Replication of Epstein-Barr virus: ultrastructural and immunofluorescent studies of P3HR1-superinfected Raji cells. We have studied by means of electron microscopy and immunofluorescence the different steps of the replication of the P3HR1 strain of Epstein-Barr virus in Raji cells. The virus entered the cell by fusion of the viral envelope with the plasma membrane, followed by the disintegration of the capsid. In some cases, the migration of nucleocapsids toward the nuclear membrane was observed. The synthesis of new virions began as early as 7 h after infection (in the case of a high multiplicity of infection [MOI]-800 particles per cell) and took place in low-electron-density areas of the nucleus. A viral envelope was acquired by budding either through the nuclear membrane or more often through membranes of the Golgi apparatus or cytoplasmic vacuoles. Comparing immunofluorescence and electron microscopic data a good correlation was found between the presence of early antigen and ultrastructurally altered cells, as well as between the presence of viral capsid antigen and virus-producing cells. With different MOIs, different types of viral cycles were observed: at a low MOI (less than or equal to 50 particles per cell), a nonproducer cycle was induced, with early antigen synthesis only; at a higher MOI (100 particles per cell), a transient production of a small amount of virions was observed, and at a high MOI (greater than or equal to 300 particles per cell), a productive cycle was the rule.


Structural protein markers in the avian oncoviruses. The proteins of purified avian oncoviruses were analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and isoelectric focusing. Certain members of the avian leukosis-sarcoma viruses (ALSV) had group-specific antigens with altered electrophoretic properties. (i) The p27 protein of Rous-associated virus 0 (RAV-0) had a lower electrophoretic mobility in SDS gels and a lower isoelectric point than the p27 of other ALSV. (ii) The p19 proteins of RAV-1, RAV-2, and the Bryan high-titer strain of Rous sarcoma virus had higher mobilities in SDS gels than did the corresponding protein of other viruses. This altered electrophoretic mobility was correlated with specific differences in the tryptic peptides of radioiodinated p19s. (iii) The p15 protein of RAV-7 had a lower mobility in SDS gels than did the p15 of other ALSV. These markers were used in a study of the structural proteins of subgroup E RAV-60 produced after infection of chicken embryo cells by exogenous ALSV. Although exogenous group-specific protein markers could often be identified in the subgroup E isolates, one RAV-60 had a p27 that comigrated with the p27 of RAV-0. The p19s of two other RAV-60 isolates had electrophoretic properties that were different than those of p19s from either RAV-0 or the exogenous viruses. These results support the hypothesis that RAV-60 is generated by recombination between endogenous and exogenous oncoviruses and indicate that at least the p27 encoded by RAV-0 is closely related to a protein specified by endogenous viral information in chicken cells.


Effects of filipin on the structure and biological activity of enveloped viruses. The interaction of the polyene antibiotic filipin with membrane-bound cholesterol in vesicular stomatitis (VS), influenza, and Rauscher leukemia virions was studied. Exposure of virions to filipin resulted in a series of depressions and ridges in the envelope of VS virions, with a periodicity of 15 to 20 nm perpendicular to the long axis of the particle; similar morphological alterations were observed in negatively stained preparations, in thin-sectioned virions, and in protease-treated virions that lack surface glycoproteins. This morphological effect was specific for filipin, since the envelopes of VS virions that had been treated with another polyene antibiotic, amphotericin B, exhibited markedly different morphology. Morphological alterations induced by filipin in influenza and Rauscher leukemia virions differed from those seen in VS virions. The infectivity of filipin-treated VS virions was reduced up to 500-fold, whereas influenza virions were resistant to filipin treatment. Incorporation of filipin into the virions was demonstrated, and no release of either lipids or proteins from virions was detected after filipin treatment. A stoichiometry of approximately 1 mol of bound filipin per mol of cholesterol was found in both intact and protease-treated VS virions. The equilibrium dissociation constant for filipin-cholesterol interaction was approximately 74-fold larger in intact than in protease-treated VS virions. The initial rate of association of filipin with cholesterol in intact virions was slower than that in protease-treated particles. The fluidity of lipids in VS viral membranes, as probed by a stearic acid derivative spin label, was markedly reduced when either intact or protease-treated virions were treated with filipin.


Uncoating and gene expression of simian virus 40 in CV-1 cell nuclei inoculated by microinjection. CV-1 cells were inoculated with simian virus 40 virus within the nucleus or cytoplasm by microinjection. Virions were uncoated with great efficiency within the cell nucleus.


Transformation-defective mutants of Rous sarcoma virus with src gene deletions of varying length. The RNAs of transformation-defective (td) deletion mutants of the Schmidt-Ruppin strain of Rous sarcoma virus were found to vary in size when compared by polyacrylamide gel electrophoresis. Three of seven td mutants appeared to recombine with a mutant of Rous sarcoma virus (Schmidt-Ruppin), which has a temperature-sensitive sarcoma (src) gene and is termed ts68, to give rise to recombinants with a reduced temperature sensitivity. The results suggested that different clones of td mutants exist: some in which the src gene appears to be deleted, and others in which the src gene is only partially deleted. A direct correlation between RNA size and the extent of src gene deletion measured by recombination was not obtained, possibly because the recombination assay could only detect src sequences homologous to the lesion(s) of ts68, whereas the electrophoretic analysis of the RNA measured src deletions as well as other possible alterations of the RNA.


Clinical evaluation of pivmecillinam in intractable urinary-tract infections with complications. A comparative study with amoxicillin by a randomized double-blind technique. A comparative study was made by the double-blind technique in order to make clear the usefulness of pivmecillinam in the treatment of intractable complicated urinary-tract infections using amoxicillin as a reference drug. Pivmecillinam was given in dosage of 400 mg (potency) per day which was one-fifth the dose of amoxicillin 2,000 mg (potency) per day. In global judgement, pivmecillinam was found superior to amoxicillin. It showed a "significant" superiority over amoxicillin for the treatment of, among others, the urinary-tract infections after prostatectomy, which are intractable diseases. When evaluated by symptoms, pivmecillinam improved bacteriuria "significantly" better than amoxicillin. When seen by causative organisms, the pivmecillinam treatment was "significantly" superior to the amoxicillin treatment against E. coli infections. Pivmecillinam was active against amoxicillin-resistant E. coli. Incidence of adverse reactions was less frequent with pivmecillinam than with amoxicillin. These results indicate that pivmecillinam is a drug of high usefulness for the treatment of intractable complicated urinary-tract infections when evaluated using amoxicillin as a reference drug.


Effect of diltiazem on insulin secretion. I. Experiments in vitro. Effect of diltiazem on glucose-induced insulin secretion was investigated in the rat islets of Langerhans isolated by a collagenase digestion technique. It was found that B-cells, main constituents of isolated islet preparations, had a well-preserved ultrastructural appearance immediately following isolation or after incubation with glucose or glucose and diltiazem. The islets released a large amount of insulin upon stimulation with glucose and CaCl2. Diltiazem (10(-6)-10(-4) M) produced a dose-related inhibition of glucose-induced insulin secretion and this effect was antagonized by the increase in extracellular concentration of CaCl2. The inhibitory effect of diltiazem on the insulin secretion was also counteracted by dibutyryl-3',5'-cyclic AMP or by theophylline. Among calcium-antagonists tested, nifedipine produced the most powerful inhibitory action on the insulin secretion, while the effect of verapamil was similar to or somewhat stronger than that of diltiazem. It was suggested that diltiazem may reduce the intracellular concentration of free calcium ion, thus causing an inhibitory effect on the glucose-induced insulin secretion by the isolated islets of Langerhans.


Serologic study on the prevalence of murine viruses in five Canadian mouse colonies. A serologic study was made of five Canadian mouse colonies to determine the prevalence of antibodies to 11 murine viruses. A total of 139 sera from the five colonies were evaluated by complement fixation or hemagglutination inhibition methods. Viral antibodies were present in all five colonies. Antibodies to pneumonia virus of mice, minute virus of mice, Theiler's encephalomyelitis virus, and reovirus type 3 were found in all five colonies. K-virus antibodies were present in four colonies. Polyoma virus antibodies were found in two colonies and Sendai virus antibodies in two other colonies. Mouse hepatitis virus antibodies were present at a low prevalence in three of the five colonies. No antibodies to adenovirus, lymphocytic choriomeningitis virus, and ectromelia were detected in any of the colonies.


The starfish (Patiria and Pisaster) oocyte: a model system for the study of hormone-surface interaction in oogenesis. Studies were made to determine if the starfish (Patiria and Pisaster) oocyte could serve as a model system for investigations of hormone-surface interactions in eucaryotic cells. Treatment of starfisch oocytes with maturation inducing hormone, 1-methyladenine, was observed by scanning electron microscopy to induce surface alteration. The action of the hormone was dependent on the presence of the cation Ca++. Although caffeine, theophylline, and theobromine supposedly inhibit maturation of oocytes, studies using the starfish oocyte showed that theobromine does not inhibit maturation and the inhibition caused by caffeine and theophylline is reversible. Other studies using the starfish oocyte and tritiated 1-methyladenine showed that the hormone does not dissociate from the oocyte once bound, and that the oocyte surface may have multiple receptors for 1-methyladenine. From these studies it was concluded that the starfish oocyte is a useful model for studies of hormone-surface interactions in eucaryotic cells.


Nonhuman primates: laboratory animals of choice for neurophysiologic studies of sleep. A systematic study was made of several nonhuman primates to learn more about their relative usefulness for studies of sleep. Species studied included the Guinea baboon (Papio papio), Kenya or yellow baboon (P cynocephalus), olive baboon (P anubis), sacred baboon (P hamadryas), vervet monkey (Cercopithecus aethiops), rhesus monkey (Macaca mulatta), bonnet monkey (M radiata), crab-eating monkey (M fascicularis), patas monkey (Erythrocebus patas), chimpanzee (Pan troglodytes), mongoose lemur (Lemur mongoz), black lemur (Lemur macaco fulvus), and bushbaby (Galago senegalensis). Comparisons were made of electroencephalographic activity, states of vigilance, and responses to sleep-waking drugs. The results showed that there were major similarities among the genera studied as well as individual intrageneric and intraspecies differences. It was concluded that the chimpanzee, olive baboon, and rhesus monkey were the best species for comparative studies, and that the rhesus monkey was the best single model because of its well defined sleep organization and ease of handling and housing.


Nerve abscess in lepromatous leprosy. A case report and a discussion of pathogenesis. An instance of nerve abscesses developing in a patient with lepromatous leprosy is reported. The pathogenesis of nerve abscess in lepromatous leprosy is briefly discussed. It appears that such abscesses may develop (i) from an ENL lesion in the nerve during ENL reaction, (ii) because of exacerbation of existing lepromatous lesion, (iii) arise as an "exacerbation nodule", (iv) due to quiet necrosis in a lepromatous granuloma, or (v) it may be iatrogenic.


Hormonal regulation of membrane phenotype. Incubation of rat hepatoma cells (HTC) in tissue culture with glucocorticoids alters several membrane properties characteristic of transformed cells, without affecting the growth rate of these cells. Variant cell lines resistant to dexamethasone inhibition of plasminogen activator production have been isolated using an agar-fibrin overlay technique to detect plasminogen activator production by individual colonies of HTC cells. The resistance of dexamethasone is not secondary to abnormal or absent glucocorticoid receptors, but due to a lesion in a later step in hormone action specific for plasminogen activator. These variants should prove useful for the study of the mechanism of steroid action as well as for the analysis of the role of proteases in the hormonal regulation of membrane function.


Relationship between thymidine transport and phosphorylation in Novikoff rat hepatoma cells as analyzed by a rapid sampling technique. Incorporation of thymidine into Novikoff rat hepatoma cells was analyzed with a rapid sampling technique which allowed collection of 12 time points in 20 sec. Transport was a rapid, saturable, nonconcentrative process with a Km of about 85 micrometer. The intracellular thymidine pool was also rapidly labeled in cells which phosphorylated thymidine, that a group translocation process involving thymidine kinase can be ruled out. Under all conditions examined, phosphorylation, not the transport, of thymidine was the rate-determining step in its incorporation into the acid-soluble pool. Estimation of transport rates from total incorporation into cells which phosphorylate the substrate is invalid in this cell system and must be questioned in all instances.


The glucagonoma syndrome and its management. The glucagonoma syndrome occurs in some but not all patients with a benign or malignant islet cell tumor and hyperglucagonemia. Manifestations may include anemia, diabetes mellitus, pruritic skin rash, glossitis, stomatitis, weight loss, diarrhea, flexible fingernails, venous thromboses, low plasma amino acid levels, and coarse folds of the jejunum and ileum. Most patients are postmenopausal women, but men and women ages 40 to 65 have been affected. The course is variable depending upon the nature of the underlying tumor. Twenty-two cases of probable glucagonoma syndrome have been reported; twelve documented with glucagon levels. The hyperglucagonemia results from elevation of the proglucagon and true glucagon immunoreactive fractions of pancreatic glucagon. Management of the rash can be accomplished rarely with topical or systemic antibiotics or corticosteroids. If the tumor is resectable, surgery reverses the syndrome. Patients with metastatic disease have responded to streptozotocin and DTIC.


[Bilateral salivary duct carcinoma of the parotid gland (author's transl)]. A case of bilateral carcinoma of the salivary duct is presented. The histological appearance of this rare tumor and the therapeutical consequences are reviewed.


p-Chlorophenoxyisobutyrate enhanced retention of homologous lipoproteins by human aortic smooth muscle cells. Human aortic smooth muscle cells (SMC) specifically bind and take up indiscriminately both the lipid and protein moieties of homologous 25I-very low density lipoproteins (VLDL) and 125I-low density lipoproteins LDL). Sixty-five to 80% of absorbed lipids are incorporated into the cell lipids, preferentially into the phospholipid fraction. Twenty to 35% of the lipid bound and the protein moiety are eliminated from the cells. Half of the eliminated protein label is recovered as TCA soluble products. Five mM of p-chlorophenoxyisobutyrate (CPIB) raise the level of intracellular radioactivity derived from the lipid moieties of VLDL and LDL by about 40% via a reduced elimination. The processing of the protein moiety and lipoprotein binding to the cell surface are not affected by 5.0 mM of CPIB. CPIB lowers the incorporation of 14C-acetate, 14C-pyruvate, and 32phosphate radioactivity into fatty acids and phospholipids of aortic SMC. Five mM of CPIB reduce the overall palmitic acid synthesis by shifting from de novo synthesis to the mechanism of chain elongation, although the further elongation to saturated C18-C24 fatty acids is also depressed. The CPIB-enhanced retention of the lipid-derived lipoprotein radio-activity is interpreted as a compensatory mechanism providing cellular fatty acids which are deficient as a result of the CPIB inhibited synthetic processes.


Effects of dibutyryl adenosine 3',5'-monophosphate on hepatic metabolism of free fatty acids. The effects of dibutyryl cyclic adenosine 3',5'-monophosphate (Bu2cAMP) on metabolism of free fatty acids by perfused livers from normal fed male rats were investigated. In one group of experiments, Bu2cAMP was added to the medium and infused at a constant rate to maintain concentrations of 0, 0.4, 1.0, 4.0, or 10. 0 X 10(-5) M nucleotide in the perfusate plasma, assuming the nucleotide was not metabolized by the liver. Oleic acid was infused as the complex with albumin at the rate of 124.3 mumoles/hr. Uptake of free fatty acid by the liver was identical in all groups. Production of ketone bodies, however, increased, and output of triglyceride decreased with increasing concentration of Bu2cAMP. The nucleotide also stimulated output of glucose. Maximal effects were observed when the concentration of Bu2cAMP was approximately 2-3 X 10(-5) M. The output of very low density lipoproteins, as judged by flotation in the zonal ultracentrifuge, was also diminshed by the nucleotide. In other experiments, 1-14C-oleate was infused (120.8 mumoles/hr) along with 2 X 10(-5) M Bu2cAMP, and the disposition of 14C into CO2, ketone bodies, and esterified lipids was evaluated. Bu2cAMP depressed the proportion of 1-14C-oleate converted to triglyceride and increased the fraction converted to ketone bodies and CO2. Not only was ketogenesis stimulated, but a larger proportion of the ketone bodies was derived from exogenous fatty acid.


Secondary type II hyperlipoproteinemia in patients with anorexia nervosa. In 18 patients with anorexia nervosa, plasma cholesterol and triglyceride concentral concentrations were repeatedly determined over a period of 14 mo. In 11 patients elevated cholesterol concentrations were found which were due to an increase of low-density lipoprotein cholesterol, whereas high-density lipoprotein and very low density lipoprotein cholesterol levels were in the normal range. The elevated cholesterol values did not correlate with clinical and laboratory parameters such as the degree of weight loss and thyroid function tests. In follow-up studies it could be shown that in patients who regained their original weight, elevated plasma cholesterol concentrations fell to normal levels parallel to weight increase. In patients who showed no change in weight, however, cholesterol levels remained high. The cause for this secondary type II hyperlipoproteinemia in anorexia nervosa is not known. Hepatic triglyceride lipase and lipoprotein lipase activities in post-heparin plasma were found to be low despite normal triglyceride concentrations.


Effects of chlorinated hydrocarbon on plasma alpha-lipoprotein cholesterol in rats. Ten chlorinated hydrocarbons (tetradifon, Chlorobenzilate, Kepone, lindane, Kelthane, toxaphene, dieldrin, hexachlorobenzene, dienochlor, and Aroclor 1254; see below) were evaluated in rats to assess agents which might selectively elevate high-density lipoprotein cholesterol (C-HDL). Single, nontoxic doses of Aroclor, dieldrin, and Kepone elevated C-HDL at day 7 (31%, 26%, and 24%, respectively, p less than 0.05). The C-HDL elevations were maintained at days 21 and 60 (28% and 31%, p less than 0.01) in the Aroclor group, while C-HDL returned to baseline in the dieldrin and Kepone groups. Liver function tests were unchanged from control, suggesting that the changes in C-HDL were not the consequence of hepatotoxicity. The hepatic microsomal cytochrome P-450 content of animals receiving Aroclor, dieldrin, and Kepone was 67%, 56%, and 44% higher than control values (p less than 0.02), indicating probable hepatic enzyme induction. If HDL (an antiatherogenic lipoprotein) is also selectively elevated by chlorinated hydrocarbons in man, then analogues might potentially be developed as antiatherogenic pharmacologic agents. Alternatively, the implications of man's exposure to halogenated hydrocarbons may be better understood.


A simple and efficient microassay method for titration of interferon. A simple and efficient microassay method for the titration of interferon was developed by the use of microtest plates for handling a large number of samples. L929 cells pretreated with interferon were infected with vesicular stomatitis virus (VSV) and cultured in the presence of 3H-uridine. The activity was expressed by the reduction of extracellular radioactive RNA released after destruction of the infected cells, which was measured in terms of the radioactivity incorporated into cold TCA-insoluble materials in the culture fluid. The interferon titer determined by this method was in the same order as that by the plaque reduction method. The activity by this method was parallel to, but lower than that expressed by the yield reduction of infectious viruses. This method requires only 0.025 ml of each test sample with higher than 1 NIH ref. unit/ml to detect its interferon activity and takes 2 to 3 days for assaying hundreds of samples.


The relationships between cyclic AMP, calcium and prostaglandins as second messengers. There is considerable dissatisfaction with present second messenger hypotheses involving cyclic nucleotides and calcium. The recent findings that methyl xanthines and adenosine are prostaglandin antagonists casts doubt on much of the evidence in favour of the cyclic AMP hypothesis. There is evidence that allosteric sites may modify the binding of calcium and cyclic nucleotides to key cellular regulators and that a range of substances including steroids, adenosine and prostaglandins may occupy those sites. This concept introduces much needed flexibility into the second messenger concept, allows many experiments to be reinterpreted and has major implications throughout the biomedical sciences.


[Cogenital goiter in a premature infant after amniography (author's transl)]. A premature infant was born with goiter and hypothyroidism 3 weeks after amniography. The correlation between excessive iodide exposure and thyroid function of discussed. The danger of using of idodine containing contrast media for amniography are emphasized.


Increased growth after long-term oral 1alpha,25-vitamin D3 in childhood renal osteodystrophy. We evaluated oral 1,25-vitamin D3 for as long as 26 months in six prepubescent children with renal osteodystrophy previously treated with vitamin D2. Therapy was given at 14 to 41 ng per kilogram per day to correct hypocalcemia and reverse bone disease. Serum levels of 1,25-vitamin D3 were initially reduced at 15 +/- 5 pg per milliliter (mean +/- S.E.M.) and after treatment rose to 54 +/- 13. Serum calcium rose from 7.5 +/- 1.6 mg per deciliter (mean +/- S.D.) to 9.8 +/- 0.6 after one month (P less than 0.02). Alkaline phosphatase activity fell from 536 +/- 298 to 208 +/- 91 IU per liter after 12 months (P less than 0.05). Serum immunoreactive parathyroid levels fell from 900 +/- 562 microliter eq per milliliter 411 +/- 377. Healing of rickets and subperiosteal erosions was found. Remineralization of bone was demonstrated by the photon absorption technic. In four patients growth velocity, evaluated for 12 months before and after therapy, increased from 2.6 +/- 0.8 to 8.0 +/- 3.2 cm per year. Growth velocity per year increased from less than third percentile in each to the 10th to 97th percentile after therapy. Height increment ranged from 27 to 113 per cent of that expected for change in chronologic age and 40 to 114 per cent expected for change in bone age after therapy. This trial demonstrates that oral 1,25-vitamin D3 can reverse renal bone disease and increase growth in uremic children.


Protein-DNA and protein-hormone interactions in prealbumin: a model of the thyroid hormone nuclear receptor? High resolution X-ray analysis of the hormone-binding protein prealbumin has shown that it has a structural complementarity to double-helical DNA. The proposed binding site is composed of two symmetry-related beta-sheets containing a pair of helically disposed arms, which can interact with the bases in the wide groove of DNA. A palindromic target sequence is indicated by the symmetry of the protein. The two identical thyroid hormone binding sites on prealbumin are located in a channel that runs completely through the molecule. These two structural features suggest prealbumin as a model for the thyroid hormone nuclear receptor, providing a number of detailed predictions of its properties.


Possible regulatory function of acetylcholine receptor in maintenance of retinotectal synapses. alpha-Neurotoxins bind to cholinergic receptor, block transmission, and induce sprouting of retinal terminals in the toad tectum. New connections retain an orderliness that suggests a selective affinity between presynaptic terminals. The results suggest that postsynaptic cells exert a control, associated with receptors, on the growth of presynaptic terminals and on the maintenance of their synaptic connections.


Recent excitement in the DNA replication problem. It is now possible to reproduce most of the reactions involved in DNA replication using prokaryotic enzymes in vitro. Such systems have revealed that DNA replication is a complex process depending on a relatively large number of proteins, and that nucleoside triphosphate hydrolysis energy is used at several discrete steps. Much of the complexity of DNA replication may arise from the need for extreme copying fidelity.


A mutation that impairs the ability of lipoprotein receptors to localise in coated pits on the cell surface of human fibroblasts. Low density lipoprotein is taken up by cultured human fibroblasts through an endocytic process that requires the binding of the lipoprotein to specific receptors located in coated pits on the cell surface. The coated pits are discrete segments of the plasms membrane that can undergo rapid invagination to form coated endocytic vesicles. In one form of the human genetic disorder familial hypercholesterolaemia, the responsible mutation produces altered lipoprotein receptors that lack the ability to become incorporated into coated pits. Instead, these mutant receptors are scattered at random over the entire plasma membrane. Becuase of their mislocation on the cell surface, the mutant lipoprotein receptors are unable to carry their bound lipoprotein into the cell. The occurrence of this 'receptor mislocation mutation' provides strong evidence for the role of the coated pit in the receptor-mediated uptake of lipoproteins. The data also have implications for the structure and assembly of plasma membranes in mammalian cells.


[EPSP's of the motor neurons of cat masticatory muscles induced by stimulation of low-threshold infraorbital nerve fibers]. The effects of the infraorbital Aalpha afferents stimulation on the masseter motoneurons were investigated in cats under chloralose-nembutal anesthesia. It is found that stimuli 1.4-2.5 times threshold (T) evoke complex EPSPs in 69% of the investigated motoneurons, their latency being 2.1 +/- 0.2 ms (1.5-3.0 ms, n = 36), amplitude up to 30 mV and duration 9-15 ms. These EPSPs consist of two simpler responses produced by activation of two separate groups of afferent fibres. The short-latent EPSPs arise after activation of the infraorbital nerve by 1.1-1.5 T stimuli and have the same latency as the complex EPSP (1.5-3.0 ms) but a smaller amplitude (up to 2.0 mV) and shorter duration (up to 6 ms). The stability of these EPSPs during high frequency stimulation (120/c) and the development of facilitation and inhibition similar to those which appear in monosynaptic EPSPs in masseter motoneurons during stimulation of the 1a muscle afferents give reason to suggest that these EPSPs are monosynaptic. The slow rising rate indicates that they appear on distal dendrites of the motoneurons. The long-latent EPSPs arise with latency of 7-9 ms after activation of the infraorbital nerve by 1.1-1.5 T stimuli. Their amplitude reaches 1.5-2.0 mV and duration 7-9 ms. The long latency of these EPSPs in combination with low ability to repeat high frequency stimulation are consistent with their polysynaptic origin. It is suggested that the excitatory input from the lowest threshold Aalpha afferents of the infraorbital nerve to masseter motoneurons creates conditions for the development of a transient jaw closing reflex in response to light tactile stimulation of the cat's perioral region.


Cyclophosphamide induced sensitivity against avian RNA myeloblastosis virus in age-resistant hosts. The effect of cyclophosphamide administration on survival of 4- to 7-week-old chickens as well as on induction of leukemia after avian myeloblastosis virus (AMV) injection was studied. The drug treatment alone did not cause any neoplastic effect in the birds during 4 months of observation. Immediate application of AMV to cyclophosphamide-pretreated age-resistant chickens induced acute myeloblastic leukemia in about 80 per cent of test animals. The sensitivity of chickens against AMV, induced by cyclophosphamide, had transient character only. When AMV was injected delayed, 3 or 10 days later, after the administration of the drug was completed, a rapid and pronounced increase of resistance was observed again.


Biochemical properties of endogenous rat C-type viruses. The cellular restriction to persistence of inducible C-type viruses of at least two rat strains can be overcome in actively in vitro replicating and/or spontaneously transforming rat embryo cells. Both, SD-RaLV and W-RaLV particles are morphologically and biochemically related to known oncorna viruses: They are of C-type, have an identical buoyant density of 1.15 g/cm3 in sucrose gradients and contain high-molecular-weight RNA. The RaLV-associated DNA polymerase exhibit both exogenous (polynucleotide-templated) and endogenous (viral RNA-templated) reactivity. The enzyme strongly preferred Mn++ over Mg++ ions (Mg++/Mn++ ratio of 0.04 to 0.07), with maximum reactivity at 0.1 mM MnCl2 concentration. The molecular hybridization experiments revealed the endogenous nature of the SD-RaLV and the W-RaLV. From the competition hybridization assay the number of proviral sequences could be calculated to be 84 per haploid cell genome. The role of endogenous rat viruses in the "spontaneous" in vitro transformation is discussed.


Mitochondrial adenosine triphosphatase of Zajdela hepatoma. III. Effect of uncouplers on the hydrolysis of intramitochondrial ATP. Hydrolysis of extramitochondrial ATP by coupled Zajdela hepatoma mitochondria is not stimulated by uncouplers of oxidative phosphorylation. The results of the present study show that the hydrolysis of intramitochondrial ATP in these mitochondria is stimulated by DNP and CCCP. It is proposed that the uncoupler insensitivity of ATPase in coupled Zajdela hepatoma mitochondria with exogenous ATP as a substrate result from an altered functional relationship between ATPase and ADP, ATP translocase.


Pars intermedia ACTH and MSH content: effect of adrenalectomy, gonadectomy and a neurotropic (noise) stress. Pars intermedia (PI) ACTH and melanocyte-stimulating hormone (MSH) content and pars distalis (PD) ACTH concentration were measured following 3 experimentally induced 'perturbations'. The feedback-induced increase in ACTH release following adrenalectomy is not accompanied by a change in PI ACTH and MSH contents, while the feedback-induced increase in gonadotropin secretion following gonadectomy is associated with significant changes in PI ACTH and MSH contents. A well-defined neurotropic (noise) stressor resulted in significant increases in PI ACTH and MSH. A striking finding in all of the studies was a positive correlation between PI ACTH and MSH contents. We conclude that PI ACTH and MSH secretion are altered with feedback-induced alterations in gonadotropin secretion and with neurotropic stressors.


[Effect of polysynaptic flexion reflex on monosynaptic reflexes of calf muscles prior to and following intramuscular injection of ethylbenzatropine in drug-induced muscle rigidity]. In patients with drug-induced parkinsonism and in healthy subjects the effect of stimulation of low-threshold skin fibres of sural nerve producing the polysynaptic flexion reflex of the short head of the biceps femoris muscle on the monosynaptic H reflex of calf muscles was studied before and after administration of ethylbenzatropine. In healthy subjects stimulation of low-threshold skin nerve was followed by facilitation of H reflex in the time of 70 to 200 msec. from the conditioning stimulation to the test stimulation. After one single intramuscular dose of ethylbenzatropine this late facilitating effect disappeared. In patients with drug-induced parkinsonism stimulation of afferent fibres of flexion reflex failed to cause late facilitation of H reflex and one dose of ethylbenzatropine brought no changes. During systematic administration of the drug an evident tendency for facilitation of H reflex was observed, similarly as in healthy subjects. The results obtained in healthy subjects and in patients with drug-induced parkinsonism are explained as evidence of inhibitory action of noradrenaline in certain chains of spinal interneurons, since noradrenaline release in the spinal cord is determined by the striatal equilibrium between the cholinergic and dopaminergic systems. The corrective action of ethylbenzatropine would depend on its central action on certain spinal chains of interneurons determining transmission of impulses from dermal afferent nerves to alpha motoneurons.


[Comparison of results of polygraphic sleep studies and cerebrospinal fluid pressure records in patients following removal of supratentorial expanding lesions]. In 4 patients operated upon for supratentorial brain tumors the authors carried out on the 3rd day after the operation polygraphic recording of nocturnal sleep with simultaneous measurements of cerebrospinal fluid pressure in the ventricular system by the method of Lundberg. A comparison of polygraphic records of nocturnal sleep with parallel measurements of intraventricular pressure demonstrated that all REM phases were associated with a rise in intraventricular pressure, while during phases 3 and 4 of sleep (NREM) the intraventricular pressure was always lower. In the remaining sleep phases 1,2 NREM and during episodes of nocturnal awakening no unequivocal consistencies were found.


Inclusion body myositis: a distinct variety of idiopathic inflammatory myopathy. We report six cases of inclusion body myositis (IBM), a distinct but infrequently recognized inflammatory disease of skeletal muscle. Clinically, IBM differs from dermatomyositis and polymyositis. It lacks features of collagen-vascular disease, has a relatively benign and protracted course, frequently involves distal muscles, is found mainly in males, and does not improve with corticosteroid treatment. Electronmicroscopic demonstration of abnormal filaments in muscle cells is necessary for definite diagnosis, but IBM may be suspected by the finding on cryostat sections of numerous hematoxylinophilic granules in "lined" vacuoles in muscle cells. These correspond to whorls of cytomembranes. Although in dermatomyositis the capillary network is partly destroyed, in IBM it is usually augmented. A viral etiology of IBM has been suggested but remains unproven.


EEG frequency spectrum characteristics of sleep states in infants of alcoholic mothers. Infants of alcoholic mothers showed prominent EEG hypersynchrony in all three stages of sleep: in quiet sleep, indeterminate sleep and active-REM sleep. Spectral analysis of the EEG using fast Fourier transform revealed significant increase in power in most frequency bands in all three stages of sleep in infants born to alcoholic mothers when compared with normal healthy infants matched for gestational age. In quiet sleep "alcoholic" infants differed from healthy babies by significantly higher power in a wide range of frequency bands (1.5--17.5 Hz) with an average 143% increase of the integrated EEG (1.5--25 Hz). In indeterminate sleep "alcoholic" infants showed significantly higher power in all analyzed frequency bands (0.1--25 Hz) with an average 196% increase of the integrated EEG (1.5--25 Hz). In active-REM sleep infants of alcoholic mothers showed significantly higher power in the frequency range from 0.1 to 17.5 Hz in comparison with healthy controls. The average increase of the integrated EEG (1.5--25 Hz) was 200%. All healthy term infants showed significantly higher power in most frequency bands during quiet sleep in comparison with active-REM sleep. In infants born to alcoholic mothers this quiet sleep--REM sleep frequency spectrum difference in the majority of cases was nonsignificant mainly due to high values of the EEG power during REM sleep. It is unlikely that the EEG hypersynchrony in infants of alcoholic mothers represents one of the symptoms of the neonatal alcohol-withdrawal syndrome since hypersynchrony has been detected as long as 6 weeks after birth. During this time any withdrawal symptoms would have been dissipated.


[Endocrine aspects of puberty. Modifications of gonadotropins (LH, FSH), testosterone and cortisol circulating after ACTH and synthetic LH-RH]. The effect of synthetic LH-RH (Relisorm Serono: 25 microgram i.v.) on plasma LH, FSH and testosterone was examined in 19 normal pubescent and prepubescent boys. Changes in cortisol and testosterone after 0.25 mg/m2 body surface ACTH (Synacthen ciba) was assessed in a further 8 prepubescent subjects. Notable amounts of FSH, LH and testosterone were noted in prepubescent subjects, pointing to hormonal production prior to sexual maturation. There was a prompt and significant increase in LH in all cases. In prepubescent subjects, this was accompanied by marked increases in FSH on some occasions, though no appreciable change in circulating testosterone. This suggests that the hypophyseal reserve of LH is sufficient in puberty. Prepubescent subjects displayed a significant rise in cortisol 30' and 60' after ACTH, whereas testosterone fell with respect to the start values. The results indicate that acute stimulation of the adrenal cortex before puberty leads to testosterone suppression.


Rat liver mitochondrial cytochrome c oxidase and cytochromes in experimental protein-energy malnutrition. Cytochrome c oxidase activity and cytochromes b, (c+c1) and a(+a3) concentrations were determined in liver mitochondria from rats fed the following diets: controls (group 1) fed ad libitum, energy-restricted (group 2) and protein-deficient (group 3). The animals were fed for two time intervals, 3--5 and 7--9 weeks. At 3--5 weeks, the cytochrome oxidase specific activity (nmol cytochrome c oxidized/mg protein/min) and cytochrome concentrations (nmol/mg protein) were not different in groups 2 and 3 as compared to group 1. At 7--9 weeks, the cytochrome oxidase specific activity and concentrations of cytochromes b, (c+c1) and a(+a3) were significantly reduced in group 2 rats as compared to well-fed controls. The Michaelis-Menten constant, Km apparent for ferrocytochrome c, was significantly higher in group 2 as compared to group 1. In group 3 rats, cytochrome oxidase specific activity and cytochrome b, a(+a3) concentrations were not different from group 1 at 7-9 weeks. However, cytochrome (c+c1) concentration was higher in group 3, resulting in an elevated ratio of cytochrome (c+c1) to cytochrome a(+a3) as compared to groups 1 and 2.


Methotrexate with citrovorum factor rescue for gestational trophoblastic neoplasms. Thirty-five patients with nometastatic gestational trophoblastic neoplasms and 3 patients with metastatic gestational trophoblastic neoplasms were treated primarily with methotrexate and citrovorum factor rescue. The antecedent pregnancy was molar in all patients. The known histologic diagnosis in 34 patients was hydatdiform mole and choriocarcinoma in 3. Up to March 1977, the duration of remissions ranged from 1 to 21 months. Complete and sustained remission was achieved in 91% of patients with nonmetastatic disease and in 2 of the 3 patients with metastases, without evidence of marrow or hepatic and with substantially reduced epithelial toxicity. Response to treatment and the number of courses required to achieve remission were determined solely on the basis of the human chorionic gonadotropin response as measured by the beta subunit radioimmunoassay.


Temperature effects on the modulation of adenylate cyclases from rat liver and Morris hepatomas. When adenylate cyclase activities in purified membranes from normal rat liver and from a series of rapid growing transplantable Morris hepatomas were examined at various temperatures, several unique features were observed. Two of the hepatomas yielded patterns similar to that of normal liver, even though glucagon did not activate either tumor adenylate cyclase but did activate the normal liver enzyme. The patterns of the third tumor line were completely different from normal. This clearly shows the heterogeneity in cancers of similar origin.


Adenosine 3',5'-cyclic monophosphate and guanosine 3',5'-cyclic monophosphate levels in human breast cancer tissue. Intracellular levels of adenosine 3',5'-cyclic monophosphate and guanosine 3',5'-cyclic monophosphate were measured in breast carcinomas. These levels were increased significantly as compared to normal breast tissues. In benign breast tumors, intermediate levels of adenosine 3'5'-cyclic monophosphate were observed, but they did not differ significantly with malignant tissues. No relation has been shown with estradiol or progesterone binding sites.


An investigation into the relationship of the immunological status of Herpes simplex virus type-2 (HSV-2) infected animals and the tumor-enhancing property of this infection. In order to compare the state of immunity of HSV-2 infected animals with their sham treated counterparts, several in vivo methods for cell-mediated immunity in addition to the stimulation in vitro of mouse splenocytes with PHA were performed. The grafting of C 3 H mice with heterologous skin and the paw test with PHA showed no noticeable difference between the two groups. The tuberculin test performed on guinea pigs infected via the footpads, permitted the detection of a slightly impaired reactivity of the virus-infected animals. The PHA assay in vitro showed reduced stimulation indices with cultures of splenocytes from virus-infected mice about 10 days p.i. These reduced stimulation indices are the result of 1. a low 3 HTdR incorporation by the PHA stimulated cultures and 2. of a relatively high 3 HTdR uptake by the unstimulated control cultures. An interpretation of these results is suggested. Finally, the possible implications of a shortlived impaired cell-mediated immunity on the tumour enhancing property of the HSV-2 infection, which has been described in previous publications, are discussed.


The phosphorylation of histones in herpes virus infected Vervet-monkey kidney cells (CV-1). The phosphorylation of histones of the F1 group and of fraction F2a2 is stimulated in monkey kidney cells (CV-1) to almost two times the control values 14 hours after their infection with herpes virus, type 2. At the same time high amounts of viral DNA are produced. It seems very likely that the stimulated phosphorylation of these histone fractions is a prerequisite for the enhanced synthesis of DNA and possibly also RNA.


Immediate endocrine and metabolic consequences of traumatic quadriplegia in a young woman. Onset of paralysis by cervical spinal cord injury led immediately to temporary adrenocortical activation and, within 2 days, to sustained skin and bone breakdown. Urine cAMP was increased, blood parathyroid hormone, renin activity, and electrolytes were normal, and fluid and electrolytes balance became negative during the initial 6 days of paralysis.


Intraspinal neoplasms in children.

Intraspinal neoplasms in children. Twenty-nine primary intraspinal neoplasms in children observed between 1936  and 1975 in Connecticut are reviewed. Most of them were gliomas: 45 per cent astrocytoma, 24 per cent ependymal neoplasm, 10 per cent glioblastoma multiforme and 7 per cent glioma. Symptoms, physical findings and therapy are reviewed.


[Evolution of cytomegalovirus antibodies of maternal origin and acquired, throughout the first year of life (author's transl)]. Cytomegalovirus antibodies have been determined by indirect hemagglutination technique in sera taken during the first year of life in two infant groups : Bantu infants in Mozambique, French infants in Alsace. IgM specific antibodies have been detected in serum fractions obtained by density gradients. The evolutions of maternal antibodies and of antibodies resulting from a primary CMV infection have been documented. The analysis of the results confirmed that the mother is the main source of infection and that contamination occurs at the time of birth.


Gold nephropathy.

Gold nephropathy. Nephropathy with proteinuria is an occasional complication of gold therapy for rheumatoid arthritis and is considered to be due to an immune hypersensitivity reaction. Three patients are described in whom the structural changes typical of gold nephropathy were demonstrated by electron microscopy of renal biopsies. There was early membranous glomerular nephropathy with multiple sub-epithelial deposits of immune complexes. In one case these were shown by immunofluorescence to contain IgG and complement. Proximal tubules contained characteristic dense granular intracytoplasmic gold inclusions, best demonstrated by electron microscopy of unstained sections.


Cerebellar calcification: a possible marker of lead poisoning. An unusual form of calcification in the cerebellum has been observed in routine autopsies in Queensland over the past 30 years. In 4 surveys carried out at intervals between 1951 and 1976 the lesion has been found microscopically in 10 to 15% of autopsies. There is a significant correlation between cerebellar calcification and raised lead levels in bone.


The effect of cyclic AMP on human colony forming cell and colony stimulating factor production. The effect of cyclic AMP (cAMP) and related nucleotides on human colony forming cells (CFC) and those cells producing colony stimulating factor (CSF) was studied in vitro. When added at physiological concentrations (10(-2) to 1 micron), exogenous cAMP stimulated maximum colony formation in cultures without feeder layers. Related nucleotides stimulated colony formation to a lesser extent and in decreasing order of free energy. All nucleotides inhibited colony formation in concentrations above 1 micron. A velocity sedimentation cell separation technique was used to obtain cell fractions rich in CFC but poor in CSF-producing cells. Such fractions did not respond to cAMP stimulation. These studies suggest that exogenous cAMP stimulates human bone marrow to form colonies in vitro by increasing the release and/or production of endogenous CSF.


Vestibular and somatosensory interaction in the cat vestibular nuclei. The vestibular nuclei of cats were explored extracellulary with micropipettes to locate units with a resting discharge rate which responded to rotation in the horizontal plane. These units were examined for somatosensory input from neck and limbs. Fewer than half responded to somatosensory stimulation. The neck region was the body area most effective in influencing unitary activity. The response pattern most often noted was an increase and decrease in discharge frequency when the body was moved towards and away from the recording electrode respectively. Change in discharge rate was observed to be primarily dependant upon neck velocity and not upon absolute neck position. Half of the somatosensory units received input from either the forelimbs or the hindlimbs, while the remaining half responded to both.


Voltage dependence of amplitude and time course of inhibitory synaptic current in crayfish muscle. The membrane of small crayfish muscle fibers was clamped to potentials between-150 and -20 mV and amplitude and time course of inhibitory postsynaptic currents (IPSCs) were studied. The IPSCs were recorded extracellularly by means of a focal microelectrode and also as total clamp current. The IPSCs lasted about 40 ms and were slowed by depolarization. The rate constant alpha of decay of the IPSC depended on membrane potential E according to the relation alpha= 44 s-1 -e-8.7 V E at 13.5 degrees C. alpha increased with temperature with a Q10 of 1.9 to 2.5. The amplitude iI of the IPSC depended nonlinearly on E and decreased with time after a potential shift. This was partly due to movement of Cl--ions, the difference (E--EI) between clamp potential and reversal potential for the IPSC decreasing to a few mV within several minutes after a shift in E. The inhibitory conductance gI increased up to 30-fold for 100 mV depolarization also changed with time. However, the the inhibitory permeability PI proved to be independent of membrane potential and time. The potential dependence of gI is thus largely due to changes in the internal Cl--concentration.


Adenosine as inhibitor of myocardial effects of catecholamines. Infusion of adenosine into the coronary arteries of isolated guinea pig hearts produced a dose-dependent inhibition of dP/dtmax caused by bolus injections of isoproterenol (4 X 10(-11) moles). Threshold concentration of adenosine was 10(-7) M and maximal inhibition (90%) occurred at 10(-5) M. Coronary dilation induced by papaverine did not influence the contractile response to catecholamines. In addition to its influence on cardiac performance, adenosine (10(-5) M) effectively inhibited the isoproterenol (10(-7)M) induced initial rise in myocardial levels of cyclic 3'5'-AMP, glucose-1-phosphate and glucose-6-phosphate. Adenosine also antagonized the effect of isoproterenol on adenylate cyclase activity in a crude membrane preparation from guinea pig ventricles; it was without effect on the activity of the membrane phosphodiesterase. Theophylline inhibited the actions of adenosine both on adenylate cyclase activity and on contractile force development. Upon infusion of isoproterenol (3 X 10(-7)M) into the coronary arteries of the isolated heart (perfusion at constant pressure), the adenosine concentration in the effluent perfusate increased within 45 s from 10(-8) M to about 10(-6) M. It thus appears conceivable that in ventricular myocardium endogenously formed adenosine may serve 2 functions: dilation of the coronary arteries and limitation of the inotropic and metabolic effects of catecholamines.


[Detection of HBs Ag in bone's metastatic malignant hepatoma (author's transl)]. A case of metastatic malignant hepatoma in bone, developed in a Vietnamen woman, is reported. Indirect immunofluorescence and orcein staining detect HBs Ag in cytoplasm of many tumor cells. This finding raises pathogenic problems, in particular that of the responsability of heaptitis B virus in the aetiology of certain hepatic tumours.


[Inappropriate secretion of ADH during acute respiratory infections due to adenovirus in children (author's transl)]. Four children aged between 7 and 19 months with severe bronchopneumonia due to adenovirus type 7, proved by virology and/or serology developed severe hyponatraemia. One of them is reported in detail: it was possible to estimate plasma ADH levels and thereby prove the existence of reversible hypersecretion of the hormone. Whilst the syndrome of hyponatraemia with inappropriate secretion of ADH has not yet been reported in association with severe pneumonia in the child, it is known in adults. The limits of the syndrome and its physiopathology are discussed. It may be due either to vagal stimulation as a result of a fall in left aressure, or to central involvement. Therapeutic implications of the problem are emphasized.


Capping structures of simian virus 40 19S and 16S mRNAs. In vivo [methyl 3H]-labeled SV40 19S and 16S mRNA species were purified and their internal methylation as well as their capping structures analyzed. SV40 viral mRNA sedimenting in the 19S region contains approximately equal proportions of m7GpppAm and m7Gppm6Am, while the 16S mRNA contains mainly m7Gpppm6Am. N6 methyl adenosine is located internally within the RNA chains of both the 19S and 16S species.


Modification of human DNA-dependent RNA polymerase activity by cyclic GMP. The effect of low concentrations of cyclic GMP (guanosine 3':5'-cyclic monophosphate) on the in vitro enzymatic activities of DNA-dependent RNA polymerases isolated from human peripheral blood lymphocytes has been investigated. In agreement with earlier studies which employed isolated nuclei as the enzyme source, an increase in the activity of partially purified RNA polymerase I is observed in the presence of cyclic GMP (10(-8) to 10(-10)M). RNA polymerase II activity is inhibited by the presence of cyclic GMP at concentrations between 10(-4) and 10(-10)M. RNA polymerase III activity is stimulated in a bimodal fashion by the presence of cyclic GMP with maximal activity noted at 10(-8) to 10(-10) M and 10(-5)M. In addition, [3H]cyclic GMP binds specifically to chromatographic fractions which are known to contain RNA polymerases I, II and III. This binding to RNA polymerases II and III is apprarently less tenacious as demonstrated by dissociation studies. The observations provide additional evidence for a role for cyclic GMP in the regulation of RNA synthesis.


High-quality, high-throughput, early whole-body bone scanning. 99mTc-IDP allows for high-quality, high-throughput and early whole-body bone scanning. Whole-body bone scans obtained at 30 min after intravenous administration of the tracer were compared with the standard and delayed 180 min p.i images. A subtraction technique allowed for the anlysis of the 30 min scans and it was found that in both types of investigation the same clinical information was present. This procedure will therefore increase considerably the throughput of whole-body bone scans in a busy department of nuclear medicine. It is however inadequate for detailed analysis of the skeleton (evaluation of benign bone disease).


A clinical situation of unusual cardiac pain. In conclusion, I believe that nurses have a unique opportunity to help patients cope with pain. Much research is already available to guide us in clinical application. Now we must generate more specific nursing research to provide the scientific rationale needed for our nursing actions. I am convinced that there are reasonable explanations for nursing actions that many of us have found so successful but so difficult to explain or teach to others. As nursing research helps us to explain these phenomena, our teaching and practice of nursing will surely improve.


Cytomegalovirus infection in the neonate and its prevention. About 0.5% of infants are infected in utero with cytomegalovirus (CMV). As many as 20% become mentally retarded and a further substantial proportion suffer lesser degrees of brain damage. The need for a vaccine is greater than in the case of rubella. A live, tissue culture-adapted strain of CMV has been shown to produce neutralizing antibody in volunteers without significant side effects or detectable virus excretion. The problems of developing such a vaccine for use in man, namely, attenuation, antigenic differences among virus strains, reactivation of latent infection and oncogenicity, are briefly discussed.


Carcinoma of the lung presenting with a myeloproliferative disorder. A report of two patients. A myeloproliferative disorder, similar to agonogenic myeloid metaplasia, has been described in patients with cancer. Two patients with carcinoma of the lung are described who presented with such a disorder but in whom there was no evidence of bone marrow fibrosis and in one of whom there was no evidence of bone marrow infiltration by tumor. The possible mechanism of this association is discussed.


MSH/ACTH4-10 influences on the CAR in human subjects: a negative finding. Twenty normal, male, paid volunteers were randomly assigned to MSH/ACTH4-10 or diluent control groups. Following a subcutaneous injection of either 30 mg MSH/ACTH4-10 or diluent, all subjects were placed in an active conditioned avoidance response situation where they learned to avoid a painful electric shock by pressing a key during a safe interval between a warning signal (light) and shock onset. MSH/ACTH4-10 did not influence any parameter of the acquisition or extinction process.  The between subjects' variability of autonomic variables was significantly less for the MSH/ACTH4-10 group. Results indicate that inhibition of extinction of conditioned avoidance responding does not generalize across species from rat to man.


Acute influences of some ACTH-related peptides of fighting and adrenocortical activity in male laboratory mice. A number of corticotrophin preparations when given acutely enhanced fighting behavior, an event which may be consequent upon elevation of circulating corticosterone. A series of short ACTH analogues (ACTH 11-24, ACTH 4-10) had slight influences on circulating plasma corticosterone values and on fighting behavior. However, 20 microgram ACTH 1-10 given 12 hr before testing did increase fighting C.F. controls. As this compound only caused a slight stimulation of plasma corticosterone titers, the possibility exists of an extra-adrenal influence. One cannot, it appears, eliminate glucocorticoid influences using these preparations in the mouse. Acutely, ACTH 1-24 had virtually no influences on Animex-assessed motor activity but did enhance fighting in castrated mice maintained with either androgen or estrogen. It is tentatively concluded that the ACTH influences on aggression in the mouse are partially mediated via glucocorticoid influences on the CNS combined with an extra adrenal influence of part of the ACTH molecule. A possible reason for the differing acute and chronic influences of ACTH on fighting in mice is presented.


Possible role of dopamine-beta-hydroxylase in the regulation of norepinephrine biosynthesis in rat brain. In Experiment 1, the dose-response effects of three dopamine-beta-hydroxylase (DBH) inhibitors (diethyldithiocarbamate, FLA-63 and U-14, 624) on the endogenous levels of norepinephrine and dopamine in pons-medulla of rat brain were determined. In Experiment 2, the effect of low doses of diethylithiocarbamate (2.5 to 120 mg/kg) on the level of norepinephrine-3H produced from dopamine-H3 was determined. The data obtained by extrapolation of the curves in both experiments provided an estimation of the in vivo level of DBH activity and suggested that it was not present in excess. Finally, in Experiment 3, the three DBH inhibitors reduced self-stimulation (a behavior dependent upon catecholamines) in a dose-related manner and intraventricular injections of 1-norepinephrine reinstated normal rates of self-stimulation. The results from the three experiments are consistent with the idea that DBH is involved in the regulation of norepinephrine biosynthesis. The relationship of this finding to our earlier report of a deficit of DBH in post-mortem brains of schizophrenics is discussed.


Paradoxical effects of calcium-glucagon interaction on cardiac muscle contractility of isolated guinea pig atria. The interrelationships of calcium and glucagon, and calcium and Isuprel were investigated in spontaneous and paced isolated guinea pig atria. Positive force responses with glucagon were in part both frequency and [Ca+2]o-dependent. Negative inotropic responses were observed with high concentrations of glucagon (5.0 microgram/ml) and calcium (10.0 mM). Persistence of a positive inotropic response of the atria to Isuprel (1.0 microgram/ml) and high [Ca+2]o (10 mM) was seen. Catecholamines stimulate c-AMP production in guinea pig atria while glucagon may not. The negative inotropism produced via calcium-glucagon interaction is consistent with the known inhibitory action of high calcium concentration on adenylyl cyclase and stimulation of phosphodiesterase. It is hypothesized that since glucagon does not activate c-AMP in this tissue then the combined action of high calcium and glucagon leads to degeneration of contractility; with Isuprel and high calcium, atrial contractility is maintained via Isuprel's c-AMP activation.


Study into the mechanism of acetylsalicylic acid-induced bronchoconstriction. In order to elucidate the mechanism of the acetylsalicylic acid-induced bronchoconstrictor responses which have been observed in asthmatic patients, experiments were done in cats, anesthetized by chloralose, vagotomized, and completely relaxed by infusion of suxamethonium. In about half of the experiments, the sensitivity of the bronchi to 5-hydroxytryptamine (5-HT) was considerably enhanced after the administration of 5--6 mg/kg acetylsalicylic acid, a dose which had previously been shown to suppress prostaglandin (PG) biosynthesis in the bronchi of the cat completely. No further increase in sensitivity to 5-HT occurred when higher doses of the drug were given. Immediate bronchoconstrictor reactions to the injection of acetylsalicylic acid were rare, in most instances the rise in sensitivity developed slowly during a period of about 30 min. The results are consistent with the assumption that PGE2 displays an antagonistic effect to the mediators of asthmatic bronchoconstriction or to their liberation and that inhibition of PG synthesis by acetylsalicylic acid or related drugs eliminate this function and may thus give rise to bronchospasm.


Effect of magnesium trisilicate and citric acid on the biovailability of tetracycline in man. Oral absorption of tetracycline hydrochloride by human subjects was compared with its absorption when coadministered with magnesium trisilicate, citric acid or magnesium trisilicate-citric acid mixture. Evaluation of the absorption rate was done by means of urinary excretion measurements. The concomitant administration of magnesium trisilicate with tetracycline hydrochloride resulted in a dramatic reduction of the excretion rate of the drug. Ingestion of citric acid with tetracycline showed no significant effect on its absorption. Citric acid administration simultaneously with tetracycline hydrocloride-magnesium trisilicate combination failed to improve the absorption of tetracycline in the presence of magnesium trisilicate contrarily to preveious in vitro results. Conclusions were made that in vitro experiments are not always successful in the prediction of in vivo findings.


Effect of clozapine on the sleep pattern in the rat. The effect of clozapine on the sleep pattern in the rat was studied after a single injection (2.5--20 mg/kg) and after daily administration for 11 consecutive days (2 X 20 mg/kg/day). After a single injection clozapine significantly suppressed REM sleep in a dose-related way, leaving slow wave sleep (SWS) unchanged. Follwing chronic administration, however, clozapine exerted its most prominent effect on SWS, the enhancement of which persisted for at least 3 days after discontinuation of treatment. REM sleep decreased with the development of tolerance and in the absence of rebound phenomena. The results are discussed in the light of disturbances in brain monoamines and the tolerance these amines developed to repeated administration of clozapine.


Effect of carboxypeptidase N, aprotinin and anti-inflammatory drugs of pyrazolidine type on experimental inflammations in rats. Inflammation was induced by application of kaolin into the hind paws of rats and its development under the influence of carboxypeptidase N and aprotinin (Antilysin) was followed and compared with the effect of the phenylbutazone type of drugs. Partly purified rat serum carboxypeptidase N, applied subaponeurally (20 mg/kg) together with a nociceptive agent (kaolin), inhibited the inflammation very effectively. Aprotinin, applied as above (10,000 U/kg), enhanced the inflammatory reaction; if applied intraperitoneally 1 h before kaolin, it displayed anti-inflammatory effects similar to that of phenylbutazone (100 mg/kg). Carboxypeptidase N, administered intraperitoneally before kaolin, had a similar anti-inflammatory effect. The effect of phenylbutazone, ketazon and trimetazon on the bradykinin-induced rat uterus contraction was followed. All these substances inhibit the response of isolated rat uterus to bradykinin. From the results it can be assumed that phenylbutazone, ketazon and trimetazon exercise their effect by blocking centres of the rat uterus which are important for the constrictor effect of bradykinin.


Effect of histamine on cyclic AMP levels in control and antigen-sensitized guinea pig lungs. The effect of histamine on cyclic AMP was investigated in control and antigen-sensitized guinea pig lungs. Although histamine elevated the cyclic AMP levels in a dose-dependent manner in both groups, the response was consistently smaller in the sensitized lungs. In vivo pretreatment of control guinea pigs with histamine decreased the subsequent effect of exogenous histamine on cyclic AMP. Phosphodiesterase activity was not different in the two groups and it was not affected by histamine. These results indicate that the process of antigen sensitization results in an altered cyclic AMP response to histamine.


Influence of structural changes in the imidazolidine ring of clonidine on hypotensive activity. 2-(2,6-Dichlorophenylimino)piperimidine with HNO3 (St-404), 2-(2,6-dichlorophenyl)-5,6-dihydroimidazo[2,1-b]thiazole fumarate (compound 44-549) and 1,2,3,5-tetrahydroimidazo[2,1-b]quinazoline with HCl (TIQ) were studied with respect to their effects on blood pressure and heart rate in the anaesthetized, normotensive rat following intravenous administration and in the chloralose-anaesthetized cat by means of infusions via the left vertebral artery. St-404 and compound 44-549 possess central hypotensive and bradycardic activities and display modes of action similar to that of clonidine. Central alpha-adrenergic receptors are presumably involved. In the anaesthetized, normotensive rat St-404 is about 3,400 times less active than clonidine, whereas compound 44-549 is 5 times more effective in lowering arterial pressure. The hypotensive effect of TIQ is brought about by a mechanism of action which is different from that of clonidine. The alpha-sympatholytic properties of TIQ suggested previously by others are not confirmed by the experiments presented in this paper.


Does REM sleep deprivation induce a supersensitivity of dopaminergic receptors in the rat brain? The possibility that REM sleep deprivation induces increased sensitivity of rats to the effects of apomorphine was investigated. Rats were deprived of REM sleep by placing them on 6.0 cm platforms for 3 days. Stress control groups consisting of rats placed on 14 cm platforms (control rats) and groups of nonmanipulated animals (normal rats) were used for comparison purposes. Several doses of apomorphine were administered to these 3 groups of animals, and aggressive behavior, stereotyped behavior, verticalization and body temperature were recorded. All measures indicated that REM-deprived rats were more responsive to apomorphine than the normal and control animals, with the exception of verticalization effect, in which both REM-deprived and control animals reacted similarly. These results suggest that REM deprivation induced supersensitivity in brain dopamine receptors. Other possible interpretations are also discussed.


Repair of defect of anterior chest wall with a turnover, dermal-fat, deltopectoral flap: case report. We present a case in which a full-thickness defect of the anterior chest wall was closed with a turnover, dermal-fat, deltopectoral flap--with split-skin grafts for the cover. This is a worthwhile alternative procedure to be added to the already numerous techniques currently available for the repair of such defects.


The adrenocortical secretion in schizophrenics during chlorpromazine treatment. 33 cases of schizophrenia received normal food and no premedication. After a clinical improvement which showed itself 3--6 weeks after the beginning of the chlorpromazine treatment, the 17-ketosteroids, the 17-ketogenic steroids and also the electrolytes were dertermined in the blood serum, the erythrocytes and the urine. 12 schizophrenics each received 25 mg of ACTH for three days puring the maximum chlorpromazine dosage. The separate results of examination were tabulated. The final results showed a restriction in the steroid synthesis, an increase in the excretion of potassium through the kidneys along with a decrease in the excretion of sodium. There was an appreciable drop in the sodium potassium concentration in the erythrocytes. It can be assumed that there is a connection between the variations in the membrane permeability to electrolytes and pharmacogenic dyskinesia.


The mechanism by which the energy is released at the level of the flavoenzyme molecule during the transfer of two electrons. The model of a mechanism producing the excitation of two electrons by the rotation of an orbital is presented functioning at the level of the flavoenzyme molecule. According to this model, during the transfer of two electrons from an NADH coenzyme to a flavoenzyme molecule, in a reaction cavity of the flavoprotein, two (Fe2+:S) covalent bonds are formed. Subsequently to the electric polarization induced by the formation of some intermolecular connections and by some fixed ions, and (Fe2+:S) orbital turns with an angle, having the nucleus of the sulfur atom as a pivot point, in such a way that an (Fe2+) ions is left out from the covalent bond, and it is replaced by a proton (released from a water molecule). A new (S:H) orbital is formed. Within this (S:H) orbital, the proton moves towards the nucleus of the sulfur atom, from 2.10 A degrees internuclear distance (position in which the proton was included by the orbital rotation) to 1.36 A degrees internuclear distance--corresponding to the length of the (S:H) covalent bond. By the proton movement, the two electrons of the (S:H) orbital arrive on an excitation state level. Then, they fall to their fundamental level and deliver two quanta of electronic excitation, which are transmitted further and help the synthesis of an ATP molecule.


Cyclic AMP in cerebrospinal fluid of manic and depressive patients. Cyclic 3',5'-adenosine monophosphate (c-AMP) was measured in cerebrospinal fluid (CSF) of manic and depressive patients with and without probenecid administration both before and during treatment with various psychotropic drugs. Oral probenecid (100 mg/kg) produced substantial c-AMP accumulations in CSF suggesting a probenecid-sensitive transport mechanism for c-AMP. Baseline and probenecid-induced accumulations of c-AMP were not significantly different in manic and depressed patients, while baseline levels in depressed patients were higher than those in neurological controls. Imipramine, amitriptyline, lithium, tryptophan, and electroconvulsant therapies did not significantly alter levels or accumulations of c-AMP in CSF of depressed patients.


Block diagrams for the study of some endocrine processes. With the help of bibliographical data and contributions of the authors two block diagrams have been built. These block diagrams are scheming two processes connected with the exposure of the organism to a moderate physical exercise: (a) the control of the release and the effect of ACTH (b) the control of the release and effect of STH. Some practical conclusion can be drawn regarding the interpretation of the endocrine modifications during exercise including those during standard functional tests.


The role of oestrogen replacement in the climacteric syndrome. Using a double-blind placebo-controlled cross-over design, the effect of equine conjugated oestrogens tablets (Premarin) was studied in 20 women with the climacteric syndrome followed during 15 months. Sixteen women were equally improved on placebo and oestrogen. Only 2 patients had an improved sense of well-being on oestrogen and not on placebo. The psychological diagnosis was unrelated to the subjective response to oestrogen or placebo. Performance in psychological tests administered before and during treatment periods was not changed by oestrogen or placebo.


Costs and benefits of behavioural psychotherapy: a pilot study of neurotics treated by nurse-therapists. A pilot study is reported of costs and benefits from behavioural psychotherapy by nurse-therapists for selected neurotic problems. Figures are based on the treatment of 42 neurotics (mainly phobics and obsessive-compulsives) who completed treatment with nurse-therapists in a mean of 9 sessions (16 hours). The year before and after treatment was studied. Apart from significant and lasting reduction in patients' distress, economic benefits to them, their families and the community yielded a worthwhile internal rate of return when benefits from the cohort continue for 3 years, a reasonable assumption on available other evidence. Though untreated phobics did not improve elsewhere over 5 years follow-up, a controlled study would seem desirable.


Dynamics of the changes in the cerebral amounts of cyclic AMP and some prostaglandins during cobalt-60 gamma-radiation-induced brain edema. The total amounts of cyclic AMP (cAMP), prostaglandin E1 (PGE1) and prostaglandin F2alpha (PGF2alpha) in cerebra have been measured in rats, at constant intervals, up to 18 days after whole body exposure to either a unique moderate dose (500 rads) or a unique lethal dose (750 rads) of cobalt-60 gamma-radiation. The experimental findings indicate that this radiation (i) results in an abrupt short-lasting increase in the amount of cerebral cAMP after a 500 rad-irradiation and a progressive long-lasting increase in its amount after a 750 rad-irradiation, and (ii) induces no change in the normally, existing correlation between cerebral PGE1 and cAMP, but affects deeply the normally existing correlation between cerebral PGF2alpha and cAMP. These biochemical alterations generally parallel the evolution of the radiation-induced brain edema.


Sonographic patterns in trophoblastic diseases. A range of sonographic patterns in patients with hydatidiform moles can be appreciated with gray scale ultrasonography, which more readily reveals areas of internal hemorrhagic degeneration and fetal growth within a molar mass. Sonography may be used to evaluate patients suspected of having recurrent or invasive trophoblastic disease; a focus of invasive trophoblastic tumor can be identified as a cluster of high-amplitude echoes within the myometrium, which is frequently associated with echo-free areas of hemorrhage. The enlargement or regression of theca lutein cysts associated with these disease processes can also be monitored.


Stimulation by cyclic nucleotides of prostaglandin E production in isolated graafian follicles. Rat Graafian follicles isolated intact responded to 8-Br-cyclic AMP and 8-Br-cyclic GMP with increased prostaglandin E (PGE) production during a 6 h incubation. By contrast, 8-Br-cyclic IMP, 8-Br-5' AMP and 8-Br-5' GMP were inactive in this respect. The effect of 8-Br-cyclic AMP and 8-Br-cyclic GMP was noted only after a lag period of about 4 h. Choleragen, LH, and the phosphodiesterase inhibitor (3-isobutyl-l-methyl-xanthine; IBMX) also stimulated PGE production. Actinomycin D and cycloheximide given simultaneously with 8-Br-cyclic AMP or LH prevented the stimulatory effect of these agents. Concomitant addition of arachidonic acid did not overcome the effect of these inhibitors. Administration of hCG in vivo or incubation with LH in vitro did not elevate endogenous ovarian free arachidonate, while PGE production was enhanced. Dexamethasone prevented this stimulatory effect of hCG. Collectively, the results suggest that stimulation of ovarian PGE production by cyclic nucleotides and LH is dependent on de novo synthesis of one or more components of the PG synthetase system rather than on substrate availability. Cyclic nucleotides may mediate the stimulatory effect of gonadotropins on PGE production.


[Effect of vincamine on sleep-wakefulness equilibrium in the cat (author's transl)]. Intra-muscular vincamine in the cat (chronic experimental preparation) causes a change in the equilibrium between sleep and wakefulness. This consists of: (1) an increase in the amount of wakefulness, the duration of which depends on the dose injected (5-10 mg/kg) and the mode of injection (more effective when given in two doses than all at once); (2) a decrease in slow wave sleep (SWS) which often has the appearance of phasic slow wave sleep; (3) a transitory suppression of paradoxical sleep (PS). At the time of recovery there is no rebound either of SWS or PS. The mechanism of vinacamine's action is not understood and some interpretations are suggested.


[The E.E.G. of herpes encephalitis in children: 3 cases including one neonate (author's transl)]. The authors report 3 cases of herpes encephalitis in children aged respectively 15 days, 12 months and 18 months. In all 3 cases E.E.G. activity of a periodic nature was noted, which was continuous or intermittent, lateralised or focal in the temporal region, appearing from the 4th to 7th day. The periodicity (of the E.E.G. complex) was short and their stereotyped appearance particularly unusual in babies. In all 3 children convulsive seizures occurred before the discovery of the periodic activity. The latter was found to change in morphology and timing during the seizures. The disease process led to death of the neonate, and recovery with serious sequellae in the other two children. Possible relationships between the transitory nature of the periodic acitivity, the age and the eventual course of the disease, were discussed.


[Pseudo-myasthenic syndromes. Clinical and electromyographic review of 4 cases, one with an unusual reaction to lidocaine (author's transl)]. The authors report 4 cases of pseudo-myasthenic syndrome followed with EMG for a period varying from a few months to 4 years. They go over the electrophysiological points which led to the diagnosis: small amplitude of the muscle potential evoked by simple shock and, in particular, the phenomenon of potentiation which may reach 2000 p. 100 during repetitive stimulation at high frequencies. Study of the behaviour of the muscle potential during ischaemia, which was carried out twice, showed an absence of fatigue of the potential which may be considered a sign of disturbance at the muscle level of the motor-end plate. On the other hand, EMG signs of peripheral neuropathy, which were found 3 times, rather suggests a motor neuropathy responsible for the presynaptic block. In 3 patients guanidine had a beneficial effect and no signs of paraneoplastic aetiology had occurred yet. The fourth case, with a bronchial neoplasm discovered at autopsy, was clearly improved by injectable lidocaine, for which no explanation can be supplied.


[Excessive 7-14-sec positive spikes during REM sleep in monozygotic non-epileptic twins with speech retardation (author's transl)]. 6-14/sec positive spikes (PS) (in our cases 7-14) were observed during 6 all-night sleep, recordings in one pair of monozygotic twins (aged 7 years), who had severe speech retardation, no epilepsy and were otherwise normal (CAT were normal). The EEG during wakefulness and sleep showed multifocal independent spikes over the left mid-temporal and right parieto-occipital area. The 7-14 PS, which were similar in both twins, occurred slightly during light sleep, were absent during slow sleep and were most prominent during REM sleep (mean=6.3 sec of PS bursts/min of REM). During REM sleep, the 7-14 PS bursts were negatively related to bursts of eye movements; PS were 7 times more frequent in the intervals between than during bursts of eye movements. In addition, long bursts of PS (up to 6 sec) might interupt the bursts of eye movements suggesting a functional antagonism between mechanisms (still unclear) responsible for PS and for REM. The predominance of PS during REM sleep and the inverse relationship with eye movements are not peculiar to our case, since similar findings have been reported in other cases (TSUZUKI 1967; OKUMA et al. 1968). During the sleep stages when Ps occurred spontaneously, PS could also be evoked by a click or a tone, with a latency of 1, 5-2 sec.


[Preliminary study of the comparative action of 1 methyl-2 (2 naphtyl) aziridine on kindling effect and on paradoxical sleep (author's transl)]. Methyl-2 (2 Naphtyl) Aziridine (MNA) is a new agent which has been demonstrated to have effects on paradoxical sleep (PS) and on the levels of cerebral noradrenaline (Yamamoto, 1975). It has been tried on a experimental model of epilepsy, the kindling effect, produced in the cat by repetitive stimulation of the amygdala. The results obtained in the cat confirmed blockage of paradoxical sleep (PS) and revealed a marked reduction in the number of stimuli required to produce the effect. The shortening of the time before the first generalised seizure did not seem attributable to privation of paradoxical sleep. The role of cerebral catecholamine reduction, however, is discussed.


Electrophysiological properties of rat heart cells in vitro and in tissue culture. Some electrical properties of right ventricles of neonatal rats and of aggregates from collagenase-dissociated cells from the same tissue are compared. The duration of the action potential does not change upon changing the stimulation frequency both in ventricles and aggregates; a decrease in temperature increases duration in both preparations to the same extent. The take-off potential and the maximal rate of rise of the action potential decrease in the same way in both preparations, while the time course of these changes is also comparable. It is concluded that the dissociation and aggregation procedure does not interfere with the membrane properties upon which the measured parameters are based; thus, aggregates are well suited for a realistic voltage-clamp analysis.


Quench-flow measurements of initial rates of Ca2+ accumulation by isolated cardiac sarcoplasmic reticulum. Using a quench-flow technique, the initial velocity of Ca2+ accumulation by isolated cardiac sarcoplasmic reticulum was estimated at free Ca2+ ion concentrations in the range encountered in the myoplasm during the cardiac contraction cycle. With cardiac microsomes exhibiting a Ca2+ accumulative capacity of 25.6 nmol Ca2+/mg protein, initial rates were found to increase from 3.7 to 33.4 nmol Ca2+/mg protein/sec, when the free Ca2+ ion concentration was raised from 0.2 to 18.0 muM. Preincubation of the cardiac microsomes with a party purified soluble cardiac protein kinase, MgATP, and cAMP led to a significant increase in the initial Ca2+ accumulation rate.


Characteristics of heart sarcolemmal calcium transport system and effect of protein kinase on sarcolemmal calcium accumulation. Properties of the ATP-dependent calcium transport system of heart sarcolemma are presented. Calcium accumulation (with oxalate) in sarcolemma was increased due to cAMP-dependent protein kinase and phosphorylase b kinase. Protein kinase increased the Vmax of the sarcolemmal calcium accumulation without any detectable effect on the affinity for Ca2+. Both kinases failed to stimulate calcium binding. Protein kinase catalyzed phosphorylation of membrane proteins of molecular weights of 100,000, 25,000, and 14,000. Phosphorylase b kinase also catalyzed phosphorylation of these proteins. Protein kinase stimulated ATPase activity of sarcolemma. Sarcolemma contained endogenous protein kinase and protein phosphatase activities.


An adenosine 3':5'-monophosphate-dependent protein kinase from human heart. Protein kinase that phosphorylated histone and lesser amounts of protamine was demonstrated in human heart. It was activated three times by 10(-6) M cyclic adenosine 3':5'-monophosphate (cAMP) and by 10(-3) M other cyclic nucleotides. Km values for cAMP, ATP, Mg2+, and Co2+ were about 2 X 10(-8) M, 4 X 10(-5) M, 2 X 10(-3)M, and 1.7 X 10(-4) M, respectively. On DEAE cellulose column, the main peak of the enzyme eluted at high NaCl concentration. On Sephadex G-200 gel filtration the majority of the holoenzyme eluted at a peak corresponding to a molecular weight of about 300,000. There was an additional peak corresponding to a molecular weight of about 400,000, with relatively high cAMP binding compared to kinase activity. Right atrium and ventricle showed significantly higher enzyme activities than left atrium and ventricle and interventricular septum. On multivariate analysis of the enzyme activity versus 12 clinical and pathological findings of 122 cases, cardiac hypertrophy and coronary sclerosis were slight but significant negative contributors to the enzyme activity. Multiple correlation coefficient was low, indicating the enzyme activity remained at a relatively stable level, despite different clinical situations. This may be suitable for control of intracellular events through the membrane adenylate cyclase system.


Significance of the membrane protein phospholamban in cyclic AMP-mediated regulation of calcium transport by sarcoplasmic reticulum. Phospholamban (molecular weight = 22,000), which serves as a regulator of Ca transport ATPase (molecular weight = 100,000) of cardiac sarcoplasmic reticulum (SR), becomes resistant to tryptic digestion upon phosphorylation by cAMP-dependent protein kinase (PK). The protective effect of phosphorylation is accompanied by persistence of the PK-induced stimulation of Ca transport. These findings indicate that structural alteration of phospholamban upon phosphorylation is closely associated with changes in the functional properties of cardiac SR. SR from fast-contracting skeletal muscle of rabbit does not contain a 22,000-dalton substrate for cAMP-dependent PK, nor is Ca transport stimulated by exogenous PK. SR preparation isolated from slow-contracting skeletal muscle of rabbit and dog contains phospholamban, and Ca transport was found to be increased by exogenous cAMP-dependent PK. In view of the distribution of phospholamban among different types of muscle, a hypothesis is presented to explain the relaxation-promoting effects of catecholamines in cardiac and slow-contracting skeletal muscle in which phospholamban is found. This may also account for the absence of a similar effect of catecholamines in fast-contracting skeletal muscle, which does not contain a similar substrate for PK.


Phosphoprotein phosphatase-catalyzed dephosphorylation of the 22,000-dalton phosphoprotein of cardiac sarcoplasmic reticulum. Similar time courses were obtained for decreases in the rate of calcium transport by cardiac sarcoplasmic reticulum vesicles previously phosphorylated by cAMP-dependent protein kinase and dephosphorylation of the 22,000-dalton phosphoprotein in these membranes. Dephosphorylation of the 22,000-dalton phosphoprotein can be attributed to a phosphoprotein phosphatase in the sarcoplasmic reticular membranes. This membrane-bound phosphoprotein phosphatase may play a role in the reversal of the relaxation-promoting effect of catecholamines on the heart.


Effect of protein kinase modulator on cAMP-dependent protein kinase-catalyzed phosphorylation of phospholamban and stimulation of calcium transport in cardiac sarcoplasmic reticulum. The heat-stable protein (protein kinase modulator), partially purified from fresh bovine heart, possessed the ability to inhibit and stimulate adenosine 3':5'-monophosphate (cAMP)-dependent protein kinase and guanosine 3':5'-monophosphate (cGMP)-dependent protein kinase activities, respectively. The inhibitory activity of protein kinase modulator on cAMP-dependent protein kinase was abolished almost completely by trypsin treatment, while the ability to stimulate cGMP-dependent protein kinase activity was resistant to trypsin. Fractionation by a linear potassium phosphate gradient on DEAE-cellulose column did not clearly separate both activities. Phosphorylation of cardiac microsomal component, "phospholamban" (molecular weight = 22,000), was inhibited almost completely by the saturating amounts of protein kinase modulator. This inhibition of phospholamban phosphorylation by protein kinase modulator was accompanied by a decreased Ca uptake rate that had been stimulated by cAMP-dependent protein kinase. These findings indicate that protein kinase modulator is functional in controlling the cAMP-dependent protein kinase-catalyzed phosphorylation of phospholamban and the rate of calcium transport, lending further support for the previously proposed mechanism, in which phospholamban is assumed to serve as a regulator of calcium transport in cardiac sarcoplasmic reticulum.


Effect of increased cAMP content on extracellular distribution of calcium and contractility in rabbit heart. Norepinephrine, dexamethasone, and papaverine hydrochloride caused a significant increase in myocardial cAMP level and an alteration of intracellular distribution of Ca, such as the reduction of Ca content on mitochondrial and microsomal fractions, with or without positive inotropic effects, in the rabbit. Positive inotropic effects evoked by norepinephrine were significantly increased under the elevated level of cAMP induced by dexamethasone and papaverine. On the other hand, verapamil showed no effects on cAMP concentration and intracellular Ca distribution of the heart. These results indicate that the accumulation of myocardial cAMP may result in the alteration of intracellular distribution of Ca, and it is suggested that there is an increase in the available Ca for contractile and metabolic mechanisms of the heart.


Effects of catecholamines on myocardial energy metabolism as studied by an organ redoximeter. With the use of an organ redoximeter, the effects of noradrenaline, adrenaline, isoproterenol, and phenylephrine on the oxidation-reduction state of the myocardial pyridine nucleotides were studied in the canine heart-lung preparation supported by a donor. Noradrenaline and adrenaline produced an initial, transient improvement, and phenylephrine a sustained improvement, of the redox state, while isoproterenol produced a depression. Pretreatment of the preparation with adrenergic alpha-blockers resulted in an abolishment of the improvement by noradrenaline, adrenaline, and phenylephrine, while the depression by isoproterenol remained unchanged. Whereas noradrenaline and adrenaline produced a sustained improvement after an adrenergic beta-blocker, propranolol, the effect of isoproterenol was abolished. These findings suggest that sympathomimetic amines can produce an improvement of the myocardial energy metabolism through activation of the adrenergic alpha-receptor. The depression of the myocardial oxidation-reduction state was taken to represent an acceleration of glycolysis.


Characteristics of energy metabolism in specialized muscle of bovine heart. Characterization of the energy metabolism pattern of the specialized heart muscle of bovine heart was studied in comparison with that of the ordinary heart muscle. Mitochondrial oxygen consumption of the specialized heart muscle was significantly lower than that of the ordinary heart muscle with succinate as the substrate. On the other hand, there was no significant difference in oxygen consumption between both heart muscles with glutamate + malate as the substrates. The activity levels of succinate dehydrogenase and lactate dehydrogenase were much lower than those of the ordinary heart muscle. The isozyme pattern of LDH of the specialized heart muscle consisted of one major component of LDH-1 (H4)  and that of the ordinary heart muscle consisted of two major components of LDH-1 (H4) and LDH-2 (H3M). The ratio of NADH to NAD of the specialized heart muscle was remarkably lower than that of the ordinary heart muscle. These results indicate that the specialized heart muscle depends not only upon anaerobic metabolism but also upon aerobic metabolism for its energy supply.


Evidence for opposing influences of cyclic GMP and cyclic AMP on force of contraction in mammalian myocardium. The inotropic effects of the 8-Br-derivatives of cyclic GMP and cyclic AMP were investigated in guinea pig or rat auricles and papillary muscles from cats. In guinea pig auricles, 8-Br-cyclic GMP had a concentration-dependent negative inotropic effect, whereas 8-Br-cyclic AMP had a concentration-dependent positive inotropic effect. A negative inotropic effect of 8-Br-cyclic GMP in rat auricles was obtained with 30 times lower doses than in guinea pig auricles; the pacemaker activity of spontaneously beating rat right auricles did not change in response to 8-Br-cyclic GMP. In cat papillary muscles, 8-Br-cyclic GMP (3 X 10(-4) M) also produced a negative inotropic effect. In contrast, 8-Br-cyclic AMP (3 X 10(-4) M) did not affect force of contraction; however, after pretreatment with the strong phosphodiesterase inhibitor papaverine (2 X 10(-5) M) 8-Br-cyclic AMP (3 X 10(-4) M) had a strong positive inotropic effect also in cat papillary muscles. The results suggest opposite influences of cyclic GMP and cyclic AMP on force of contraction in mammalian myocardium.


Studies on myocardial mitochondria in failing dog hearts: studies by electron spin resonance (ESR) spectrometry. The formation of free radicals, which was represented by the concentration of free radicals in state 4 respiration, and the respiratory control ratio were studied in the myocardial mitochondria isolated from dog hearts that were failing due to aortic constriction. Both formation and respiratory control ratio were lowered, compared to each control value, in acute and chronic heart failure. A positive correlation was recognized between free radicals in state 4 respiration and respiratory control ratio. From these results it was concluded that there was uncoupling of oxidative phosphorylation in the mitochondria isolated from failing dog hearts. Measurement of free radicals in state 4 respiration, as well as respiratory control ratio, is a useful method for evaluation of the function of oxidative phosphorylation.


Effects of acetylcholine and cyclic nucleotides on the bullfrog atrial muscle. Effects of ACh on the slow inward current and tension components of bullfrog atrial muscle were examined with voltage-clamp technique. ACh decreased the phasic component of contractile tension and the slow inward current with a reduction of gCa. Contracture tensions were augmented by ACh and decreased by adrenaline. Intracellularly injected cGMP through microelectrode decreased the overshoot and the plateau level of action potential without affecting the resting membrane potential, indicating a decrease in the slow inward current. cAMP enhanced the overshoot and the plateau. ACh seems to reduce the slow inward current through an elevation of intracellular cGMP concentrations.


Arrhythmogenic effects of acute free fatty acid mobilization on ischemic heart. The mechanism of action of the acute abnormal rise of plasma free fatty acids (FFA) in the provocation of arrhythmia in ischemic heart was studied by means of electron spin resonance (ESR) spectrometer. A sudden and abnormal rise of plasma FFA caused a significant fall of the respiratory control index (RCI) of the amount of free radical myocardial mitochondria in state 4 respiration. Based on these findings, a sudden and abnormal rise of plasma FFA seems to further facilitate the uncoupling of oxidative phosphorylation in the myocardial mitochondria of the ischemic portion of the heart. These observations indicate that it may play an important role in the provocation of arrhythmia by high plasma FFA on the ischemic heart. Nicomol (2,2,6,6,-tetrakis (nicotinoyloxymethyl) cyclohexanol), an inhibitor for the rapid rise of plasma FFA, was effective in the treatment and prevention of arrhythmia in ischemic heart disease and diabetes mellitus.


Adenylate cyclase, cyclic nucleotide phosphodiesterase, and phosphorylase activity of cardiomyopathic hamster hearts. Activities of adenylate cyclase in homogenates were reduced, whereas those of phosphodiesterases were elevated in hearts of myopathic hamsters (BIO 82.62). Affinities for either Mg2+ or ATP of cyclase were unaffected in myopathy. Ca2+ stimulation particulate phosphodiesterases was not observed in myopathy. Although the cardiac phosphorylase content was reduced at the advanced stages of myopathy,-AMP/+AMP ratios remained similar to those found in normal hearts.


Differential responses of canine myosin ATPase activity and tissue gases in the pressure-overloaded ventricle dependent upon degree of obstruction: mild versus severe pulmonic and aortic stenosis. Mild pulmonic stenosis, induced in dogs by banding the pulmonary artery, elevated right ventricular peak systolic pressure to 60% above the control and elevated right ventricular K+- and Ca2+- activated myosin ATPase activities. In contrast, severe pulmonic stenosis, which elevated right ventricular peak systolic pressure to 300% above the control, did not produce an increase in myosin enzymatic ATPase Vmax values but caused a decrease in myosin activity. Mild aortic stenosis, induced by banding the ascending aorta, forcing a transaortic pressure gradient of 25 mm Hg, caused an elevation in left ventricular muosin ATPase, whereas severe aortic banding, brought about by creating a transaortic pressure gradient of 55 mm Hg, never caused an elevation in left ventricular myosin enzymatic Vmax values, but, like severe pulmonic banding, caused a decrease in K+- and Ca2+- activated myosin activities. Normal left ventricular myosin Vmax values in mumol of PO4/mg-min at 37 degrees C were: K+ = 2.84 +/- 0.22, and Ca2+ = 0.97 +/- 0.14. For right ventricular myosin they were: K+ = 2.15 +/- 0.16, and Ca2+ =0.74 +/- 0.10. Analyses of tissue gases, based on mass spectrometry data, showed that the hypertrophied ventricles had an elevated tissue pCO2 and an elevation in the cGMP/cAMP ratio.