Experimental protein malnutrition in primates: electronmicroscopic observations on the cerebellum of healthy and malnourished squirrel monkeys (Saimiri sciureus). Eight young adult squirrel monkeys were maintained on a protein deficient (2% calories from proteins) diet for a period of 15 weeks during which the animals lost wieght steadily and developed varying degrees of sensory and motor impairments as evidenced by lack of interest in the surrounding environment and changes in posture and gait. An additional eight animals were maintained on a high protein diet (25% calories from proteins) to serve as controls. Two animals each from the experimental and control groups were sacrificed at 9, 11, 13 and 15 weeks of special feeding schedule. The Purkinje cells of the cerebellum of malnourished animals showed significant changes in the ribosomes, endoplasmic reticulum and Golgi zone indicating greatly disturbed protein metabolism. The granule cells, especially of the pale variety show prominent watery swellings with scant cell organelles. The most prominent change appeared in various types of neurons related to the multiplication of various types of lysosomes, multivesicular lamellar bodies and the lipofuscin pigment bodies. In the animals sacrificed at different intervals, the relatively more malnourished animals showed significantly larger number of lipofuscin pigment in the various types of neurons of the cerebellar cortex. The dendritic changes included honey-comb formation in the lumen. The astrocyte glial fibers were greatly increased. The pericytes of the blood vessels showed irregular outlines and were greatly enlarged with numerous osmiophilic bodies (presumably lipofuscin) in the experimental animals as compared to the controls.


A test for the assessment of disinfectant/sanitizers used for napkin storage. A test is described for assessing the effect of hypochlorites in reducing the bacterial load on soiled napkins during storage, between removal from the infant and laundering.


Identification of pregnancies at risk from cytomegalovirus infection. The fluorescent antibody technique was used for the identification of specific cytomegalovirus IgM in the sera of twenty-four of 1065 unmarried pregnant women. Seventeen of them were followed to term and five infected infants were identified. Two other infants had CMV IgM in neonatal serum samples but virus excretion was not demonstrated. The congenital infection rate in this study was 5.3 per 1000 births by virus excretion and 7.9 per 1000 if cases with specific IgM are included; from previous studies a rate of 8.8 per 1000 was expected. The reasons for the lack of relationship between specific IgM in the mothers' serum and infected babies is discussed.


Distribution and titres of rotavirus antibodies in different age groups. Three hundred and fifty-seven sera selected at random from hospital patients of all ages were examined for rotavirus antibodies using indirect immunofluorescence (FA) and complement fixation levels (CFT). Three hundred and fourteen of these were also tested for neutralizing antibodies to human rotavirus. Sera from patients admitted with a diagnosis of acute gastroenteritis were excluded from this survey. FA antibodies were found in newborn infants but fell to undetectable titres at 3 months. The highest titres were found in children between the ages of one and three years. In older age groups, the model titre fell gradually with increasing age until, in sera from those above 70 years of age, FA antibodies were almost undetectable. The same pattern was observed with neutralizing antibodies. A high model titre of CF antibodies was only found in sera from those aged one to three years.


The metabolic inhibition test for mycoplasmas based on phosphatase production. A metabolic inhibition test based upon phosphatase production is described. This is of value for those Mycoplasma species which produce phosphatase but which cannot be examined by the previously published metabolic inhibition techniques.


Scanning electron microscopic study of hamster tracheal organ cultures infected with Bordetella pertussis. Hamster tracheal organculture was employed as a model for the study of the pathogenesis of infection due to bordetella pertusis. Scanning electron microscopy provided a three-dimensional view of the surface infection of the tracheal explants. Phase I B. pertussis attached only to the ciliated epithelial cells, and a sequence of events involving the injury, expulsion, and destruction of these differentiated cells occurred. This in vitro model provides insights into the mechanisms by which B. pertussis mediates host cell injury at the site of infection.


Diagnostic skin test reactions with varicella virus antigen and clinical application of the test. Varicella virus antigen was prepared and used in skin testing of 75 children one to 17 years old. Results of skin tests were analyzed in relation to history of varicella, presence of absence of neutralizing antibody, and susceptibility to clinical varicella. All 22 children with no history of varicella and no neutralizing antibody were negative in the skin test when an erythematous change of greater than or equal to 5 mm in diameter was taken as a positive result. Of the 53 children with neutralizing antibody, only three were negative in the skin test. The skin test was applied clinically in an institution for mentally retarded children immediately after varicella infection occurred in a patient. A total of 65 patients were examined by the skin test, and 24 children with negative results were given live varicella virus vaccine. As a result, no spread of varicella was observed except for three cases, which developed two weeks after onset of illness in the index case. Thus the skin test seems clinically useful in the determination of susceptibility of individuals to clinical varicella.


Cervical excretion cytomegalovirus: correlation with secretory and humoral antibody. Cytomegalovirus (CMV) was isolated from cervical secretions of 10 of 121 outpatients at a venereal disease clinic. The buoyant densities of the DNA of five of the 10 CMV isolates were determined and found to be the same as those reported for strains isolated from other anatomical sites. Nine of 102 specimens of cervical secretions were reactive for IgG antibody to CMV, and five of 61 specimens tested were reactive for secretory IgA antibody, whereas two excretors had only IgG and secretory IgA antibody, whereas two excretors had only IgG antibody. Antibody to early antigen was identified in the sera of excretors whose cervical secretions were reactive for IgG and secretory IgA antibody.


A rapid and sensitive radiochemical assay for phosphatidic acid phosphohydrolase activity. A technique is described for the radiochemical assay of phosphatidic acid phosphohydrolase activity in rat brain. Radiochemically pure 32P-labeled phosphatidic acid of known specific radioactivity and structure, which was biosynthesized in vitro by the diacylglycerol kinase of E. coli, was used as the substrate. As little as 5 microgram of microsomal or mitochondrial protein can be used for the assay, and product formation in the picomole range can be determined accurately. This procedure should be useful in situations where only limited amounts of tissue are available.


Effect of dibutyryl cyclic AMP and oestradiol on incorporation of [3H]leucine into the proteins of luminal fluid of the rat uterus. An injection of 12-5 nmol dibutyryl cyclic AMP (dbcAMP) into each uterine horn of ovariectomized rats maintained on progesterone alone, enhanced incorporation of [3H]leucine into uterine luminal proteins. A value of 11-73 X 10(3) c.p.m.  [3H]leucine incorporated/rat was found after injection of dbcAMP compared with 3-69 X 10(3) c.p.m./rat after injection of saline. An injection of 1-0 microgram oestradiol, in contrast, gave 58-05 X 10(3) c.p.m./rat. Electrophoretic analysis showed that the radioactively labelled proteins found after an injection of dbcAMP or oestradiol were qualitatively similar to those detected on day 5 of pregnancy, the day of blastocyst implantation. The nucleotide thus apparently elicits a uterine response similar to that observed after an injection of oestradiol.


Effect of human chorionic gonadotrophin on adenylate cyclase activity and testosterone content in rat testicular mitochondria. The adneylate cyclase activity and the concentration of testosterone in testicular mitochondria from immature rats were measured after administration of human chorionic gonadotrophin (HCG) or dibutyryl cyclic AMP in vivo or in vitro. Intratesticular injection of HCG produced an increase in adenylate cyclase activity which preceded the rise in the level of testosterone, whereas addition of the trophic hormone in vitro resulted simultaneous increases. Administration of dibutyryl cyclic AMP in vivo enhanced the testosterone content of the mitochondria. However, the cyclic nucleotide added in vitro at concentrations up to 5 mmol/l had no effect. Cycloheximide injected intraperitoneally before the administration of HCG abolished the stimulatory effect of the trophic hormone on the level of testosterone in the mitochondria, whereas chloramphenicol had no effect. These results, although they confirm the role of cyclic AMP as an intermediate in the stimulatory effect of HCG on the concentration of testosterone in rat testis, do not support a role for mitochondrial adenylate cyclase in this action. A protein regulator(s) formed extramitochondrially appears to be involved in the stimulatory effect of gonadotrophins on steroidogenesis.


Testicular receptors for luteinizing hormone after immunoneutralization of gonadotrophin releasing hormone in the male rat. The effects of immunoneutralization of endogenous gonadotrophin releasing hormone (GnRH) on the serum concentrations of testosterone and gonadotrophins and the binding of 125I-labelled human chorionic gonadotrophin (HCG) to testicular membrane fractions were studied in adult male rats. Four days after the administration of 1 ml anti-GnRH serum, the level of testosterone in the serum decreased to 44% of the concentration before the injection, whereas administration of normal rabbit serum had no effect. Multiple injections of anti-GnRH serum for 4 days dramatically suppressed the secretion of gonadotrophins in rats orchidectomized 2 months earlier. In intact male rats treated identically, immunoneutralization of GnRH decreased the level of serum testosterone to 32% of the concentration present in saline-treated controls, but did not decrease the number of testicular binding sites for HCG (LH). Administration of testosterone or oestradiol for 3 or 6 days caused a marked reduction in the concentration of serum gonadotrophins but did not decrease the number of LH receptors. This study provides further support for the concept that one releasing hormone governs secretion of both FSH and LH. In addition, these studies indicate that selective reduction of gonadotrophins for 3-6 days has no effect on the number of testicular LH receptors. This suggests that pituitary hormones other than gonadotrophins may be important in the maintenance of testicular receptors for LH.


Steroid and prostaglandin concentrations in the plasma of pregnant ewes during infusion of adrenocorticotrophin or dexamethasone to intact or hypophysectomized foetuses. Catheters were implanted into 16 ewes and their foetuses between days 110 and 124 of gestation. Hypophysectomy was attempted in eight of these foetuses. Continuous infusion of synthetic ACTH (10 microgram/h) or dexamethasone (1mg/24 h) into the foetus, starting between days 124 and 129, induced premature parturition. The concentration of progesterone in the maternal peripheral plasma decreased before parturition in all animals while the level of oestradiol increased in ewes with intact foetuses or in those in which hypophysectomy was incomplete. When hypophysectomy was complete, no increase in the maternal level of oestradiol occurred before delivery. The concentration of 13,14-dihydro-15-oxo-prostaglandin F2alpha increased in the peripheral plasma of ewes with intact or hypophysectomized foetuses infused with ACTH. It is suggested that an intact foetal pituitary gland is required for the rise in the level of oestrogen prepartum, but that this rise is not essential for increased prostaglandin production of parturition.


Mammary tumors and mammary tumor virus expression in hybrid mice of strains C57BL and GR. Mammary tumorigenesis in genetic crosses between the high mammary tumor incidence GR and the low incidence C57BL mouse strains is highly correlated with murine mammary tumor virus expression in milk. Although the F1 and first backcross females had a mammary tumor incidence which was consistent with a single dominant gene segregation, the tumor incidence in the critical second backcross segregants disproved the single gene hypothesis. Genetic factors were clearly involved in regulation of virus expression which in turn correlated with both tumor incidence and tumor latency; these complex phenotypes are however best explained as threshold or quasicontinuous characters. As predicted from this model, the age specific incidence of mammary tumors showed a broad peak at 14-19 mo of age with no evidence of an early or late phase. Hematopoietic tumors showed no correlation with virus expression or mammary tumorigenesis suggesting different etiologies for these tumors.


The interaction of Herpes Simplex Virus with murine lymphocytes. I. Mitogenic properties of herpes simplex virus. Herpes simplex virus (HSV) stimulates DNA synthesis in mouse spleen cultures prepared from normal, macrophage-depleted, and T-cell-depleted spleen cells, but not from thymocytes. In addition, a polyclonal antibody response is observed in HSV-infected spleen cultures. These findings indicate that the cells stimulated to undergo DNA synthesis after HSV infection appear to be the bone marrow-derived lymphocytes. The newly synthesized DNA is host cell and not of viral origin. Heat treatment and ultraviolet irradiation of HSV before addition to spleen cultures prevents the induction of DNA synthesis. We consider the use of this system as assay for the study of cell transformation by HSV and also for the study of host cell control of the expression of the viral genome.


Secretion of plasminogen activator by human polymorphonuclear leukocytes. Modulation by glucocorticoids and other effectors. Purified human PMNs secrete plasminogen activator. This secretion is stimulated by Con A and low concentrations of PMA, and is inhibited by low concentrations of glucocorticoids, and by cAMP, actinomycin D, and cycloheximide. In contrast, the release of granule-bound enzymes, such as elastase, is achieved only at higher concentrations of PMA, and is not affected by any of the inhibitors that block plasminogen activator production. These results show that the production of plasminogen activatory by PMNs is controlled by agents that affect inflammations, and that this control is not shared by other lytic enzymes known to be associated with these cells. This suggests a particular role for plasminogen activator in the response pattern of PMNs and also supports the concept, previously developed for macrophages, that the secretion of this enzyme is correlated with cell migration in vivo.


Recovery of avian sarcoma virus from tumors induced by transformation-defective mutants. Transformation-defective (td) mutants of the Schmidt-Ruppin strain of Rous sarcoma virus (RSV), which contains deletions in the gene responsible for transformation (src gene), are unable to transform chicken embryo fibroblasts in vitro. Injection of some of these td mutants into newborn chickens resulted in the formation of sarcomas from which sarcoma virus was unfailingly recovered. The possibility that transforming RSV was present in the td virus preparations was excluded by further purification of the td viruses. Morphology of the foci induced by the newly recovered sarcoma virus was distinct from that of foci induced by the parental Schmidt Ruppin strain of RSV. It is suggested that the new sarcoma virus was generated as a result of the genetic interaction between the genomes of td virus and chicken cells.


Inhibition of lymphocyte proliferation stimulated by lectins and allogeneic cells by normal plasma lipoproteins. Lipoproteins, isolated by sequential flotation at densities 1.006, 1.019, 1.063, and 1.21, were examined for their ability to inhibit human lymphocytes stimulated by allogeneic cells and by lectins (phytohemagglutinin-P and concanavalin A). All the classes of normal plasma lipoproteins inhibited lymphoproliferation when peripheral blood lymphocytes were cultured in autologous, heterologous, or lipoprotein-deficient plasma (d greater than 1.21). The rank order of inhibitory potency was intermediate density lipoprotein (IDL) greater than very low density lipoproteins (VLDL) greater than low density lipoproteins (LDL) greater than high density lipoproteins (HDL), regardless of the mode of stimulation. The concentrations of IDL, VLDL, and LDL required for complete inhibition of stimulated lymphoproliferation were considerably below the levels of each of these lipoproteins normally found in human plasma. In addition, the concentration of HDL required for 50-90% inhibition was in the range of HDL levels normally found in human plasma. Moreover, at relatively higher concentrations, lipoproteins suppressed the incorporation of [3H]thymidine into DNA below the levels seen with reseting, unstimulated lymphocytes. The results suggest that circulating lymphocytes may normally be highly suppressed by the combined effects of all the endogenous lipoproteins and that the lipoproteins may play important roles in vivo in modulating lymphocyte functions and responses.


The determination of nonesterified fatty acids in blood serum using a stable cupric reagent. A modification of the method for the determination of nonesterified fatty acids in blood serum using a stable cupric reagent stabilized with sodium citrate, has been developed. 33 g of NaCl are dissolved in 100 ml of the described reagent, and its pH should not be additionally adjusted. The reagent is stable for at least one year if kept in the refrigerator at 5 degrees C. The determination may be done in 0.1--0.2 ml of blood serum. The relationship between the absorbance and concentration of palmitic acid is still linear at a concentration of 2500 mumol/l. Error of the method is +/- 3%.


Virus-induced diabetes mellitus: VIII. Passage of encephalomyocarditis virus and severity of diabetes in susceptible and resistant strains of mice. The diabetogenic capacity of the M-variant of encephalomyocarditis (EMC) virus was markedly diminished after passage in mouse kidney cell cultures. One passage in mice fully restored this capacity. Virus harvested after five passages in either susceptible (SWR/J) or resistant (C57BL/6J) strains of mice was capable of producing diabetes in susceptible SWR/J mice but not in resistant C57BL/6J mice. Resistance was not overcome by inoculating mice with high concentrations of virus. Immunofluorescence studies showed that islets from strains of mice (i.e. CBA, AKR, C57BL/6J, A/J) that did not develop diabetes after infection with EMC virus, nonetheless, contained virus antigens. The percentage of cells in the islets containing virus antigens varied from 3-6% in CBA to 13-5% in A/J. In contrast 38% of the islet cells in susceptible SWR/J mice contained virus antigens. It is concluded that both the genetic background of the host and the passage history of the virus influence the development of diabetes.


Alterations of actin-containing structures in BHK21 cells infected with Newcastle disease virus and vesicular stomatitis virus. The distribution pattern of actin-containing structures in BHK21 cells and the changes which they undergo upon infection with Newcastle disease virus (NDV) and vesicular stomatitis virus (VSV) were studied by means of immunofluorescence. Double labelling with antibodies conjugated with fluorescein (for actin) and rhodamine (for virus antigens) has shown that the progressive cytopathic effects after virus infection are accompanied by extensive alterations of the structures demonstrable by antiactin antibodies. In NDV-infected BHK21 cells the number of actin filaments increases, some zones which contain virus antigens apparently being in close association with the actin structures. By contrast, infection with VSV results in a strong reduction of actin-containing fibres. The results indicate that in the genesis of morphologically detectable alterations of a cell after virus infection--the 'cytopathic changes'--alterations of those structural elements are involved which are also probably responsible for maintenance of cell shape and motility.


An assessment by competition hybridization of the sequence homology between the RNAs of the seven serotypes of FMDV. A comparison has been made of the RNAs of the seven serotypes of foot-and-mouth disease virus (FMDV) by competition hybridization. Homology among the three European serotypes A, O, C and the Asia I serotype was 60 to 70%. Similar homologies were found among the three Southern African Territories serotypes (SAT I, SAT 2, SAT 3), but homology between the two groups was much lower (25 to 40%). Homology between the RNAs of subtypes within serotypes A and O was greater than 70%. Double competition experiments with the Eropean serotypes indicate sharing of nucleotide sequences.


Characterization of two distinct molecular populations of type I mouse interferons. The molecular heterogeneity of acid-stable (Type I) mouse interferons induced in C243 cells by Newcastle disease virus was analysed by SDS-polyacrylamide gel electrophoresis under non-reducing and reducing conditions, and the profiles of antiviral activities obtained were characterized biologicaly in mouse cells and in heterologous (guinea-pig) cells. Two bands of activity, A and B, were consistently present in all interferon preparations tested: under reducing conditions, the activity in all fractions of band A (with a peak of activity at about 38000 daltons) was uniformly increased, while that of band B (with a peak at about 22000 daltons) was uniformly diminished. All the active fraction in band A had only slight activity (less than 10% of homologous titres) on guinea-pig cells, whereas all those in band B were significantly more active an guinea-pig cells than on homologous L cells. Thus, mouse type I interferon preparations contain two molecular populations of interferons that can be distinguished physically (by size), biochemically (by the effect of reduction on reactivation from SDS) and biologically (by activity in heterologous cells).


Structural studies of encephalomyocarditis virus RNA both in situ and in free solution. The secondary structure of encephalomyocarditis (EMC) virus RNA has been studied in situ and in free solution by absorbance-temperature relationships and by circular dichroism (CD). Extracted EMC virus RNA melts reversibly and has a hypochromicity of about 20%; analysis of CD spectra and formaldehyde treatment suggests that approx. 60% of the nucleotides are involved in base-pairing at 25 degrees C. It is shown that the RNA within the virus particle is less structured than when it exists in free solution, being partially stabilized by capsid protein against melting until the virion is disrupted to release the intact RNA. Upon clarification to remove denatured capsid protein, the released RNA gives a melting profile identical with that of phenol-extracted virus RNA. The results suggest that the intact structure of the virus is dependent upon intimate non-covalent bonds between RNA and protein together with hydrophobic bonds between the protein subunits.


Avian tumour virus interactions with chicken fibroblast membranes: partial characterization of initial attachment site activity. Exposure of chicken embryo fibroblasts (CEF) to trypsin solubilizes cell surface components, including the initial attachment site for avian tumour viruses (ATV). The soluble attachment site activity appears to interact directly with the ATV in vitro thereby interfering with the binding of at least two ATV subgroups to both intact CEF and CEF plasma membranes. A result of this interaction in vitro is reduced ATV infectivity, demonstrated by reduced transforming capacity of RSV (RAV-I).


Scanning electron microscopical observations on the cytopathology of porcine enteroviruses in PK-14 cells. Examination by scanning electron microscopy (SEM) of PK-15 cells infected with each of the two cytopathogenic types of porcine enterovirus revealed changes similar to those observed by light microscopy. However, differences between the two types could be detected at an earlier stage of the infection. Cells infected with T80 virus (type I) were rounded, with surface microridges, while V13 (type II) infected cells were characterized by cytoplasmic protrusions.


Immunosuppression reactivates and disseminates latent murine cytomegalovirus. When searched for by standard virological methods, murine cytomegalovirus (MCMV) becomes undetectable by 4 months after inoculation of mice. However, a 2-week regimen of immunosuppression with antilymphocyte serum and corticosteroid results in reactivation and dissemination of the virus in virtually all animals. This system should be useful in defining the pathogenesis of generalized cytomegalic disease resulting from reactivation of latent virus in immunocompromised individuals.


Replication of herpes simplex virus DNA after removal of hydroxyurea block from infected cells. Hydroxyurea (HU) treatment of HSV-infected cells markedly inhibits the synthesis of virus DNA. Only 0-3.6 of the 3H-thymidine label was incorporated into virus DNA in the presence of HU as compared to untreated infected cells. Removal of HU resulted in a renewed synthesis of virus DNA as determined by the gradual increase in the incorporation of 3H-thymidine into HSV DNA. The labelled virus DNA molecules were isolated and chromatographed on benzoylated napthoylated DEAE (BND)-cellulose columns to separate the replicative intermediates that have single-stranded (ss) sequences from the mature double-stranded (ds) DNA genomes. Mature radioactive dsDNA molecules were found to appear at 22 min after removal of HU and gradually increased in amount thereafter. The virus DNA molecules synthesized during the initial 20 min after removal of HU, constitute the replicative intermediates of HSV DNA. It was calculated that the synthesis of HSV DNA proceeds at the rate of about 5 X 10(6) daltons per min.


Observations on the morphology of two rotaviruses. The negative staining technique was used to study the morphology of two rotaviruses, the epizootic diarrizootic diarrhoea of infant mice (EDIM) and the simian virus SA II. It is proposed that the inner capsid of the virion has icosahedral symmetry and consists of 180 morphological units arranged in an open lattice formation with the 12 spaces at the apices being surrounded by 5 capsomeres and the other 80 spaces each surrounded by six capsomeres. The outer capsid of the virion consists of a honeycomb-like lattice which corresponds to the lattice arrangement of the inner capsid.


A micro-electrode study of peripheral neuropathy in man. Part 1. Responses to single graded stimuli. Multi-unit micro-electrode recording were obtained from sensory fascicles of the sural and median nerves of 12 control subjects and of 28 patients with peripheral neuropathy. Spontaneous activity and mass responses to mechanical and electrical stimuli were examined. Mechanoreceptor function appeared normal but there was a reduced number of responsive receptors in peripheral neuropathy. The electrical activation threshold of nerve fibres of all conduction velocities was increased in neuropathy and a greater number of fibres needed to be activated for preception to occur. Clinical sensory impairment was associated with a reduction in size of the initial compound action potential of the maximal evoked neurogram and with dispersion of fibre responses. Pathological slowing of fibre conduction velocity was demonstrated in demyelinating neuropathy but in most cases of axomal degeneration the changes in velocity could have been due either to a reduced number of fast conducting fibres, or to conduction block. No changes were observed in C-fibre activity in these patients.


A micro-electrode study of peripheral neuropathy in man. Part 2. Responses to conditioning stimuli. Surface, needle and micro-electrode recordings were obtained from sensory nerves of patients with various types of peripheral neuropathy. Changes in amplitude and conduction velocity of nerve action potentials were measured after a single conditioning stimulus and after tetanic stimulation for 2 min. In patients with hereditary forms of axonal degeneration (AD), recovery processes of nerve fibres of all conduction velocities were normal; in acquired forms of AD fibres with conduction velocity less than 30 m/sec had greater and more prolonged post-tetanic depression than control nerves of similar conduction velocity. Where neuropathy was associated with segmental demyelination (SD), fibres of all conduction velocities had prolonged recovery processes after both single and tetanic stimulation. The changes were especially marked at higher skin temperature, and were greater than the changes seen in nerves with acquired forms of AD. Finally, 2 sural nerves were studied during the process of Wallerian degeneration after a biopsy had been obtained proximally, and recovery processes did not change during the period of degeneration. Perceptual abnormalities were similar in AD and SD. It is suggested that changes in recovery processes of nerve fibres with segmental demyelination or regeneration after injury contribute to the perceptual abnormalities which occur in clinically encountered peripheral neuropathies.


Spontaneous phagocytosis of boutons on spinal motoneurons during early postnatal development. An electron microscopical study in the cat. The boutons making synaptic contact with different regions of spinal motoneurons have been studied during the early postnatal period. The observations suggest that a spontaneous glial phagocytosis of boutons of different types occurs mainly during the second postnatal week. This finding is discussed in relation to the previously demonstrated disappearance of boutons from the initial axon segment of spinal motoneurons during the same period of development and also in relation to other degenerative and eliminatory phenomena associated with the normal development of the nervous system.


Localization of 5-hydroxytryptamine and protein(s) in the secretion granules of the rudimentary photoreceptor cells in the pineal of Lacerta. In order to localize 5-hydroxytryptamine (5-HT) stores, the pineal organ of Lacerta was examined by electron microscopy following argentaffin and chromaffin reactions. The results obtained with these ultracytochemical reactions were correlated with those obtained previously using the Falck and Hillarp method and electron microscopic radioautography. The secretion granules (diameter: 60-340 nm) of the secretory rudimentary photoreceptor (SRP) cells are the locus of the argentaffin and chromaffin reactions. The reactivity of the granules disappeared after p-CPA or reserpine administration and increased after nialamide injection. From the previous and present results it can be concluded that 5-HT in the pineal organ of Lacerta is stored in the secretion granules of SRP cells and coexists with a proteinaceous and/or possibly a glycoproteinaceous component. Whether this secretion is neurohormonal in nature is still not clear.


Immunobiology of primary intracranial tumors. Part 3: Microcytotoxicity assays of specific immune responses of brain tumor patients. Fifty-six in vitro microcytotoxicity assays were conducted on 30 patients with intracranial tumors at various times during the postoperative course. Significant specific cellular cytotoxic responses were found in nine of 56 assays, humoral cytotoxic responses in nine of 54 assays, and host effector cell-dependent, antibody-dependent cytotoxic responses in four of 28 assays. Variables that might influence the occurrence of cytotoxicity were studied, and the relationship of these findings to other immune parameters was discussed.


Effect of diet restriction on some key enzymes tryptophan-NAD pathway in rats. Dietary intake of rats was restricted by feeding varying amounts of a 20% protein diet. After 6 weeks of feeding, some key enzymes of the tryptophan and nicotinic acid-NAD pathway, liver nicotinamide nucleotide concentration, and urinary metabolites of tryptophan and nicotinic acid were studied. With an increase in diet restriction, liver tryptophan oxygenase (EC 1.13.1.12) activity increased. Quinolinate phosphoribosyltransferase (EC 2.4.2.a) activity, on the other hand, was found to decrease with moderate diet restriction up to 50% restriction, but increased again with more severe diet restriction in rats fed 25% of ad libitum intake. Liver nicotinate phosphoribosyltransferase (EC 2.4.2.11) activity was also observed to decrease with moderate diet restriction and did not further change when the restriction was severe while picolinate carboxylase (EC 4.1.1.45) activity increased significantly only in severe diet restriction. In rats fed 25% of ad libitum intake, urinary quinolinic acid excretion was low whereas N'-methylnicotinamide excretion was elevated. Alterations in the enzyme activities accompanied by changes in the levels of urinary metabolites, observed in the present study, suggest that the potential efficiency of conversion of tryptophan to nicotinamide nucleotides is not constant and is influenced by dietary intake.


The secretion of adrenal androgens and growth patterns of patients with hypogonadotropic hypogonadism and isiopathic delayed puberty. Plasma concentrations of adrenal androgens (DHEA and DHEA-S) and the growth patterns of four male patients with hypogonadotropic hypogonadism have been compared and contrasted with those of eight male patients wit idiopathic delayed puberty. As a group the patients with HH presented at an older age with delayed puberty and normal heights; the growth rate and pattern have been normal except for an absence of the pubertal growth spurt. Adrenal androgens were usually normal for chronologic age and high for bone age. The patients with IDP presented at a younger age, usually with short stature, low adrenal androgens relative to the chronologic age, but normal relative to the bone age. Patients with IDP appear to exhibit a delay in maturation of both the adrenal cortex and the hypothalamo-pituitary gonadal axis usually with short stature and retardation of the bone age. The importance in measurements of adrenal androgens in the diagnosis of HH and of IDP is emphasized.


Chronotropic and cyclic adenosine monophosphate response of fetal rat heart in organ culture to isoproterenol, quinidine, and a dysrhythmogenic agent. The fetal rat heart in organ culture was used to investigate rate chanes by various cardioactive agents. Concomitantly, the effect of these pharmacologically induced rate changes on steady-state cyclic adenosine monophosphate levels was determined. Isoproterenol increased the fetal rat heart rate and cyclic adenosine monophosphate in a concentration-related fashion. Quinidine produced a concentration-related decrease in heart rate and no change in cyclic adenosine monophosphate level. The dysrhythmogenic agent produced concentration-related negative chronotropism in the fetal rat heart preparation and significant elevations in cyclic adenosine monophosphate at concentrations without chronotropic action. No correlation between chronotropic effect of a drug and cyclic adenosine monophosphate levels was observed.


Preparation, identification, and quantitative NMR determination of silyl derivatives of 6-aminopenicillanic acid, 7-amino-3-methyl-delta3-cephem-4-carboxylic acid, and 7-amino-3-acetoxymethyl-delta3-cephem-4-carboxylic acid. A rapid and accurate method for the quantitative determination of the extent and ratio of amino and carboxyl group trimethylsilylation of 6-aminopenicillanic acid, 7-amino-3-methyl-delta3-cephem-4-carboxylic acid, and 7-amino-3-acetoxymethyl-delta3-cephem-4-carboxylic acid is presented. The method utilizes NMR spectroscopy and is based on the difference in chemical shifts between N-trimethylsilyl and O-trimethylsilyl groups or, in cephalosporin derivatives, between the methyl group in the 3-position and free amino resonances. The spectra of the N,O-bis(trimethylsilyl) derivatives are discussed.


GLC determination of 6-aminopenicillanic acid and 7-amino-3-methyl-delta3-cephem-4-carboxylic acid. A quantitative GLC determination of 6-aminopenicillanic acid and 7-amino-3-methyl-delta3-cephem-4-carboxylic acid is presented. The results obtained are in good agreement with those of known chemical procedures. The method is free from interference by related substances.


Antidiarrheal and central nervous system activities of SC-27166 (2-[3 - 5 - methyl - 1, 3, 4 - oxadiazol - 2 - yl) - 3, 3 - diphenylpropyl] - 2 - azabicyclo [2.2.2]octane), a new antidiarrheal agent, resulting from binding to opiate receptor sites of brain and myenteric plexus. Pharmacological studies were performed to investigate the interaction of SC-27166 (2-[3-(5-methyl-1,3,4--oxadiazol-2-yl)-3,3-diphenylpropyl]-2-azabicyclo[2.2.2]octane), a new antidiarrheal agent, with opiate receptor sites in vitro and in vivo. Morphine, loperamide and SC-27166 inhibited the binding of [3H]naloxone to homogenates of guinea-pig brain and myenteric plexus and the inhibition was diminished in the presence of 100 mM Na+. Unlike that of morphine and [3H]naloxone itself, the binding of loperamide and SC-27166 was complex and Scatchard plots indicated the presence of low and high affinity sites for both compounds. Morphine, loperamide and SC-27166 inhibited the contractions of electrically driven longitudinal muscle from guinea-pig ileum and naloxone antagonized these effects. In the anesthetized dog, i.v. administration of morphine and SC-27166 enhanced the contractile activity of circular muscle in proximal and distal duodenum and distal ileum but duodenal longitudinal muscle was relaxed; these effects were completely reversed by subsequent administration of naloxone. In the rat, p.o. administration of loperamide and SC-27166 inhibited intestinal propulsion at doses considerably lower than were necessary to produce activity in the hot plate test; this specificity of action was not seen with morphine. In the rat, p.o. administration of loperamide and SC-27166 inhibited diarrhea at doses considerably lower than were necessary to produce withdrawal symptoms. The authors concluded that both loperamide and SC-27166 are specific antidiarrheal agents that produce both their central and antidiarrheal effects by binding to opiate receptor sites.


Control of cyclic adenosine 3':5'-monophosphate-elevating effect of adenosine in C-1300 murine neuroblastoma in tissue culture. The elevation of cyclic adenosine 3':5'-monophosphate (cyclic AMP) in response to adenosine in C-1300 murine neuroblastoma (clone N2a) in surface culture is increased in magnitude in cultures pretreated overnight with theophylline or adenosine deaminase. This "potentiating" effect of theophylline takes time to develop and is blocked by cycloheximide. The cyclic AMP-elevating effect of adenosine decreases in magnitude as the cultures approach confluence. This reduced responsiveness is reversed by the overnight treatment with theophylline. It is hypothesized that adenosine is continually released by the cells to the growth medium and that this adenosine acts extracellularly to modulate the sensitivity of the cells to the cyclic AMP-elevating effect of adenosine.


Protection from habituation of the crayfish lateral giant fibre escape response. 1. Habituation of the lateral giant fibre escape response in the crayfish to repetitive tactile stimuli is believed to result from homosynaptic depression at the first synapse of the reflex, between tactile afferents and interneurones. Normally, habituation of escape responses to repeated innocuous stimuli is presumed to be adaptive. Experiments reported here were undertaken to determine whether habituation would occur under circumstances when it would presumably be maladaptive - in particular, when tactile receptors are stimulated by an animal's own tail-flip movements.2. Experiments were carried out on the crayfish isolated abdominal nerve cord, which contains the lateral giant reflex pathway.3. Compound e.p.s.p.s elicited in the lateral giant by electrical stimulation of tactile afferents decline by from 25 to 36% over a series of eleven trials at 1/5 sec (control series).4. To determine whether such a decline would occur when sensory afferents are stimulated during a ;tail-flip', stimuli were given as in the control series but each stimulus occurred 20 msec after direct electrical stimulation of a medial giant or lateral giant escape-command fibre at which time tail flexion movements of an intact animal would be in progress. Under these conditions% e.p.s.p. decline over 11 trials at 1/5 sec was only 16-45% of that occurring on the control series.5. This protective effect starts at about 10 msec after escape command neurone firing, is maximal at 20 msec, and thereafter declines, remaining weakly detectable at 100 msec. This time course is commensurate with that required for execution of a tail-flip movement. Thus, sensory afferent-to-lateral giant transmission is protected from depression if stimuli occur when a tail-flip movement is or should be occurring.6. Giant fibre spikes do not superimpose facilitation upon a depressed reflex pathway, nor accelerate rate of recovery from depression; rather, protection is attributable to actual prevention of development of the depressed state.7. Protection was also examined at the first synapse of the reflex, where the depression responsible for habituation is believed to occur, by recording intracellularly in the largest of the first-order interneurones (interneurone A) of the pathway. In absence of protection, ten stimuli presented at 1/4 sec caused a mean decline of 32% in the e.p.s.p. in interneurone A. When such stimuli followed directly evoked escape command neurone firing by 20 msec this decline was reduced by 59-100%.8. We suggest that protection serves to prevent crayfish from habituating to stimuli produced by their own tail-flip movements.


Presynaptic inhibition: the mechanism of protection from habituation of the crayfish lateral giant fibre escape response. 1. Mechanism of protection from habituation of the lateral giant escape reflex of the crayfish was studied. Experiments were designed to determine whether presynaptic inhibition of primary afferents for the reflex occurs following escape command neurone firing, and if so, whether it could account for protection of the first synapse from depression. 2. Synaptic transmission between afferents and interneurone A of the escape reflex is strongly inhibited following giant fibre spikes. 3. Giant fibre firing results in post-synaptic inhibition of interneurone A. However, inhibition of afferent input to interneurone A consistently outlasts both i.p.s.p.s and post-synaptic conductance increases in the neurone; the inhibition, therefore, is probably not exclusively post-synaptic. 4. Giant fibre firing results in excitability changes in sensory afferent terminals as measured by the amplitude of antidromic compound action potentials to focal electrical stimuli applied in the region of afferent terminals in the last abdominal ganglion. The time course of this effect parallels those of protection and inhibition of the first synapse. 5. The magnitude and time course of protection and inhibition of transmission to interneurone A parallel each other closely. Both processes considerably outlast measurable signs of post-synaptic inhibition. 6. We conclude that following giant fibre activity the first synapse of the lateral giant reflex is presynaptically inhibited and the presynaptic inhibition is responsible for the protection effect described in the preceding paper.


Transmission from photoreceptors to ganglion cells in turtle retina. 1. Synaptic transfer between photoreceptors and impulse-generating cells was studied in isolated eyecups from turtles. Single red-sensitive cones or rods were stimulated by current passed through an intracellular electrode, and impulses generated by the resulting synaptic action were recorded with an external micro-electrode. This technique permits study of retinal transmission without the operation of the visual transduction mechanism. Antidromic stimulation of the optic nerve indicated that most of the impulse-generating cells were ganglion cells.2. Individual ganglion cells responded transiently to changes in the membrane potential of a receptor and could be classified into three groups on the basis of the direction of the effective change in potential. Off centre ganglion cells responded selectively to depolarizations of a receptor, while on centre ganglion cells responded selectively to hyperpolarizations. On-off ganglion cells responded to both depolarizations and hyperpolarizations of a receptor.3. Ganglion cells gave the same pattern of response to electrical hyperpolarization of a receptor and to light in the centre of their receptive fields. Subthreshold depolarizing currents passed in a receptor antagonized the ganglion cell's response to light, and subthreshold hyperpolarizing currents reinforced the response. These observations are consistent with the view that the hyperpolarization generated by visual transduction is responsible for regulating the release of transmitter at the first retinal synapse.4. When a receptor was stimulated with weak current pulses of fixed intensity the number and latency of the ganglion cell impulses fluctuated randomly in successive trials. The relation between the fraction of trials yielding a response and the stimulus intensity was broad. These results indicate that the link between retinal input and output is noisy.5. In the most sensitive pairs of cells, a response of one or more impulses could be obtained in half the trials with a current of about 2 x 10(-11) A, which changed the potential of the receptor by 1-2 mV. A current of similar magnitude would be developed by about 130 photoisomerizations in a red-sensitive cone or 50 photoisomerizations in a rod.6. Dim background light producing a steady hyperpolarization of a few millivolts in the rods raised the threshold for electrically-evoked transmission from a rod to a ganglion cell. In experiments on red-sensitive cones, background light raised the threshold in the off pathway, in which depolarization was the effective stimulus, and lowered the threshold in the on pathway, in which hyperpolarization was the effective stimulus. These changes in sensitivity were not accompanied by obvious changes in the input resistance of the stimulated receptor. Regulation of retinal sensitivity in background light thus involves changes in synaptic transfer as well as changes in the sensitivity of the visual transduction mechanism.


Kinetics of synaptic transfer from receptors to ganglion cells in turtle retina. 1. Synaptic transfer between the retinal input and output was studied in turtle eyecups by injecting rectangular current pulses into a single cone or rod while recording externally from a ganglion cell.2. When a receptor was activated with weak steps of polarizing current, the probability of obtaining a ganglion cell impulse rose after an S-shaped delay to a peak at about 0.1 sec and then declined. This suggests that the transmission chain behaves like an electrical band-pass filter containing delay and differentiating elements.3. To further characterize the kinetics of excitation in the subthreshold region, the duration and polarity of the polarizing current pulses were varied while determining the magnitude of the threshold current and the delay to the ganglion cell impulses. The results of these experiments were described with linear models which assume that synaptic transfer occurs over a cascade of first-order delay stages and a single differentiating stage.4. The pathways which relay off responses to light from rods and red-sensitive cones were formally similar, but the time scale in the rod path was several times slower. The path carrying off responses from the red-sensitive cones was faster than the on path. These kinetic differences indicate that independent pathways mediate each of the three categories of response and suggest that the kinetics of each path are ;matched' to the input signals generated by light.5. The strength-latency relations for the responses of on-centre ganglion cells to flashes and steps of light were approximately predicted from the description of synaptic transfer developed here and the description of visual transduction in red-sensitive cones from a previous study.6. It is suggested that the retinal paths have kinetics which might be useful in transmitting light-evoked signals whilst attenuating noise present near the input.


Changes in renal cyclic nucleotide content as a possible trigger to the initiation of compensatory renal hypertrophy in rats. 1. Cyclic adenosine 3', 5'-monophosphate (cyclic AMP) and cyclic guanosine 3', 5'-monophosphate (cyclic GMP) have been estimated in the kidneys of rats.2. Ten minutes after unilateral nephrectomy there was a threefold increase of cyclic GMP in the remaining kidney, which was accompanied by a moderate fall of cyclic AMP.3. The changes in cyclic nucleotides in the remaining kidney after unilateral nephrectomy were of short duration.4. When an anephric rat was cross-circulated with a normal litter-mate, there was an increase of cyclic GMP concentration in the kidneys of the latter, which reached its maximum 10 min after the establishment of the cross circulation.5. In experiments where one kidney of a litter-mate was transplanted to the neck of another rat, unilateral nephrectomy was not followed by changes of the level of cyclic nucleotides in either the transplanted or the remaining kidney. Bilateral nephrectomy, however, resulted in a marked increase of cyclic GMP in the transplanted kidney.6. The clamping of the blood vessels to one kidney for periods up to 10 min had the same effect as unilateral nephrectomy on the concentration of cyclic GMP in the remaining kidney. When the clamp was removed and the circulation restored, the concentrations of cyclic nucleotides returned to preoperative levels in both kidneys.


Synaptic excitation and inhibition resulting from direct action of acetylcholine on two types of chemoreceptors on individual amphibian parasympathetic neurones. 1. Synaptic transmission was studied in visually identified parasympathetic ganglion cells that modulate the heart beat of the mudpuppy Necturus maculosus).2. The brief pulse of acetylcholine (ACh) released from terminals of the vagus nerve after each impulse can produce two distinct post-synaptic responses in individual principal cells of the ganglion: (i) within a milli-second of release, ACh generates a rapid and strong excitatory post-synaptic potential (e.p.s.p.) that normally initiates a post-synaptic impulse; (ii) this excitation is usually followed by a slow hyperpolarizing inhibitory post-synaptic potential (i.p.s.p.) that lasts for several seconds. The magnitude and time course of the i.p.s.p. depends on the frequency and number of vagal stimuli. When the hydrolysis of ACh is inhibited by prostigmine, a train of nerve stimuli may be followed by an i.p.s.p. lasting half a minute or longer.3. The rapid e.p.s.p. and slow i.p.s.p. result from the direct action of ACh on two different types of chemoreceptors in the post-synaptic membrane of the principal cell. The e.p.s.p. can be preferentially blocked by the nicotinic antagonist dihydro-beta-erythroidine (5 x 10(-7)M), while the i.p.s.p. is selectively blocked by the muscarinic antagonist atropine (5 x 10(-9)M).4. Potentials resembling nerve-evoked e.p.s.p.s and i.p.s.p.s can be produced by iontophoretic release of ACh from micropipettes onto the post-synaptic membrane. Application of the muscarinic agonist bethanechol generates exclusively inhibitory responses.5. The reversal potential for the i.p.s.p. is about -105 mV, which is approximately the equilibrium potential for potassium (E(K)). When the external K(+) concentration is altered, the reversal potential for inhibition is shifted to the new value of E(K) as expected from the Nernst equation. Changes in the external Na(+) and Cl(-) concentrations have no appreciable effect on the reversal potential. Thus, the i.p.s.p. is the result of a conductance increase for K(+).6. The conductance change producing the i.p.s.p. is voltage sensitive. When the membrane potential is shifted from -40 to -60 mV, the i.p.s.p becomes larger and longer. Beyond -60 mV the inhibitory response decreases in proportion to the driving force on K(+) without any further change in time course.7. The inhibitory response produced by an iontophoretically applied pulse of bethanechol has a delayed onset of about 150 msec at 24 degrees C. The early portion of this response, including the delay, is proportional to t(3), where t is time. The proportionality factor (the apparent rate constant) decreases elevenfold when the temperature is lowered by 10 degrees C. This suggests that a multi-step process is involved in the activation of the conductance increase that leads to the inhibitory response. Inhibitory responses with similar kinetics were produced in heart muscles of the mudpuppy upon application of ACh.


[Norepinephrine interference in adrenal hormone balance related to blood calcium effects of PTH (author's transl)]. Decreased hypercalcemic effects of PTH are observed after medulloadrenalectomy, (table I), metoprione (table II) or cortisone (table IV) treatments in guinea pigs. 2. Norepinephrine restores the blood calcium effects of PTH in these cases but epinephrine is inactive. 3. Immediate activation of kidney adenylate cyclase by PTH (increased urinary AMPc) is also reduced in medulloadrenalectomised, cortisone or metopirone treated guinea pigs (table V). But in this test, norepinephrine reduces the effect of PTH (fig. 1).


[Placental and fetal growth in the rabbit: influence and mechanism of action of ovarian hormones (author's transl)]. During the second half of gestation in normal rabbits, hormonal overdoses result in similar modifications of placental and fetal weights, as in castrated animals identically treated (fig. 1). Nucleic acid and protein assays show that hormonal mechanisms are very different for the maternal (table I) and fetal portions of the placenta (table II): progesterone alone promotes a hypertrophy of the fetal placenta and hyperplasia of the maternal placenta. Estradiol at physiological levels counteracts these effects. Moreover, progesterone (fig. 3) and estradiol (fig. 2) receptors are found only in the maternal placenta, although the fetal placental is the most sensitive to the hormonal environment.


Problem solving revisited.

Problem solving revisited. In this paper the increasing problems and need for novel solutions in today's world are discussed. A framework consisting of the combination of the problem-solving process and the creative thinking process is considered. Inherent in each process is a predominant mode of thought: reasoning or vertical thinking in the problem-solving process, and imagination or lateral thinking in the creative thinking process. These processes directed in a complementary manner are suggested as a means of solving problems creatively in situations which call for unique and novel solutions.


A collaborative model for university nursing education and agency staff development. A model is proposed for the integration of the education of senior nursing students and staff at a mental hospital, maximizing utilization of time and resources. A pilot study employed small group techniques with faculty as facilitators and resource persons, with the dual purpose of increasing competence and reducing communication barriers. Four students and four nursing staff participated in weekly meetings, alternating supervision groups (2 groups, each composed of 1 faculty, 2 students and 2 staff) with discussion groups (2 faculty, 4 students and 4 staff). Evaluation tools used did not yield significant differences between pre- and post-tests. However, objective and subjective indices point to positive changes of attitudes between all unit staff and students, improved therapeutic skills of students and staff, both on individual and group level. Ongoing group meetings of patients run by students and staff resulted.


Microwave fixation of brain tissue as a neurochemical technique- a review. Microwave devices have been developed for rapidly inactivating brain enzymes by focusing the power output into the heads of small laboratory animals. The rapid inactivation achieved prevents postmortem changes and permits the measurement of neurochemicals such as acetylcholine at concentrations close to those obtained in vivo. The technique promises the assay of neurochemical parameters not possible before.


Leukocyte migration in ovarian carcinoma: comparison of inhibitory activity of tumor extracts. Inhibition of migration of leukocytes from patients with serous cystadenocarcinoma of the ovary was studied by the use of several different types of ovarian carcinoma extract as antigen. KCl extract of an ovarian carconoma was found to be the most effective antigen preparation in comparison with saline, deoxycholate, and perchloric acid extracts. Low concentrations of KCl ovarian carcinoma extract significantly inhibited migration of leukocytes from 11 of 17 patients with ovarian carcinoma (migration index, less than 0.74). Leukocytes from patients with breast, colon, or endometrial carcinoma showed minimal reactivity with ovarian carcinoma KCl extract, and leukocytes from patients with ovarian carcinoma showed minimal reactivity with KCl extracts of breast, colon, and endometrial carcinoma. These results suggested that the 3 M KCl procedure is superior for the isolation of antigens active in the leukocyte migration inhibition test and that this test may be of use for the isolation of tumor-associated antigen and the immunodiagnosis of ovarian carcinoma.


Cytoplasmic leucine naphthylamidase activity expressed in signet-ring cell carcinoma of the stomach. Histochemical studies were performed on signet-ring cells in surgically resected gastric carcinomas from 64 patients. On the basis of periodic acid-Schiff reactivity of the intracellular mucin, the signet-ring cells were classified as type A (immature), type B (intermediate), or type C (mature). Different staining reactions to Alcian blue at pH 2.5 enabled type C cells to be subclassified as C-1 or C-2. In 63 cases (98.4%), intense leucine naphthylamidase (LNAase) activity was found histochemically in the cytoplasm of nearly all cancer cells except type C-2. Localization of LNAase activity in the cytoplasm was confirmed by conventional biochemical assay. From the present results, histochemical characteristics of signet-ring cell carcinoma are discussed.


Cyclic nucleotide phosphodiesterase and protein activator in human cancer cell lines and Brown-Pearce carcinoma. 3':5'-Cyclic-AMP phosphodiesterase (EC 3.1.4.17) and the activating factor of cyclic nucleotide phosphodiesterase were detected in cultured human cell lines from patients with lymphoblastic leukemia and retinoblastoma and in the Brown-Pearce (rabbit) carcinoma. The homogenate of lymphoblasts contained levels of the activating factor in excess of that required to produce maximal activation of the endogenous phosphodiesterase. The activating factor found in these malignant cells appears to be similar to the calcium-binding protein activator of bovine brain phosphodiesterase on the basis of the molecular weight obtained from gel filtration, electrophoretic patterns, calcium requirement for the activity, and the effect of calcium on the proteolysis. In addition, the tumor-derived activator was able to restore the activity of activator-deficient phosphodiesterase from the bovine brain.


Malignant tumors in rats fed nafenopin, a hepatic peroxisome proliferator. Nafenopin (2-methyl-2-[P-(1,2,3,4-tetrahydro-1-naphthyl)phenoxy] propionic acid; Su-13,437), a potent hypolipidemic hepatic peroxisome proliferator, was fed to male F344 rats at a dietary concentration of o.1% until the end of the experiment at 25 months. Between 18 and 25 months, 12 of 15 rats (80%) developed tumors. Approximately 73% (11/15) developed hepatocellular carcinomas, and 10% (3/15) developed pancreatic acinar cell tumors, including 1 metastasizing carcinoma. The hepatocellular carcinomas as well as the acinar cell carcinoma of the pancreas were transplantable successfully through 6 generations.


Cellular immunity to murine sarcoma virus-induced tumors as measured by macrophage migration inhibition assays. C57BL/6N mice immunized with regressor murine sarcoma virus (MuSV) were studied at different times after inoculation for their cellular immune responses as measured by migration inhibition assays. We used both the direct capillary and indirect agarose dropiet methods and, as the source of antigens, viable tumor cells and 3 M KCl-solubilized extracts. Migration inhibitory factor (MIF) production coulctivity becoming undetectable after 21 days. However, the level of activity and the kinetics of production of this lymphokine were strongly influenced by the source of the antigen and the form in which it was presented to the immune spleen cells (ISC). Using RBL-5 ct 9-10 days, a decrease to low levels at 14 days, and a second peak of activity between 17 and 21 days. However, uith MBL-2 cells or with 3 M KCl-solubilized antigen from fresh RBL-5 ascites cells, MIF production was observed as early as 9-10 days after tumor induction, peaked at 14 days, and decreased substantially by 21 days. T-cells appeared to be required for migration inhibition reactivity, since ISC depleted of T-lymphocytes by treatment with antibody plus complement were unable to produce MIF after antigen stimulation. The results obtained from specificity studies on the response of ISC in migration inhibition to 11 different tumor lines agreed with the results previously obtained from cytotoxicity studies. With the use of RBL-5 cells as the antigen, there appeared to be an inverse relationship between the development of specific cytotoxic effector cells in 51Cr-release assay and the development of specific effector cells needed for MIF production. However, after removal of adherent cells, the level of cytotoxicity observed correlated with MIF release by immune lymphocytes.


In vitro inhibition of tritiated thymidine uptake in Morris hepatoma cells by normal rat liver extract: a possible liver chalone. A phosphate-buffered saline extract of normal rat liver tissue almost completely inhibited DNA synthesis in Buffalo rat Morris hepatoma 7777 cells in vitro. This effect was tissue-specific because it did not occur with extracts of kidney, spleen, heart, lung, muscle, and hepatoma. The liver extracts did not inhibit in vitro human prostate carcinoma or phytohemagglutinin-stimulated human and rat peripheral blood lymphocyte cultures. The addition of 10% fetal calf serum in the incubation medium had no effect on this inhibition. We determined that doses of liver extract that inhibit thymidine incorporation were not toxic inasmuch as treated cells remained viable, as indicated by trypan blue exclusion. The liver extract may thus contain a cell-specific mitotic inhibitor, a chalone for hepatoma cells.


Effect of strain differences and tumor presence on microsomal drug metabolism in the guinea pig: brief communication. In vitro drug metabolism in the Hartley guinea pig was compared with that in two inbred guinea pig strains used as carriers for the line 10 hepatoma. We observed minor differences in enzyme specific activity among the three strains. Three weeks after intradermal inoculation of Strain 2 guinea pigs with line 10 hepatoma cells, cytochrome P450 levels and aminopyrine demethylase activity were significantly decreased. Seven to 10 days after inoculation with the ascites form of the tumor, the activities of aniline and biphenyl hydroxylases, p-aminobenzoic acid N-acetyltransferase, and dichloronitrobenzene glutathione S-aryltransferase, in addition to those of cytochrome P450 and aminopyrine N-demethylase, were probably also described.


Induction of hepatic tumors in a teleost (Oryzias latipes) after treatment with methylazoxymethanol acetate: brief communication. Liver neoplasms including trabecular hepatoma and cholangioma were induced in medakas (Oryzias latipes) by the addition of methylazoxymethanol acetate to their aquarium water at levels of 0.1-3 ppm for periods ranging from 1 to 120 days. More than 80% of the surviving fish developed tumors at 3 or 5 months after commencement of the treatment, irrespective of levels of the drug, when appropriate times of exposure were chosen.


Abrogation of antitumor effects of Corynebacterium parvum and BCG by antimacrophage agents: brief communication. The consistently demonstrable antitumor effect of Corynebacterium parvum and BCG against a 7,12-dimethylbenz[a]anthracene-induced rat fibrosarcoma, growing either as a localized subcutaneous tumor or in ascites form, was abrogated by treatment of rats with antimacrophage agents such as silica or carrageenan.


Distribution and excretion of 2,4,5,2',4',5'-hexabromobiphenyl, the major component of Firemaster BP-6. The intestinal absorption, distribution, and excretion of the major component of Firemaster BP-6,2,4,5,2',4',5'-hexabromobiphenyl, has been studied in the male rat. This polybrominated biphenyl was readily absorbed from the intestine, initially distributed throughout the body, and eventually stored primarily in the adipose tissue, was not subject to appreciable metabolism, and was excreted almost exclusively in the feces and at a very slow rate. Approximately 90% of an oral dose was absorbed from the intestine, and extrapolation of the rate of excretion to infinity indicates that less than 10% of the total dose would ever be excreted.


Radiation-induced stenosis of the renal artery causing hypertension: case report. A case is presented of renal artery stenosis causing renin-dependent hypertension 11 1/2 years after irradiation for Wilms tumor. The literature was reviewed briefly for other cases of large artery damage after irradiation therapy.


Hepatoma in a child with methotrexate-induced hepatic fibrosis. During autopsy, an 11-year-old girl with acute lymphocytic leukemia was found to have hepatic fibrosis with a 2.5-cm nodule of hepatocarcinoma. For 5 1/2 of the six years of treatment, chemotherapy consisted of daily orally administered methotrexate with monthly doses of vincristine sulfate and prednisone. The cumulative dose of methotrexate was 2.5 g. To our knowledge, this report constitutes the first association of hepatoma with methotrexate-induced hepatic fibrosis. A careful follow-up of patients receiving methotrexate for malignant neoplasms is urged; caution is suggested in the prolonged use of methotrexate when treating benign disease.


[Myocardial function in patients with congenital pulmonary valve stenosis]. The studies of metabolism and morphology of the myocardium, and of ECG parameters in 72 patients with valvular pulmonary stenosis conducted with biopsy and autopsy material has demonstrated that while the workload on the right ventricle grows and the severity of the pathological process increases, the oxidative capacity of the myocardium also increase, the indices of its energy supply decrease, the degree of myocytes hypertrophy grows, and the number of mitochondria and the coefficient of their energy efficacy decrease. A correlation was established between the degree of these changes and the severity of electrocardiographic signs of hypertrophy and disorders in the repolarization process in the right ventricle.


[Study of the mechanisms of adrenaline effect on the coupling of respiration with phosphorylation and K+ content in the cardiac mitochondria]. The binding of Ca2+ with fatty acids during the isolation of mitochondria from a conine heart was shown not to prevent the dissociation of respiration and physphorylation produced by adrenalin. The proteinkinase activated by cyclic AMP participates in the dissociation of respiration and phosphorylation in the mitochondria when adrenalin is administered in vivo. The reduced K+ content in the mitochondria under the effect of adrenalin is attributed to a reduction of the membrane potential of the mitochondria, the passive permeability of the mitochondria for K+ ions remaining unchanged.


[Hydroxyproline content in blood and urine in ischemic heart disease]. The content of protein-bound oxyproline in blood and the total oxyproline content in urine were studied in 70 healthy individuals between the ages of 20 and 60 and in 124 patients with chronic ischemic heart disease whose ages ranged from 32 to 74. No age dependence of the protein-bound oxyproline level in blood plasma was revealed in healthy individuals. The concentration of oxyproline excreted in the urine was higher in individuals between the ages of 20 and 30 than among older individuals. In patients with chronic ischemic heart disease the level of protein-bound oxyproline in blood plasma  was normal, excretion of oxyproline with the urine was increased. In patients with myocardial infarction a period of increased content of oxyproline in blood and urine was observed, the duration of which was determined by the extent and depth of infarction and the clinical course of the disease.


[The diagnostic value of corneal arcus as symptom of hyperlipoproteinemia (author's transl)]. In the normal population a physiologic arcus senilis develops in the 6th and the following decades. Hyperlipoproteinemia (type II a  and type II b) however may be the cause of an earlier onset of a corneal arcus. 63 patients with hyperlipoproteinemia type II a and type II b were examined with slit-lamp. In more than 75% of these patients corneal arcus were found already in the 5th decade. Normal population of the same age in contrast shows an arcus corneae only in 25%. Prospective examinations of the Western Collaborative Group found out, that men, 39-49 years old, had with corneal arcus a significantly higher incidence of coronary heart disease, compared with those without arcus. The average annual incidence of symptomatic myocardial infarction was 6.8 per 1000 with arcus as compared to 3.2 per 1000 without. In the 6th decade however there was no more any difference between subjects with arcus and those without arcus. These results, together with the findings of our study, show that one ought to examine the blood-lipids, if by chance of the first refraction a fully developed arcus corneae is found.


[Perimetry in chiasmal syndrome -Liminal and surpraliminal stimuli (author's transl)]. The value of static and kinetic perimetry with white and colored stimuli in chiasmal syndrome seems to be equal. The presentation of evenly moving supraliminal targets allows for an evaluation of sensory disturbance. This procedure is even more subjective, variable and difficult to perform accurately. It may reveal, however, more discrete sensory pathology than conventional perimetry can do. Red is often better than other colors, spectral colors are better than pigment colors. The sector-shaped pattern of disturbance of sensation might relfect the arrangement of nerve-fibers in the optic chiasm.


[The use of photon absorptiometry in renal osteodystrophy (author's transl)]. In 58 patients with end-stage renal failure before commencement of regular hemodialyses treatment (RTD) and 58 patients under RTD bone mineral content (BMC) was determined by the use of the photon absorptiometry. Further the effect of a treatment with bitamin D3 and 5,6-trans-25-OH-vitamin D3 on BMC was studied. There existed a negative correlation between the duration of chronic renal failure or of RDT as well as of the serum parathyroid hormone level to BMC. No correlation was found between BMC and serum calcium level. During a 14 months-course treatment with vitamin D3 or 5,6-trans-25-OH-vitamin D3 BMC increased. The method of photon absorptiometry presented itself as an easy and well reproducible technique for routine examinations.


Evaluation of the classical methods for the diagnosis of type III hyperlipoproteinemia. Familial type III hyperlipoproteinemia is characterized by the presence of elevated plasma levels of very low density lopoproteins (VLDL) which contain an increased amount of cholesterol and by the presence of a significant amount of lipoproteins with an intermediate density between that of VLDL and low density lipoproteins (LDL); the intermediate density lopoproteins, designated IDL or Lp III, have a slower electrophoretic migration rate than VLDL, and are found in the ultracentrifugal top fraction as a contaminant. Classically, the diagnosis of type III is based on the demonstration of beta-migrating lipoproteins in the ultracentrifugal top fraction (density less than 1.006), thus "floating beta-lipoprotein". More recently, it has been proposed that an elevated VLDL-cholesterol to triglyceride ratio is diagnostic of the disorder. In the present report, we have compared the two methods for their diagnostic value and have concluded that the chemical index definition is the more reliable method for the diagnosis of type III hyperlipoproteinemia.


Hypercholesterolemia and triglyceride secretion rates in monkeys fed different dietary fats. The influence of hypercholesterolemia on the triglyceride secretion rate was studied in both squirrel and cebus monkeys fed coconut oil, corn oil, or safflower oil. The triglyceride secretion rate (TGSR) was determined in vivo following the administration of Triton WR1339, which blocks the clearance of very low density lipoprotein (VLDL). Thus, the increase observed in circulating triglyceride after Triton administration presumably reflecte hepatic triglyceride (VLDL) secretion in the fasted state. The VLDL-TGSR was lowest in hypercholesterolemic monkeys and highest in those fed unsaturated fat diets and having a low serum cholesterol. In all instances, TGSR was inversely correlated with the plasma cholesterol concentration. While a definitive explanation for these observations must await further investigation, the possibility that circulating low density lipoprotein (LDL) acts to feed back on VLDL secretion is discussed. The decreased TGSR associated with the diet-induced cholesterolemia also implies clearance of VLDL is impaired under these conditions.


Autoxidation of fatty acid monolayers adsorbed on silica gel: I. Nature of adsorption sites. An unsaturated fatty acid monolayer deposited on a silica gel surface has been chosen as a model for studying non-enzymatic autoxidation of membrane lipids. Studies to determine the suitability of this system as a model for biomembranes were conducted to define the nature of the monolayers, particularly with respect to the factors determining the concentration of the fatty acid molecule on unit area of the surface. The results from adsorption isotherm, high temperature dehydroxylation, and infrared spectra studies show that adsorption of a monomolecular layer of fatty acids occurs and that the number of molecules absorbed corresponds to the number of isolated, non-hydrogen-bonded silanol groups. It is presumed the binding is by hydrogen bonding of the carboxyl groups to silanol groups. The packing density of the fatty acid molecules is 1.25 molecule/100 A2 which is similar to the density of the isolated silanol sites on the surface.


Autoxidation of fatty acid monolayers adsorbed on silica gel: II. Rates and products. Unsaturated fatty acid monolayers on silica gel have been autoxidized, and the rate of fatty acid disappearance and products obtained from those membrane-like assemblies have been studied. Fatty acid monolayers consisting of pure linoleic acid, linolelaidic acid, and oleic acid were autoxidized at 60 C. The rates of autoxidation of linoleic acid and linolelaidic acid monolayers followed by the disappearance of substrates are considerably faster than that in bulk phase, and the rates conform to apparent first order kinetics. Autoxidation of linoleic and linolelaidic acid monolayers, unlike bulk phase, produced only a small amount of diene conjugation, and the major products formed were identified as 9,10-epoxy and 12,13-epoxyoctadecenoic acid in roughly equal quantities. The epoxidation is stereospecific, with cis and trans olefins giving cis and trans epoxides, respectively. Oleic acid was autoxidized to only a small extent during 27 hr and produced no detectable amount of epoxide.


Effect of polymyxin B on outer membranes of Serratia marcescens: morphological alterations of the outer membranes and their lipopolysaccharide components. The in vitro or the in vivo treatment of outer membranes and their lipopolysaccharide (LPS) components from Serratia marcescens with the antibiotic polymyxin B appeared to alter their normal morphology in a sequential manner. The normal spherodial morphology was destablized into a flattened structure after the in vitro treatment of either the resistant strain 08 or the sensitive strain Bizio. The more severe in vivo treatment of the outer membranes from the resistant strain converted the flattened forms further into spheres with undefined periphery and diminished sizes. On the other hand, the same treatment of the outer membranes from the sensitive strain resulted in numerous incomplete spheres and short rods, which were similar to the various morphological forms of the LPS components after polymyxin B treatment. The difference in the morphological changes of the outer membranes and their LPS components of the resistant and sensitive strains after polymyxin B treatment may be explained by the variation of the susceptiblity of the membrane components to the degradative effects of the antibiotic.


Leukemia following Reye's syndrome treated with exchange transfusions: search for type-C viral infection. A 5-year-old child developed acute lymphoblastic leukemia during convalescence from an episode of Reye's syndrome that was treated with multiple exchange transfusions. Routine laboratory, histology, and viral serology were unable to establish a common etiology for the 2 illnesses. Cultural and immunologic methods to search for evidence of infection with type-C viruses or viral genes in lymphoblasts from the buffy coat and bone marrow failed to reveal these agents. Although no common infectious etiology was defined for the close temporal occurrence of 2 rare diseases, the possibility of an iatrogenically induced malignancy was considered.


Ertosterol biosynthesis in Saccharomyces cerevisiae: mutants deficient in the early steps of the pathway. Thermosensitive mutants, auxotrophic for ergosterol synthesis, have been isolated, analyzed genetically and their enzymatic deficiencies investigated. These mutants were classified into seven unlinked complementation groups. These groupes lack the following enzymatic activities: squalene epoxidase (erg 1), 2,3-oxidosqualene-lanosterol cyclase (erg 7), phosphomevalonic kinase (erg 8), mevalonic kinase (erg12) and squalene synthetase (erg 9, erg 10, erg 11).


On the structure of the deo operon of Escherichia coli. A characterization of a specialized transducing lambda phage for the deo operon (lambdaddeo), and some composite colE1-deo plasmids is given in this paper. This includes localization of the RSmaI, RHind/III, RBamI, and REcoRI sensitive sites. The deo genes have been localized by construction of composite colE1-deo plasmids. Using the DNA fragments, obtained by digestion with REcoRI and RHindIII, respectively, as templates in an in vitro protein synthesizing system, it has been possible to give the direction of transcription and the exact location of the deo genes, relative to the endonuclease sites. Furthermore, the cytO,P and deoO,P regions have been mapped relative to the structural genes. Supercoiled co1E1-deo DNA has been used as template in the in vitro system; this DNA gives essentially the same results as the endonuclease-fragmented DNA. The use of the different types of templates is discussed.


Uridine-cytidine kinase. III. Competition between uridine and cytidine for a single enzyme. The combined phosphorylation of uridine and cytidine by a partially purified preparation of uridine-cytidine kinase has been studied with dual-substrate kinetics. The kinetic patterns obtained are consistent with the theoretical analysis for two competing, alternate substrates interacting with a single enzyme. Thus, despite feedback regulation of the kinase by both UTP and CTP, the results allow a clear conclusion that both nucleosides are phosphorylated by the same enzyme, and probably at a single site, rather than by two closely related isozymes, each specific for one pyrimidine.


[Classification and supervision of hyperlipemias (author's transl)]. Different methods for determination and classification systems of serum lipoproteins were used and compared in 300 patients with hyperlipidemia. The procedures and methods were examined to find out which would provide a useful classification for diagnosis and therapy. Lipid electrophoresis on agarose gel gave no reliably reproducible typing. With acetate and polyacetate foils, Type llb was not recognizable in the majority of cases. Neither the determination of beta-cholesterol after polyanion precipitation nor quantitative lipid electrophoresis calculating the values as lipoprotein lipid gave type determinations which completely agreed with the ultracentrifuge, but the best coincidence with the ultracentrifuge was obtained with these two methods. The second method is chiefly suitable for supervision.


Glycosphingolipid hydrolases: properties and molecular genetics. This is a review of the properties and molecular genetics of six lysosomal hydrolases: beta-galactosidase, hexosaminidases A and B, alpha-galactosidase, beta-glucosidase and alpha-fucosidase. Each enzyme is discussed with regards to isoenzymes and substrate specificity, subunit structure, genetic relationship of isoenzymes and genetic variants. The molecular genetics of human diseases caused by deficiencies of each enzyme are discussed.


[Operative possibilities for tumors in single kidneys with particular reference to in situ perfusion (author's transl)]. Surgical removal of malignant tumors from a single kidney is definitely a problem which can be solved by modern techniques of urological surgery. The different operation techniques in 5 of the patients operated on in the Urologische Universitätsklinik Giessen are presented and discussed. In the individual case, besides extracorporeal tumor surgery, the excision of the tumor especially using in situ perfusion with the aid of the floating catheter can facilitate the surgical intervention considerably, and in particular reduce the danger of a postoperative renal failure.


Selective deficiency of hepatic triglyceride lipase in uremic patients. To investigate the pathogenesis of hypertriglyceridemia in patients with renal disease we measured plasma lipoprotein composition as well as hepatic triglyceride lipase and lipoprotein lipase in post-heparin plasma. Three groups with renal disease were studied: conservatively treated chronic uremia; patients undergoing maintenance hemodialysis; and renal-allograft recipients. A selective decrease of hepatic triglyceride lipase with normal lipoprotein lipase was found in conservatively treated uremia and in patients undergoing hemodialysis. Elevated levels of very-low-density lipoproteins and increased triglycerides in low-density lipoproteins occurred in these patients. In contrast, hepatic triglyceride lipase and lipoprotein lipase were both normal in patients after renal transplantation who had Type II hyperlipoproteinemia as a common lipoprotein pattern with increased low-density-lipoprotein cholesterol and decreased high-density-lipoprotein cholesterol concentrations. The accumulation of a triglyceride-rich low-density lipoprotein in the majority of patients with renal disease may be the consequence of low hepatic triglyceride lipase.


Malakoplakia: evidence for monocyte lysosomal abnormality correctable by cholinergic agonist in vitro and in vivo. We studied monocyte function in a case of malakoplakia in an attempt to characterize the immune defect in this condition. Our patient's intracellular cyclic-GMP levels were abnormally low (mean +/- S.D. of 0.17 +/- 0.05 pmol per 10(7) malakoplakia cells, versus 0.79 +/- 0.12 in normals) p less than 0.001). After phagocytosis, his monocytes failed to release beta-glucuronidase. In the bactericidal assay, incubation of the patient's monocytes with Escherichia coli allowed growth of 542 +/- 46 colonies, normal monocytes allowed 95 +/- 22 (p less than 0.001). The percentage of monocytes with large lysosomal granules was 23 +/- 4 in the patient and 4 +/- 2 in normal controls. After in vitro incubation of the patient's cells or in vivo treatment with bethanechol chloride, the cyclic-GMP levels, bactericidal ability and lysosomal granules of the cells returned to normal levels. Low levels of cyclic-GMP could impair lysosomal function and bacterial killing in this condition. Cholinergic agonists correct the in vitro abnormalities and are beneficial in vivo.


Synchronised transmembrane insertion and glycosylation of a nascent membrane protein. Studies of the synthesis and incorporation of the vesicular stomatitis virus glycoprotein into membranes in a synchronised cell-free system demonstrate a tight coupling between polypeptide synthesis and membrane insertion, as a result of which the nascent chain crosses the membrane. The studies reveal a surprisingly precise sequence by which the nascent chain of this membrane glycoprotein is glycosylated in two steps. These findings have important implications for the mechanisms of membrane assembly.


[Molecular biology of tumor viruses].

[Molecular biology of tumor viruses]. All classes of vertebrates harbor tumor viruses that are capable of inducing either tumors or leukemias. After infection, their genomes become integral parts of the host cell's genetic material (DNA). Many biological functions such as the capacity to code for the synthesis of new proteins and, in particular, the oncogenic property (oncogen) have already been assigned to specific regions (on physical maps) of their DNA.


Effects of type A botulinum toxin on the cholinergic transmission at spinal Renshaw cells and on the inhibitory action at Ia inhibitory interneurones. 1. The central action of botulinum toxin A (BTA) on the cholinergic transmission at Renshaw cells (RCs) and on the RC-induced inhibition of Ia inhibitory interneurones (IaINs) was studied in anaesthetized cats. BTA was administered by application directly into the spinal cord, injection into a ventral root (L7) and/or injection into the triceps surae (GS) muscle. 2. A direct application of BTA into the spinal cord led to a decrease of the early and the late response of RCs. 3. When the neurotoxin was injected into the GS muscle, the RC activity remained unaffected during the test period (33-46 h after application). 4. No effect appeared up to 10 h after an injection into the ventral root L7. 5. The RC-induced inhibition on IaINs, when tested in animals with local botulismus, remained intact during the test period. 6. From the present results it is suggested that on the spinal level the central action of botulinum toxin predominantly passes on the motoneurones.


[Electrotonic and chemical EPSPs in lamprey motor neurons following stimulation of the descending tract and posterior root afferents]. In experiments carried out on the isolated spinal cord of the lamprey (Lampetra fluviatilis) post-synaptic responses produced in spinal motoneurons by stimulation of the descending tract and dorsal roots were investigated by means of the intracellular recording technique. It is found that, in addition to giant reticulospinal (Müller's) axons, many other descending fibres as well as dorsal root afferents establish synaptic linkage with both chemical and electrical mode of transmission as revealed by their sensitivity to calcium-deficient and magnesium-rich perfusing solutions. Complex electrotonic EPSPs could have a very fast time course characteristic for the elementary responses but could also produce slow depolarization of the post-synaptic membrane, thus suggesting an integrative function of electrical synapses.


Enzymes relevant to cancer chemotherapy II. Heterogeneity of rat kidney uridine kinase following chromatography on Sepharose 6B coupled to N4-(5-amino-Pentyl)-cytidine. Using Sepharose 6B coupled to N4-(5-aminopentyl)-cytidine at least three different peaks of rat kidney uridine kinase activity have been found. Prior to affinity chromatography the enzyme was partially purified by ammonium sulfate precipitation. The elution was carried out with 0.15 M NaCl or with 0.012 M ATP. The importance of different forms of uridine kinase in eukaryotic cells for cancer chemotherapy with uridine and cytidine analogues is discussed.


Mitochondrial ATPase of Zajdela hepatoma. II. Mitochondria of Zajdela hepatoma contain less adenosine triphosphatase than mitochondria of rat liver. The specific activity and the content of ATPase in mitochondria of rat liver and Zajdela hepatoma were compared. The specific activity of ATPase in sonicated mitochondria and in mitochondrial membrane fraction of rat liver was almost two times higher than the specific activity in the corresponding fraction of Zajdela hepatoma. Accordingly, the autovertin binding capacity of rat liver mitochondrial membrane fraction as well as the yield of F1-ATPase from this fraction were about two times higher than those of the mitochondrial membrane fraction of Zajdela hepatoma. The results show that mitochondria of Zajdela Hapatoma possess about half amount of ATPase present in rat liver mitochondria.


Effect of hematin in porphyric neuropathy. Early, intravenous administration of hematin in a patient with acute intermittent porphyria and severe quadriparesis may have produced partial but remarkable improvement of neuropathy, and resulted in simultaneous decline of porphyrin precursors in the blood. Intermittent, biweekly hematin infusions given 1 month after the onset of the porphyric relapse had no effect on recovery of the residual neuropathy. We believe hematin may be effective in the treatment of porphyric neuropathy, if administered before irreversible neuronal damage has occured.


Adrenomyeloneuropathy: a probable variant of adrenoleukodystrophy. I. Clinical and endocrinologic aspects. We have studied four unrelated males with a heritable disorder that we term adrenomyeloneuropathy; limited clinical information is available on a fifth case. All had adrenal insufficiency beginning in childhood and developed progressive spastic paraparesis in the third decade. Hypogonadism of variable severity was present in all four cases appropriately examined. Neurologic features included peripheral neuropathy, impotence, and sphincter disturbances. Late manifestations were cerebellar dysfunction in one case and dementia and hemiparesis in another. A family history of adrenal disease or spastic paraparesis was present in two cases and absent in one; in the other two, no family history was available. Although males are predominantly affected, the mode of inheritance is uncertain. Adrenomyeloneuropathy probably represents a clinically and genetically distinct variant of childhood adrenoleukodystrophy.


Adrenomyeloneuropathy: a probable variant of adrenoleukodystrophy. II. General pathologic, neuropathologic, and biochemical aspects. Histopathologic features were studied in postmortem examination of two men with adrenomyeloneuropathy, and biochemical studies were performed on one of these individuals. The histopathologic picture of one case included dying-back features in the nervous system and lamellar cytoplasmic inclusions in the brain, adrenal gland, and testis similar to those in adrenoleukodystrophy. Biochemical studies of the cerebral white matter of this individual revealed increased amounts of long-chain saturated fatty acids in cholesterol esters, an abnormality characteristic of adrenoleukodystrophy. Despite differences in clinical presentation and neuropathology, adrenomyeloneuropathy probably represents a distinct variant of adrenoleukodystrophy.


Cytomegalovirus complement fixation antibody in Guillain-Barré syndrome. Cytomegalovirus, measles, and adenovirus antibodies were measured in the sera of 92 Guillain-Barré patients and 120 controls. Thirty patients (33 percent) had markedly elevated levels of complement-fixing antibody to cytomegalovirus and in 21, a fourfold or more alteration in titer was demonstrated. Diagnostic falls in titer were seen in most instances and no significant elevation to the other viral agents was found. The serologic findings reported here suggest that cytomegalovirus may be a common agent involved in the pathogenesis of the Guillain-Barré syndrome.


[Radiological picture Turner's syndrome]. A review of the relevant literature is followed by presentation of the radiological picture, particularly that of the skeleton, in 8 patients with the clinical and endocrinological features and the chromosome pattern of Turner's syndrome. An indication is given of the weight to be attached to the main and lesser radiological signs, especially in cases where the absence of somatic features and detectable hypogonadism impede early diagnosis.


Unexpected disseminated herpes simplex infection in a newborn. Disseminated herpes simplex virus type 2 was unexpectedly discovered in a baby delivered by primary cesarean section at 36 weeks' gestation prior to rupture of the membranes. The mother was free of orogenital lesions at each prenatal examination. Examination of the placenta revealed chorioamnionitis implicating an ascending infection from the mother's infected genital tract.


Clostridial myonecrosis of the postpartum uterus with radiologic diagnosis. A case of clostridial uterine infection following prolonged labor and vaginal delivery is presented. Uterine myonecrosis with gas formation was diagnosed by radiologic studies. Antimicrobial therapy with megadosage penicillin was initiated and total abdominal hysterectomy performed. Three episodes of cardiac arrest occurred during surgery. Resuscitation was carried out with recovery.


Adenoid cystic carcinoma of the vulva. Malignant cylindroma. A case of adenoid cystic carcinoma (cylindroma) of the vulva is presented, reviewing the clinical course and response to therapy over a 5-year period. Only 9 previous cases had been reported by 1963. Several sources of cell origin had been previously suggested. Findings and histology in this case support a relation to a similar tumor occurring in other sites of the upper respiratory tract and salivary gland.


[Chloroquine neuromyopathy. One case in prophylactic maleriatherapy (author's transl)]. A case of neuromyopathy due to chronic chloroquine intoxication is reported. The neuromyopathy developped 9 months after malaria suppression therapy with chloroquine was started. The clinical picture was that of a peripherical neuropathy in the lower limbs and of a generalized myasthenic syndrome. Muscular biopsy showed typical pictures of "vacuolar myopathy". The metabolism of the drug was normal. The patient improved soon after chloroquine was discontinued.


Compact form of SV40 viral minichromosome is resistant to nuclease: possible implications for chromatin structure. We report two new findings bearing on the "supranucleo-somal" level of the structure of the Simian Virus 40 minichromosome. I) Isolated SV40 minichromosome which contains all five histones including HI/I/ exists in solution under approximately physiological ionic conditions as a compact roughly spherical particle approximately 300 A in diameter which is capable of fitting within the virus capsid. In spite of such a compact conformation of the minichromosome individual nucleosomes can be readily visualized within the particle. Compact state of SV40 minichromosome depends on both the presence of histone HI and maintenance of approximately physiological ionic strength of solution (micron approximately 0.15). Removal of HI results in a conversion of the compact minichromosomes into an extended (circular beaded) structure. 2) The compact form of the SV40 minichromosome in contract to its circular beaded form is virtually completely resistant to staphylococcal nuclease, strongly suggesting that in particular nuclease-sensitive parts of the internucleosomal DNA regions are not exposed on the outside of the compact SV40 minichromosome. On the other hand, DNase I which is known to attack both inter-and intranucleosomal DNA in the chronatin /2,3/ readily digests the compact form of the SV40 minichromosome. Possible models of the compact minichromosome and implications for higher order structures of the cellular chromatin are discussed.


Histone synthesis during infection of monkey kidney cells with Simian Virus 40. The synthesis of histones during lytic infection of BSC-1 (African Green Monkey kidney) cells with SV40 has been investigated. The synthesis of all five classes of histones was stimulated, and all classes appeared to be stimulated to the same extent. The increase in rate of histone synthesis in response to SV40 infection was detectable several hours before SV40 DNA synthesis was measureable, and the rate of histone synthesis decreased at a time when SV40 DNA synthesis was occuring at a maximal or relatively high rate. In addition, the changes in rates of histone synthesis did not correlate well with the rates of host DNA synthesis during infection. Thus it appears that DNA synthesis and histone synthesis may not be strictly coupled in SV40 infected cells.


A rapid method for preparation of calf spleen exonuclease. Chromatography on Concanavalin A sepharose is a simple procedure for partial purification of spleen phosphodiesterase. This procedure removes much, but not all, of the ribonuclease activity found as contaminant in most spleen phosphdisterase preparations.


Normal growth and development: current concepts. Human growth and development are a continuum starting at conception. Prenatal growth can only be estimated at present but is an important consideration in an evaluation of a child's developmental status. Postnatal growth can be assessed for size attained (distance curve) or change over time (velocity curve). Velocity curves truly reflect the pattern of growth in stature.