Effects of uridine and inosine on glucose metabolism in skeletal muscle and activated lipolysis in adipose tissue. In a first series of experiments, the effects of uridine and inosine on glucose metabolism in rat diaphragm muscle incubated in Krebs-bicarbonate buffer were studied. Uridine in concentrations of 10(-4) to 10(-6) M stimulated the uptake of glucose and  increased the content of glycogen, but had no effect on the production of lactate. When diaphragm muscles were incubated in the buffer without glucose, uridine (10(-4)-10(-6) M) had no effects on the content of glycogen and on the production of lactate. On the other hand, inosine in concentrations of 10(-4) to 10(-6) M stimulated the uptake of glucose and the production of lactate, but had no effect on the content of glycogen in the muscle. In a second series of experiments, uridine (10(-4)-10(-5) M) and inosine (10(-4)-10(-7) M) inhibited the relase of glycerol from isolated rat epididymal adipose tissue in Krebs-bicarbonate buffer. Uridine and inosine in concentrations of 10(-4) M inhibited the epinephrine (10(-5) M)-, the norepinephrine (10(-5) M)- and the theophylline (10(-3) M)-stimulated lipolysis. Dibutyryl 3',5'-adenosine monophosphate-stimulated lipolysis was further activated in the presence of 10(-4) M uridine or inosine. Dose-response curves studies suggested that inosine, but not uridine, has a common receptor site with epinephrine in adipose tissue. These results demonstrated that both nucleosides stimulated the glucose uptake, but only uridine increased the synthesis of glycogen in the muscle. Both nucleosides also inhibited lipolysis in adipose tissue. The mechanism of antilipolytic action of these nucleosides is unknown, but one of the receptor sites for inosine might be adenylate cyclase.


L-glutamate as an excitatory transmitter at the Drosophila larval neuromuscular junction. The possibility that L-glutamate is the excitatory transmitter at the Drosophila larval neuromuscular junction and the ionic basis of its action on the muscle membrane are examined. 2. Iontophoretically applied L-glutamate causes muscle depolarization (L-glutamate potential) if and only if the L-glutamate pipette is within a few mum of the nerve ending. D-glutamate, substance P, ACh and GABA are ineffective. 3. Bath-applied L-glutamate produces similar changes in the time course and amplitude of miniature excitatory junctional potential (m.e.j.p.), excitatory junctional potential (e.j.p.) and the L-glutamate potential. 4. Neuromuscular transmission and excitation-contraction coupling are operative in a haemolymph-like solution containing 1 mM L-glutamate. 5. The reversal potentials of the e.j.p. and the L-glutamate potential are identical to each other, changing similarly with changes in the ionic compositions of the external medium (twelve solutions). 6. The ionic dependence of the reversal potentials is predicted from an extended constant-field equation using a ratio of sodium:potassium permeabilities of PNa/PK=1-3, and a ratio of magnesium:potassium permeabilities of PMg/PK=4-7. 7. It is concluded that L-glutamate is, or is an agonist of, the excitatory transmitter at certain Drosophila larval neuromuscular junctions.


The release of endogenous amino acids from the rat visual cortex. The release of endogenous taurine, GABA, glycine, aspartate, glutamate, glutamine and alanine from the rat visual cortex was measured using a cortical cup technique. The electrocorticogram (ECoG) was monitored throughout most experiments. 2. Spreading depression, evoked by the dropwise placement of 10% KCl solution on to the brain outside the cup was associated with a significant increase in the release of GABA and glutamine but a marked fall in that of glutamate. The evoked release of GABA and glutamate but not of glutamine was Ca2+ dependent. 3. A solution containing 50 mM-K+ placed within the cup elicited a significant increase in the release of taurine and GABA, whereas 100 mM-K+ additionally released aspartate and glutamate. The K+-evoked release of these amino acids with the exceptions of taurine and glutamine was Ca2+-dependent. 4. Three series of experiments were carried out in which the preparations were stimulated electrically. Bipolar stimulation (100 Hz, 1 msec pulse width, 2-5 mA for 5 min) with the electrode within the cup was followed by significant increases in taurine, GABA and glutamate release; using a 5 mA current, there was an additional release of aspartate and alanine. Only the evoked release of GABA and glutamate was Ca2+ dependent. 5. In the second and third series of experiments, the electrode was sited adjacent to the cup or on the contralateral cortex respectively. Following stimulation (100 Hz, 1 msec pulse width, 2-5 mA for 5 min) there was a significant increase in taurine and GABA release and a significant fall in the release of aspartate and glutamate. With the exception of taurine, these changes in release were Ca2+ dependent. Reducing the stimulus current to 1-5 mA or the period of stimulation to 2-5 min initiated similar but statistically insignificant changes in release. A range (10-100 Hz) of stimulation frequencies was examined: the evoked release of GABA was linearly related to frequency whereas that of taurine was frequency-independent. The fall in aspartate and glutamate release was maximal at a frequency of about 50 Hz. 6. The results are discussed in relation to (a) the possible sites of release of the amino acids and (b) the proposed neurotransmitter roles of the physiologically active amino acids.


Phosphorylation reactions in islets of Langerhans. The possible significance of phosphorylation reaction in islets of Langerhans in relation to the secretion of insulin is discussed. The secretagogues, glucose and its metabolites, cAMP and Ca++ and their influences on protein-kinase activity are given particular attention. The data obtained by the authors, as well as by other groups, are in agreement that cAMP is a potent stimulator of protein kinase activity. Glucose and its metabolites influenced protein kinase activity in one instance. Ca++ in supraphysiological amounts inhibited protein phosphorylation. The links between phosphorylation reactions and insulin secretion are, at the present time, conjectural.


Studies on the function of pancreatic islet cell membranes. Pancreatic islets rich in beta-cells were isolated from non-inbred ob/ob-mice and used for studying various aspects of the function of the plasma membrane. A review is given of the authors' work along the following lines: the role of transmembrane transport or membrane binding in the recognition of insulin-releasing sugars, amino acids, sulfonylureas, and sulphydryl-blocking agents; the role of cyclic 3',5'-AMP and cations in the coupling of stimulus recognition to insulin discharge; alloxan beta-cytotoxicity in vitro and its prevention by sugars; the isolation of a subcellular fraction enriched by plasma membranes. 1. It is suggested that D-glucose is recognized as an insulin secretagogue by being metabolized in the beta-cells; the teleological purpose of the transmembrane transport system being to allow fluctuations of the extracellular glucose concentration to be rapidly transmitted to the cell interior. Insulin-releasing sulfonyluraes and sulphydryl reagents are thought to act directly on the beta-cell plasma membrane, however. 2. Although cyclic 3',5'-AMP may amplify the expression of a secretory signal induced by D-glucose, studies with cholera toxin suggest that activation of the adenylate cyclase does not per se elicit secretion. The increase of islet cyclic 3',5'-AMP observed in response to several secretagogues, including D-glucose, may be secondary to membrane depolarization. 3. The possible role of an electrodiffusional mechanism in controlling the electrical potential is emphasized; a decrease of K+ permeability, rather than an increase of Na+ permeability, is suggested to be involved in the depolarizing action of D-glucose. Studies with the lanthanum-wash technique indicated that D-glucose causes a net flux of Ca2+ from the outside to the inside of the beta-cells. Although this uptake may relate to the enhancement of insulin secretion, the detailed mechanisms are unclear. 4. Inhibition of the Na+/K+ pump may be one of the earliest events in damage to the beta-cell by alloxan, on the basis of Rb+ studies. Protective effects of glucose against alloxan toxicity appear to be close related. 5. Studies of enzyme markers, the binding of wheat germ agglutinin, and electron microscopy indicate the presence of plasma membranes in a smooth-membrane fraction obtained by fractionating islet homogenates at consecutive sucrose gradients.


Teen-age alcohol abuse.

Teen-age alcohol abuse. In summary, we have seen that there isn't sufficient data available to properly assess this problem, but from what exists we are fairly sure that teen-age alcohol abuse is on the rise. We have also noted that there are many area agencies for alcoholics, but that there is not much coordinated effort, and essentially none for teen-agers. This is probably due to its being a relatively new trend and that many people are apparently refusing to admit that it exists. Also covered were possible causes for this new problem and the various roles of nurses in prevention. These include school nurses, public health nurses, and others in community and state politics, promotion of new educational methods, family teaching both at home and in the hospital, new responsibilities of the school nurse, and research. These are challenging roles, but also very important considering that these teen-agers who are flirting with trouble today will be rearing families and defining social behavior tomorrow.


Some barriers to effective alcoholism research. Barriers to effective alcoholism research include the search for unitary etiological factors, the difficulties in assembling homogeneous samples of subjects for study, the lack of widespread multidisciplinary efforts, and uncertainty about the goals of treatment for alcoholism. Innovative research approaches are needed for alcoholism prevention in order to preclude the proliferation of patterns of alcohol abuse.


[Post-operative radiotherapy of hemisphere gliomas in the adult. A statistical study of 134 cases (author's transl)]. A series of 134 adults patient with hemisphere gliomas who received post-operative radiation after surgical excision, either by conventional radiotherapy or high energy radiation, between 1966 and 1972 in the Léon-Bérard Centre. Overall survival for grade I and II astrocytomas at two years was 15/29, or 51%. Overall survival for glioblastomas was 30/91, i.e. 33% at one year, and 3/91 or 3.3% at two years. High energy radiation gave slightly superior results to conventional radiotherapy. The best results were obtained with concentrated irradiation when two series of irradiation were possible : 8/17 at one year (47%). Analysis of the literature indicates that radiotherapy, whilst remaining controversial for grades I and II astrocytomas, is recognised to be useful by all in the case of glioblastomas. The authors favour the post-operative irradiation 6,00 to 6,500 Rads, localised) for grade I and II astrocytomas with pejorative characteristics and routin- post-operative irradiation for glioblastomas over a wide field taking in almost the entire brain. This may be spread (6,000 Rads) or in a "split course" (two series of 1,800 each). The latter technique has the advantage of a simple protocol, better adapted to lesions in which the final prognosis remains very poor. Finally, the indications for radiotherapy in the case of supra-tentorial ependymomas (3 cases) and oligodendrogliomas (6 cases) are presented, together with an analysis of the complications of radiotherapy for cerebral tumours.


[A quantitative and quanlitative examination of bone metastases by whole body scanning with 99mTc-pyrophosphate (author's transl)]. The purpose of the present work is to "quantify" bone scanning, in order to calculate indices of localized uptake that will make it possible to differentiate between normal and pathological uptake, and approach an etiologicial diagnosis. The authors use an Elscint dual head whole body scanner and an offline video display calculator. The patient is given 8 mc technetium 99m pyrophosphate 4 hours prior to scanning. The whole body count is expressed in thousands of counts multipled by two so that the knee to whole body ratio is close to one, to facilitate comparison of the indices. Results show that the presence of metastases is demonstrated more clearly by using a ratio bone segment to whole body. Quantitative scanning with 99mTc-pyrophosphate makes it possible to: 1) detect bone metastases easily, 2) follow the metastases during treatment, 3) avoid the gross errors that can be made in nomquantitative scanning.


Effect of prolactin and growth hormone on prolactin and LH receptors in the dwarf mouse. Dwarf mice (DW/J;dw/dw) which exhibit a deficiency of prolactin and GH secretion were treated for 8 days with ovine prolactin and/or human GH (10 or 20 mug/day) and the effect on hepatic and testicular prolactin receptors was investigated. In both sexes there was a significant increase in body weight after all hormone treatments, but an increment in testicular weight was observed only after prolactin administration. Prolactin treatment increased the specific binding % of prolactin in liver membranes in females but not males, and in testicular homogenates (together with an increase in LH receptors). The results suggest that lack of prolactin but not of GH retards sexual development in these mice. Treatment with prolactin partly counteracts this deficiency, and the effect may be mediated by the induction of hepatic and testicular prolactin and LH receptors.


Dysfunctional uterine bleeding.

Dysfunctional uterine bleeding. The etiology and pathogenesis of dysfunctional uterine bleeding are discussed. For both age groups, adolescents and perimenopausal women, gonadotropin-steroid patterns as well as endometrial changes are presented. Methods of evaluation and therapy are described, including curettage and judicious of estrogen-progestogen combinations. Finally, long-term follow-ups of these patients are presented, with plans for reevaluation as well as palliative treatment based on the bleeding patterns and gonadotropin levels.


Structure of the peptide antibiotic polypeptin. Polypeptin, a basic peptide antibiotic isolated from Bacillus circulans, was separated into two components by countercurrent distribution. The two components, polypeptin A and polypeptin B, had identical amino acid compositions but varied in the structure of the hydroxy acid constituent attached to the alpha-amino group of the peptide chain. Polypeptin A contained 3-hydrosy-4-methylhexanoic acid and polypeptin B contained 3-hydrosy-5-methylhexanoic acid. T-HE STEROCHEMISTRY OF THESE HYDROXY ACIDS WAS NOT DETERMINED. Studies involving partial acid hydrolysis and chemical synthesis are consistent with the lactone sturcture for polypeptin A. Polypeptin B differs only in the position of the methyl group in the hydroxyacyl moiety.


Adrenocorticotropin. 49.1 Synthesis and biological activity of [2-delta-aminovaleric acid, 5-arginine-a1adrenocorticotropin-(2-19). An adrenocorticotropin analogue, [2-delta-aminovaleric acid, 5-arginine]adrenocorticotropin-(2-19), has been synthesized by the solid-phase method and its biological activity has been determined. It was found that substitution of arginine for glutamic acid at position 5 of [2-delta-aminovaleric acid]adrenocorticotropin-(2-19) increased the steroidogenic potency in idolated rat adrenal cells and the lipolytic potency in isolated rat fat cells but decreased the lipolytic potency in isolated rabbit fat cells. The synthetic analogue had only 2% of the melanotropic potency of the parent molecule.


Action of insulin and cell calcium: effect of ionophore A23187. We have measured the effects of the carboxylic Ca++ ionophore A23187 on muscle tension, resting potential and 3-O-methylglucose efflux. The ionophore produces an increase in tension that is dependent on external Ca++ concentration since (a) the contracture was blocked by removing external Ca++ and (b) its size was increased by raising outside Ca++. Neither resting potential nor resting and insulin-stimulated sugar efflux were modified by the ionophore. These data imply that the action of insulin is not mediated by increasing cytoplasmic [Ca++]. Additional support for this conclusion was obtained by testing the effects of caffeine on sugar efflux. This agent, which releases Ca++ from the reticulum, did not increase resting sugar efflux and inhibited the insulin-stimulated efflux. Incubation in solutions containing butyrated derivatives of cyclic AMP or cyclic GMP plus theophylline did not modify the effects of insulin on sugar efflux. Evidence suggesting that our experimental conditions increased the cytoplasmic cyclic AMP activity was obtained.


Evaluation of BCG administered by scarification for immunotherapy of metastatic hepatocarcinoma in the guinea pig. In inbred guinea pigs, administration of Mycobacterium bovis strain BCG by scarification at a site distant from an excised skin tumor, but in the regional lymph node drainage, was evaluated for its immunotherapeutic effect on the development of lymph node metastases. Scarification was performed after surgical excision of intradermally transplanted syngeneic (line-10) hepatocarcinoma at a time when microscopic foci of tumor cells were present in regional lymph nodes. Various strains of BCG were evaluated for their immunotherapeutic potential: fresh-frozen Phipps, Pasteur, and Tice; and lyophilized Pasteur, Tice, and Connaught. Scarification commenced 3 days after surgical removal of the tumor and continued once a week for 5 weeks. Only lymph nodes from fresh-frozen Phipps- and Pasteur-scarified animals were significantly smaller than those in the control groups. Differences in lymph node weight correlated histologically with less detectable metastases. This cytostatic effect was short lived; eventually, the metastatic tumor growth was not significantly different from that of control animals. No significant differences were observed in mean survival time: All animals died as a result of metastases 3 months after tumor inoculation. These results demonstrated that limited scarification with BCG of certain strains temporarily inhibits the growth and proliferation of metastases in regional lymph nodes after removal of the primary tumor.


Antibody-induced modulation and shedding of mammary tumor virus antigens on the surfaces of GR ascites leukemia cells as compared with normal antigens. The distribution, antibody-induced redistribution, and shedding of murine mammary tumor virus (MuMTV) antigens and the surfaces of GR mouse ascites leukemia (GRSL) cells were studied by the immunoferritin technique and compared with the same activities of thy 1.2 and H-2.8 antigens. MuMTV antigens were redistributed easily and then largely shed from the cell surface; in contrast, H-2.8 antigen moved easily and probably was partially released from the plasms membrane and Thy 1.2 antigen moved slowly and was some what interiorized. The complement-dependent cytotoxicity test was used to study the possibility of antigenic modulation for these cell-surface antigens from the surface of the GRSL cells could be modulated by preincubation with anti-MuMTV serum, in contrast to H-2.8 and Thy 1.2 antigens. The results obtained with the immunoferritin technique and the cytotoxicity test correlated well and sug-ested that the shedding of MuMTV antigens from the cell surfaces may occur in vivo, providing the tumor a way to escape from the immune defense of the host. Thy 1.2 and H-2.8 antigens were present on the envolope of B and C particles, which suggested that these viruses do not select a Thy 1.2 or H-2.8-negative area of the GRSL cell surface as amaturation site.


Immunofluorescence and histologic studies of virus-induced murine lymphocytic leukemias. Mice were inoculated with either the lymphatic leukemia virus associated with Friend and Rauscher viruses or with the Graffi or BALB/Tennant leukemia viruses. A total of 48 leukemic mice were examined histologically and by immunofluorescence. All four viruses induced a histologically similar disease that particularly involved tha thymus-dependent lymphoid regions. Lymphocytes from the spleens and thymuses of leukemic animals were examined for Thy-1 antigen and immunoglobulins on the cell surface; all the leukemias were composed of T- cells and/or nonreactive cells lacking both the Thy-1 antigen and immunoglobulins. By immunofluorescence, the spleen and thymus from the same leukemic animal frequently showed different cell-surface markers, though no morphologic differences were seen in leukemias involving the different classes of lymphocytes.


Oncornavirus-like particles in baboon type C virus-infected chimpanzee lung cells (SFRE:CL-1). Intracytoplasmic type A particles were observed in a fetal chimpanzee lung culture (SFRE:CL-1) inoculated with type C virus-containing supernatants from a coculture of baboon placenta and SFRE:CL-1 cells. Budding, immature, and mature type C particles were also noted. In thin section, spike-like structures were rarely detected on budding intracytoplasmic type A particles but were occasionally observed on some immature and mature virus particles. Unlike mouse mammary tumor virus or Mason-Pfizer monkey virus, infected SFRE:CL-1 cells contained no eccentric or rodshaped nucleoids.


Chronic Herpesvirus saimiri infection in an owl monkey. An adult owl monkey (Aotus tricirgatus) used for immunologic studies of Herpesvirus saimiri (HVS) developed early, late, membrane, and neutralizing antibodies to HVS approximately 3 weeks after the beginning of the experiment. HVS was isolated by the cocultivation of peripheral blood for over 1 year. No clinical, gross, or histopathologic findings of malignancy were exhibited by the animal. The HVS isolate from the animal was indistinguishable biologically and serologically from the original HVS strain of Meléndez and from an isolate of an experimentally HVS-induced tumor. Inoculation of this isolate into 2 young white-lipped marmosets (Saguinus fuscicollis) produced typical malignant lymphoma and lymphocytic leukemia. Our findings suggested that the virus from the chronically infected animal was oncogenic and that host factors were primarily responsible for determining the disease manifestation of the virus infection. Another owl monkey chronically infected with HVS for over 2 years has remained asymptomatic.


Effect of allelic substitutions at the hairless locus on endogenous ecotropic murine leukemia virus titers and leukemogenesis. Leukemia-prone hairless (HRS/J; hr/hr) mice had significantly higher leukemia virus titers than did the leukemia-resistant nonmutants (hr/ + and +/+). This difference was ascribed to the allelic substitution at a single gene locus; at 6 months of age it averaged 13-fold, immediately preceding the large divergence in leukemia incidence between the mutant and nonmutant mice.


Cell-mediated immunity in mice against papain-solubilized histocompatibility and tumor-specific antigens by a macrophage migration inhibition microassay. The cell-mediated immune status of B10.D2 (H-2d) mice immunized with spleen cells from a congenic strain, B10.A (H-2a), differing at the H-2 locus and of BALB/c mice immunized with a syngeneic simian virus 40 (SV40)-induced sarcoma (mKSA-TU5) was evaluated by an agarose microassay for migration inhibition factor. The inducing antigens in this experiment were papain-solubilized and partially purified chromatographic preparations of spleen cells from A/J mice (H-2a) and a papain-solubilized antigen extract prepared from a tissue culture-adapted cell line (TU-5), derived from the SV40-induced mKSA tumor. The assay used microliters of normal or immune peritoneal exudate cells (PEC) resuspended in a 2-mul droplet of agarose and cultured in the presence or absence of antigen. Specific migration inhibition of PEC from immunized mice was observed with concentrations of solubilized antigen preparations as low as 2.0 mug/ml (3.67 mug/chamber).


Immunostimulation of cytomegalovirus-transformed cells and its inhibition by blocking sera. Spleen cells cytotoxic at high effector cell: target cell ratios specifically enhanced target cell survival at low ratios. Both cytotoxicity and immunostimulation could be specifically blocked by sera from tumor-bearing or virus-sensitized hosts. The results suggested that serum factors participate in the regulation of the cell-mediated immune response.


Characteristics of a human cell line successively transformed by Rous sarcoma virus and simian virus 40. Human enbryo cells were successively transformed by the Schmidt-Ruppin strain of Rous sarcoma virus (SR-RSV) and simian virus 40 (SV40) in vitro, and the double transformant HuE 13 RS was established. From this cell line the two clonal cell lines RSa and RSb were isolated. In both, presence of SV40 T antigens was demonstrated by the fluorescent antibody technique, and the presence of RSV genomes was verified in one RSb clone by focus formation after fusion with chick embryo cells. Growth of these cells was affected by dibutyryl cAMP without marked morphologic changes. Cells were extremely sensitive to the anticellular action of human leukocyte interferon.


Spontaneous tumors and common diseases in two colonies of Syrian hamsters. II. Respiratory tract and digestive system. Spontaneous respiratory tract neoplasms in Syrian hamsters occurred in almost equal frequencies in two colonies: 3% in the Eppley Colony (EC) and 3.6% in the Hannover Colony (HC). Neoplasms were in the nasal cavity, trachea, and lungs, and most were benign; however, 2 adenocarcinomas of the nasal cavity (EC) and 1 adenocarcinoma of the larynx (HC) were found. The incidence of digestive tract tumors showed a more marked difference than that of the respiratory tract: 7% in the EC and 23% in the HC. Digestive tract tumors accounted for 15 and 41% of all tumors in the EC and HC, respectively. All EC digestive tract neoplasms were benign and occurred mostly in males; 19 (83%) were forestomach papillomas and the remaining 4 )17%) were liver hemangioendotheliomas (2) and pancreatic duct adenomas (2). In the HC, almost 50% of the digestive tract neoplasms were malignant and most frequent in females. These tumours include 19(42%) intestinal adenocarcinomas, 18(40%) liver neoplasms (hemangioendotheliomas, cholangiomas, cholangiocarcinomas), 5 (13%) forestomach papillomas, and 2 (4%) gallbladder polyps. The morphology of these neoplasms was reported.


Transformation of tonsil lymphocytes by Epstein-Barr virus. Tonsil lymphocytes obtained from children and adults at tonsillectomy were examined for susceptibility to infection with Epstein-Barr virus (EBV) in vitro. Of 37 specimens, 24 (65%) were transformed by EBV. Rate of transformation was higher in the cells from younger children than in those from older children and adults, but the susceptibility was not directly correlated to the titer of antibody against EBV in donor serum. All 22 transformed tonsil cell lines tested were carrying EBV-associated nuclear antigen, and 8 of them produced cytoplasmic IgA in 15-45% of cells. The results suggest that tonsil lymphocytes are targets of EBV.


Cellular immunity in rats with primary brain tumors: inhibition of macrophage migration by soluble extracts of avian sarcoma virus-induced tumors. Rats bearing primary tumors of the brain induced by avian sarcoma virus (ASV) were studied with the migration-inhibition factor (MIF) assay for the presence of cell-mediated immunity to tumor-associated antigens. Astrocytomas and sarcomas of the brain were induced in 34 neonatal F344 rats by the intracerebral inoculation of Bratislava-77 ASV. At weekly intervals from 4 to 9 weeks after the inoculation with virus, peritoneal exudate cells (PEC) from rats bearing brain tumors were tested an an MIF assay against soluble and KCl-treated extracts of a syngeneic, ASV-induced sarcoma. Incubation of the PEC with a soluble extract of syngeneic liver or with a KCl extract of a syngeneic, chemically induced tumor served as controls. Of 14 rats tested against the soluble tumor extract, 6 (43%) had statistically significant inhibition of migration (P less than or equal to 0.05). Of 23 animals tested against the KCl extract, 16 (70%) had significant inhibition. Immunity to the KCl extract was significant in most rats at each  period. Ten rats were tested against a KCl extract of a hamster ASV-induced tumor; 7 gad significant inhibition of migration. None of 3 tested against a soluble extract of a syngeneic, chemically induced tumor had significant inhibition. Rats bearing ASV-induced brain tumors displayed cell-mediated immunity to tumor-associated antigen or antigens of ASV-induced tumors, which could be solubilized.


Feline oncornavirus-associated cell membrane antigen. VI. Cytotoxic antibody in cats exposed to feline leukemia virus. Feline leukemia virus (FeLV)-infected specific pathogen-free (SPF) cats, normal uninfected SPF cats, and healthy cats from leukemic households were tested for antibody reactive to the feline oncornavirus-associated cell membrane antigen (FOCMA)-containing target cell line FL-74 by microcytotoxicity and indirect membrane immunofluorescence. Of the infected SPF animals, 81% showed concordant reactivity for the two tests. In contrast, only 55% of the healthy cats known to be naturally exposed to FeLV for long periods showed such concordance. FOCMA antibody could not be detected in normal SPF cats by either indirect membrane immunofluorescence or microcytotoxicity. Most cats in the FeLV-infected SPF group developed antibody detectable by both procedures by the fifth week post inoculation. Antibody detectable by membrane immunofluorescence persisted in a high percentage (75-90%) of the animals throughout the observation period of 19 weeks; after 9 weeks, fewer cats had antibody that was also detectable by microcytotoxicity.


Transplantation of human tumors in nude mice. Ninety-one human tumors, including various common carcinomas, low-grade malignant tumors, and benign tumors, were transplanted into athymic nude mice. Tumor take was confirmed histologically for 22 neoplasms at the initial transplantation, and 14 serially transplantable tumors were established, including some hitherto unestablished or unreported, such as lung and hepatic cell carcinomas. Among the 91 tumors were 21, 14, and 13 carcinomas of the lung, stomach, and breast, respectively. Transplantability was highest in lung carcinomas (10/21), followed by gastric carcinomas (2/14) and breast carcinomas (1/13). Morphology of original tumors was retained well in most transplanted tumors, but desmoplastic or scirrhous tumors, such as gastric and breast carcinomas, tended to become medullary with a decrease in amount of tumor stroma. The ability to produce mucin in gastric carcinomas or melanin in malignant melanoma was maintained in serially transplantable tumors. In addition, ectopic production of adrenocorticotropin and beta melanocyte-stimulating hormone continued in a transplanted small cell carcinoma of the lung. Preliminary results were obtained on hormone dependency of the transplantable breast carcinoma and on alpha1-fetoprotein in the transplantable hepatic cell carcinoma.


Synthesis and half-life of a serum factor in normal and tumor-bearing animals. The synthesis and disappearance of a serum factor from the circulating blood of rats bearing Morris hepatomas 7777 or 7800 were described. We provided evidence that the factor was synthesized by the livers of normal rats and continued to be synthesized in tumor-bearing animals. Despite very low levels in the sera of tumor-bearing animals, the factor was synthesized at twice the rate in these animals compared to that in normal rats. The half-life of the total serum protein was 2 days in normal rats and 2.9 days in hepatoma 7777-bearing animals. The half-life of the serum factor, on the other hand, was 23 hours in normal rats but only 9 hours in hepatoma-bearing rats. This rapid disappearance of the factor from the circulating blood, despite an increased synthesis, accounts for the complete absence of the factor in the sera of hepatoma-bearing animals with tumor weights larger than the liver weights.


Formation of larger cell aggregates by transformed cells: an in vitro index of cell transformation. Virus-transformed rat and hamster embryonic cells formed larger cell aggregates than those formed by normal counterpart cells within 1-3 days. This aggregate property correlated with growth in soft agar and tumorigenicity.


Hormone synergism in the in vitro production of the mouse mammary tumor virus. The production of mouse mammary tumor virus (MMTV) in primary cell cultures of BALB/cfC3H mammary tumor cells was measured by radioimmune assay and RNA=dependent DNA polymerase activity. Maximum virus production was dependent on cell density, nutritional milieu, and hormone supplementation. The addition of insulin (u), estradiol-17 beta (E2), progesterone (P), prolactin (PRL), or thyroxine (T3) alone had little or no effect on MMTV production. Hydrocortisone (F) had a primary stimulatory effect. The combination of I, F, and T3 increased MMTV levels. The combinations containing I, F, and E2 had the greatest stimulatory effect. The stimulation of MMTV production was dose dependent. These experiments demonstrate that a variety of hormones act in a synergistic manner to stimulate MMTV production.


Steroid-binding proteins of the mammary gland and their clinical significance in breast cancer. Differentiation and development of the mammary gland are influenced by estrogens, glucocorticoids, and progesterone. Specific binding sites for each of these classes of steroid hormones have been characterized in vivo and in cell-free preparations of lactating mammary gland of the rat. These properties were compared with those of steroid-binding proteins in the R3230AC and dimethylbenz[a]anthracene-induced mammary tumors of the rat and of human breast carcinomas. The binding components that were protein in nature were more concentrated in tissue from lactating than from virgin or pregnant rats. The steroid-binding capacities of mammary tumors were highly variable. Using a dextran-coated charcoal procedure, the rate constants of 3H-ligand association with specific binding sites were temperature dependent. Ligand specificity indicated that [3H]estradoil-17beta(3H-E) binding was inhibited only by estrogens, whereas [3H]triamcinolone acetonide (3 H-TA) was blocked by glucocorticoid/s and progesterone. Likewise, [3H]progesterone (3H-P) binding was inhibited by progestins and certain glucocorticoids. Incubation of minced mammary gland with 3H-steroid in the presence and absence of unlabeled competitor demonstrated specific binding sites in cytoplasm that were translocated to nuclei in a tempreature-dependent fashion. Using a steroid-exchange procedure both DEX and TA inhibited 3H-P binding to exchangeable sites that were loaded previously with unlabeled progesterone. These data suggest that certain glucocorticoid and progesterone binding sites were translocated simultaneously to nuclei. Administration in vivo of either 3H-E or 3H-TA indicated binding in cytoplasm and subsequent translocation to nuclei.


Proteolytic enzyme activity in the granulation tissue of the human burn wound. A procedure for adsorbing enzymes from the human burn wound onto solid sheets of substrate is described. Using this technique, low levels of enzyme activity with chymotrypsin-like specificity can be demonstrated in the wound approximately 2 weeks after injury. This activity disappears at about 5 weeks after the burn. The enzyme activity corresponds with the clinical experience for the time course of natural loss of the burn eschar. A trypsin-like enzyme of very low level activity is present in the wound. No collagenase was detected in the human burn wound. Preliminary evidence shows an additional leucine-specific enzyme in the human burn wound. A more detailed analysis of enzymes in the human burn wound should permit the development of a useful artificial debriding agent. Presently these preparations must be used with caution.


Neoplasms of the temporal bone.

Neoplasms of the temporal bone. A series of 209 patients with chemodectomas and malignant neoplasms of the external auditory canal and middle ear seen during a 20-year period at the Mayo Clinic was reviewed. Ninety-nine patients had chemodectomas, and 110 had malignant lesions; 57 of the 110 patients with malignancies had squamous cell carcinoma. The study revealed that patients with chemodectomas had a normal expected length of survival, whereas patients with malignant lesions (especially those with squamous cell carcinomas and embryonal rhabdomyosarcomas) did poorly.


Clinically relevant physiology of the vestibulo-ocular reflex. This review attempts to explain those aspects of the physiology of the vestibulo-ocular reflex (VOR) which could be of future clinical value. The literature cited has been selected for its didactic worth to readers with limited time, preferring concise reviews to detailed reports wherever possible. Physiological data provide the background for the following possible improvements in clinical diagnosis: 1) In gaze analysis, coordination of the VOR with other motor patterns can be analyzed. 2) Precision of vestibular tests can be improved by selecting stimuli within the range of natural movements.3) Resolution of the caloric test can be imporved when the change of temperature at the semicircular canal mimics endolymph pressure changes during natural movements. 4) A direct test of the three neuron VOR pathways is possible, but not practicable. 5) Integration of the input signal (transformation from head acceleration to eye position information) can be directly tested. 6) Plasticity (the adaptation to visual requirements) of the VOR can be examined. 7) It is possible to quantify vestibular damage and detect the side of lesion in one test analyzing gain and binocular symmetry of the vertical VOR.


Content and expression of integrated viral DNA in hamster cells transformed by nondefective adenovirus type 2- simian virus 40 hybrid viruses. Hamster cells transformed by adenovirus 2 (Ad2) and five nondefective Ad2-simian virus 40 (SV40) hybrid viruses are all of the Ad2-transformed phenotype. All lines accumulate Ad2 RNA and Ad2 T antigen; two hybrid-transformed lines accumulate SV40 RNA, but only one contains detectable amounts of SV40 antigens. We examines selected lines from this group of transformed hamster cells for Ad2 DNA content and viral RNA expression by using hydroxyapatite chromatography and separated strands of labeled viral DNA as probes. All lines studies contain 1 to 2 Ad2 genome equivalents per haploid equivalent of cell DNA. The expression of Ad2 RNA exhibits two distinct patterns. In one pattern, transcription of the heavy strand of Ad2 DNA is less than that of the light strand, whereas in the second pattern of Ad2 RNA expression the extent of heavy-strand transcription is greatly increased. In the two lines containing SV40 RNA, the entire SV40 (-) strand is transcribed; the (+) strand is not expressed in these cells. These patterns of viral RNA expression suggest the possibility that host promoters may play a role in regulating transcription of integrated viral DNA.


Distinguishing cytomegalovirus, mycoplasma, and cellular DNA polymerases. Cytomegalovirus-induced DNA polymerase can be distinguished from infected-cell enzymes by activity in 100 mM (NH4)2SO4. Virus polymerase is stimulated to 145% of control, whereas mock-infected cell polymerase is inhibited to 12% of control without added salt. Mycoplasmas induce a DNA polymerase in cell extracts that is stimulated to 130 to 180% by 25 mM (NH4)2SO4. Mycoplasma DNA polymerase may be mistaken for a virus-induced polymerase when virus stocks are contaminated. Identification of virus, cellular, and mycoplasma DNA polymerases in total cell extracts is described using sedimentation rate and effect of inhibitors on DNA polymerase activities.


RNA synthesis in cells infected with herpes simple virus. XIII. Differences in the methylation patterns of viral RNA during the reproductive cycle. Herpex simplex virus 1 (HSV-1) RNA labeled with with [methyl-3H] methionine at various times during the infectious cycle and purified by hybridization to viral DNA was analyzed for the presence of methylated nucleotides. The data indicate the following. (i) RNA labeled from 0 to 14 h postinfection and accumulating in the cytoplasm contained internal base-methylated nucleotides and terminal oligonucleotides consistent with the structure 7mG(5')ppp-(5')XmpYmpNp. Similar methylated nucleotides and oligonucleotides were also found in viral RNA accumulating in the cytoplasm of cells treated with cycloheximide from the time of infection. Previous studies (M. Kozak and B. Roizman, 1974) have shown that, whereas the RNA accumulating in the 14-h infected cells contains all of the sequences functioning as mRNA throughout infection, the RNA accumulating in the cytoplasm of cycloheximide-treated cells is associated with polyribosomes synthesizing the earliest (alpha) group of polypeptides specified by the virus. (ii) Cytoplasmic viral RNA from cells labeled 11 to 14 h postinfection as well as the total adenylated RNA in the cytoplasm and polyribosomes labeled in the same fashion contained the terminal oligonucleotide but not the internal base-methylated nucleotide.


Genetic evidence for a product of the Fv-1 locus that transfers resistance to mouse leukemia viruses. Extracts of mouse cells have been shown to transfer to N- or B-trophic host range types of mouse leukemia viruses. The genetic specificity of the inhibition was tested in two ways: (i) by correlating the Fv-1 genotype of a number of mouse strains with the restriction-transferring activity of extracts of the respective embryo cell cultures, and (ii) by correlating the Fv-1 genotype of BLC3F2 (C57BL/6 female [Fv-1bb] by C3H male [Fv-1nn] parental strains) mouse embryos, which segregate the Fv-1 alleles in a 12:1 ratio, with the inhibitor activity of extracts of the cells from each embryo. Five independent matings, totaling 45 individual embryos, were tested. Each embryo was cultured, and the Fv-1 genotype was determined independently by titration of N- and B-tropic viruses; the extracts of replicate secondary cultures were tested for their effect on infection of permissive cells by N- and B-tropic viruses. The specific-restriction-transferring activity of the embryos was found to segregate with the appropriate Fv-1 genotype. These res-lts confirm the suggestion that the inhibitor of the leukemia virus host range types in the cellular extracts is a product of the Fv-1 locus.


Effect of hypertonic medium on protein synthesis of Mahoney and LSc poliovirus. The initiation of synthesis of Mahoney virus protein is more sensitive to high osmolarity than initiation of synthesis of LSc virus protein. Production of Mahoney virus protein appears to be only slightly more resistant to high osmolarity than synthesis of HeLa-Rhino cell protein.


Analysis of structural polypeptides of purified human cytomegalovirus. Human cytomegalovirus strain C87 was purified by the following procedures. (i) Extracellular virus was concentrated by centrifugation at 100,000 X g for 90 min and passed through a Bio-Rad Bio-Gel A-15m column. Most of the virus was recovered in the void volume. (ii) After two consecutive isopycnic potassium tartrate gradient centrifugations (20 to 50%), coinciding peaks of plaque titer, protein, and radioactivity were found at a density of from 1.20 to 1.21 g/cm3. To characterize the structural polypeptides of human cytomegalovirus and to establish relative purification criteria, virus was purified from two mixtures: (i) [35S]methionine-labeled extracellular virus mixed with an equal volume of unlabeled normal culture fluid; (ii) unlabeled extracellular virus mixed with an equal volume of [357a1methionine-labeled normal culture fluid. The extent of purification, as judged by the ratio of cellular to viral radioactivity, was 39-fold; i.e. about 2.5% of the protein in the purified virus preparation could be accounted for by host protein contamination. Electrophoresis of purified [35S]methionine-labeled virus on a polyacrylamide gel slab showed that there were at least 33 viral structural polypeptides (VPs), and their molecular weights ranged from 11,000 to 290,000. Autoradiograms obtained from electropherograms of purified [14C]glucosamine labeled virus showed six bands. Four of these were so broad that several VPs corresponded to each of the glycosylated bands. When heavy (two fractions close to 1.21 g/cm3) and light (two fractions close to 1.20 g/cm3) fractions of the PFU peak from the second potassium tartrate gradient were analyzed separately, the number of polypeptides observed was the same, but the relative amounts of some polypeptides differed. The major polypeptide, VP17, was found in greater amounts in the heavy fraction (35%) than in the light fraction (22%). The amount of DNA as a percentage of the weight of protein was 2% for the light fraction and 1% for the heavy fraction.


Association of vesicular stomatitis virus proteins with HeLa cell membranes and released virus. The association of vesicular stomatitis virus proteins with intracellular and plasma membranes was examined by pulse and pulse-chase labeling of virus-infected HeLa cells with [35S]methionine and separation of cell homogenates into three major membrane fractions in discontinuous sucrose gradients. The glycoprotein G was primarily associated with rough endoplasmic reticulum-like membranes after short radioactive pulses (2 to 4 min) but accumulated in the plasma membrane-enriched fraction and the smooth internal membrane fraction with longer pulse or chase periods. The nucleocapsid protein N and the matrix protein M accumulated in the rough endoplasmic reticulum and plasma membrane-like fractions but not in the smooth internal membrane fraction. Only a fraction (35 to 40%) of the viral protein synthesized during a short pulse in the mid-cycle of infection was apparently utilized in released virus. The newly synthesized virus proteins first appeared in released virus in the order: M, N and L, and G.


Glycosylation of vesicular stomatitis virus glycoprotein in virus-infected HeLa cells. Glycosylation of the envelope glycoprotein of vesicular stomatitis virus was examined using virus-infected HeLa cells that were pulse-labeled with radioactive sugar precursors. The intracellular sites of glycosylation and the stepwise elongation of the carbohydrate side chains of the G protein were monitored by membrane fractionation and gel filtration of Pronase-digested glycopeptides. The results with short pulses of sugar label (5 to 10 mtein linkage (glucosamine and mannose) are added to G which was associated with the rough endoplasmic reticulum-enriched membrane fraction, whereas the more distal sugars (galactose, sialic acid, fucose, and possibly more glucosamine) are added in the light-density internal membrane fraction. Accumulation of mature G was observed in the plasma membrane-enriched fraction. The gel filtration studies indicated that the initial glycosylation event may be the en bloc addition of a mannose and glucosamine oligomer, followed by the stepwise addition of the more distal sugars.


Asymmetric distribution of phosphatidylethanolamine in the membrane of vesicular stomatitis virus. The membrane-impermeable reagent trinitrobenzenesulfonate has been shown to react only with the surface components of vesicular stomatitis virus (VSV) membranes. When the amount of phosphatidylethanolamine (PE) available to modification by trinitrobenzenesulfonate in intact virions was determined, it was found that 36% of the total membrane PE was converted to the trinitrophenyl derivative. The same proportion of the total membrane PE was reactive after removal of the surface glycoprotein by trypsin digestion, but disruption of the virus membrane by sonication rendered all of the PE reactive. These results indicate that PE is asymmetrically distributed in the VSV membrane; 36% is present in the outer lipid leaflet, whereas 64% is found on the inner layer.


Ultrastructural studies of hepatitis A virus by electron microscopy. Well-defined, core-like structures were visualized in hepatitis A virus particles by a modified microelectron microscopy technique.


In vitro studies on the mechanism of acquired resistance to tuberculous infection. II. The effects of the culture supernatants of specifically stimulated-sensitized lymphocytes on the growth of tubercle bacilli within macrophages. Immune lymph node cells were obtained from mice immunized with bovine gamma globulin (BGG) in complete Freund's adjuvant or allogeneic MH134 tumor cells. They showed the capacity of conferring bactericidal activity on macrophages infected with Mycobacterium tuberculosis, H37Rv, when they were incubated on macrophage monolayers together with the corresponding antigen, i.e., BGG or solubilized cellular antigen of the tumor cells. However, such capacity was lower than that of tubercle bacilli-immune lymph node cells. Culture supernatants were harvested after incubation of tubercle bacilli-immune, BGG-immune or allogeneic tumor-immune lymph node cells with the corresponding antigen for 24 hr. Macrophages were altered so as to suppress intracellular bacillary growth when macrophage monolayers were exposed to the supernatants for more than 2 days. When normal lymph node cells were incubated on normal macrophage monolayers together with a mitogen such as PHA or concanavalin A, growth of tubercle bacilli within the macrophages was slightly but difinitely suppressed. The mechanism of elicitation of cellular immunity to the infection with tubercle bacilli is discussed on the basis of results presented in this and the preceding paper.


Immune response against hamster erythrocytes in the low-responder mouse strains. XI. Strain difference in the effects of various microbial adjuvants. Enhancing and suppressing effects of microbial adjuvants were studied in female mice of the C3H/He, AKR and SL strains. Propionibacterium acnes, Bordetella pertussis, BCG and yeast cell wall (YCW) were chosen as adjuvants. As antigens, we chose hamster erythrocytes (HRBC) which proved to be a weak antigen for mice. Adjuvants were given on day --7, day 0 or day 3, and HRBC were injected on day 0. The results were as follows. 1) P. acnes facilitated IgM and IgG antibody production in AKR mice and suppressed IgM antibody production in SL mice, when given on day --7. When P. acnes was given on day 0, they suppressed IgM antibody production in all of the strains used. 2) When B. pertussis was given on day 0, it exhibited enhancing effects on IgG antibody production in all of the strains and a suppressing effect on IgM antibody production in SL mice. 3) BCG suppressed IgM antibody production in all strains when given on day 0. 4) YCW showed no influence on antibody production in any combination used in this work. 5) SL mice were very sensitive to suppressing effects by adjuvants. Strain differences in the expression of enhancing and suppressing effects by adjuvants appear to be under some control independent of antigen-specific immune response genes.


Persistent high numbers of Clostridium perfringens in the intestines of Japanese aged adults. TSN agar was applicable for enumeration of Clostridium perfringens in fecal samples of adults but not in those of infants. It was demonstrated using TSN agar that some healthy aged adults had persistently carried C. perfringens at levels ranging from 10(7) to 10(9), while some others ranged from 10(3) to 10(6) per ml volume of fecal sample although all of these adults had the same diets. In the test for agglutinability of isolates of C. perfringens collected from two elderly adults, a younger adult and a baby, it was demonstated that most of the isolates obtained from an aged adult of high levels for 19 months belonged the same serotype, while rapid alteration of serotypes could be observed in three other persons with high or low levels. In spite of as many as 10(9) C. perfringens per ml of feces, no trace of a-toxin could be detected in the fecal samples. In in vitro tests, fecal suspension suppressed the production of a-toxin although it allowed the organism to grow sufficiently.


Rapid protein uptake and digestion in proximal tubule lysosomes. The aim of the present study was to determine the initial time course for the handling of protein by the proximal tubule cells in rat kidney, using 125I-labeled cytochrome C as a tracer. The uptake of the protein was followed by electron microscope autoradiography in kidneys from rats fixed by perfusion after i.v. injection of cytochrome C. Protein degradation was studied by incubating cortical slices, taken from rats, injected with the same label, and measuring the release of trichloroacetic acid (TCA)-soluble radioactivity from the slices. It was shown by autoradiography that lysosomes in proximal tubule cells start to accumulate labeled cytochrome C within 3 min after injection of the protein, and that the concentration of label in lysosomes increases during the first 30 min after injection, whereas it decreases in endocytic vacuoles. The catabolism of protein as measured in cortical slices began within 13 min after the i.v. injection. Other experiments showed that the accumulation of cytochrome C in the kidneys is very fast. The maximum accumulation, 37% of the injected dose, was reached seven minutes after injection. The results show that protein uptake in proximal tubule cells, transport into lysosomes and the digestion of protein is a more rapid process than previously reported in ultrastructural studies.


Intracellular collagen fibrils in prolapsed ("floppy") human atrioventricular valves. Ultrastructural study of prolapsed atrioventricular valves from five patients, one of whom also had the Marfan syndrome, disclosed granular cells that were characterized by numerous membrane-bound, electron-dense inclusions resembling those in the Hurler syndrome. Collagen fibrils were present within some of these inclusions. The intracellular collagen deposits may form as a consequence of abnormal interactions between newly secreted collagen and acid mucopolysaccharides.


Pathogenesis of visna. II. Effect of immunosuppression upon early central nervous system lesions. It was hypothesized that the lesions of visna might represent an immunopathologic process. To test this hypothesis, a 1-month schedule of immunosuppressive treatment was devised, using horse anti-sheep thymocyte serum. In Hampshire sheep, this regime was shown to protect against experimental allergic encephalomyelitis, to inhibit development of tuberculin hypersensitivity, to retard rejection of skin homografts by 3 weeks, and to markedly reduce the number and mitogenic responsiveness of peripheral blood lymphocytes. Two groups of Icelandic sheep received intracerebral inoculations of visna virus, and one group was treated with horse antisheep thymocyte serum supplemented by a short terminal course of cyclophosphamide. Central nervous system lesions were seen in only one of eight suppressed animals at sacrifice 25 days after infection, whereas definite lesions were present in eight of eight infected control animals. The frequency of central nervous system virus isolation was similar in the two groups, indicating that treatment suppressed the cellular proliferative response without preventing the central nervous system phase of infection. Sheep receiving horse antisheep thymocyte serum had a reduced number of virus isolations from peripheral lymphoid tissue, presumably reflecting the lympholytic effect of treatment. These observations are consistent with the immunopathologic hypothesis and suggest several different ways in which suppression could modify the immune response to visna virus.


Pathogenesis of visna. III. Immune responses to central nervous system antigens in experimental allergic encephalomyelitis and visna. Experimental allergic encephalomyelitis (EAE) was induced in sheep in pursuit of the hypothesis that an immune response against central nervous system antigens might play a role in the pathogenesis of visna. Nine to 12 days after sensitization with whole sheep brain and complete Freund's adjuvant, approximately 50% of sheep developed a fulminating lethal form of EAE. Following a second sensitization, another 20% of animals developed EAE whereas a residual 30% failed to develop any signs or histologic evidence of disease. A histologic comparison of EAE and visna indicated considerable similarity in the nature of the pathologic process. However, the distribution of lesions was quite different, suggesting cellular responses to two different antigens, Cell-mediated immunity to myelin basic protein, as measured by lymphocyte blast transformation, was minimally elevated in sheep sensitized with whole brain suspension in complete Freund's adjuvant, whereas no response could be detected in visna-infected sheep. Complement-fixing antibody titers to basic protein and to a lipid antigen of brain, probably galactocerebroside, rose briskly after sensitization. In visna-infected sheep, on the other hand, there was no increase in either antibody. A large proportion of both Hampshire and Icelandic sheep had low levels of complement-fixing antibody to central nervous system antigens prior to induction of EAE or infection with visna virus. The origin of this antibody is undetermined, but it appeared to have no effect on the course of either disease. Immunosuppression of sheep with antilymphoid serum prevented induction of EAE. Acute EAE was, thus, successfully induced in sheep and used as a model to measure immune responses to central nervous system antigens and as an index of immunosuppression. However, these comparative studies did not provide any evidence for the role of an autoimmune response, to the two central nervous system antigens tested, in the pathogenesis of visna.


In vivo assembly and maturation of vesicular stomatitis virus. Previous studies on vesicular stomatitis virus (VSV) maturation in infected cells have utilized in vitro cell cultures. The present study is, to our knowledge, the first in vivo analysis of VSV-cell interaction in the central nervous system of weaning outbred Swiss mice. Intracerebral inoculation of wild-type VSV resulted in rapid viral replication in brain and spinal cord. By immunoflourescence, viral antigens were first seen in ependymal cells of brain and spinal cord and soon thereafter in surrounding neurons. The large anterior horn neurons of spinal cord appeared to be in the most heavily infected. Ultrastructurally, VSV-neuron interaction evolved in three phases. The first phase consisted of viral entry into the cell by fusion and viropexis. The second phase was characterized by nucleocapsid accumulation and resulted in the appearance of large cytoplasmic inclusions. The third phase was maturation and release from the infected cell and was accomplished by viral budding from plasma membranes. Degenerative changes in infected neurons were generally absent. Cells in the area of the central canal seemed to present a different pattern of virus-cell interaction especially at the level of maturation and release. Some of these cells in advanced stages of degeneration showed viral particles free in nucleocapsid material with no virus-membrane association. Viral budding was not observed and, because these cells do eventually die, it is possible that virus was released in the intercellular space at the moment of cellular disruption. These results suggest that VSV-cell interactions may vary depending upon the nature of the infected cells.


The effect of changes in dietary sodium on lung and serum antiotensin-I-converting enzyme in the rat. Three groups of rats were fed a sodium-deficient diet, a normal sodium diet, and a diet high in sodium for up to 8 weeks. Animals on a sodium-deficient diet had a significant decrease of serum sodium levels with a concomitant increase of renal renin granules. The reverse was true in rats on a high sodium diet whereas no significant changes in renin granulation were seen in rats fed a normal sodium diet. The changes in serum sodium levels and in number of renal renin granules were not associated with changes in lung or serum angiotensin-I-converting enzyme activity which did not vary significantly in any of the three groups of rats throughout the experiment. It is, therefore, suggested that such activity is independent of changes in sodium balance sufficient to affect the renal renin content.


A study of adenosine 3'-5' cyclic monophosphate binding sites of human erythrocyte membranes using 8-azidoadenosine 3'-5' cyclic monophosphate, a photoaffinity probe. An earlier report (1a) has shown the utility of 8-N3cAMP (8-azidoadenosine-3', 5'-cyclic monophosphate) as a photoaffinity probe for cAMP binding sites in human erythrocyte membranes. The increased resolution obtained using a linear-gradient SDS polyacrylamide gel system now shows that: 1) both cAMP and 8-N3cAMP stimulate the phosphorylation by [gamma-32P]-ATP of the same red cell membrane proteins; 2) the protein of approximately 48,000 molecular weight whose phosphorylation by [gamma-32P]-ATP is stimulated by cAMP and 8-N3cAMP migrates at a slower rate than the protein in the same molecular weight range which is heavily photolabeled with [32P]-8-N3cAMP;3) other cyclic nucleotide binding sites exist besides those initially reported; 4) the variation in the ratio of incorporation of [32P]-8-N3cAMP into the two highest affinity binding sites appears to be the result of a specific proteolysis of the larger protein.


Change in the stability of tendon collagen during ageing in the reptile, Calotes versicolor. A brief note. Age related changes in the digestibility of tendon collagen of the garden lizard, Calotes versicolor, were traced using type I collagenase from Clostridium histolyticum. Collagen from older lizards showed less susceptibility for collagenase digestion than the younger lizards suggesting increased number of cross-linkages comparable with mammals.


A system for measuring the noradrenaline receptor contribution to the flexor reflex. A system is described for recording psoas muscle contractions in response to stimuli delivered to the sole of the foot of the acute spinalized unanaesthetized rat. Clonidine, an alpha adrenergic receptor agonist, caused a major increase in this flexor reflex amplitude. The increase was blocked by phenoxybenzamine at a dosage level of 20 mg/kg which did not, of itself, reduce flexor reflex amplitude. Older, larger rats responded significantly more forcefully to clonidine than did younger, smaller rats. However, these differences between rats of different size tended to disappear when the data were expressed in terms of 0/0 of original baseline amplitude. L-dopa, 75 mg/kg, increased the flexor reflex amplitude more so than did L(+)erythro dihydroxyphenylserine (DOPS) at twice the disage in rats pretreated with reserpine and nialamide. Other DOPS isomers were less effective, and L(-)threo DOPS was toxic. These results support the view that spinal noradrenaline receptors exist that increase the psoas flexor reflex. The advantages of the present system are twofold: first, the results may be quantitated; second, small amounts of potential central noradrenaline receptor agonists and antagonists may be tested for their effect on the flexor reflex using relatively few unanaesthetized rats.


Clinical significance of viral latency.

Clinical significance of viral latency. Evidence is accumulating to show that a number of viruses have the ability to adapt to man's defense mechanisms and survive in a latent state for what appears to be the life of the human host. Unfortunately, latent viral presence, which may appear clinically benign initally, may manifest itself later as severe and often fatal disease. Some members of the herpes virus family have latent potential and are discussed in detail. Clinical competence would suggest a thorough understanding of these late manifestations of occult viral presence.


Effect of ethanol ingestion on choline phosphotransferase and phosphatidyl ethanolamine methyltransferase activities in liver microsomes. The effect of ethanol ingestion on choline phosphotransferase and phosphatidyl ethanolamine methyltransferase activities, the two enzymes involved in phosphatidyl choline biosynthesis in liver microsomes, has been investigated. Female rats were fed a 5% ethanol-liquid diet containing amino acids, minerals, vitamins, with and without choline, for 2, 6 and 10 weeks. Control animals were pair-fed the same isocaloric diet with 5% sucrose with and without choline. Ethanol administration with or without dietary choline stimulated significantly (P less than 0.001) the specific activities of phosphatidyl ethanolamine methyltransferase in liver microsomes in the animals fed 5% ethanol for 2, 6, and 10 weeks, when compared to those control animals pair-fed the isocaloric diet with or without choline. Ethanol administration with or without dietary choline for 2 weeks stimulated significantly (P less than 0.02) the specific activities of choline phosphotransferase. The specific activities of phosphatidyl ethanolamine methyltransferase continued to increase in the liver microsomes from the animals in which dietary choline was omitted for 2, 6, and 10 weeks in the sucrose controls and alcohol-fed animals. Ethanol administration stimulates significantly (P less than 0.001) the phosphatidyl ethanolamine methyltransferase specific activities in liver microsomes of animals fed the liquid diet with dietary omission of choline and methionine for 2 weeks.


The optimization of Porous glasses for immobilized enzyme (IME) systems. Critical parameters involved in the selection of porous glass for use as support materials in the immobilization of biologically active materials are discussed. The importance of physical properties such as pore morphology, surface area and particle size relative to the final activity of the enzyme is shown using glucoamylase IME as a model system. The process of selection of specific materials with respect to the environmental conditions of the enzyme and the chemical durability of the support is considered, as are the mechanical properties required in the scale-up of an application.


Physiologic significance of glucocorticoids and insulin in the regulation of hepatic gluconeogenesis during starvation in rats. The physiologic significance of glucocorticoids and insulin in the regulation of hepatic gluconeogenesis was investigated during a 48-hr starvation period by studying the factors presumed to control the rate of glucose synthesis in the final gluconeogenetic pathway. Rats were used, in which glucorticoids were removed by adrenalectomy before starvation, and in which serum insulin was kept constant before and after food withdrawal by pre-feeding with a proteinfree diet. It was found that adrenalectomized rats at constantly low serum insulin levels responded to starvation as rapidly, and to the same degree, as intact control subjects (1) by a significant increase in plasma glucagon and, consequently, in hepatic cAMP concentration; (2) by a coordinate elevation of the activities of hepatic pyruvate carboxylase, P-enolpyruvate carboxykinase, and fructose-1,6-diphosphatase; (3) by systematic alterations in the concentration of effectors of gluconeogenetic key enzymes; (4) by a shifting of the cytoplasmic NAD system towards the reduced state; (5) by a decrease in the intrahepatic concentration of glycogenic precursor substrates. These results suggest that the hepatic gluconeogenic response to starvation with respect to the regulatory factors 1-5 occurs independently from changes in the concentration of plasma glucocorticoids and insulin. The crossing over of the gluconeogenetic intermediates between pyruvate and P-enolpyruvate (PEP), which was observed in intact but not in adrenalectomized rats, supports the assumption that during starvation, glucocorticoids enhance the rate of glucose production by the liver predominantly by permitting hepatic cAMP to stimulate the yet undefined mechanism, which has been demonstrated in the isolated perfused rat liver to control the substrate flow between pyruvate and PEP.


Effect of acute uremia on triglyceride kinetics in the rat. Plasma triglyceride (TG) levesl were elevated 24 hr after the production of acute uremia in rats. The effect of acute uremia on TG production rate was estimated by determining the rate of TG accumulation following Triton WR 1339 inhibition of lipoprotein removal, by measuring hepatic TG secretion rate during in situ liver perfusion, and by quantifying hepatocyte very low density lipoprotein content with the electron microscope. The results of all three of these approaches indicated that TG synthesis and secretion were decreased in acute uremia, suggesting that the associated increase in plasma TG levels had to result from a removal defect. This hypothesis was tested directly by injecting pre-labeled very low density lipoprotein TG into acutely uremic and control rats, and measuring its rate of disappearance from plasma. The t1/2 of removal in acutely uremic rats was found to be approximately twice that of control, confirming the hypothesis that the rise in plasma TG levels in acute uremia is due to a defect in removal of TG from plasma.


Sequential alterations in the hepatic content and metabolism of cyclic AMP and cyclic GMP induced by DL-ethionine: evidence for malignant transformation of liver with a sustained increase in cyclic AMP. There is evidence than adenosine 3',5'-monophosphate (cAMP) and guanosine 3',5'-monophosphate (cGMP) may have antagonistic actions on cell growth, with cAMP inhibiting and cGMP stimulating this process. However, reductions in cAMP and increases in cGMP are not charactersitic of all neoplastic tissues. Thus, benign and malignant tissues from hepatoma-bearing rats exposed to the hepatic carcinogen DL-ethionine have elevated rather than depressed cAMP, compared to control liver, and parenteral administration of this drug increases hepatic cAMP within hours. In the present study, the effects of ethionine ingestion on the hepatic content and metabolism of both cAMP and cGMP were examined sequentially in rats at 2 and then 6 wk intervals, from the initiation of drug administration until the development of hepatomas. After 2 wk, cAMP content of quick-frozen liver from rats receiving ethionine (E) was significantly increased (826 +/- 91 pmole/g wet weight) above that of liver from pair-fed controls (C, 415 +/- 44), whether calculated by tissue wet weight, protein, or DNA content. In benign tissue from E, higher cAMP was still evident after in vitro incubations of slices with 2 mM 1-methyl-3-iso-butylxanthine (MIX) and was associated with enhanced adenylate cyclase and unchanged high or low Km cAMP-phosphodiesterase activities. These findings are compatible with accelerated cAMP generation in liver from E. Protein kinase activity ratios were significantly increased in frozen liver from E (0.52 +/- 0.04 versus 0.36 +/- 0.03 in C), and the percent glycogen synthetase in the I form was clearly reduced (19% +/- 2% in E versus 47% +/- 5% in c). incubation of hepatic slices from E or C with MIX and/or 10 muM glucagon further increased cAMP and protein kinase activity ratios, data which imply higher effective, as well as total, cellular cAMP in E. Changes in cAMP metabolism and action observed at 2 wk persisted throughout the 38-wk period of drug ingestion. Adenylate cyclase activity, cAMP content, and protein kinase activity ratios of ethionine-induced hepatomas exceeded those of both the surrounding liver from tumor-bearing rats and that of control liver, but alterations in these parameters were qualitatively similar in both tissues from E. By contrast, while cGMP in quick-frozen surrounding liver from tumor-bearing rats (36 +/- 4 pmole/g wet weight) did not differ from that of control liver (30 +/- 3), cGMP in the hepatomas was increased. This change was evident in both frozen tumor (89 +/- 10) and in tumor slices incubated in vitro with MIX (C, 90 +/- 11; surrounding liver, 85 +/- 10; hepatoma 231 +/- 29). These results indicate that malignant conversion can occur in liver with a sustained elevation of both total and effective cAMP during the premalignant phase. The increase in cGMP detected in ethionine-induced hepatomas could also be a key determinant of malignant transformation in the model, although premalignant changes in cGMP were not apparent.


Enzymic activities of cadmium- and zinc-tolerant strains of Klebsiella (Aerobacter) aerogenes growing in glucose-limited chemostats. The activities of phosphatases and some enzymes of glucose metabolism were determined in K. aerogenes NCIB 418 and in two strains derived from it, resistant to 50 mug Cd2+ ml-1 and 16 mug Zn2+ ml-1 respectively, during growth at D = 0.38 h-1 in medium containing beta-glycerophosphate as sole phosphorus source and supplemented with Cd2+ and Zn2+, as appropriate, for the resistant strains. The pH-activity profiles of the phosphatases differed from strain to strain but all showed maximum activity at an acid pH and this activity was very much lower in the Zn2+-resistant strain than in the control and even lower in the Cd2+-resistant strain. Resistance to either metal was associated with decreased glucose-6-phosphate dehydrogenase activity and increased phosphoglucose isomerase activity, suggesting an increased flow of carbon through the Embden-Meyerhof pathway relative to the pentose phosphate pathway, but the efficiency of the conversion of glucose into biomass was largely unaffected. Glucose phosphoenolpyruvate phosphotransferase activity was also lower in the resistant strains.


Malignant disease of the nasopharynx presenting as pain in the head and neck. Two patients with an unusual pattern of pain in the head and face, who presented a diagnostic problem for one and four years respectively, subsequently proved to have a carcinoma of nasopharyngeal origin. The neurological complications of this condition are described.


Treatment with gentamicin monitored by serum antibiotic assay. Gentamicin is very widely accepted as the most effective antibiotic for treatment of a number of serious Gram-negative infections, but one disadvantage associated with its use is the risk of toxicity particularly affecting the eighth nerve. An account is given of eight years' experience with this antibiotic in which obvious ototoxicity was avoided in hospitalized patients in whom gentamicin treatment was monitored by assay of the residual amounts of antibiotic present in the serum. The relationship between accumulation of gentamicin in patients with renal impairment and ototoxicity is discussed. Attention is drawn also to the presence of an increased risk of ototoxicity in patients, particularly the elderly, in whom mild-to-moderate degrees of renal impairment may be overlooked. In such patients modification of the frequency of the dose of gentamicin is necessary and a schedule of treatment based on the results of assay of residual serum levels of gentamicin is given.


Multiple regulation of nucleoside catabolizing enzymes in Escherichia coli: effects of 3:5' cyclic AMP and CRP protein. The regulation of the synthesis of nucleoside metabolizing enzymes has been studied in cya and crp mutant strains of Escherichia coli. The synthesis of the cyt-enzymes, cytidine deaminase and uridine phosphorylase regulated by the cytR gene product, is activated by the cAMP-CRP complex. On the other hand the synthesis of the deoenzymes: deoxyriboaldolase, thymidine phosphorylase, phosphodeoxyribomutase and purine nucleoside phosphorylase, appears to be increased if an active cAMP-CRP complex cannot be formed. It also seems that nucleosides serve as poor carbon sources for cya and crp mutants; this could not solely be explained by low levels of nucleoside metabolizing enzymes nor by a deficiency in nucleoside uptake. Addition of casamino acids stimulated the growth of cya and crp mutants, with nucleosides as carbon sources. When grown on glucose and casamino acids growth could be stimulated by adenine and hypoxanthine nucleosides; these results suggest an impaired nitrogen metabolism in cya and crp mutants.


Mutliple hepatitis viruses in multiple attacks of acute viral hepatitis. We evaluated the causes of 30 episodes of acute viral hepatitis in 13 patients who had multiple attacks. Two (seven per cent) of 30 bouts were caused by hepatitis A virus, and 12 (40%) by hepatitis B virus. No patient, however, had more than one attack with the serologic characteristics of Type A or Type B disease. Thus, there were 16 bouts (53 per cent) not attributable to either of the two recognized hepatitis viruses. None of these "non-A, non-B" episodes, evaluated for infectious mononucleosis and cytomegalovirus infection, could be ascribed to either. From this evidence, therefore, it appears that the clinical syndrome of viral hepatitis is produced not only by the two viruses (hepatitis A virus and hepatitis B virus) recognized since the 1940's but also, in all probability, by two non-A, non-B agents.


Effects of adenine and cytokinins on growth and protein kinase activity of Verticillium albo-atrum. Growth of Verticillium albo-atrum in liquid Czapek-Dox broth was stimulated about four-fold by added 10 mM adenine, N6-benzyladenine, or kinetin. Less stimulation was evident at lower concentrations. With none of these included in the basal growth medium, detectable protein kinase activity in cell-free extracts was low and responded minimally to cAMP (adenosine 3',5'-cyclic monophosphate) in the reaction mixture. With each of these compounds as an additive to the growth medium, protein kinase activity was not only greater but also responded markedly (several fold) to cAMP. These results demonstrate a strong correlation between exogenously supplied adenine and related compounds, increased growth, increased protein kinase activity, and kinase response to cAMP in this organism.


[Behavior of certain parameters of lipid and energy metabolism. 3. Relationships between cyclic 3',5'-adenosine monophosphate and acetyl coenzyme A in liver of growing rats deduced from model experiments with diets differeing in fat content]. Schemes of the lipid and the pyruvate metabolism serve to show that a great part of the enzymes which intervence in the metabolic pathways and are associated with the formation and the consumption of acetyl coenzyme A may be regulated by cyclic 3',5'-adenosine monophosphate (cAMP) in the sense of activation or inhibition. The cAMP increase in the liver, which has been demonstrated in the present study for a diet containing 25% of fat, opens the metabolic pathway to the formation of acetyl coenzyme A by means of fatty acid degradation and simultaneous inhibition of lipogenesis. The deficiency of insulin (which has been evidenced in previous paper) characterizes, together with the facts mentioned, a state like diabetes or the fasting state. Acetyl coenzyme A is mainly used for energy supply. The close negative correlation between cAMP and acetyl coenzyme A (which is shown in the present paper) permits to conclude that the extent and trend of the increase and decrease in the liver is subjected to intensive hormonal control in which cAMP in involved.


Quantal parameters of transmission at the frog neuromuscular junction. Estimates of quantal release parameters at frog neuromuscular junctions showed that alterations in [Ca2+]o affected m (number of quanta), p (probability of quantal release) and n (stores of quanta available for release). The effect of [Ca2+]o depended upon the initial value for p. When p was low, raising [Ca2+]o increased m and p, but not n. However, when p was large, raising [Ca2+]o had no further effect on p but increased m and n. During prolonged repetitive nerve stimulation to cause a decrease in m, n was decreased and p was increased. This finding was attributed to a failure of transmitter mobilization to maintain the stores of transmitter available for release.


Relationship between the level of cAMP and the contractile force under stimulation of alpha- and beta-adrenoceptors by phenylephrine in the isolated rabbit papillary muscle. The time course of changes of the level of 3',5'-cyclic AMP (cAMP) and of the tension developed under stimulation of alpha- and beta-adrenoceptors by phenylephrine was investigated in the isolated rabbit papillary muscle. Furthermore the dose-response relationships for increases of cAMP and of developed tension elicited by phenylephrine were determined. 1. A submaximally effective concentration of phenylephrine (10(-5) M) increased significantly the level of cAMP of the papillary muscle at 15 and 30 s by 45 and 36% respectively; the level of cAMP returned to the control value at 60 s after the administration. The developed tension increased significantly not before 45 s and reached its maximal level at 180 s. 2. When alpha-adrenoceptors were blocked by phentolamine (10(-6) M), the positive inotropic effect of phenylephrine was decreased significantly but the increase of cAMP induced by phenylephrine was not reduced. In the presence of phentolamine the increase of cAMP induced by phenylephrine lasted longer than in the control experiments. 3. The effects of phenylephrine (10(-5) M) both on the level of cAMP and the developed tension mediated via stimulation of beta-adrenoceptors in the presence of phentolamine were enhanced by the phosphodiesterase inhibitor papaverine throughout the course of responses. 4. Phenylephrine produced an increase in developed tension as well as in cAMP. The corresponding dose-response curves run parallel to each other but differed by about 1.5 log units whereby the developed tension was evoked by lower concentrations. Phentolamine (10(-6) M) shifted the curve for the positive inotropic action by about 1.5 log units but did not affect that for increase in cAMP. Therefore, in the presence of the alpha-adrenolytic drug phentolamine the difference between both curves became smaller so that both curves were superimposed. Papaverine (10(-5) M) shifted the whole curve for cAMP upwards and enhanced the maximal contractile response to phenylephrine mediated by stimulation of beta-adrenoceptors. 5. The present results indicate that the positive inotropic action of phenylephrine in lower concentrations (less than 10(-5) M) induced by stimulation of alpha-adrenoceptors is independent of the level of cAMP. The positive inotropic action of the higher concentrations of phenylephrine induced via stimulation of beta-adrenoceptors was preceded by an accumulation of cAMP; the inhibition of the cAMP phosphodiesterase activity by papaverine enhanced the actions of phenylephrine both on the level of cAMP and on the contractile force.


Binding of botulinum neurotoxin to the synaptosome fraction of rat brain. The radioactive 125I-labelled neurotoxin of C. botulinum type A, when incubated with rat brain homogenate, is bound selectively to the synaptosome fraction. Intact toxin was liberated from the synaptosome fraction by treatment with Triton X-100, SDS, trypsin or neuraminidase.


[Adenoid cystic carcinoma extended to Gasserian ganglion-report of two cases (author's transl)]. Adenoid cystic carcinoma is an uncommon tumor, especially in cases of its extension into the cranial cavity. Two cases of this tumor involving Gasserian ganglion were reported. The first case was a 47 year old female who developed paresis of the right side fifth cranial nerve. The plain skull X-ray of the axial view demonstrated enlargement of the right sided foramen ovale. The tumor was removed exploring the middle fossa extradurally, however its extention through the foramen ovale remained behind. A small induration was found at the base of the oral cavity postoperatively, and the biopsy revealed characteristic findings of adenoid cystic carcinoma histologically. Radiation therapy was followed by considerable clinical improvement.


Autopsy correlations of computerized tomography: experience with 6,000 CT scans. Seventy-nine autopsy correlations of CT scans showed (1) excellent correlations in normal brains, but the size of the lateral ventricles consistently larger during life than after death; (2) a distinctive pattern differentiating obstructive from nonobstructive hydrocephalus; (3) infarctions appearing as areas of decreased densities of parenchyma in vascular distributions; (4) distinctive high density appearances of hemorrhages that differentiated them from infarctions and, in general, all other pathologic processes; (5) supratentorial, intraventricular, and posterior fossa tumors appearing as masses that displaced, distorted, collapsed, and enlarged normal spaces and structures such as ventricles and pineal gland; (6) 11 false-negative CT scans in some cases of brain stem infarction, brain stem hemorrhage, and small metastasis; and (7) an overall accuracy of 86.2 percent of CT scanning in correctly identifying pathology of the brain.


[Motor nerve conduction velocity in arterial hypertension]. Motor nerve conduction velocity was studied in a group of 44 hypertensive patients; velocity was measured at the level of the median and external popliteal sciatic nerves. From the series were excluded those hypertensive subjects with renal insufficiency, diabetes mellitus and peripheral arterial disease. No significant differences were observed in MNCV values compared to controls. Within the hypertense group, no appreciable variations were observed depending on the duration and degree of hypertension, and signs of visceral damage (assessed at myocardial and retinic levels). The results do not confirm previous published data suggesting the existence of MNCV reduction during arterial hypertension, the reduction being considered an subclinical expression of peripheral neuropathy and an index of the gravity of visceral damage during hypertensive disease.


[Polyneuropathy syndrome in a case of spinal meningeal carcinomatosis]. The authors report a case of polyneuropathy of lower extremities developing in carcinomatosis of spinal meninges in a patient after gastrectomy for mucus-producing carcinoma.


Isolated ACTH deficiency and pregnancy.

Isolated ACTH deficiency and pregnancy. Isolated deficiences of anterior pituitary hormones are rare. A patient is presented whose hypoglycemia during pregnancy resulted from ACTH deficiency. Glucocorticoid treatment enabled her to complete pregnancy successfully. In addition, she did not menstruate during an interval in which she did not receive glucocorticoid therapy. The dynamics of fetal and maternal glucose metabolism are discussed as well as menstruation and fertility in adrenal insufficiency.


A prevalence study of oral mucosal lesions in an adult Swedish population. The prevalence of oral mucosal lesions in Sweden is virtually unknown. Available prevalence figures concern but a few types of lesions and relate to highly selected, special populations. Investigations on general populations are sparse also in other countries. The aim of the present study was to survey the occurrence of oral mucosal lesions in a relatively large general population in Sweden. The study was performed in collaboration with a health screening organization in the middle of Sweden. The populations of two municipalities in the County of Uppsala were investigated. A mobile health screening group visited several places in the selected area and during the time of the present investigation a total of 30,118 persons, aged 15 years or above, were summoned. Of these, 62% or 18.659 individuals attended and were examined. The non-participants became the subject of a special investigation for which every fifth individual, or 2.292 persons, was randomly sampled. After a second summons, 933 persons presented for examination. Through personal contact, an additional 741 individuals were examined in private homes, places of work, hospitals, old age homes and other institutions. The final non-participation, representing 10.3% of the total population, was considered to have but a marginal influence on the results of the study. The diagnostic procedure was based on clinical criteria set forth especially for the investigation. The validity of the final diagnoses was evaluated by, for instance, inter-examiner tests and comparisons between clinical diagnoses and histologic descriptions and was found to be acceptable. Tests of the reliability were, among else, undertaken through re-examinations and indicated, that underregistration apparently occurred for a few of the lesions investigated. The prevalences of about 60 oral mucosal lesions were recorded and compared with previous findings. Notably high prevalences were found for focal epithelial hyperplasia (0.11%), leukoedema (49.07%), geographic tongue (8.45%) and lichen planus (1.85%). For some lesions prevalence figures are difficult to compare with findings from previous studies. This includes lesions which are directly or indirectly related to local etiologic factors such as denture status and tobacco habits. As regards these factors basic information has been collected and will be used for further analyses. The collected total material has also been designed to form the basis for longitudinal studies of, for example, precancerous lesions.


Studies on kinetics of DNA synthesis in CV-1 cell cultures infected with different multiplicities (MOI) of Herpes simplex virus type 2 (HSV-2). Infection of CV-1 cell cultures with HSV-2 at MOI 5 respectively 1PFU/cell resulted in markedly different patterns of DNA-synthesis. Isolation and separation of cellular and viral DNA on CsCl equilibrium density gradients during the "late" phase after infection with 5PFU/cell revealed a rapid increase in synthesis of virus-specific DNA (which banded at density q25 = 1.729 g/ccm) while synthesis of host cell DNA (banding at 1.705 g/ccm) was constantly inhibited. 15 hours after infection and incubation with labelled medium, around infection and incubation with labelled medium, around 75% of total isolated DNA was virus-specific. On lowering the MOI to 1 PFU/cell, however, synthesis of host cell DNA continued and was even partially stimulated during the late phase whereas synthesis of virus-specific DNA advanced only slowly. 16 hours after infection, approximately 25% of the total assayed after infection, approximately 25% of the total assayed DNA was virus-specific and the amount of host cell DNA approached values of uninfected control cells. 24 hours after infection and incubation, virus specific DNA rose to approximately 45% of overall DNA assayed, and synthesis of host cell DNA was completely inhibited. However, infectivity rose constantly and reached 106.8 TCD 50/ml at 22 hours p.i. These findings were independent of passage number of CV-cell culture, and there were no alterations in karyotype and morphological behaviour of the cells.


[Blood lipoprotein transfers in the normal arterial wall. Study auto-historadiography in optic microscopy]. In order to study the mecanisms of serum lipoproteins transfers and binding sites in arterial wall, auto-historadiographies in optical microscopy were realised with rats aortas. Animals received infusion of in vivo labelled on the cholesterol and purified serum lipoproteins. The results confirmed the serum lipoproteins transfers in arterial wall and slow that the inflow is all the more quicker and higher as the molecule is smaller. The very particular aspect of these transfers rises several hypothesis and leeds to suggest the possibility of preferential way of passage inside the arterial wall.


Historadioautographic and biochemical studies on the transport of atherogenic lipoproteins into the rabbit aortic wall. Historadioautographic and biochemical studies were undertaken on the transport of 14C-labelled serum very low density and low density lipoproteins (VLDL and LDL respectively) into the aortic wall. Aortas from normal and atherosclerotic rabbits were perfused in vitro with a medium containing VLDL and LDL for 6 hours. Historadioautography revealed the label in all the layers of the perfused aorta. The gradient in the distribution of this label from the inner layers of the aortic wall dawn to the outer part of the media demonstrated that lipoprotein infiltration of the vessel wall is due to uptake of lipoproteins from the aortic lumen. The non-uniform pattern of the distribution of lipoproteins taken up by the atherosclerotic aorta was considered as a result of atherogenesis induced changes in the aortic connective tissue. Both morphological and biochemical studies revealed the increased permeability of the atherosclerotic aortic wall to VLDL and LDL, which appeared to penetrate the walls of blood vessels in the form of intact particles.


Atopic dermatitis: etiology and pathogenesis. Atopic dermatitis is a recognizable phenotype that most likely results from several mechanisms. An increasing number of children with atopic dermatitis are recognized as having immune defects, although the exact incidence of such immunodeficiency states among patients with atopic dermatitis is unknown. Children with atopic dermatitis who suffer recurrent or persistent infections should undergo immunologic evaluation. Ideally, this should include evaluation of cell-mediated immunity as well as neutrophil and monocyte chemotaxis. Further investigation into the beta-blockade theory may enhance our understanding of atopic dermatitis. Careful attention to factors exacerbating atopic dermatitis is essential in understanding this problem. Abnormal sweating, dry skin, sensitivity to contactants, and emotional stress should always be considered in evaluation of children with atopic dermatitis.


Dietary lipid and postnatal development. II. Palmityl coenzyme A oxidation in heart and liver. There is only indirect evidence to suggest that the sudden postpartum appearance of dietary lipid regulates the perinatal development of the enzyme pathways required for fatty acid oxidation. To test this idea directly, rabbit pups were maintained on diets containing lipid to equal either 14.2% (LF) or 77.6% (HF) of the total caloric intake. Palmityl coenzyme A oxidation rates in the presence of excess ADP and carnitine were measured polarographically in heart and liver homogenates. No significant difference in oxidation rates between HF and LF groups was observed even at 10 days of age. Palmityl coenzyme A oxidation in both groups was carnitine dependent and was in general the same as that of mother-fed animals. Similarly, an evaluation of cytochrome oxidase activity and glutamate + malate-supported respiration in heart and liver homogenates revealed no difference attributable to diet. To consider the possibility that fatty acid oxidation might be specifically increased or decreased over other mitochondrial respiratory activity as a function of diet, palmityl coenzyme A oxidation rates were normalized with respect to glutamate + malate oxidation rates. A similar comparison was made relative to cytochrome oxidase activity. Still no differences were observed between HF and LF groups. By studying the maximum rate of oxygen utilization in the presence of excess carnitine and palmityl-coenzyme A we would have detected any change in a rate-limiting step for fatty acid oxidation beyond acyl activation. We must conclude, therefore, that large differences in the proportion of postnatal dietary lipid do no influence the cellular capacity to oxidize palmityl coenzyme A.


Encephalitis in infectious mononucleosis: diagnostic considerations. Four atypical cases of presumed infectious mononucleosis (IM) encephalitis are presented. To establish an etiologic diagnosis, Paul-Bunnell-Davidsohn heterophil titers (PBD), antibody titers to the antigens of the Epstein-Barr virus (EBV), and oropharyngeal excretion of EBV were determined. Criteria for a primary EBV infection are (1) an antiviral capsid antigen titer of 1:160 or greater, (2) the presence of antibody to the diffuse component of the early antigen, (3) absence of antibody to the nuclear antigen, and (4) excretion of the virus from the oropharynx. Three of the four cases met these criteria; of the three, one did not have a positive heterophil titer. The fourth case turned out not to be IM; there was a positive PBD heterophil, but there was no evidence of primary EBV infection. Although the PBD heterophil is usually a reliable test to diagnosis IM, it is not always present in children, and it is sometimes nonspecifically elevated. Some EBV titers can be nonspecifically elevated as well; however, the above criteria are diagnostic of primary EBV infection.


The dynamics of insulin release from mouse pancreatic islet cells in suspension. The overall dynamics of glucose-induced insulin release was strikingly similar in dispersed cells and intact islets perifused in parallel. Both preparations exhibited a latency of 1-2 min, after which period there was a brisk rise of insulin release followed by a sustained second phase. During the second phase, insulin release from dispersed cells attained a stable plateau rate, whereas the release from intact islets continued to rise. Epinephrine (1 muM) inhibited the release in both preparations, but the return to basal rate was faster in the dispersed cells than in the intact islets. The dispersed cells oxidized glucose at a constant rate for at least 60 min; the glucose oxidation was markedly sensitive to changes of the glucose concentration in the range of 3-20 mM.


Loss of ATP in micromolar amounts after perchloric acid treatment. Treatment of fluid samples containing known amounts of ATP with 6.0 N perchloric acid (PCA) results in a total loss of 65-71% when the initial concentrations of ATP ranged between 0.5 to 50 muM. Half of this loss was attributed to desensitization of firefly extract (luciferin-luciferase reaction) while the remaining loss was presumably due to adsorption of ATP to perchlorate precipitate upon neutralization. Similar treatment of solutions with higher initial concentrations (100-1000 muM) resulted in apparent total losses averaging 22%. These losses were due solely to desensitization of firefly extract by neutralized PCA. Both the adsorption and desensitization phenomena must be taken into account when the ATP content is measured from tissue extracts and fluid samples subjected to this procedure.