopathies incompatible with survival are attributed to dominant lethal mutations induced in developing male germ cells by methyl methanesulphonate and the isomeric forms of dimethyl-myleran.


Update of naltrexone treatment.

Update of naltrexone treatment. To summarize, then, our work with naltrexone in a multi-modality treatment program indicates that it appeals to about 5-10 percent of narcotic addicts--those who conciously want to obtain drug-free status. It does significantly attentuate the effect of narcotics to the extent that they lose their reinforcing properties, and it has minimal side effects. Although we treated a self-selected population, it is noteworthy that about 10 percent of those who remained on naltrexone for more than a week were still opiate free 6 months after cessation of treatment. For some it appeared to be a turning point in their lives. It was the first time in years that they could live in their neighborhood and not be either intoxicated or occupied with the pursuit of narcotics. Our behavioral studies have identified a number of conditioned psychophysiological responses associated with the self-injection ritual. So far, although we have succeeded in extinguishing some of these responses, this has not resulted in an improved clinical outcome. We are currently working on more comprehensive behavioral treatments, but the clinical significance of these conditioned responses is uncertain at present.


[Chemical mechanism of the effect of alcohol]. The formation of tetrahydro-isoquinoline alkaloids in human and animal metabolisms subsequent to alcohol consumption is reported. The spectrum of identified products ranges from simple structures such as salsolinol to the complicated skeleton of protoberberines. These substances, hitherto found exclusively in plants, interfere with the biosynthesis, biodegradation and neurotransmission of the structurally closely related biogenic amines from which they are formed, and thus affect the vegetative nervous system.


Prostacyclin (PGI2) decreases the cyclic AMP level in coronary arteries. The effects of prostacyclin (PGI2) on vascular tension and cAMP content were measured in isolated bovine coronary artery strips. 3 nM PGI2 did not alter the tension but diminished the cAMP content by 56% of the control level (P less than 0.005). 30 and 300 nM PGI2 diminished the tension and further reduced the cAMP content, which amounted to only 5% of the control at 300 nM PGI2. These results are in contrast to the increase in cAMP level by PGI2 in blood platelets and might indicate a different mechanism of action of PGI2 in platelets and vascular tissue.


Dehydro-digitoxosides of digitoxigenin and digoxigenin: binding to beef heart (Na+ + K+)-ATPase in relation to unchanged digitoxosides. Dehydro-digitoxosides are metabolites of digitalis glycosides. In order to study their possible biological activity their affinity to (Na+ + K+)-activated ATPase was determined and compared with unchanged glycosides. Based on the dissociation constants of glycoside-enzyme-complexes, the affinity of the dehydro-digitoxosides ranged in the same order of magnitude as that of the native glycosides. Comparing mono-, bis-, and tris-digitoxosides of digitoxigenin (dt-1, dt-2, dt-3) and of digoxin (dg-1, dg-2, dg-3) with the corresponding dehydrodigitoxosides (3'-dehydro-dt-1, 9'-dehydro-dt-2, 15'-dehydro-dt3, 3'-dehydro-dg-1 and 9'-dehydro-dg-2, respectively) the dehydro-digitoxosides had lower affinities to the enzyme. The highest dissociation constants (KD) were found for 3'-dehydro-dt-1 and 3'-dehydro-dg-1. The half maximal inhibition of (Na+ + K+)-ATPase activity (I50) coresponded to affinity measurements in all but two cases: dehydro-dt-3 and dehydro-dt-2 showed very low I50 values.


Presynaptic inhibitory actions of 2-substituted adenosine derivatives on neurotransmission in rat vas deferens: effects of inhibitors of adenosine uptake and deamination. In the isolated rat vas deferens, various 2-substituted adenosine derivatives and adenosine inhibited contractions elicited by field stimulation but had little effect on responses to exogenous noradrenaline. 2-Chloroadenosine, 2-bromoadenosine, 2-hydroxyadenosine and 2-fluoroadenosine were all more potent than adenosine. Theophylline antagonized the action of the 2-substituted derivatives. The inhibitory action of adenosine was potentiated by dipyridamole, 2-amino-6-[(2-hydroxy-5-nitro)benzylthio]-9-beta-D-ribofuranosylpurine (HNBTG) or 2'-deoxycoformycin while that of 2-choloroadenosine was not altered by any of these drugs or by phenoxybenzamine, atropine or indomethacin. Pretreatment of the vas deferens with both HNBTG and 2'-deoxycoformycin eliminated the difference in inhibitory potency between adenosine and 2-chloroadenosine. These results indicate that 2-substituted adenosine derivatives, like adenosine, produce inhibition of transmission by acting on a presynaptic site which can be blocked by theophylline. The high apparent potency of 2-chloroadenosine compared to adenosine appears to be due to the former being neither taken up nor deaminated, while the apparent potency of adenosine is masked by uptake and deamination in this tissue.


A characterization of the nature and control of cellular allograft rejection. The mechanism of cellular rejection has been analyzed. The cells infiltrating rat cardiac allografts have been recovered and identified. An important role for cytotoxic T lymphocytes in cellular rejection has been discerned. The entrapment of cytotoxic T lymphocytes within rejecting grafts is not rigorously dependent upon antigen recognition. The cellular processes by which cytotoxic T lymphocytes destroy target cells have been determined in part. Cellular levels of the 3',5' cyclic nucleotides modulate this process. Elevations in cellular levels of cyclic AMP inhibit cytotoxicity whereas increased cyclic GMP stimulates augmented cytotoxicity.


Post-obstructive nephropathy in the rat: relationship between NA-K-ATPase activity and renal function. Na-K-ATPase activity and renal function were compared in rats studied after relief of 24 h of unilateral or bilateral ureteral ligation (UUL or BUL), that is, in the absence or presence of post-obstructive diuresis. Na-K-ATPase activity in the outer medulla of the rat kidney after relief of UUL was not significantly altered immediately but was markedly reduced 1 and 3 days post-obstruction. The decrease in medullary Na-K-ATPase activity was not significantly different from that observed after relief of BUL. These results indicate that decreased Na-K-ATPase activity in the post-obstructive kidney is not responsible for post-obstructive diuresis and is not due to uremia, but is a local phenomenon which is probably secondary to altered renal structure or function. It may be due to decreased filtered sodium load or direct tubular damage, but other data suggests that the decreased medullary solute concentration gradient in the post-obstructive kidney (UUL or BUL) may influence Na-K-ATPase activity which, in turn, contributes to the decreased ability to conserve sodium and water.


The influence of brain cytosol on RNA synthesis and RNA products of isolated mouse brain nuclei. The incubation of isolated nuclei obtained from 10-day-old mouse brain in the presence of brain cell cytosol resulted in an increase in the synthesis of RNA. Under conditions of saturating concentrations of nucleoside triphosphates, the influence of cytosol could not be duplicated by the addition of cyclic nucleotides. The stimulatory activity of cytosol on brain nuclear RNA synthesis could not be attributed to either alterations in the permeability of the nuclear envelope or an increased uptake of radioactively-labeled precursors. Sucrose gradient analysis demonstrated that the RNA products synthesized by nuclei isolated from 10-day-old and adult mouse brain were of a relatively low molecular weight. However, the addition of cytosol resulted in a significant increase in the size of the RNA transcripts. In contrast to the observations with 10-day-old and adult brain nuclei, the RNA from 2-day-old mouse brain nuclei was larger in size and relatively unaffected by the presence of cytosol. Although cytosol caused an increase in the amounts of poly[A]-RNA in nuclei of 2-day-old and adult animals, no comparable effect could be measured in nuclei from 10-day-old brain tissue.


Met-enkephalin: rapid separation from brain extracts using high-pressure liquid chromatography, and quantitation by binding assay. Met-enkephalin can be rapidly separated with high recovery from leu-enkephalin, beta-endorphin, and enkephalin metabolites by high-pressure liquid chromatography on a Dupont SCX column. The technique permits separation of endogenous enkephalins from brain extracts for measurement by opiate-receptor binding assay, immunoassay, or for study of the incorporation of radiolabeled amino acids into enkephalin. Using this technique, met-enkephalin was separated from striatal extracts of rats killed by focused microwave irradiation and quantitated by the opiate-receptor binding assay. The concentration of met-enkephalin in the column eluate was 1.3 micron/g tissue. This value is comparable to that obtained by radioimmunoassay without purification of the extracts.


ACTH-induced inhibition of endogenous rat brain protein phosphorylation in vitro: structure activity. ACTH1--24 inhibits the endogenous phosphorylation in vitro of distinct SPM protein bands. Using N-terminal fragments of ACTH, the structure-activity requirements for this effect were studied. A rather complex interaction of the ACTH fragments with endogenous SPM phosphorylation was observed. The effects were not only dependent on the primary structure of the peptide used, but also on the protein band studied and the ATP/SPM ratio used in the incubation system. ACTH1--24 did not interfere with the ATP-hydrolyzing activity of the SPM preparation, nor did it influence the endogenous phosphatase activity. Therefore, a direct interaction of ACTH with SPM protein kinase(s) is likely to be responsible for its effect on phosphorylation.


Relationship between ACTH secretion and corticoid binding to specific receptors in perifused adenohypophyses. This study reports on the kinetics of glucocorticoid-induced inhibition of ACTH release, together with steroid binding to specific pituitary receptors. It was shown that corticosterone (CORT) inhibited ACTH output provoked by either corticotropin-releasing factor (CRF) extracts or dbcAMP, in a manner which was both dose- and time-dependent. A close correlation appeared to exist between the degree of ACTH blockade and the percentage of filled steroid-binding sites. However, exposure of hypophyses to CORT for a critical period of time was a prerequisite for such a relationship to develop. Furthermore, it was found that dexamethasone (DEX) was more potent than CORT in inhibiting ACTH secretion and, in addition, bound to a greater extent to nuclei of pituitary cells. These data suggest the existence of a close correlation between occupancy of pituitary glucocorticoid receptors and modulation of ACTH secretion.


[Results of polygraphic nocturnal sleep recording in patients with supratentorial brain tumors prior to and following surgery]. The purpose of the study was to establish the influence exerted by a supratentorial brain tumour on the EEG correlates of sleep, and to compare the results before and after operation. Five patients were studied (aged 38 to 59 years) who were treated surgically for supratentorial expanding lesions: two cases of meningioma (in left parietal area), two cases of glioma (right parietal area and occipital area) and one case of intracerebral haematoma. The control group comprised 5 patients operated upon for lesions of the peripheral nerves of extremities. In the patients with supratentorial brain tumours and after their removal the following sleep parameters were unchanged: total sleep duration, number of awakening during sleep, latency time of the first REM phase and stage 3 NREM (deep and superficial sleep). The amount of REM sleep increased after tumour removal. REM-sleep density was selectively and permanently disturbed. In presence of tumour REM-sleep density was found in traces only, after tumour removal it increased but showed a permanent high-grade deficit in relation to controls.


Studies of cultured human and simian fetal brain cells. II. Infections with human (BK) and simian (SV40) papovaviruses. Both simian virus 40 (SV40) and BK viruses infected and lysed not only oligodendroblasts and astrocytes, but also neuroblasts and epithelioid or mesenchymal cells in cultures of fetal brain cells derived from human, rhesus, and cynomolgus hosts. Lytic infections of these four cell types differed ultrastructurally in the amount and arrangement of virions in the nucleus, the extent of nuclear membrane redundancy, the presence of nuclear virion arrays, and the amounts of virions in the cytoplasm. However, major differences were not noted between SV40 and BK viruses, nor between different species of origin or region of brain explanted. Modified astrocytic cells persisted in cultures from all three sources after infection by either virus. These cells stained for glial fibrillary acidic protein (GFAP) and polyomavirus tumour (T) antigen, but did not subculture indefinitely.


The accuracy of predicting histologic grades of supratentorial astrocytomas on the basis of computerized tomography and cerebral angiography. This report concerns a group of 49 patients who had been diagnosed as having gliomas on the basis of CT scans plus angiograms and for whom histologic proof was available. Ninety percent were found to be gliomas, 4% were metastases, and histology was unclear in the rest. Combining the CT criteria of inhomogeneous enhancement and surrounding edema with one or more of three angiographic signs (early venous drainage, stain, abnormal vessels) allowed a more confident diagnosis of glioblastoma.


Progression of herpes encephalitis in rabbits following the intra-ocular injection of type I virus. The injection of HSV type I into the vitreous body of the eye in the 18-day-old albino rabbits consistently induces herpes encephalitis with 90% survival. The longterm observations up to 64 days post-inculation indicate that HSV travels slowly by cell-to-cell infection of neuroglia. The lesions follow a defined anatomical pathway producing a progressive disease not dissimilar to the natural human disease. Semiserial (1 micron) sections show that as the infection progresses tissue repair follows. It appears that HSV does not become latent, repeated episodes of viral activity cause further damage with which the repair does not keep pace. By the sixteenth day virus particles and lesions are found in contralateral side of the chiasma, lateral geniculate body and both optic nerves, but not in the ipsilateral geniculate body. This suggests that the virus does not follow the axonal route.


[Embolization and balloon occlusions in tumoral processes: seven years' experience (author's transl)]. The authors report 169 therapeutic embolizations in cases of tumoral lesions in the craniocerebral, ENT, and vertebrospinal territories. Most endovascular occlusions performed for tumoral processes are presurgical indications, with the aim of reducing hemorrhage at operation. Embolization becomes practically mandatory for nasopharyngeal angiofibromas, considerably reducing the peroperative bleeding. Indications of embolization for meningiomas must be discussed according to the size and location of the tumors: embolization is especially useful in large tumors or those inserted on the base of the skull, mainly in the middle fossa. In chemodectomas, embolization can be used presurgically or on its own when surgery becomes impossible; angiographic follow-up shows that secondary repermeabilization is frequent in spite of clinical improvement. In malignant tumors of the craniofacial region, indications of embolization must be considered with care because failure of vascularization tends to make radiotherapy less efficient; in these cases, embolization can be useful in reducing pain or to contend with cataclysmal hemorrhages.


Embolization and surgical removal of nasopharyngeal angiofibroma. The usefulness of embolization of one or several feeding arteries before surgical removal of nasopharyngeal angiofibroma has been proven. However, there are two questions which remain: (a) At what point should the embolization stop? (B) What is the optimal delay between embolization and operation? These questions are discussed but not resolved.


[Comparison of CT, scintigraphy, and angiography in the topographic and pathological diagnosis of 300 supratentorial tumors (author's transl)]. The results of CT, scintigraphy (CSS), and angiography in 300 supratentorial cerebral tumors were compared with regard to the topographic and pathological diagnosis proposed in the initial reports. CT elucidated the topography in 98% of the cases and the pathology in variable percentages, depending on the type of tumor. The use of scintigraphy provided a high accuracy of pathological diagnosis. Except for meningiomas, arteriography was less accurate than the two other methods of examination. A radiological management of supratentorial cerebral tumors is proposed based on clinical and CT data.


Impact of CT in the neurosurgical management of intracranial tumors. A total of 103 brain tumor patients examined with CT, radionuclide brain scan, and angiography or pneumoencephalography, and all surgically verified, were studied to evaluate the impact of CT on the neurosurgical handling of brain tumors. CT alone was usually sufficient for optimal handling of astrocytoma patients, angiography in most meningioma cases, and pneumoencephalography in cases with sellar, suprasellar, and some other midline tumors. Information obtained only through CT sometimes altered the therapy. Sometimes it led to biopsy instead of a meaningless attempt at a 'radical' excision: in other cases it permitted a 'radical' excision otherwise not possible.


Parainfluenza 1 (6/94) virus-induced white matter degeneration: enhancement by adoptive transfer of virus-sensitized immunocytes. Intracerebral (i.c.) inoculation of parainfluenza 1 (6/94) virus into weanling Lewis rats produced meningitis, choroiditis, ependymitis, and a rare noninflammatory white matter degeneration (WMD). Adoptive transfer of immune spleen cells (ISC), 3 to 5 days after virus infection, significantly increased the incidence of WMD. Conversely ISC given 2 days prior to virus infection not only protected rats from developing WMD, but also from developing meningitis, choroiditis, or ependymitis. Thus the temporal relationship of 6/94 virus inoculation to the virus-specific immune response appears critical in determining whether WMD will develop in association with 6/94 virus infection. Although sensitized immunocytes may prevent pathologic changes if given prior to virus inoculation, such immunocytes enhance the production of WMD if given after such inoculation. These studies underline the dual role of the immune response in 6/94 virus infection, and indicate that 6/94 virus-induced WMD is, in part, immunemediated.


Subacute sclerosing panencephalitis with unusual ocular movements: polygraphic studies. A 5-year-old child with subacute sclerosing panencephalitis had continuous involuntary ocular movements while awake. At this stage of illness, stereotyped periodic EEG complexes over the two hemispheres were asynchronous. Polygraphic studies showed three diffrent types of ocular movements, each apparently related to the degree of phase shift between the EEG complexes from the two hemispheres. The involuntary ocular movements in one direction were always preceded by periodic EEG complexes from the contralateral hemisphere. Overnight polygraphic studies revealed early disruption of non-rapid eye movement (NREM) sleep patterns and preservation of rapid eye movement (REM) sleep, even in a late stage of illness.


Narcolepsy: regional cerebral blood flow during sleep and wakefulness. Serial measurements of regional cerebral blood flow were made by the 135Xe inhalation method during the early stages of sleep and wakefulness in eight normal volunteers and 12 patients with narcolepsy. Electroencephalogram, electro-oculogram, and submental electromyogram were recorded simultaneously. In normals, mean hemispheric gray matter blood flow (Fg) during stages I and II sleep was significantly less (-9.2 percent) than waking values (84.3 +/- 13 ml per 100 gm brain per minute). Maximum regional blood flow decreases during sleep occurred in the brainstem-cerebellar (-25.1 percent), right inferior temporal (-23.1 percent) and bilateral frontal (-18.9 percent) regions (p less than 0.05). In patients with narcolepsy, mean hemispheric Fg while awake was 80.5 +/- 13 ml per 100 gm brain per minute. During REM sleep (n = 2), mean hemispheric Fg increased by 9.8 percent concurrently with large increases (+34.6 percent) in brainstem-cerebellar region flow. During stages I and II sleep without REM (n = 6), there were significant increases in mean hemispheric Fg of +/-20.2 percent (p less than 0.01) and brainstem-cerebellar Fg of 38.0 percent (p less than 0.01), just the opposite of changes in normals. In narcolepsy, there appears to be a reversal of normal cerebral deactivation patterns, particularly involving the brainstem, during stages I and II sleep.


Single-fiber electromyography in myasthenia gravis. One-hundred-sixty single-fiber EMG studies of the extensor digitorum communis muscle were performed on 127 patients with myasthenia gravis; 131 demonstrated defective neuromuscular transmission. Jitter determinations in the biceps, deltoid or frontalis muscles increased the diagnostic yield significantly. Evoked-potential EMG studies were abnormal in less than 50 percent of patients in whom they were performed. The most sensitive criterion of abnormality was the percentage of fibers with increased jitter; the sensitivity of the test was enhanced, however, if the mean jitter of the tested muscle was also used as a criterion of abnormality. Since increased jitter may also be seen in primary muscle and nerve disease, these disorders must be excluded by other means before diagnosing myasthenia gravis on the basis of the single-fiber studies.


Metachromatic leukodystrophy: a comparative study of the ultrastructural findings in the peripheral nervous system of three cases, one of the late infantile, one of the juvenile and one of the adult form of the disease. The ultrastructural findings in the peripheral nerve of three cases of M. L. D., namely of the late infantile, the juvenile and the adult form are described. The pathological changes consist of two phenomena: segmental demyelination and lysosomal storage of sulfatides, both probably separate results of a deficiency of the enzyme Arylsulfatase A. Some differences are found in the degree of segmental demyelination and the type of lysosomal storage products. A possible relation between these differences and the supposed difference in the defect of the enzyme Arylsulfatase A, responsible for the different subtypes of the disease, is postulated.


Estrogen replacement therapy I: a 10-year prospective study in the relationship to osteoporosis. A 10-year, double-blind prospective study was undertaken to evaluate the effects of estrogen replacement therapy. The sample population consisted of 84 pairs of randomly chosen postmenopausal patients who were matched for age and diagnosis. One half of the patients received conjugated estrogens and cyclic progesterone, while the other half received placebo. Estrogen-treated patients whose therapy started within 3 years of menopause showed improvement or no increase in osteoporosis. Control patients demonstrate an increase in their osteoporosis.


Genital herpes simplex virus (HSV) isolation during pregnancy. Eight patients with laboratory-confirmed herpes simplex virus (HSV) infection were cultured repeatedly during their pregnancies. Two of the 8 patients did not have typical HSV lesions. All patients became culture negative and all delivered normal infants vaginally. Because asymptomatic women can transmit HSV to neonates at parturition, it is important to do viral cultures as well as to clinically assess the mother.


In vitro cultivation of human tumor tissues. II. Morphological and virological characterization of three cell lines. Nineteen human tumors, mostly of sarcomatous nature, were cultured in vitro. Three cell lines were isolated and further characterized: MG-57 derived from a giant cell tumor, MG-63 derived from an osteosarcoma and MG-72 derived from a xanthohistiocytoma. The cell lines varied in morphology and growth pattern. An abnormal karyotype with marker chromosomes was present in Mg-63 and MG-72. None of the cell lines spontaneously produced detectable C-type virus particles. Stimulation with IUDR and dexamethasone also failed to induce detectable particle release.


A clinical trial of topically applied 3 percent vidarabine against recurrent herpes labialis. Seventy-six participants were enrolled in a clinical trial to determine therapeutic effectiveness of 3 percent vidarabine applied topically to recurrent perioral herpetic lesions. Following a 6- to- 12-month natural history phase, a 12-month clinical trial was conducted. Seventy participants developed 463 lesions during 361 episodes. Three percent vidarabine in a water-miscible gel was applied six times daily for 7 days to each lesion in the experimental group. Identically packaged placebo was used by the control group. Group assignment was by computer-generated randomization. Lesion size was reduced when vidarabine, rather than placebo, was applied. The difference was statistically significant (Student's t test, P = 0.02). Vesiculation followed tingling more rapidly when vidarabine, rather than placebo, was applied prior to vesiculation (P = 0.05). No significant difference between the two groups was found in episode frequency or lesion duration. Adverse reactions to vidarabine were not experienced.


[Renal receptors of parathyroid hormone and calcitonin (author's transl)]. The two first steps of the renal cellular action of parathyroid hormone and of calcitonin are the hormonal binding onto specific receptors and the stimulation of adenylate cyclase by the hormone-receptor complex producing an increase in the intra-cellular concentration of 3'-5' cyclic adenosine monophosphate (cyclic AMP). Specific glomerular and tubular receptors for parathyroid hormone have been demonstrated using either tritiated parathyroid hormone or an indirect technique with 125 I labelled specific antibodies. Tubular receptors are localized both in the proximal and distal segments of the nephron. Parathyroid hormone stimulates glomerular and tubular adenylate cyclase. The main unsolved problem is the difficulty for demonstrating high affinity binding sites and stimulation of adenylate cyclase at low physiological concentrations of parathyroid hormone. In man, administration of parathyroid hormone produces a marked increase in the urinary excretion of cyclic AMP chiefly concerning its nephrogenous fraction. The peak of excretion is early and precedes the decrease in phosphate tubular reabsorption. Tubular receptors for calcitonin have been demonstrated using 125 I labelled salmon calcitonin. Calcitonin stimulates renal adenylate cyclase in only some segments of the nephron allowing receptors for calcitonin to be localized in the wide ascending branch of Henle's loop and the initial part of the convoluted distal tubule. In the presence of guanylnucleotides, binding of calcitonin onto its receptors and activation of adenylate cyclase are observed in the range of physiological concentrations of calcitonin in the rat. In man, administration of calcitonin produces a moderate increase in the urinary excretion of cyclic AMP coming from a non renal tissue.


Polyoma infected cells contain at least three spliced late RNAs. Poly(A)-containing polyoma cytoplasmic RNA was hybridized with linear double-stranded polyoma DNA and RNA displacement loops (R-loops) were formed. The structures visualized in the electron microscope are consistent with the conclusion that there are at least three late polyoma specific RNAs and that the leader sequences at the 5' ends of these viral RNAs are not coded immediately adjacent to the bodies of the RNAs. Measurements carried out on the R-loop structures have provided the locations on the physical map of polyoma DNA, for the bodies and leaders of the RNAs and the length of the bodies, leaders and the corresponding intervening DNA sequences.


An electron microscope study of the proteins attached to polio virus RNA and its replicative form (RF). A recently described method (Wu, M. and Davidson, N. (1978), Nucleic Acids Research 5, in press) for visualizing proteins attached to nucleic acids in the electron microscope has been applied to study proteins attached to poliovirion RNA and to the viral double-stranded intracellular RF form. A protein is found at the 5' end of the plus strand virion RNA, and protein components are found at both ends of the duplex RF. In the RF as usually extracted, there is frequently a larger or compound protein aggregate at the end which contains the 3' end of the plus strand and the 5' end of the minus strand. Banding in CsCl-guanidinium hydrochloride in the presence of sarkosyl causes dissociation of some components of this aggregate, leaving both ends labeled with the covalently bound VPg. These results confirm and extend previous biochemical studies of proteins bound to poliovirion RNA and to the RF form.


Selection of methionine tRNAs by avian oncornaviruses. The free 4S RNA of avian RNA tumor viruses is greatly enriched in one of the four methionine tRNAs of the host cells, tRNA4Met. On the assumption that viral tRNAMet forms are identical to the corresponding tRNAs of mouse or chick cells, the following conclusions were drawn concerning the tRNAMet content of oncornaviruses: (1) tRNAMet species may be compartmentalised within the host cells, and the viral tRNA pool could reflect the cellular compartment in which viral maturation takes place since tRNAMet forms distribute unevenly between different fractions of a cell homogenate. (2) tRNA4Met appears to have no special role in the modulation of protein synthesis in as much as no functional difference between tRNA2Met and tRNA3Met, tRNA4Met could be demonstrated in in vitro protein synthesising systems. (3) tRNA4Met differs in nucleotide sequence from all other host cell tRNAMet forms except possibly tRNA2Met. The nucleotide sequences of two tRNAMet species, tRNA1Met and tRNA4Met, have already been determined and the sequence of another host cell tRNAMet, tRNA3Met, was derived from the analogy of its sequence to that of tRNA4Met since the two molecules differ in only 6 nucleotides out of 76. (4) Avian myeloblastosis virus reverse transcriptase has been shown to bind specifically tRNA4Met and tRNATrp in whole cell tRNA and therefore the free tRNA4Met in the virion particle may exist substantially bound to virion-associated transcriptase.


Quantitative analysis of specific labelled RNA'S using DNA covalently linked to diazobenzyloxymethyl-paper. Substantial amounts of DNA (at least 25 microgram per cm2) can be stably bound to diazobenzyloxymethyl (DBM)-paper. Complementary RNA will hybridize to the DNA paper almost completely in 24 hours. Using several different conditions of hybridization and washing, the background of RNA bound non-specifically is very low (between 0.01 and 0.02%) and the efficiency of hybridization is very high (75 to 50% of complementary RNA is bound and retained through the washing procedure). Because the DNA is bound to the paper convalently, it is retained through all the washing and elution steps, and the DNA papers can be re-used many times.


A method for the enzymatic synthesis and purification of [alpha-32P] nucleoside triphosphates. A simplified method is described for the enzymatic synthesis and purification of [alpha-32P]ribo- and deoxyribonucleoside triphosphates. The products are obtained at greater than 97% radiochemical purity with yields of 50--70% (relative to 32Pi) by a two-step elution from DEAE-Sephadex. All reactions are done in one vessel as there is no need for intermediate product purifications. This method is therefore suitable for the synthesis of these radioactive compounds on a relatively large scale. The sequential steps of the method involve first the synthesis of [gamma-32P]ATP and the subsequent phosphorylation of nucleoside 3' monophosphate with T4 polynucleotide kinase to yield nucleoside 3', [5'-32P]diphosphate. Hexokinase is used after the T4 reaction to remove any remaining [gamma-32P]ATP. Nucleoside 3',[5'-32P]diphosphate is treated with nuclease P-1 to produce the nucleoside [5'-32P]monophosphate which is phosphorylated to the [alpha-32P]nucleoside triphosphate with pyruvate kinase and nucleoside monophosphate kinase. Adenosine triphosphate used as the phosphate donor for [alpha-32P]deoxynucleoside triphosphate syntheses is readily removed in a second purification step involving affinity chromatography on boronate-polyacrylamide. [alpha-32P]Ribonucleoside triphosphates can be similarly purified when deoxyadenosine triphosphate is used as the phosphate donor.


Characterization of the genomic RNA from a Rous sarcoma virus mutant temperature sensitive for cell transformation. We have found that the LA23 t/s mutant of Rous sarcoma virus (phenotype Prague B), even when passaged repeatedly at high multiplicity of infection, does not give rise to transformation defective deletion mutants comparable to those derived from RSV. In view of this fact and of the high rate of production of this mutant at 41 degrees C, we have undertaken a detailed analysis of the genome of this virus by ordering all large T1 oligonucleotides and by determining their nucleotide sequences. The results indicate a high degree of mutation in the onc gene as compared to that of Pr-A or Pr-B.


Aminonucleosides and their derivatives. IV. Synthesis of the 3'-amino-3'-deoxynucleoside 5'-phosphates. A new procedure has been developed for the synthesis of 3'-amino-3'-deoxyribonucleosides of adenine, cytosine and uracil by condensing the trimethylsilylated bases with peracylated 3-azido-3-deoxyribose derivative. The azido group could subsequently be reduced to amino. The 5'-phosphates of these nucleosides have been prepared and the analogues have been tested for their ability to stimulate the ribosome-catalyzed reaction of 3'(2')-O-(N-formylmethionyl) adenosine 5'-phosphate with phenylalanyl-tRNA.


A combined neurofibroma-granular cell tumor of the middle cranial fossa. A case of a combined neurofibroma-granular cell tumor in a 52 years old gunsmith is presented. The tumor developed after an intracranial trigeminal nerve operation 25 years previously. The solid tumor in the left middle cranial fossa had displaced and infiltrated the temporal lobe. It had expanded via the optic nerve into the left orbit, and further the apex partis petrosae was destroyed. The neurofibroma part shows histological aspects of malignancy, the granular cell tumor, considering its infiltrating and destructive growth, may be regarded as malignant as well. In intermingling portions of the tumor, transitional types of fiber-like and granular cells are prominent. In the peripheral zone of the tumor apparently reactively proliferated polynuclear astrocytes are seen with occasionally intracytoplasmatic lymphocytes (emperipolesis?). A short review of the literature and the theories concerning the histogenesis of the granular cell tumor is given. Whereas most authors in recent years suggest a Schwann cell origin, based on electron microscopic findings, this intermediate tumor type motivates us to postulate a mesodermal origin of the granular cell tumor. The question of viral influence is discussed briefly.


[Mechanism of the diuretic action of epiphyseal extracts]. Experiments on dogs demonstrated that intravenous administration of the epiphysis extracts prevented the antidiuretic action of both the endogenous (traumatic stress) and exogenous (pituitrin) antidiuretic hormone (ADH). Injection of the extract into the renal artery gave a unilateral diuretic effect. The diuretic action of the preparation was eliminated by the block of the alpha-adrenergic receptors, whereas beta-adrenoblockers were ineffective. A conclusion was drawn that the mechanism of the diuretic action of the epiphysis extracts consisted in the competitive suppression of the ADH action at the renal tubules level on account of excitation of the alpha-adrenoceptors and inhibition of the adenylcyclase activity.


[Adrenal cortex and vitamin PP. The effect of corticosteroids on the NAD level in the rat liver after the administration of excess nicotinic acid or nicotinamide]. The NAD content in the liver was experimentally studied in intact, adrenalectomized rats and those given hydrocortisone following nicotinic acid and nicotinamide administration. The NAD concentration in the liver rose linearly and equally during the first 4 experimental hours after the nicotinamide administration (200 mg/kg). A conclusion was drawn that in administration of exogenous precursors the coenzyme biosynthesis did not depend on the initial hormonal background.


[Effect of thyroxine on oxidative enzymatic activity in the rat aorta]. The activity of hexokinase, dehydrogenases of pentose cycle, total lactate dehydrogenase, and cytochrome-C-oxidase level was determined in the aorta of rats with hyperthyroidism caused by the intraperitoneal administration of L-thyroxin. In the administration of hormone doses approaching the physiological ones there was an increase in the activity of cytochrome-C-oxidase and a tendency to the increase of 6-phosphogluconate dehydrogenase. Elevation of the activity of all the enzymes under study, excluding lactate dehydrogenase, and in increase of the amount of extractable protein occurred in the administration of moderate thyroxin doses. An increase of hexokinase and particularly of cytochrome-C-oxidase activity was revealed in hyperthyroidism with phenomena of thyrotoxicosis. The activity of the rest of the enzymes approached control. The data obtained are discussed from the point of view of the role played by thyroid hormones in the pathogenesis of atherosclerosis.


Rapid inactivation of cyclooxygenase activity after stimulation of intact platelets. Trypsin, thrombin, and ionophore A23187 activate phospholipid breakdown of platelets that have been labeled with [(14)C]arachidonate, releasing their cyclooxygenase and lipoxygenase products. Intact platelets can also very effectively directly degrade low concentrations of exogenous, free [(14)C]arachidonate. Pretreatment of platelets with trypsin, thrombin, or ionophore A23187 for a minimum time of 30 sec leads to complete inactivation of cyclooxygenase activity, as demonstrated by subsequent exposure to [(14)C]arachidonate. Lipoxygenase activity is lost after 5 min. The thrombin-induced inactivation of cyclooxygenase and lipoxygenase is prevented by cyclic AMP (which inhibits the stimulated activity of phospholipase A(2)), although cyclic AMP does not affect the degradation of exogenous [(14)C]arachidonate. Exposure of platelets labeled with [(14)C]arachidonate to unlabeled arachidonate under conditions that lead to use of the latter also results in a similarly rapid inhibition of cyclooxygenase activity, as determined by subsequent challenge with thrombin. Under these conditions lipoxygenase activity is much less markedly inactivated. The arachidonate-induced inhibition of cyclooxygenase activity is not prevented by cyclic AMP. Trypsin does not induce platelet aggregation, and platelets whose cyclooxygenase activity has been inactivated are intact insofar as they are still able to undergo aggregation. These studies demonstrate that operation in intact platelets of the cyclooxygenase pathway, through use of endogenous or exogenous substrate, leads to a very rapid, irreversible inactivation of this enzyme. The lipoxygenase pathway is also progressively impaired, but much less rapidly than the cyclooxygenase enzyme and much less markedly on use of exogenous compared to endogenous substrate. The possible consequences of these physiological processes of spontaneous inactivation are considered.


DNA binding activity from cultured human fibrolasts that is specific for partially depurinated DNA and that inserts purines into apurinic sites. A protein of molecular weight approximately 120,000 was isolated from cultured human fibroblasts and HeLa cells on the basis of its ability to bind specifically to apurinic DNA. After separation from apurinic endonuclease activity, the protein was found to incorporate purine, but not pyrimidine, bases specifically into depurinated DNA so as to protect the apurinic sites from alkali. Purine base insertion activity was sensitive to heating and freezing as well as to caffeine and EDTA; it required K+ but not a divalent cation. Guanine, but not adenine, was incorporated into depurinated poly(dG-dC), whereas adenine, but not guanine, was incorporated into poly(dA-dT). After incorporation into depurinated DNA, guanine could be reisolated as dGMP. Although this activity suggests an alternative pathway for DNA repair that is independent of nucleotide excision, other functions for such an enzyme are possible.


Definition of cytochrome c binding domains by chemical modification: kinetics of reaction with beef mitochondrial reductase and functional organization of the respiratory chain. An assay has been developed to study the steady-state kinetics of the reduction of cytochrome c by purified beef heart mitochondrial cytochrome c reductase (cytochrome bc(1) complex, complex III). An analogue of coenzyme Q(2) (2,3-dimethoxy-5-methyl-6-decylhydroquinone) was employed as an antimycin-sensitive reductant. The kinetics of reaction of ten different mono(4-carboxy-2,6-dinitrophenyl) derivatives of horse cytochrome c were determined. The modified proteins showed higher apparent K(m) values than the native protein and greater sensitivity to ionic strength, defining an interaction domain on cytochrome c for purified cytochrome c reductase. This interaction site is located on the front surface of the molecule (which contains the exposed heme edge) and surrounds the point at which the positive end of the dipole axis crosses the surface of the protein. The site is similar to that previously determined for mitochondrial cytochrome c oxidase and yeast cytochrome c peroxidase, suggesting that the primary interaction with redox partners is directed by the dipolar charge distribution on cytochrome c. The extensive overlapping of the interaction domains for the mitochondrial cytochrome c oxidase and reductase indicates that cytochrome c must be mobile in order to transfer electrons between them, depending on their relative positions in the membrane. Whether such mobility is necessary in intact mitochondria depends on whether the interactions with the complete membrane-bound system are the same as with the purified components.


Low and high density lipoproteins and chylomicrons as regulators of rate of cholesterol synthesis in rat liver in vivo. The steady-state levels of plasma cholesterol carried in high and low density lipoproteins and in chylomicrons were varied over a wide range by use of a constant-infusion technique. After 40 hr, the rates of hepatic cholesterol synthesis and levels of hepatic cholesterol esters were measured and were related to the plasma level of each of the lipoprotein fractions. From the rates of infusion and the steady-state plasma levels attained, the whole animal clearance rates for cholesterol carried in low density and high density lipoproteins and in chylomicrons were calculated to be 0.53, 0.61, and 42.6 ml/hr, respectively. Hepatic cholesterol ester content increased by 0.8 mug/g for each 1.0 mg/dl increase in the steady-state level of plasma low density lipoprotein cholesterol and by 1.4 mug/g for a similar elevation in plasma high density lipoprotein cholesterol. In contrast, the increase in ester content was 300-fold greater when chylomicrons were infused (330 mug/g). The rate of hepatic cholesterol synthesis was inhibited by a factor of 0.004 and 0.007, respectively, per 1.0 mg/dl increase in the steady-state level of plasma cholesterol carried in either low density or high density lipoprotein but the inhibition was by a factor of 0.255, 50-fold greater, when chylomicrons were infused. Thus, in the steady state, cholesterol carried in either low density or high density lipoproteins is apparently taken up by the liver and regulates the rate of hepatic cholesterol synthesis; however, cholesterol carried in chylomicrons is at least 50-fold more effective in this regard. This marked difference can be attributed to the much higher rates of transport of chylomicron cholesterol into the hepatocyte than of cholesterol carried in either low density or high density plasma lipoproteins.


An oligosaccharide-containing factor that induces cell differentiation in Dictyostelium discoideum. When Dictyostelium discoideum amoebae (strain V12 M2) are spread at high density on agar containing 1 mM cyclic AMP, essentially all the cells differentiate into stalk cells after 2-3 days. We showed previously that isolated amoebae plated at low density on agar containing cyclic AMP do not differentiate, but can be induced to do so by a layer of high density helper cells from which they are separated by a thin cellophane membrane. We now show that the high density cell population releases a dialyzable factor that, in the presence of cyclic AMP, is capable of inducing isolated amoebae to differentiate into stalk cells. Sugar inhibition, sensitivity to glycosidases, and lectin affinity suggest that the differentiation-inducing activity requires the integrity of an oligosaccharide group containing sialic acid, L-fucose, and N-acetylgalactosamine. In addition, a phosphate group appears to be necessary for biological activity.


Involvement of spectrin in membrane fusion: induction of fusion in human erythrocyte ghosts by proteolytic enzymes and its inhibition by antispectrin antibody. In contrast to intact human erythrocytes, human erythrocyte ghosts can be agglutinated but not fused by Sendai virus. Membrane fusion can, however, be induced in virus-agglutinated erythrocyte ghosts by addition of proteolytic enzymes such as trypsin, papain, or Pronase. When erythrocyte ghosts were reacted with antispectrin antiserum, the antiserum inhibited both the induction of fusion and the proteolysis of the membrane spectrin. The correlation between the membrane fusion process and the membrane cytoskeleton is discussed.


Regulation of alkaline phosphatase expression in human choriocarcinoma cell lines. The coincident expression of two structurally distinct isoenzymes of human alkaline phosphatase was demonstrated in two independently derived gestational choriocarcinoma cell lines. These proteins were shown to have enzymatic, antigenic, and physical-chemical properties resembling those of isoenzymes from term placenta and adult liver. The regulation of these isoenzymes has been studied during the exposure of both cell lines to 5-bromodeoxyuridine and dibutyryl cyclic AMP. The responses of the alkaline phosphatase isoenzymes to these agents have also been compared with the response of another protein phenotypic to placenta, the alpha subunit of chorionic gonadotropin. The results show that (i) the separate structural genes coding for placental and liver alkaline phosphatases are regulated in a noncoordinate fashion; (ii) both alkaline phosphatase genes respond independently of the alpha subunit; and (iii) the induction of the placental type isoenzyme occurs via at least two independent pathways.


Binding site on macrophages that mediates uptake and degradation of acetylated low density lipoprotein, producing massive cholesterol deposition. Resident mouse peritoneal macrophages were shown to take up and degrade acetylated (125)I-labeled low density lipoprotein ((125)I-acetyl-LDL) in vitro at rates that were 20-fold greater than those for the uptake and degradation of (125)I-LDL. The uptake of (125)I-acetyl-LDL and its subsequent degradation in lysosomes were attributable to a high-affinity, trypsin-sensitive, surface binding site that recognized acetyl-LDL but not native LDL. When (125)I-acetyl-LDL was bound to this site at 4 degrees C and the macrophages were subsequently warmed to 37 degrees C, 75% of the cell-bound radioactivity was degraded to mono[(125)I]iodotyrosine within 1 hr. The macrophage binding site also recognized maleylated LDL, maleylated albumin, and two sulfated polysaccharides (fucoidin and dextran sulfate) indicating that negative charges were important in the binding reaction. A similar binding site was present on rat peritoneal macrophages, guinea pig Kupffer cells, and cultured human monocytes but not on human lymphocytes or fibroblasts, mouse L cells or Y-1 adrenal cells, or Chinese hamster ovary cells. Uptake and degradation of acetyl-LDL via this binding site stimulated cholesterol esterification 100-fold and produced a 38-fold increase in the cellular content of cholesterol in mouse peritoneal macrophages. Although the physiologic significance, if any, of this macrophage uptake mechanism is not yet known, we hypothesize that it may mediate the degradation of denatured LDL in the body and thus serve as a "backup" mechanism for the previously described receptor-mediated degradation of native LDL that occurs in parenchymal cells. Such a scavenger pathway might account for the widespread deposition of LDL-derived cholesteryl esters in macrophages of patients with familial hypercholesterolemia in whom the parenchymal cell pathway for LDL degradation is blocked, owing to a genetic deficiency of receptors for native LDL.


Is the product of the src gene a promoter? Addition of a potent promoter, 12-O-tetradecanoylphorbol 13-acetate (TPA), to primary avian tendon or chicken embryo fibroblast cells infected with a temperature-sensitive mutant of Rous sarcoma virus produced a complete transformed phenocopy at the nonpermissive temperature by the criteria tested. While normal, uninfected cultures also shifted towards a transformed phenotype after TPA addition, they did not achieve the same degree of morphological and biochemical alteration seen in virus-infected, TPA-treated cells. It is proposed that viral carcinogenesis, despite its rapidity, may occur in two stages: an "initiation" step caused by expression of a part of viral genome other than src (or by integration) and a promotion step (itself a multistep process) caused by the activation of the src gene. The src gene product could be enhanced or replaced by other promoting agents.


Recognition of differentiation antigens of spermatogenesis in the mouse by using antibodies from spleen cell-myeloma hybrids after syngeneic immunization. Two differentiation antigens of spermatogenesis in the mouse are defined by monoclonal antibodies from hybridomas produced between the myeloma P3-X63Ag8 and spleen lymphocytes immunized with syngeneic testis cells. These antigens are on testis cells, and not liver, kidney, brain, spleen, or whole ovary. They are species but not strain specific and trypsin sensitive but collagenase insensitive. On the basis of their appearance during the onset of spermatogenesis they appear to be differentiation antigens expressed during different but overlapping time windows during spermatogenesis.


In vitro suppression of UAG and UGA mutants in the thymidine kinase gene of herpes simplex virus. We have studied the in vitro translation, in nuclease-treated reticulocyte lysates, of mRNA from cells infected with several thymidine kinase-deficient mutants of herpes simplex virus type I. The addition of suppressor tRNAs from yeast resulted in suppression of the mutant property in the case of two mutants. Synthesis of enzymatically active viral thymidine kinase was restored by serine-inserting amber suppressor tRNA in the case of HSV TK4- and synthesis of the intact, but inactive, thymidine kinase protein was restored by serine- and leucine-inserting UGA suppressor tRNAs in the case of HSV TK43-. Read-through of the normal termination at the end of the thymidine kinase gene was promoted by UGA suppressor tRNAs. We conclude that HSV-TK4- is an amber (UAG) mutant and that HSV-TK43- is an opal (UGA) mutant.


Generation of cytolytic T lymphocytes after reovirus infection: role of S1 gene. Cytotoxic T lymphocytes (CTLs) can be generated if spleen cells from reovirus-infected mice are stimulated in vitro with syngeneic reovirus-infected cells. These cytolytic effector cells demonstrate: (i) serotype specificity (i.e., maximal cytolytic activity is observed on target cells infected with the serotype used to induce the CTLs) and (ii) H-2 restriction. The SI gene was shown to be the predominant viral gene determining the specificity of the cytotoxic T cells. This genome segment has previously been demonstrated to encode the viral hemagglutinin and determines reovirus cell tropism in the nervous system.


Rapid turnover of acetyl groups in the four core histones of simian virus 40 minichromosomes. The four core histones (H2a, H2b, H3, and H4) bound to simian virus 40 minichromosomes isolated from infected cells contain rapidly labeled acetyl groups in internal positions of the histone polypeptide chain. Upon chase, these acetyl residues decay with a half-life of less than 15 min. The acetyl groups are incorporated in histones bound to mature chromosomes and not in newly synthesized histones bound to replicating viral chromosomes. The rate of acetate incorporation is not related to the degree of steady state acetylation of the individual viral or cellular histones. This rate is 4-fold higher for the viral chromatin than for its cellular counterpart isolated from the same nuclei. The possible role for histone acetylation in viral genome expression is discussed.


Enkephalin inhibits release of substance P from sensory neurons in culture and decreases action potential duration. Sensory neurons grown in dispersed cell culture in the absence of non-neuronal cell types contain immunoreactive substance P that is chemically similar to synthetic substance P. When depolarized in high-K+ media (30-120 mM), the neurons release this peptide by a Ca2+-dependent mechanism. An enkephalin analogue, [D-Ala2]enkephalin amide, at 10 micron inhibits the K+-evoked release of substance P. At the same or lower concentrations, [D-Ala2]enkephalin amide and enkephalin decrease the duration of the Ca2+ action potential evoked and recorded in dorsal root ganglion cell bodies without affecting the resting membrane potential or resting membrane conductance. This modulation of voltage-sensitive channels may account for the inhibition of substance P release.


Semisynthetic horse heart [65-homoserine]cytochrome c from three fragments. Horse heart cytochrome c was treated with methylsulfonylethyloxycarbonyl succinimide (Msc-ONSu) to give fully N(epsilon)-protected cytochrome c. Treatment of this derivative with a hard base for 15 sec regenerated the native tetrahectapeptide chain. CNBr degradation of the protected compound produced three fragments bearing only protective Msc functions on epsilon-amino groups. The fragment comprising the sequence 81-104 was isolated from the mixture and acylated with N-hydroxysuccinimidyl-t-butyloxycarbonyl-L-methioninate. The resulting pentacosapeptide derivative was partially deprotected by treatment with acid and condensed in good yield (65%) with fully synthetic N(alpha66), N(epsilon72,73,79)- tetra-Msc-cytochrome-c-(66-79)-tetradecapeptide azide. This pathway is preferred because the pentadecapeptide azide derivative 66-80 acylated the N(epsilon)-protected tetracosapeptide sequence 81-104 in an unpredictable manner. Subsequent treatment of the product with a base produced unprotected semisynthetic cytochrome-c-(66-104)-nonatriacontapeptide, which is known to undergo acylation by unprotected [Hse(65)]cytochrome-c-(1-65)-pentahexacontapeptide lactone. The high specificity of this condensation is ascribed to "conformation direction." Semisynthetic [Hse(65)]cytochrome c thus prepared reacts like native cytochrome c with a succinate cytochrome c reductase preparation and with cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1). This semisynthetic strategy may provide a rapid route for the production of cytochrome c analogs modified in the highly conservative sequence 66-80.


Novel protein kinase, AUT-PK 85, isolated from adrenocortical carcinoma: purification and characterization. We describe the purification to apparent homogeneity of a protein kinase (designated AUT-PK 85) from adrenocortical carcinoma 494, as evidenced by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The enzyme binds cyclic AMP (cAMP) and autophosphorylates but does not use histone, casein, or polysomes as substrates in the presence or absence of cAMP. Stoichiometry of phosphate incorporation was 0.71 mol/mol of enzyme. The enzyme was found to have a molecular weight of 85,000 based on gel filtration. The protein was composed of polypeptides having the same molecular weight 42,000, and thus it appears to consist of two subunits of equal size. The enzyme bound two cAMP molecules, indicating that each subunit binds one molecule of cAMP. The homogeneous enzyme did not inhibit the protein kinase activity of the free catalytic subunit of normal adrenal cAMP-dependent protein kinase under conditions such that recombination with the free regulatory subunit occurred. cAMP bound specifically to the enzyme with an apparent dissociation constant (cfKd) of 1.2 X 10(-8) M. Scatchard plot data indicated one type of binding sites for cAMP. The enzyme did not bind adenosine. This novel autophosphorylating, cAMP-binding, protein kinase may be a characteristic of certain adrenal neoplasms.