H2O2 release from human granulocytes during phagocytosis. Relationship to superoxide anion formation and cellular catabolism of H2O2: studies with normal and cytochalasin B-treated cells. Normal and cytochalasin B-treated human granulocytes have been studied to determine some of the interrelationships between phagocytosis-induced respiration and superoxide and hydrogen peroxide formation and release into the extracellular medium by intact cells. By using the scopoletin fluorescent assay to continuously monitor extracellular hydrogen peroxide concentrations during contact of cells with opsonized staphylococci, it was demonstrated that the superoxide scavengers ferricytochrome c and nitroblue tetrazolium significantly reduced the amount of H(2)O(2) released with time from normal cells but did not abolish it. This inhibitory effect was reversed by the simultaneous addition of superoxide dismutase (SOD), whereas the addition of SOD alone increased the amount of detectable H(2)O(2) in the medium. The addition of sodium azide markedly inhibited myeloperoxidase-H(2)O(2)-dependent protein iodination and more than doubled H(2)O(2) release, including the residual amount remaining after exposure of the cells to ferricytochrome c, suggesting its origin from an intracellular pool shared by several pathways for H(2)O(2) catabolism. When cells were pretreated with cytochalasin B and opsonized bacteria added, reduced oxygen consumption was observed, but this was in parallel to a reduction in specific binding of organisms to the cells when compared to normal. Under the influence of inhibited phagosome formation by cytochalasin B, the cells released an increased amount of superoxide and peroxide into the extracellular medium relative to oxygen consumption, and all detectable peroxide release could be inhibited by the addition of ferricytochrome c. Decreased H(2)O(2) production in the presence of this compound could not be ascribed to diminished bacterial binding, decreased oxidase activity, or increased H(2)O(2) catabolism and was reversed by the simultaneous addition of SOD. Furthermore, SOD and ferricytochrome c had similar effects on another H(2)O(2)-dependent reaction, protein iodination, in both normal and cytochalasin B cells. When oxygen consumption, O(2.) (-), and H(2)O(2) release were compared in the presence of azide under identical incubation conditions, the molar relationships for normal cells were 1.00:0.34:0.51 and for cytochalasin B-treated cells 1.00:0.99:0.40, respectively. Nonopsonized, or opsonized but disrupted, bacteria did not stimulate any of these metabolic functions. The results indicate that with normal cells approximately 50% of H(2)O(2) released during phagocytosis is derived directly from O(2.) (-) by dismutation, the remainder appearing from an (intra)cellular source shared by azide-inhibitable heme enzymes. With cytochalasin B treatment the evidence is consistent with the derivation of all H(2)O(2) from an O(2.) (-) precursor which is released from the cell surface. Furthermore, when activated by phagocytic particle binding, the neutrophil O(2.) (-) generating system appears to make more of this compound than can be accounted for by dismutation to H(2)O(2). This establishes conditions for the direct participation of both compounds in the microbicidal and cytocidal activity of these cells.


Vasopressin-stimulated prostaglandin E biosynthesis in the toad urinary bladder. Effect of water flow. Prostaglandin E biosynthesis and its effect on water permeability were investigated in the toad urinary bladder. Arginine vasopressin (1 mU/ml) increased prostaglandin E (PGE) biosynthesis from 0.5+/-0.1 to 5.0+/-0.4 pmol/min per hemibladder (mean +/-SEM, n= 8, P less than 0.001). Maximal vasopressin-stimulated PGE biosynthesis, 6.4+/-0.2 pmol/min per hemibladder, occurred at vasopressin concentrations in excess of 3 mU/ml. Half-maximal stimulation of PGE biosynthesis occurred at a vasopressin concentration of approximately 0.7 mU/ml, whereas half-maximal stimulation of water flow occurred at a vasopressin concentration of approximately 5 mU/ml. Vasopressin-stimulated PGE biosynthesis did not depend on water flow along an osmotic gradient or upon sodium transport. Thin-layer chromatographic analysis of the lipids released from hemibladders labeled with tritium-arachidonic acid revealed that vasopressin stimulates the release of arachidonic acid from intracellular lipid stores without affecting the percentage of free arachidonic acid converted to PGE. Neither cyclic AMP nor theophylline stimulated PGE biosynthesis although they mimic arginine vasopressin (AVP) in stimulating water permeability. Biosynthesis of PGE was inhibited by mepacrine, a phospholipase inhibitor, and by agents that inhibit arachidonic acid oxygenase. The inhibition of PGE biosynthesis resulted in augmented vasopressin- and theophylline-stimulated water flow, but had no effect on cyclic AMP-stimulated water flow. We interpret these results to mean that endogenous PGE inhibits basal and vasopressin-stimulated adenylate cyclase activity. In contrast to the effects of AVP on permeability and transport, AVP stimulates PGE biosynthesis by a mechanism that does not depend on an increase in cellular cyclic AMP levels. The water permeability response of the toad urinary bladder to vasopressin is inhibited by PGE synthesized by the bladder in response to vasopressin.


Inhibition of vasopressin-stimulated prostaglandin E biosynthesis by chlorpropamide in the toad urinary bladder. Mechanism of enhancement of vasopressin-stimulated water flow. Chlorpropamide is known to enhance the water permeability response of the toad urinary bladder to vasopressin and to theophylline. In other studies, we have shown that prostaglandin E synthesis by the toad bladder inhibits the water permeability response to arginine vasopressin and to theophylline. In this study, the effect of chlorpropamide on vasopressin-, theophylline-, and cyclic AMP-stimulated water flow and on prostaglandin E biosynthesis was investigated in the toad urinary bladder in vitro. Chlorpropamide inhibited prostaglandin E biosynthesis during vasopressin-, theophylline- and cyclic AMP-stimulated water flow. Tolbutamide and glyburide, two other sulfonylurea compounds, also enhanced vasopressin-stimulated water flow and inhibited vasopressin-stimulated prostaglandin E biosynthesis. We conclude that the mechanism of enhancement on vasopressin-stimulated water flow by the sulfonylureas is the inhibition of prostaglandin E biosynthesis.


Human rotavirus enteritis induced in conventional piglets. Intestinal structure and transport. To better understand the pathogenesis of infantile viral gastroenteritis, we studied Na+ and Cl- fluxes in vitro in short-circuited jejunal epithelium from 8-10-day-old piglets after infection with a standard dose of human rotavirus given via nasogastric tube. 11 infected piglets, all of whom became ill, were compared with 9 uninfected, healthy litter-mates. When killed 72 h after infection, intestinal villi were shorter and crypts deeper (P less than 0.025) in duodenum, upper jejunum, and mid-small intestine, but not ileum in infected piglets. Virus antigen was seen by fluorescence microscopy in occasional jejunal villus tip cells in only four infected piglets and no controls at 72 h. Net Na+ and Cl- fluxes did not differ from noninfected litter-mate controls under basal conditions, but response to glucose was blunted in infected piglets (P less than 0.001). Theophylline stimulated net Cl- secretion in both infected and control animals, and cyclic AMP concentration in isolated jejunal villus enterocytes did not differ significantly. In isolated jejunal villus enterocytes of infected piglets, thymidine kinase activity increased (P less than 0.001), and sucrase activity decreased (P less than 0.001). We conclude that in this invasive enteritis caused by a major human viral pathogen, glucose-coupled Na+ transport is impaired in the jejunum at a time when the villus epithelium shows enzyme characteristics of crypt epithelium, and when little or no virus is present. These findings are identical to those occurring in an invasive coronavirus enteritis of piglets but differ markedly from those seen with enterotoxigenic diarrhea.


An electron microscope observation of the right and the two left portions of the habenular nuclei of the frog. The right dorsal habenula (RDH), the left dorsal habenula, lateral portion (LDH-LP) and the left dorsal habenula, medial portion (LDH-MP) of the adult frog Rana esculenta have been studied by the electron microscope with the use of three types of fixatives: osmium tetroxide, aldehydes and potassium permangarate. The study has demonstrated in the habenulae of both sides a variety of syraptic vesicles, special synaptic patterns (en passant, serial, axo-somatic) containing small spherical transparent vesicles, never mixed with other types of vesicles, and large pale processes containing big granules. However, a particular type of synaptic vesicles characterized by small clear spherical vesicles with a tiny dark granule revealed by potassium permanganate fixition has been demonstrated only in the neuropil of the LDH and not in the RDH. Further, a peculiar type of intracytoplasmic inclusion appears in the form of myelin-like and crystal-like formations only in the LDH-MP. On the basis of these consistent ultrastructural differences, the dorsal habenular nuclei of the frog can be considered as composed of three individual nuclei, each with its own morphological characterization, dislocated, for some unknown reason, one on the right and two on the left side of the epithalamus. The results suggest that the LDH-MP is a nucleus metabolically different from the others and that the projections to the dorsal habenular nuclei are different on the right and left sides of the brain.


Granular cell myoblastoma.

Granular cell myoblastoma. Electron microscopic observations in three cases of granular cell myoblastoma revealed that this tumor is made up of two types of cells. 1) Granular cells grouped into nests, surrounded by a basement membrane, and displaying characteristic inclusions. Numerous processes were observed close to these cells, showing filamentous and microtubular components lying parallel to their axes. 2) Satellite fibroblasts which included angulated bodies. The relationship between these two types of cells is discussed.


Effects of sodium nitroprusside, nitroglycerin, and sodium azide on levels of cyclic nucleotides and mechanical activity of various tissues. Three agents that activate guanylate cyclase, sodium nitroprusside, nitroglycerin and sodium axide, were examined for their effects on cyclic GMP and cyclic AMP accumulation and muscle motility with several tissues. All of these agents, except nitroglycerin with ventricle preparations, increased cyclic GMP levels and did not alter cyclic AMP in incubations of preparations of bovine tracheal smooth muscle, guinea pig tracheal chains, taenia cecum, atria and ventricle, and rat liver and cerebral cortex. Increases in cyclic GMP with these agents occurred with relaxation of smooth muscle preparations and without alteration in the contractility of atrial preparations. These observations support the hypothesis that cyclic GMP accumulation in smooth muscle may be related to relaxation rather than contraction as proposed previously. Relaxation with these agents is not associated with alterations in cyclic AMP levels. Increases in cyclic GMP levels in atrial preparations can also occur without changes in contractile force or rate of contraction.


Distortion of cyclic AMP responses to catecholamine due to destruction of the hormone. The acute actions of low and moderate concentrations of catecholamines on cyclic AMP metabolism in SV40-transformed human lung fibroblasts (VA13) were seriously distorted by non-enzymatic destruction of the agonist. Catecholamine destruction, as measured directly with an isotopic method, was slowed by a variety of anti-oxidants and chelating agents. A combination of two anti-oxidants, ascorbate and thiourea, was very effective in protecting isoproterenol in the cell culture system. That is, there was a 10-fold increase in the sensitivity of VA13 to isoproterenol and the duration of action of the catecholamine was greatly prolonged. However, the anti-oxidants did not alter the responses of the cells to prostaglandins. We conclude that any quantitative studies of cyclic AMP responses to catecholamines must address the question of agonist destruction if meaningful results are to be expected. The use of anti-oxidants, especially the combination of ascorbate and thiourea, would appear to be advisable, particularly in situations where the catecholamine concentrations are less than supramaximal.


Immunohistochemical localization of cyclic GMP in rat cerebellum. The technique of cyclic nucleotide fluorescence immunohistochemistry has been applied for the specific localization of cyclic GMP in rat cerebellum. We report immunofluorescence associated with fibres and membranes, contrasting with previously reported cytoplasmic localization of cyclic AMP in different cell populations, using a similar technique. We have been unable to detect changes in cyclic GMP staining in response to post-mortem changes, harmaline and pentobarbitone administration. A role of cyclic GMP is suggested in membrane ion transport.


Differences in thermal sensitivities of the regulatory-catalytic and catalytic forms of cyclic AMP-dependent protein kinases from various tissues. The cAMP-dependent protein kinase from various tissues was more thermally sensitive when activated by cAMP than the non-activated enzyme. For example, when the activity ratio (the activity of protein kinase assayed -cAMP/+cAMP) was 0.40, 80% and 76% of total hepatic cAMP dependent protein kinase activity was recoverable after incubations at 45 degrees C for 15 and 30 minutes, respectively. However, when the activity ratio was elevated to about 0.80 - 0.90 by increasing cAMP levels in vivo or adding exogenous cAMP to soluble liver extracts, the total protein kinase activity recoverable after incubations at 45 degrees C for 15 minutes was 34-44% and 19-22%, respectively. This observation was used to estimate the degree of activation of the enzyme in vivo and in vitro, since the loss of enzyme activity at 45 degrees C was directly related to the degree of activation of the enzyme in tissue extracts. The regulatory-catalytic form of cAMP-dependent protein kinase was thermally resistant at 45 degrees C unless activated by incubation with exogenous cAMP, histones or NaCl, while the catalytic form of the enzyme was highly thermally sensitive at this same temperature. These data describe a new property of the cAMP-dependent protein kinase and suggest an alternative method which measure the degree of activation of the enzyme.


Effect of exogenous corticotropin and climatic conditions on bovine adrenal cortical function. Interactions between three climatic conditions and exogenous adrenocorticotropin (200 IU) or saline on glucocorticoids of plasma were tested in six pregnant Holstein heifers. Animals were under cold (5 C, 30% relative humidity), thermoneutral (18 C, 50% relative humidity), and hot (35 C, and 80% relative humidity) conditions. Effects of treatment, interactions of environment with treatment, and environment with treatment with interval on glucocorticoids of plasma were measured through timed intervals of blood collection at pre- and postintravenous injection of a treatment solution. Adrenocorticotropin elevated corticoid concentrations in plasma, but interactions of environment with treatment and environment by treatment by sampling interval were nil for responses of corticoids in plasma.


Gallium imaging in pediatrics.

Gallium imaging in pediatrics. Gallium-67 citrate imaging was carried out in 59 children from 3 mo to 20 yr of age. Indications for the study included the search for occult inflammatory disease and the detection and staging of malignant disease. The Ga-67 citrate scan had a 96% reliability in confirming or excluding the site of purulent material as a cause of sepsis, and 76% reliability in detecting malignant disease. Tissue distributions in children differ from those in adults primarily in that the epiphyseal plates, spleen, and thymus may show increased activity normally and in the presence of sepsis. These variations are illustrated as possible sources of incorrect interpretation. "Cold" defects may be seen in sterile collections and avascular masses. Experience in this age group seems sufficiently encouraging to continue the use of gallium-67 citrate for the purposes described.


A case of myocardial abscess evaluated by radionuclide techniques: case report. A patient with infective endocarditis was evaluated by Ga-67 citrate imaging, Tc-99m pyrophosphate imaging, equilibrium gated blood pool imaging, and Tl-201 imaging of the chest. The diagnosis of ventricular abscess was first suggested by an abnormal gallium scan. At surgery, an abscess was identified in the area where the scan was abnormal, and postoperatively a repeat scan was normal.


Clinical evaluation of the Cleon imager. We have compared bone images from a number of patients using three instruments. In 38 patients, Cleon body images were compared with whole-body rectilinear scans. Cleon images were also compared with scintillation camera images of the trunk or extremities of 31 patients and of the skull of 70 patients. The Cleon was superior to the rectilinear scanner in resolution, lesion detectability, and speed of scanning. The Cleon and gamma camera were comparable in lesion detection, but the Cleon was consdierably faster. Clinical studies and comparative evaluation are presented.


The calcium and vitamin D status in an elderly female population and their response to administered supplemental vitamin D3. Recent reports have implicated the importance of impaired calcium absorption, inadequate dietary vitamin D, and low serum vitamin D levels in the genesis of metabolic bone diseases in the elderly. This study evaluated the dietary intakes of calcium and Vitamin D, calcium absorption, and serum 25 hydroxycholecalciferol (25 OH D) levels in a population of women (mean age 83 yrs) compared to healthy volunteers (mean age 35 yrs). Dietary intakes of both calcium and vitamin D and serum 25 OH D levels were found to be comparable in both groups. Calcium absorption was normal in the subjects studied and did not change significantly after two 2-week periods of oral supplementation with vitamin D3 in doses of 500 I.U./day and 10,000 I.U./day, although the serum 25 OH D levels rose significantly and comparably in both groups. Thus other etiological factors may play relevant roles in the causation of bone diseases in the elderly and unless deficient dietary or serum vitamin D levels are demonstrated, empirical supplementation with vitamin D should not be undertaken. The differences of the above findings from previously published data are discussed.


[Indications for hysterectomy in placental tumors]. Indications for hysterectomy in placental tumours are now most often to carry out hysterectomy as a second attack designed to deal with residual lesions of choriocarcinoma that prove resistant to chemotherapy. Of 80 patients treated, 24 were first seen for chemotherapy after hysterectomy and this is too great a number. On the remaining 56 patients we carried out hysterectomy in 8 cases of whom 7 were secondary to chemotherapy. Seven times we found a residual lesion. These patients are now apparently cured.


[Radioimmunoassay of HCG and HCG beta in blood during pregnancy and trophoblastomas]. This work was carried out on 151 estimations of the radioimmune levels of hCG-hCG beta in the first half of normal pregnancy and 390 levels estimated in 10 molar pregnancies, in 4 choriocarcinomata following moles and 1 primary choriocarcinoma of the ovary using homologous hCG beta. In normal pregnancy the level of hCG-hCG beta rises rapidly. The levels are highest between the 8th and 9th weeks of pregnancy, and then gradually decrease. In active molar pregnancies the levels of hCG-hCG beta in the plasma are significantly higher than those found in normal pregnancies of the same gestational age. When the outcome is going to be favourable the levels drop progressively. Complete absence in peripheral blood is usually found about 14 weeks after a mole has been expelled or evacuated. In choriocarcinomata, if the level of hCG-hCG beta rises it is a certain sign of reactivation of the tumour.


Protein A-peroxidase: a valluable tool for the localization of antigens. Protein A of Staphylococcus aureus has been conjugated to horseradish peroxidase and used in an indirect immunolabeling technique to visualize membrane and viral antigens. The same Protein A-peroxidase conjugate was used with antisera from five different species. Using this indirect test, membrane markers for T and B lymphocytes were labeled with a greater specificity than when peroxidase conjugated anti-immunoglobulin was used in the second step. Viral antigens on cells infected with measles, vesicular stomatitis, herpes or visna virus, respectively, were also stained in the protein A-peroxidase indirect test with a greater specificity than indirect method using anti-immunoglobulin. Paired preparations were examined in the light and electron microscope. Ultrastructural analysis showed that the protein A-peroxidase conjugate penetrated well through fixed viral membranes and resulted in fine resolution of antigenic sites.


Thymus-derived lymphocyte-dependent rejection of syngeneic papovavirus (SV40) and methylcholanthrene tumors in inbred hamsters. Treatment of specifically sensitized MHA hamster lymphoid cells with rabbit antisera specific for hamster thymus-derived lymphocytes, in the presence of C, eliminated those cells capable of inhibiting the growth of syngeneic SV40 and methylcholanthrene tumors in vivo. Thymectomized, lethally-irradiated, bone marrow-reconstituted hamsters, shown to be devoid to T cell function, were, after attempted specific sensitization to the two syngeneic tumor cell lines, unable to reject either tumor by direct challenge in vivo. In addition, lymphocytes from such animals were incapable of inhibiting the growth of either tumor cell line in normal syngeneic recepients in the tumor cell neutralization assay. These data strongly support the conclusion that specifically sensitized thymus-derived lymphocytes are required for the rejection of syngeneic SV40 and methylcholanthrene tumors in inbred hamsters.


Modulation of Fc-mediated phagocytosis by cyclic AMP and insulin in a macrophage-like cell line. Studies on genetically selected mutants in phagocytosis of E[IgG] indicated that the defect in some mutants could be corrected by addition of 8 Br-cAMP, and suggested that cyclic AMP may be involved in the mechanism of phagocytosis through the Fc receptor. In order to elucidate the role of cyclic AMP in phagocytosis in the parental, nonmutant macrophage-like cell line, J774.2, it was necessary to employ restrictive conditions which rendered phagocytosis suboptimal. When 4774.2 cells were cultured in nontissue culture Petri dishes phagocytosis was markedly reduced. Addition of 8 Br-cAMP or inducers of intracellular cyclic AMP such as isoproterenol restored the phagocytic ability of these cells. Similarly, treatment of the parental J774.2 cells with insulin reduced the level of phagocytosis, and once again this suppression could be corrected by addition of 8 Br-cAMP. In no case did AMP mimic the effects of 8 Br-cAMP. The effect of cyclic AMP action in this system was not instantaneous, but rather reached optimal levels at 5 to 10 hr, suggesting that cyclic AMP is not the immediate signal for phagocytosis. The genetic analysis of macrophage variants may provide a useful model for studies on the mechanisms of phagocytosis, and also the effects of insulin and cyclic AMP on an easily measurable biologic function in a specialized cell type.


Resistance of Bordetella pertussis phase I to mucociliary clearance by rabbit tracheal mucous membrane. Cultivated mucous membrane of rabbit trachea was point inoculated with Bordetella pertussis phase I or III. Phase I (virulent) bacteria were found to be infective at the point-inoculated site, but phase III (avirulent) bacteria rarely showed such behavior. After inoculation, homogenized segments of mucous membrane were spread on plates. Large numbers of phase I bacteria were recovered from the inoculated segment; however, the laryngeal segment was the site of recovery of large numbers of phase III bacteria. The difference was not due to the ciliostatic bacterial toxin. After coinoculation with phase I and III bacteria, phase III bacteria were recognized at the laryngeal end, whereas phase I bacteria were recognized at the inoculated site. Therefore, the adherence of the bacteria to the ciliated epithelium is considered to be the most probable mechanism of the resistance to the mucociliary stream. Scanning electron microscopy facilitated visualization of phase I bacteria adhering to the inoculated site.


Regional localization of activity of Clostridium perfringens type A enterotoxin in the rabbit ileum, jejunum, and duodenum. Rabbit ileal, jejunal, and duodenal loops were exposed to purified enterotoxin from Clostridium perfringens type A and then perfused for comparative analysis of effects of the enterotoxin on each region of the intestine. Ileal loops responded with enhanced net secretion of fluid and sodium, inhibition of chloride and glucose uptake, and substantial sloughing of epithelial cells. The jejunum responded with fluid secretion, enhancement of sodium secretion only during the first 20 min, inhibition of chloride and glucose uptake, and substantial sloughing of epithelial cells. In the duodenum, transport of fluid, sodium, and chloride was significantly altered only during the first 20 min of perfusion, and significant inhibition of glucose uptake varied from one period to another. Epithelial damage was much less than that seen in the jejunum or ileum. Levels of fluid protein in all three sections corresponded closely to extent of tissue damage. In general, it was found that the severity of response to fixed doses of enterotoxin varied as follows: ileum greater than jejunum greater than duodenum.


Interaction of cytomegalovirus with leukocytes from patients with mononucleosis due to cytomegalovirus. Cytomegalovirus (CMV) was isolated from hemic cells from seven of seven patients with acute CMV mononucleosis and from one of six patients with mononucleosis during convalescence. CMV was isolated from the mononuclear leukocyte fraction, the polymorphonuclear leukocyte fraction, or both cell fractions. Virus was not detected in washed erythrocytes, plasma, or leukocyte lysates. Mononuclear leukocytes from patients with acute CMV mononucleosis displayed an elevated level of incorporation of [3H]thymidine on the day of donation compared with that in cells from convalescent patients or normal donors. Responses to pokeweed mitogen and concanavalin A were significantly lower in patients with CMV mononucleosis than in normal donors. There were no significant differences among the acute, convalescent, and normal donor groups in response to phytohemagglutinin or in the one-way mixed leukocyte reaction. These findings suggest that in patients with CMV mononucleosis virus infects and may persist within peripheral blood leukocytes and that the lymphocytes of these patients are selectively hyporesponsive to certain mitogens.


Herpetic whitlow as part of genital virus infection. Lesions on fingers or hands caused by Herpesvirus hominis have in the past been attributed to inoculation with virus-containing oropharyngeal secretions. Often these "herpetic whitlows" were caused by H. hominis type 1 and occurred in medical personnel. A recent series of 13 recurrent "herpetic whitlows" yielded 11 isolates of H. hominis type 2 and only two of H. hominis type 1. None of the subjects was engaged in patient care, and none had oral or facial herpetic lesions. Nine of the 11 infections due to H. hominis type 2 were in females, and more than half were associated with genital herpetic lesions. Herpetic finger or hand lesions in adults appear to be a common part of genital virus infections caused by H. hominis type 2 and are not associated with infections acquired in hospitals or during primary gingivostomatitis.


Clinical and histological malignancy of adenoid cystic carcinoma. The authors report on 25 cases of adenoid cystic carcinoma of the head and neck region. The tumours were divided into 4 histological malignancy groups. Treatment generally consisted of surgery performed with as large a margin as possible, in combination with split course million volt therapy, about 6,000 rads, usually given preoperatively. It was found that the patients in the groups of lower malignancy generally managed for a comparatively long time without a clinically demonstrable recurrence, while the patients in the groups of higher malignancy relapsed rather quickly and often had metastases. The patients in the groups of higher malignancy also died relatively soon after the histological tumour preparations.


Cryptophthalmos: symptoms and treatment of a rare deformity. A case report. Since 1872 more than 50 cases of the cryptophthalmos syndrome have been reported. In addition to other deformities, the typical anomalies of this syndrome are the missing palpebral fissure, eyebrow and the rudimentary eyeball. This paper deals with the possibilities of operative correction in a 13-year-old female. We are of the opinion that by means of plastic surgery, social integration into the community of the patient is possible.


[Metabolism and action of intracellular magnesium (author's transl)]. The activity of Mg2+-activated enzymes shows a bell shaped pMg dependency, with a pMg optimum at 3. Intracellular Mg2+ concentrations, determined by different methods, are about 10(-3) mol/1. Thus Mg2+-dependent enzymes are optimally activated, or nearly so, with Mg2+. If the substrates of an enzyme form complexes with Mg2+ of different stabilities, and if free substrate and the substrate-Mg2+ complex react differently with the enzyme, the equilibrium will change with the concentration of Mg2+. When the extracellular concentration of Mg2+ is increased, Mg2+ becomes bound practically exclusively to the cell membrane. In Mg2+ deficiency, the intracellular concentrations of Na+, K+, Ca2+ and cycl. AMP are changed, and the rates of synthesis of DNA, RNA and protein are decreased.


Cation and anion sequences in dark-adapted Balanus photoreceptor. Anion and cation permeabilities in dark-adapted Balanus photoreceptors were determined by comparing changes in the membrane potential in response to replacement of the dominant anion (Cl-) or cation (Na+) by test anions or cations in the superfusing solution. The anion permeability sequence obtained was PI greater than PSO4 greater than PBr greater than PCl greater than Pisethionate greater than Pmethanesulfonate. Gluconate, glucuronate, and glutamate generally appeared more permeable and propionate less permeable than Cl-. The alkali-metal cation permeability sequence obtained was PK greater than PRb greater than PCx greater than PNa approximately PLi. This corresponds to Eisenman's IV which is the same sequencethat has been obtained for other classes of nerve cells in the resting state. The values obtained for the permeability ratios of the alkali-metal cations are considered to be minimal. The membrane conductance measured by passing inward current pulses in the different test cations followed the sequence, GK greater than GRb greater than GCs greater than GNa greater than GLi. The conductance ratios obtained for a full substitution of the test cation agreed quite well with permeability ratios for all the alkali-metal cations except K+ which was generally higher.


Replication of enterovirus 70 in non-primate cell cultures. Replication of the strain J 670/71 of enterovirus 70 (EV70) in non-primate cell cultures at 33 degrees C was studied using strain L (mouse), BHK21 (hamster), RK13 (rabbit), RK17 (rabbit), PK15 (porcine), IB-RS-2 (porcine), ESK (porcine), MDBK (bovine), and BK1 (primary bovine) together with that of the LSc, 2ab strain of poliovrus type 1 (PV1) as a control. All the cells tested adsorbed from 54 to 90% of EV70. The replication with complete c.p.e. was evident in RK13, RK17 and BK1 cells; replication without c.p.e. was shown in L, BHK21, IB-RS-2 and ESK; but PK15 and MDBK were non-permissive despite a high virus adsorption rate. On the contrary, none of these non-primate cells allowed the adsorption and growth of PV1. One-step growth of EV70 in RK13 was almost identical with that in the primate cells. Two other strains of EV70 were found having similar host range in vitro. Therefore, it is concluded that EV70 has a wider host range in vitro than ordinary human enteroviruses, and its implication is discussed.


The effect of temperature on the synthesis of rhinovirus type 2 RNA. The reduced yields of rhinovirus type 2 at temperatures above 37 degrees C were shown to result from the degradation of virus-induced RNA, leaving little RNA available for inclusion into mature infectious virions. The degradation occurred about 6 h p.i., and appeared to be selectively effecting the single-stranded species. Lysosomal nucleases do not appear to have a role in this supra-optimal degradation.


Growth of cytomegalovirus at supra-optimal temperatures. An isolate (Mira) of cytomegalovirus is shown to replicate in human embryonic lung fibroblasts at supra-optimal temperature (40 degrees C). The ability of the Mira isolate to grow at 40 degrees C decreased as a function of age of cells in which the virus was grown. The unusual morphology of the lesions in late passage cells infected and maintained at 40 degrees C is illustrated.


Some properties of recombinants between type 1 and type 2 herpes simplex viruses. Four intertypic recombinants of herpes simplex virus have been shown to possess genetic information for functions characteristic of each of the two parental types. The functions were identified by (a) polyacrylamide gel electrophoresis of purified virus particles and of polypeptides synthesized in cells infected with the recombinants and (b) analysis of antigenic sites interacting with type specific neutralizing antibody. The analysis shows that each recombinant possess a different combination of these type specific markers. Finally we have been unable to detect recombination between herpes simplex type 1 and pseudorabies viruses.


Studies on the structure of a coronavirus-avian infectious bronchitis virus. When avian infectious bronchitis virus (IBV) is fixed in formaldehyde, negative stain is able to penetrate the particle and an internal component is visualized. This component is seen as a tongue or flask shaped structure attached at one point to the outer virus membrane. A model yielding transmission patterns similar to the virus has been made. Gradient centrifugation studies on IBV reveal that the RNP is associated with the internal sac.


Biological properties of avian coronavirus RNA. RNA with a sedimentation coefficient of 64S was isolated from infectious bronchitis virus, an avian coronavirus. The SNA contained a polyadenylic acid tract and was found to be infectious.


The circular, segmented nucleocapsid of an arenavirus-Tacaribe virus. The nucleocapsid structures of Tacaribe virus, a member of the Arenaviridae, were purified from detergent-treated virus particles by equilibrium density gradient centrifugation. Negative-contrast electron microscopy indicated that they were coiled, circular filaments. They had a mean diam. of 5 to 10 nm and two predominant length classes of 640 nm and 1300 nm were found.


Infectious DNA from cells infected with feline syncytium-forming virus (Spumavirinae). DNA isolated from cells infected with FSFV (a foamy virus) is infectious when tested on susceptible cells. The virus produced by this infectious DNA is identical to the original infecting virus in terms of plaque and virion morphology and serology.


Resistance to murine cytomegalovirus linked to the major histocompatibility complex of the mouse. Studies of the resistance patterns to infection with a murine cytomegalovirus in inbred strains of mice revealed the existence of resistant and susceptible strains. Resistance was found to be associated with possession of the H-2k allele at the major histocompatibility locus of the mouse. The F1 hybrid between a resistant strain (C3H/HeJ) and a susceptible strain (BALB/c) was found to have a resistance intermediate between that of both parents, indicating that the gene(s) controlling resistance is partly dominant. Susceptible BALB/c mice could be made resistant to lethal infection by pre-treatment with thioglycollate broth but not by pre-treatment with endotoxin or BCG. Resistant C3H/HeJ mice could be made susceptible to lethal infection by pre-treatment with cyclophosphamide.


Localization of the restrictive event of EMC virus replication in semi-permissive monkey and monkey-mouse hybrid cells. Encephalomyocarditis (EMC) virus replication was investigated in permissive mouse MKS cells, semi-permissive monkey CV1 cells, and in somatic monkey-mouse MKCVIII hybrid cells whose permissiveness is under the negative control of the simian genome. We found that in CV1 cells the synthesis of both single- and double-stranded virus RNAs was restricted. In contrast, in semi-permissive hybrid Cl4/3 cells only the single-stranded virus RNA was synthesized in small amounts, whereas the double-stranded virus RNA accumulated late after infection. The synthesis of virus polyribosomes and virus polypeptides was lowered in semipermissive conditions. In the presence of quaternary ammonium ions, the synthesis of EMC virus was partially relieved in CV1 cells. Thus, it can be postulated that a defective function in the replication complex is involved in the restrictive event.


Separation and infectivity of two particle types of human rotavirus. Two morphological types of human rotavirus particles were separated in caesium chloride density gradients. The particles of higher density ('incomplete' particles without an outer capsid layer) banded at a density of 1-38 g/ml, while the less dense ('complete' particles with an outer as well as an inner capsid layer) banded at a density of 1-36 g/ml. Some particles were found with an incomplete outer layer of capsomeres. The particle/infectivity ratio for tissue cultures of the fraction containing complete particles was more than 3 log10 higher than that of the fraction containing incomplete particles. But, as there was a small number of complete particles in the fraction containing mostly incomplete particles, it was impossible to determine whether the incomplete particles had a low infectivity or whether they had none at all.


Effects of cycloheximide and puromycin on the antiviral and anticellular activities of human interferon. Human leucocyte interferon has antiviral activity and anticellular effects on the transformed human cell lines, RSa and RSb. Treatment of the cells with cycloheximide or puromycin at 0-5 to 5-0 microgram/ml suppressed the antiviral action of interferon but increased its anticellular effects. Interferon also has antiviral activity in IFr cells, but this is relatively resistant to its anticellular action. Nevertheless, both drugs suppressed the antiviral activity of interferon and increased its anticellular action.


Induction of interferon in human lymphoblastoid cells by Sendai and measles viruses. Sendai and measles viruses were tested for their interferon-induced capacity in human lymphoblast cells. Sendai virus reproducibly induced considerable amounts of interferon (I research reference interferon unit/10(3) cells), but no increase in infectious virus titre was observed. Two Edmonston-Enders strains of measles virus grew very well. The attenuated (A) strain was a good interferon inducer (4 units/10(3) cells), while the virulent (V) strain induced only minimal amounts at a high multiplicity of infection. Pre-treatment of the cells with 5-iododeoxyuridine (IdUrd) had no effect on the growth of Sendai or EE measles virus and on interferon yields induced by Sendai virus. It slightly potentiated interferon induction by EE measles virus. Partial u.v.-light-inactivation of Sendai virus infectivity resulted in a parallel loss in interferon-inducing capacity.


Circular and circular-linear DNA molecules of herpes simplex virus. Circular and circular-linear DNA molecules of herpes simplex virus were isolated from infected cells. Two types of circular-linear DNA molecules are described, one with a circular component of about 8 micrometer and a linear component of 45 micrometer and the other with a circular component of 45 micrometer and a linear component of 8 micrometer. The circular DNA molecules were either the size of linear DNA molecules or were shorter and corresponded to the length of the L (long) component of linear virus DNA.


Cell DNA replication as a function in the synthesis of human cytomegalovirus. The rate of virus and cell DNA synthesis was studied in human embryonic lung cells pre-treated with 5-iodo-2'-deoxyuridine (IdUrd) and exposed to cytomegalovirus (CMV) or medium. Analysis of DNA in CMV-infected cells following sequential 4 h pulses with 3H-thymidine indicated that a temporal relationship existed in the pattern of virus and cell DNA synthesis. The pattern of DNA replication in infected cells resembled that of a typical cell cycle, whereas the rate of cell DNA synthesis in uninfected cells remained low throughout the study. Increased rates of cell and virus DNA synthesis began concomitantly at 16 h post-infection and reached a maximum at 36 h post-infection. The rate of DNA synthesis then declined and remained at lower levels until 48 h post-infection. This was subsequently followed by a second increase in the rate of cell and virus DNA synthesis. The rates of cell and virus DNA replication were similar throughout the study in that increased and decreased rates of synthesis occurred simultaneously. It was of interest to note that CMV induced cell DNA replication in IDUrd arrested cells; in contrast, addition of fresh serum did not induce a similar increase in the rate of DNA synthesis in IdUrd arrested, but uninfected, cells.


Phophorylation of polyoma and SV40 virus proteins. The polypeptides of polyoma and SV40 virions are phosphorylated. An estimate of the amount of phosphorylation of the major virus capsid protein (VPI) has been made using two-dimensional gel electrophoresis to resolve phosphorylated from non-phosphorylated forms. The results suggest that in both polyoma and SV40 virions about 12% of VPI molecules are phosphorylated. In unassembled VPI molecules immunoprecipitated from extracts of infected cells the proportion is greater, about 33%. The possibility that phosphorylated VPI may form the penton proteins of the virus capsid is discussed.


Sarcoid-like disorder of the intracranial optic nerve. Two cases with a sarcoid-like disorder apparently restricted to the intracranial optic nerve and the adjacent dura mater are described, doublilng the number of reported cases. In both instances there were unilateral visual loss and unusual radiographic changes in the optic canal area. Biopsy samples showed localised infiltration of tissues with non-caseating tubercles containing epithelioid and multinucleated giant cells.


Sleep apnoea syndrome: states of sleep and autonomic dysfunction. Eleven patients with upper airway apnoea during sleep (one with SHY-Drager syndrome) were monitored polygraphically for wakefulness, sleep, and cardiovascular variables. Systemic hypertension and most of the severe arrhythmias recorded during sleep were secondary to repetitive obstructive apnonea and were mediated through the autonomic nervous system. Sleep related elevations of pulmonary arterial pressure were not influenced by atropine or impaired autonomic functions. Upper airway sleep apnoea is sleep related; the type of sleep (REM or NREM) is critical in the appearance of abnormalities. The distinction between two patient subgroups (total sleep dependent and NREM sleep dependent) has haemodynamic, and possibly long-term, implications. Sleep apnoea syndrome should be looked for in pateints with the Shy-Drager syndrome.


Immunofluorescence studies in a case of rheumatoid neuropathy. Findings in a sural nerve biopsy from a patient with rheumatoid neuropathy are reported. Inflammatory changes in different stages were observed in epineurial arterioles. Arterioles with fibrinoid necrosis contained IgG, IgM, rheumatoid factor and complement. The same substances were found at the inner site of the perineurial sheaths. It is suggested that these proteins leaked out of endoneurial vessels and became trapped in the perineurial sheaths. With anti-herpes simplex virus serum fluorescence was seen in monuclear cells of infiltrated vessel walls. This virus could not be isolated in tissue culture.


Respective importance of different electrophysiological parameters in alcoholic neuropathy. An electrophysiological study of alcoholic and normal subjects is presented. The aim was to evaluate the respective importance of the various parameters. The subjects were divided into 4 groups: (I) normal subjects; (II) chronic alcoholics without clinical evidence of neuropathy; (III) chronic alcoholics with sensory symptoms; (IV) chronic alcoholics with both motor and sensory symptoms. The electrophysiological parameters tested were: conduction velocity (CV) in Ia sensory fibres, motor fibres, and cutaneous sensory fibres of the popliteal nerve, CV in the sural nerve, amplitude of the cutaneous sensory action potentials (SAP) in the sural and popliteal nerves, H reflex and M response of the soleus muscle, and electromyograms from the extensor digitorum brevis muscle. In the 3 groups of alcoholics, the electrophysiological findings were more abnormal than the clinical symptoms could have predicted. The more sensitive parameters were: (1) CV in the Ia sensory fibres of the popliteal nerve, which is slowed very early (in Group II) and (2) measurement of the amplitude of sural and popliteal SAP's which are also reduced early (in Group II). The nature of the mechanisms involved are discussed.


Electron-microscopic appearance of the DA virus, a demyelinating murine virus. The DA virus is a neurotropic murine virus which can induce acute encephalomyelitis in suckling mice and a chronic myelopathy in weanlings. The agent has been attenuated by serial passage in baby hamster kidney (BHK-21) cells. When attenuated virus is inoculated in 8-week-old C3HeJ mice a myelopathy of delayed onset with prominent demyelination of lateral and anterior columns occurs. The DA virus is believed to be related to the Theiler murine encephalomyelitis (TME) viruses because of the similar clinical and pathological conditions which it causes, and because neutralization tests indicate shared antigens between it and GD7, a TME virus. This paper reports electron-microscopic studies of BHK-21 cells infected with DA virus. The cells were prepared 24 and 48 hr after inoculation. Cytopathic effects were observed and infected cells contained plaques consisting of numerous 25 nm virus particles in crystalline array. The virions were exclusively intracytoplasmic and were morphologically indistinguishable from human poliomyelitis virus. These observations appear to establish DA as a picorna virus, related to the TME virus group. The chronic myelopathy caused by DA may prove relevant to chronic demyelinative myelopathies in man, such as multiple sclerosis, and also to amyotrophic lateral sclerosis.


Myotonic dystrophy with nerve hypertrophy. Report of a case with electrophysiological and ultrastructural study of the sural nerve. A striking enlargement of peripheral nerves was observed in a patient with myotonic dystrophy (Steinert's disease). Clinical findings disclosed a polyneuropathy which was confirmed by electrophysiological and morphological investigations. Myopathic as well as neurogenic alterations were found on electromyography in wasted hand muscles. The prominent finding on ultrastructural study of sural nerve biopsy was an important collagen infiltration with rather few "onior bulb" formations. This extensive endoneural collagen formation appeared to be responsible for the nerve hypertrophy. Evidence of segmental demyelination and remyelination was also found. Glycogen deposits were present in some myelinated axons and is some Schwann cells. Peculiar crystalline inclusions were observed in fibroblasts processes. The relation between myotonic dystrophy and polyneuropathy remains a matter of discussion.


Vitamin E binding proteins in human serum. The distribution of vitamin E in human serum lipoprotein fraction was investigated by measuring the amount of vitamin E in various protein fractions prepared by ultracentrifugation, gel filtration and electrophoresis. The larger part of the total serum vitamin E (up to 44%) was present in the alpha1-lipoproteins, while the beta-lipoproteins contained smaller concentrations (up to 26%).


Application of 13C-NMR spectroscopy to in vitro analysis of enzyme kinetics. The conversion of D,L-alpha-13C-histidine to similarly labeled alpha-13C histamine by bacterial and mammalian histidine decarboxylase was studied by 13C-NMR spectroscopy and GLC-mass spectrometry. The results obtained with the partially purified bacterial enzyme were in essentially perfect agreement with results obtained simultaneously with a standard  radioisotopic  method using carboxyl-labeled-14C-L-histidine. For a crude tissue preparation of the mammalian enzyme, the radioisotopic method indicated an activity three times that based on 13C-NMR measurement of alpha-13C-histamine. The difference in results was accountable in terms of additional 13C-NMR signals attributable to  products other than histamine due in part to enzymatic degradation of the latter.


A role for endogneous prostaglandins in the short-circuit current responses to osmolal changes in isolated frog skin. 1. Reduction in osmolality of the Ringer solution bathing the morphological inside of frog skin (by lowering the NaCl concentration) caused a significant increase in sodium transport as measured by the short-circuit current. Pretreatment of the skin with acetylsalicylic acid (2.5 x 10(-4)M) abolished the short-circuit current and open-circuit potential responses to osmolal change.2. The output of prostaglandin-like material from isolated frog skin was increased by incubating the skin in hypotonic Ringer solution.3. The cyclic AMP levels of isolated frog skin were also increased by a reduction in the osmolality of the Ringer fluid bathing the skin.4. Prostaglandin-like material was released both by the separated epithelial and dermal layers of frog skin and the output from both layers, on a unit wet weight basis, did not differ.5. The output of prostaglandin-like material from the separated layers of the skin was substantially greater than from whole skin. Indomethacin (6 x 10(-6)M) reduced the output of this material by more than 90% from both layers.6. The release of prostaglandin-like material from the separated layers of the skin was not altered by a reduction in osmolality of the bathing medium.7. It is concluded that a reduction in the osmolality of the solution bathing frog skin stimulates prostaglandin production and that the increased level of prostaglandins stimulates transepithelial sodium transport by stimulating cyclic AMP accumulation. It is also concluded that the response to osmolal change can only occur in intact skin since separation of the epithelial and dermal layers abolished the increase in the release of prostaglandin-like material to osmolal change. The site of the increased prostaglandin production and the exact nature of the stimulus remain to be determined.


[A study on the gustatory effects of tetrodotoxin in rat (author's transl)]. Effects of tetrodotoxin (TTX) on neural responses of the chorda tympani to four basic taste stimuli were investigated electrophysiologically in rats. When the TTX (10 mg/ml) was applied directly to the tongue surface for 3 minutes, magnitude of the integrated responses of the chorda tympani was diminished to about 60% of that of the control response. This diminution of response was recovered within 30 minutes by degrees and the effect of the TTX was antagonized by guanylate. This result gives a suggestion that guanidyl group in the TTX may play an important role for the inhibitory actions to the responses of the chorda tympani. On the other hand, when the TTX (0.25 mg/100 g b. wt.) was applied intravenously, magnitude of the responses of the chorda tympani to four basic taste stimuli decreased gradually to 20 approximately 30% of that of the control responses within 60 minutes and did not recover more than 10 hours. This is assumed due to the blocking of the sodium pump of nerve fibers in the chorda tympani by the TTX.


Binding of LH and FSH to porcine granulosa cells during follicular maturation. The uptake of 125I-labelled LH by equal numbers of granulosa cells from small, medium or large follicles was greater by cells from large follicles. In contrast, granulosa cells obtained from small follicles bound much more 125I-labelled FSH per cell than did cells obtained from medium and large follicles. Competition studies with unlabelled hormones indicated that porcine granulosa cells have specific receptors for LH and FSH. The addition of diethylstilboestrol enhanced the binding of 125I-labelled LH and inhibited the binding of 125I-labelled FSH to granulosa cells harvested from small and medium-sized follicles, but had no effect on those from large follicles.


Radioreceptor assays: plasma membrane receptors and assays for polypeptide and glycoprotein hormones. Receptors for peptide, protein and glycoprotein hormones, and the catecholamines are located on the plasma membranes of their target cells. Preparations of the receptors may be used as specific, high-affinity binding agents for these hormones in assay methodology akin to that for radioimmunoassay. A particular advantage of the radioreceptor assay is that it has a specificity directed towards the biologically active region of the hormone, rather than to some immunologically active region that may have little (or no) involvement in the expression of hormonal activity. Methods for hormone receptor preparation vary greatly, and range from the use of intact cells (as the source of hormone receptor) to the use of purified or solubilized membrane receptors. Receptors isolated from plasma membranes have proved to be of variable stability, and may be damaged during preparation and/or storage. Moreover, since they are present in relatively low concentration in the cell, their preparation in sufficient quantity for use in a radioreceptor assay may present technical problems. In general, there is good correlation between radioreceptor assays and in-vitro bioassays; differences between results from radioreceptor assays and radioimmunoassays are similar to those noted between in-vitro bioassays and radioimmunoassays. The sensitivity of the method is such that normal plasma concentrations of various hormones have been assayed by this technique.


Synthesis of beta-spiro[pyrrolidinoindolines], their binding to the glycine receptor, and in vivo biological acitivity. A series of beta-spiro[pyrrolidinoindolines], 3a-d, was prepared and evaluated for their ability to bind to the glycine receptor. These compounds were also tested in vivo to determine if they would produce convulsant or anxiolytic effects. The target indolines were chosen because they represent rings A, B, E, and a portion of ring C of strychnine. Results of this study indicate that, in this series, an acetylindoline in the endo configuration and a tertiary amine, such as that of the pyrrolidine ring nitrogen, are required for biological activity. In all of the cases studied, the activity was of a convulsant rather than a relaxant nature. Excellent correlation was found to exist between the binding affinities to the strychnine site of the glycine receptor and clonic convulsions (ED50) and death (LD50) in the mouse.


Heterocyclic and piperonylic acid esters of 1-methyl-4-piperidinol as analgesics. Heterocyclic and piperonylic acid esters of 1-methyl-4-piperidinol were synthesized and evaluated for analgesic activity. 1-Methyl-4-piperidinol 4-piperonylate (14) exhibited activity in the codeine range (mouse hot plate). In monkeys, 14 acted neither as a typical narcotic agonist nor as a typical antagonist and it showed no physical dependence liability of the morphine type. The nonquaternary C-4 peperidinol esters 1a, 4, 8, and 14 exhibited marginal to virtually no binding to the opiate receptor in rat brain homogenates. The interaction of various functional groups of this series with potential binding sites of a nonopiate type receptor is discussed.


Synthesis and biological activity of two metabolites of 1-methyl-5-(1-methylethyl)-2-nitro-1 H-imidazole, an antiprotozoal agent. 5-(-Hydroxyl-1-methylethyl-)-1-methyl-2-nitro-1 H-imidazole (6) and 5-(1,2-dihydroxyl-1-methylethyl)-1-methyl-2-nitro-1 H-imidazole (9) are the principal metabolites found in urines of animals (mice, rats, and dogs) treated with 1-methyl-5-(1-methylethyl)-2-nitro-1 H-imidazole (1), an effective antitrichomonas agent. These two metabolites have been synthesized. Compound 6 was found to be less toxic than the parent compound 1 and to possess essentially the same activity against Trichomonas vaginalis in experimental infections. Compound 9 showed a low degree of in vivo activity.


Radioimmunoassay of herpes-simplex and measles virus antibodies in serum and cerebrospinal fluid of patients without infectious or demyelinating diseases of the central nervous system. A solid-phase radioimmunoassay was used to detect IgG antibodies against herpes-simplex virus antigens (capsid, envelope and excreted) and against measles virus antigen in serum and cerebrospinal fluid (CSF) specimens of 61 patients with no evidence of infectious or demyelinating disease of the central nervous system. Quantitative determinations of IgG and albumin in serum and CSF were also performed. Of the 61 serum and 61 CSF samples tested, 57 and 56 respectively contained antibodies against subunit antigens of herpes simplex virus. Antibody against measles virus was found in 59 serum and 47 CSF specimens. A positive correlation (P less than 0-001) was found between each of the four serum to CSF antibody ratios and the serum to CSF total IgG ratios. This indicated that the distribution of antiviral IgG antibodies in serum and CSF normally follows the distribution of total IgG. The ratios between viral antibody in serum and CSF were also correlated with albumin ratios (P less than 0-05). An inverse relation (P less than 0-001) was found between the age of the patients and their serum to CSF albumin ratios, but not their IgG ratios, suggesting that the albumin ratio is a useful indicator of a blood brain barrier lesion and that the IgG ratio should be used in evaluating disturbed antibody ratios.


Cytomegaloviru; antibody in the urine of renal transplant patients. Small amounts of cytomegalovirus (CMV) antibody were detected in the urine of renal transplant patients excreting the virus. The antibody was probably produced locally, as a result of active CMV infection of the urinary tract.


Interaction of Simian Virus 40 chromatin with Simian Virus 40 T-antigen. We have studied the binding of the tumor antigen (T-antigen) of simian virus 40 to simian virus 40 chromatin (minichromosomes). The minichromosomes isolated from infected cells by a modification of standard techniques were relatively free of contaminating RNA and cellular DNA and had a ratio (by weight) of protein to DNA of approximately 1; their DNA was 50 to 60% digestible to an acid-soluble form by staphylococcal nuclease. Cleavage of this chromatin with restriction endonucleases indicated that the nuclease-resistant regions were randomly distributed in the population of minichromosomes, but were not randomly distributed within minichromosomes. Only 20 to 35% of these minichromosomes adsorbed nonspecifically to nitrocellulose filters, permitting binding studies between simian virus 40 T-antigen and chromatin to be performed. Approximately two to three times as much T-antigen was required to bind chromatin as to bind an equivalent amount of free DNA.  When T-antigen was present in excess, both chromatin and free DNA were quantitatively retained on the filters. On the other hand, when DNA or chromatin was present in excess, only one-third as much chromatin as DNA was retained. We suggest that T-antigen-chromatin complexes may be formed by the cooperative binding of T-antigen to chromatin, whereas T-antigen-DNA complexes may be formed by simple bimolecular interactions.


Herpes simplex virus DNA polymerase as the site of phosphonoacetate sensitivity: temperature-sensitive mutants. Temperature-sensitive (ts) mutants in a number of complementation groups of herpes simplex virus type 1 (HSV-1) are deficient in DNA polymerase induction at the restrictive temperature. Twenty-two mutants in 15 complementation groups were tested for sensitivity to phosphonoacetate (PAA), a compound that inhibits HSV replication in vivo and the DNA polymerase in vitro. One mutant, tsD9, was resistant to PAA (Pr), whereas all others were sensitive. Revertants of tsD9 to the ts+ phenotype simultaneously lost PAA resistance. Additional Pr mutants were isolated from ts mutants belonging to several complementation groups of HSV-1. Double mutants (ts Pr phenotype) were used in three-factor recombination analyses to locate the PAA locus on the genetic map at a position indistinguishable from the ts lesion in tsD9. In all cases, resistance or sensitivity to PAA in vivo was correlated with resistance or sensitivity of DNA polymerase in vitro. These data are compatible with the temperature-sensitive lesion of tsD9 and the determinant of PAA sensitivity both residing in the structural gene for DNA polymerase.


Characterization of RNA from equine infectious anemia virus. The genome of equine infectious anemia virus, a nononcogenic retrovirus, has been characterized by velocity sedimentation, electrophoresis in polyacrylamide gels, buoyant density in CS2SO4, and susceptibility to nuclease digestion. The nucleic acid of purified virus was resolved by sedimentation analysis into a fast-sedimenting genome component, which comprises about two-thirds of the virion RNA, and a slow-sedimenting RNA, which is probably comprised of host-derived tRNA and a trace amount of 5S RNA. The fast-sedimenting RNA had a sedimentation coefficient of 62S and a molecular weight of 5.4 X 10(6) to 5.6 X 10(6), as determined by sedimentation velocity and electrophoretic mobility. Upon heat denaturation, [3H]uridine-labeled 62S RNA dissociated into material comprised of 90 to 95% single-stranded species, sedimenting predominantly at 34S, with a molecular weight of 2.7 X 10(6) to 2.9 X 10(6) and 5 to 10% 4S RNA. The 62S RNA was predominantly single-stranded but contained double-stranded regions, as indicated by partial resistance to RNase IA and SI nuclease and by a lower buoyant density in CS2SO4 than that of the single-stranded 34S RNA derived by heat denaturation. These data indicated that the viral genome consisted of two 34S subunits of single-stranded RNA held in a high-molecular-weight complex with 4S RNA by a mechanism involving a small degree of base pairing. Thus, the structure of equine infectious anemia virus RNA is similar to that of other retroviruses.


Measurement of proviral genes in uninfected and avian myeloblastosis virus-infected cells by hybridization with 3H-labeled complementary DNA probe excess. Viral RNA (vRNA) from avian myeloblastosis virus or DNA from virus-infected and uninfected cells was hybridized with [3H]DNA complementary to viral RNA ([3H]cDNA) under conditions of [3H]cDNA excess. When [3H]cDNA was used to drive the hybridization reaction with vRNA, a rate constant of 33.2 liters/mol-s was obtained. The same rate constant was obtained when vRNA excess was used as the driver. The specific activities of the [3H]DNA probe, estimated from kinetic measurements of the hybridization reaction and from the amount of [3H]cDNA in hybrid form at equilibrium, were 9.1 and 8.6 cpm/pg, respectively. DNA isolated from uninfected cells contained five or six copies of proviral DNA per cell genome. DNA isolated from erythrocytes infected with avian myeloblastosis virus had an additional five or six viral genes added to the cell genome, and the virus-infected target cell (myeloblasts) contained about 15 additional copies of proviral DNA per cell. The use of excess [3H]cDNA probe is an easy and accurate method to quantify the frequency of proviral DNA sequences in cell DNA and to measure a small amount (40 to 200 pg) of vRNA. Probe excess hybridization offers a number of advantages over other procedures and these are discussed.


Nucleotide sequences derived from pheasant DNA in the genome of recombinant avian leukosis viruses with subgroup F specificity. Recombination between viral and cellular genes can give rise to new strains of retroviruses. For example, Rous-associated virus 61 (RAV-61) is a recombinant between the Bryan high-titer strain of Rous sarcoma virus (RSV) and normal pheasant DNA. Nucleic acid hybridization techniques were used to study the genome of RAV-61 and another RAV with subgroup F specificity (RAV-F) obtained by passage of RSV-RAV-0 in cells from a ring-necked pheasant embryo. The nucleotide sequences acquired by these two independent isolates of RAV-F that were not shared with the parental virus comprised 20 to 25% of the RAV-F genomes and were indistinguishable by nucleic acid hybridization. (In addition, RAV-F genomes had another set of nucleotide sequences that were homologous to some pheasant nucleotide sequences and also were present in the parental viruses.) A specific complementary DNA, containing only nucleotide sequences complementary to those acquired by RAV-61 through recombination, was prepared. These nucleotide sequences were pheasant derived and were not present in the genomes of reticuloendotheliosis viruses, pheasant viruses, and avian leukosis-sarcoma viruses of subgroups A, B, C, D, and E. They were partially endogenous, however, to avian DNA other than pheasant. The fraction of these nucleotide sequences present in other avian DNAs generally paralleled the genetic relatedness of these avian species to pheasants. However, there was a high degree of homology between these pheasant nucleotide sequences and related nucleotide sequences in the DNA of normal chickens as indicated by the identical melting profiles of the respective hybrids.


Role of potassium in the synthesis and decay of two specific bacteriophage T4 messages. The role of K+ in the in vivo metabolism of specific phage T4 messengers was studied. By using a mutant of Escherichia coli defective in its ability to accumulate K+ from the growth medium, it was possible to rapidly deplete cells of their intracellular K+ and in this way determine K+-dependent reactions in vivo. The rate constants for accumulation, synthesis, and decay of the early enzymes deoxynucleotide kinase and alpha-glucosyl transferase were determined. It was shown that there is a very close association between mRNA synthesis and its decay, indicating that a mechanism may be present in the cell that can regulate the concentration of these RNAs. Since the mRNA's for these enzymes are very stable in cells depleted of K+, K+ depletion may be a useful method for the isolation of functional T4 mRNA.


Recombination between endogenous and exogenous simian virus 40 genes. I. Rescue of a simian virus 40 temperature-sensitive mutant by passage in permissive transformed monkey lines. Passage of the simian virus 40 (SV40) temperature-sensitive (ts) mutant tsD202 at the permissive temperature in each of three permissive lines of SV40-transformed monkey CV1 cells resulted in the emergence of temperature-insensitive virus, which plated like wild-type SV40 at the restrictive temperature on normal CV1 cells. In independent experiments, the amount of temperature-insensitive virus that appeared after passage on transformed cells was from 10(3)- to 10(6)-fold greater than the amount of ts-revertant virus that appeared after an equal number of passages in nontransformed CV1 cells. The virus rescued by passage on transformed cells bred true upon sequential plaque purification, plated on normal CV1 cells with single-hit kinetics at the restrictive temperature, and displayed no selective growth advantage on transformed cells compared to non-transformed cells. Hence, the reversion of the ts phenotype is neither due to complementation effects nor to the selection of preexisting revertants, which grow better on transformed cells. In the accompanying article (T. Vogel et al., J. Virol. 24:541-550, 1977), we present biochemical evidence that the rescue of tsD202 mediated by passage on transformed cells is due to recombination with the resident SV40 genome. Parallel experiments in which tsA, tsB, and tsC SV40 mutants were passaged in each of the three permissive lines of SV40-transformed monkey cells resulted in either only borderline levels of rescue (tsA mutants) or no detectable rescue (tsB and tsC mutants). Evidence is presented that the resident SV40 genome of the transformed monkey lines is itself a late ts mutant, and we suggest that this accounts for the lack of detectable rescue of the tsB and tsC mutants. We furthermore suggest that the borderline level of rescue observed with two tsA mutants is related to a previous finding (Y. Gluzman et al., J. Virol. 22:256-266, 1977) which indicated that the resident SV40 genome of the permissive transformed monkey cells is defective in the function required for initiation of viral DNA synthesis.


Recombination between endogenous and exogenous simian virus 40 genes. II. Biochemical evidence for genetic exchange. The genome of the simian virus 40 (SV40) temperature-sensitive (ts) mutant tsD202 rescued by passage on transformed permissive monkey lines (see accompanying paper [Y. Gluzman et al., J. Virol. 24:534-540, 1977]) was analyzed by restriction endonuclease cleavage mapping to obtain biochemical evidence that the rescue of the ts phenotype results from recombination with the resident SV40 genome of the transformed cell. It was demonstrated that the endonuclease R. HaeIII cleavage site, which is located at 0.9 map unit in the standard viral genome (and which is in the proximity of the known map position of the tsD lesion), is missing in the DNAs of the parental tsD202 virus and of three independent revertants of tsD202. In contrast, this cleavage site was shown to be present in the DNAs of four out of five independently derived rescued D202 populations and in the DNA of the SV40 strain, 777, used to transform the monkey cells. Comparison of the endonuclease R. Hin(II + III) cleavage patterns of SV40 strain 777 DNA and tsD202 DNA revealed differences in the electrophoretic mobilities of Hin fragments A, B, and F. However, the corresponding Hin fragments from all four rescued D202 genomes were identical in their mobilities to those of tsD202 DNA, indicating that these regions of the rescued D202 genome are characteristic of the tsD202 parent. We conclude, therefore, that the genome of the rescued D202 virus is a true recombinant, since it contains restriction endonuclease cleavage sites characteristic of both parents, the endogenous resident SV40 genome of the transformed monkey cells and the exogenous tsD202 mutant.


Characterization of Gazdar murine sarcoma virus by nucleic acid hybridization and analysis of viral expression in cells. Gazdar murine sarcoma virus (Gz-MSV) and Moloney murine sarcoma virus (M-MSV) are closely related. The complete M-MSV-specific nucleic acid sequences constituted a major portion of Gz-MSV-specific sequences. The MSV-specific sequences in both Gz-MSV and M-MSV genomes shared homology with hamster leukemia virus nucleic acid sequences. Both rat cells (S+L+) and hamster (S+L-) cells expressed two viral proteins of 68,000 and 70,000 daltons. These proteins were immunologically related to p60 purified from m1 virions of M-MSV.


Alterations of neutral glycolipids in cells infected with syncytium-producing mutants of herpes simplex virus type 1. The isolation of syncytium-producing mutants of herpes simplex virus type 1 (KOS strain), which cause extensive cell fusion during otherwise normal infections, has been reported previously (S. Person, R. W. Knowles, G. S. Read, S. C. Warner, and V. C. Bond, J. Virol. 17:183-190, 1976). Seven of these mutants, plus two syncytial strains obtained elsewhere, were used to compare the incorporation of labeled galactose into neutral glycolipids of mock-infected, wild-type-infected, and syncytially infected human embryonic lung cells. Five predominant cellular glycolipid species were observed, denoted GL-1 through GL-5 in order of increasing oligosaccharide chain length; for example, GL-1 and GL-2 correspond to glycolipids that contain mono- and disaccharide units, respectively. Wild-type virus infection caused an increase in galactose incorporation into GL-1 and GL-2 relative to GL-3 through GL-5. For a single labeling interval from 4 to 10 h after adsorption, syncytial infections generally resulted in a relatively greater incorporation into more complex glycolipids than did wild-type infections. One mutant, syn 20, was compared with wild-type virus throughout infection by using a series of shorter labeling pulses and appeared to delay by at least 2 h the alterations observed during wild-type infections. These alterations are apparently due to defects in synthesis, since prelabeled cellular glycolipids were not differentially degraded during mock or virus infection.


RNase T1-resistant oligonucleotides of an N- and a B-tropic murine leukemia virus of BALB/c: evidence for recombination between these viruses. We used two-dimensional gel electrophoresis to obtain fingerprints of 32P-labeled RNase T1-resistant oligonucleotides derived from the genomes of an N- and a B-tropic murine leukemia virus of BALB/c. These viruses share approximately 30 large T1-resistant oligonucleotides. In addition, there are eight large oligonucleotides unique to the N-tropic virus, and there are six B-trophic virus-specific oligonucleotides. Viruses, designated XLP-N, which appear by biological criteria and analysis of virion proteins to be recombinants between these N- and B-tropic viruses, possess some but not all of the N or B virus-specific oligonucleotides.


RNase T1-resistant oligonucleotides of Akv-1 and Akv-2 type C viruses of AKR mice. We used two-dimensional gel electrophoresis to obtain fingerprints of RNase T1-resistant oligonucleotides derived from the genomes of Akv-1 and Akv-2 type C viruses of AKR mice. The fingerprints of these two viruses look identical. The products of pancreatic RNase digestion of corresponding oligonucleotides of the two viruses were indistinguishable. These observations are consistent with, but not proof of, the possible identity of the genomes of the Akv-1 and Akv-2 viruses and, thus, of the viral genetic material believed to comprise the Akv-1 and Akv-2 loci of AKR mice.


Measurements of the genome sizes of simian virus 40 and polyoma virus. We have measured the genome sizes of simian virus 40 and polyoma virus and found them to be 5,010 +/- 125 base pairs and 5,080 +/- 125 base pairs, respectively.


BALB/3T3 cells infected by the ts3 mutant of polyoma virus fail to accumulate virus-specific early RNA at the nonpermissive temperature. We measured the accumulation of virus-specific early RNA in BALB/3T3 cells infected by the ts3 mutant of polyoma virus by annealing cytoplasmic RNA from infected cells to purified, radiolabeled, "early" strand of polyoma DNA. Cells infected by the ts3 mutant fail to accumulate virus-specific early RNA at the nonpermissive temperature.


The use of evoked electromyographic responses in diagnosing lesions of the cauda equina. An electrophysiologic technique to evaluate the anatomic integrity of the peripheral pathways to the bladder and sphincters is described. Evoked electromyographic responses of the anal sphincter are produced by stimulating the bladder wall and urethra. Impulses travel via the pelvic nerves and cauda equina to the conus medullaris, activating the pudendal nerve nucleus and resulting in contraction of the external anal sphincter. Lesions along this pathway produce either prolonged latencies and depressed responses or complete loss of response. Correlation of the results of this technique in 110 patients with clinical myelographic and operative findings indicates that the technique is a useful clinical tool.


Paget's disease of the scrotum presenting as an inguinal lymph node. Extramammary Paget's disease is a rare malignant entity. A case with a unique clinical presentation is described and emphasis is placed on the need for early biopsy of skin lesions in the genital area.


Evidence for the association of cytomegalovirus with carcinoma of the prostate. A human genital isolate of cytomegalovirus is shown to have transformed human embryonic lung cells in vitro. These cells produce tumors when injected into athymic nude mice. Two cell lines derived from tissue from human prostatic carcinoma have survived more than 20 passages in vitro and demonstrate cytomegalovirus-specific membrane antigen. Significant humoral antibody titers against cytomegalovirus have been demonstrated. Cell-mediated lymphocytotoxicity against these transformed cells has been demonstrated in patients with urinary tract tumors. This evidence indicates that an association between cytomegalovirus and human prostatic cancer may be more than coincidental.


Pheochromocytoma with prominent calcification and associated pancreatic islet cell tumor. A case of bilateral pheochromocytoma with an associated pancreatic islet cell tumor is reported. Prominent calcification was noted in the left adrenal tumor.


Poxvirus fibromas on African hares.

Poxvirus fibromas on African hares. Small dermal tumors were found on three African hares (Lepus capensis) in the Laikipia District, Kenya. Gross and histopathologic studies revealed similarities to the Shope's fibroma of wild rabbits in North America and fibromas of European hares. Histological examination of the African hare fibromas revealed intracytoplasmic inclusion bodies characteristic of poxviruses and poxvirus virions were demonstrated by electron microscopy of ultrathin sections. Attempts to propagate the virus in rabbit skin, embryonated chicken eggs and cell cultures were unsuccessful.


Primary liver tumors and oral contraceptives. Results of a survey. Data on 378 (in females) and 165 (in males) cases of primary liver tumors reported by 477 hospitals in the United States during 1970 to 1975 show that in males, 91.5% of the tumors were malignant, and in females, 43.9% were malignant and 59.1% were benign. Of the 212 benign tumors in females, 96 were hepatic cell adenomas and 58 were focal nodular hyperplasias. A history of oral contraceptive use was found in nearly half of all women: 65% with benign tumors, 74% with hepatic cell adenomas, and 74% with focal nodular hyperplasias. Symptoms were more severe among users. No case of intraperitoneal bleeding was observed in nonusers. The findings confirm the suggested association between oral contraceptive use and hepatic cell adenomas and focal nodular hyperplasias.


Plasma high-density lipoprotein cholesterol level. Influence of risk factor intervention. Determinants of circulating high density lipoproteins (HDL) were sought in 301 men studied during a year of participation in a coronary prevention program. Mean plasma HDL-cholesterol concentration rose by 2.8 mg/dl (6%) in the group receiving multifactor intervention, but the change did not differ significantly from that in the comparison group. Larger changes in both directions were seen in many individuals. Multiple regression analysis of these changes indicates that increased plasma HDL levels occur when plasma triglyceride level is decreased, cigarette smoking is reduced, and habitual alcohol intake is increased. Increases in the concentration of HDL-cholesterol also tended to accompany adherence to the fat-controlled diet, reduction in LDL-cholesterol level, and loss of body weight. Conventional coronary prevention programs are unlikely to have an adverse influence on this new, risk-lowering factor.


Peripheral diabetic neuropathy treated with amitriptyline and fluphenazine. The pain of diabetic peripheral neuropathy responds poorly to current modes of treatment. We treated eight patients with this disorder whose pain was refractory to standard regimens but who experienced remarkable pain relief within two to five days after treatment with fluphenazine hydrochloride, amitriptyline hydrochloride, or a combination of the two. In four patients whose regimens were discontinued, pain recurred within two days and again remitted on reinstitution of the drug regimens. These findings suggest that fluphenazine alone or in combination with amitriptyline may be of benefit in treating the painful peripheral neuropathy associated with diabetes.


Cytomegalovirus encephalitis in immunologically normal adults. Successful treatment with vidarabine. Acute cytomegalovirus (CMV) encephalitis developed in two immunologically normal adults. The diagnosis was confirmed by isolation of CMV from the CSF and urine in one case and from temporal lobe biopsy tissue, CSF, and urine in the second case. Both patients were treated with vidarabine and showed dramatic clinical improvement. Virus excretion, which had been chronic in one case, cleared after therapy. To our knowledge, these are the first persons with CMV encephalitis evidenced by isolation of the virus from the CNS. The response to vidarabine was impressive and warrants further evaluation.


Probucol (Lorelco) in treatment of hyperlipemia. The chemical structure of probucol differs from that of other hypolipemic agents, and its mechanism of action is unknown. This agent lowers elevated serum cholesterol (and low-density lipoprotein) levels and appears to be effective when used as an adjunct to a low-cholesterol, low-saturated-fat diet for treatment of type IIa hyperlipoproteinimia in adults; however, it is less effective than cholestyramine resin in patients with familial type IIa disorder. Although probucol has no consistent effect on elevated triglyceride levels, it may be useful as an adjunct to other drugs that lower these concentrations in patients with types IIb, III, and IV hyperlipoproteinemia when hypercholesterolemia persists. Probucol is generally well tolerated.


Properties of thirteen kinds of adsorbents for removal of hydrogen sulfide, methanethiol, methyl sulfide, trimethylamine, and ammonia. Adsorption of hydrogen sulfide, methanethiol, methyl sulfide, trimethylamine, and ammonia on thirteen kinds of adsorbents (5 kinds of silicate, 4 kinds of activated carbon, and 4 kinds of zeolite) was measured by gravimetry, at 30 degrees C and 50 Torr, using an adsorption apparatus with a spring balance in order to find the most suitable adsorbent for the removal of these gases by dry process. The relations between the amount of these gases adsorbed (mmol/cm2) on the adsorbent and the surface properties or the porous structure were examined to clarify the mechanism of adsorption of these gases on them through surface pH, pore size distribution, and area of an adsorbed particle of these gases. Among the thirteen adsorbents, the activated carbon Nos. 6 and 7 were the most suitable adsorbent for methanethiol, methyl sulfide, and trimethylamine whose area of an adsobed particle (wrho) was larger than about 17 A2, and zeolite Nos. 12 and 13 were most suitable for removal of hydrogen sulfide and ammonia (wrho less than about 17A2). The amount of these gases adsorbed (mmol/cm2) on these adsorbents was mainly determined by their porous structure rather than by their surface properties.


Studies on the respiratory system of Aspergillus oryzae. V. Some properties of the respiratory system of mitochondria from mycelia grown in the presence of chloramphenicol. Presence of chloramphenicol in the growth medium for mycelia of Aspergillus oryzae was without effect on the oxidative activity, respiratory control, or P/O ratio of isolated mitochondria. The mitochondria oxidized Krebs cycle intermediates even in the presence of cyanide at the concentration markedly inhibiting the normal mitochondrial oxidation. However, the P/O ratio during the mitochondrial oxidation decreased by about 1.0 on addition of cyanide. The c-type cytochromes, shown to occur in large amounts than in normal mitochondria (Wakiyama and Ogura, 1972), were suggested to act as electron carriers in this cyanide-resistant oxidation. A novel pigment, demonstrated only in the mitochondria prepared from chloramphenicol-treated mycelia by a CO-difference spectrum, was presumed to be the terminal oxidase of the respiration in the presence of cyanide.


Depressive effect of epinephrine mediated via adrenergic beta-receptor in isolated rat colon and duodenum. Differences in sensitivity to catecholamines between colon and duodenum were examined in tissues from the rat, monitoring the depressive effect of catecholamines on contractile response to acetylcholine (ACh). The sensitivity of colonic tissue to ACh was higher than that of duodenal. Epinephrine (Ep, 10(-7) g/ml) depressed the contractile response to ACh in the colonic tissue, but not in the duodenal. The depressive effect of Ep on the contractile response to ACh is attributed to the stimulation of adrenergic beta-receptors in the colonic tissue as the depression disappeared by pretreatment with propranolol (10(-6) g/ml). There was no difference on the depressive effect of papaverine on the contractile response to ACh, except when low concentrations were used. Dibutyryl cyclic AMP (10(-4) g/ml) depressed the contractile responses of both tissues to ACh. After treatment with Ep (10(-7) g/ml), cyclic AMP content was increased in the colonic tissue but not in the duodenal. However, papaverine (3 X 10(-6 g/ml) and a higher dose of Ep(10(-6) g/ml) increased cyclic AMP content in both tissues. The increase of cyclic AMP and the decrease of tension caused by Ep were not correlated in these tissues. However, a positive correlation was observed between the depressive effect of Ep on the contractile response to ACh and the increase of cyclic AMP content in these tissues.


Effects of p-chlorophenylalanine (p-CPA) on sleep in olfactory bulb lesioned rats. The effect of p-CPA and 5-HTP followed by p-CPA on sleep was studied in rats with olfactory bulb lesions (O.B. lesioned rats). In these rats, electrodes were chronically implanted to record the EEG (frontal cortex and dorsal hippocampus), the cervical electromyogram and eye movements. The REM sleep stage was selectively decreased from 24 to 32 hours after 200 mg/kg of p-CPA in the sham lesioned rats, whereas both the slow wave sleep and REM sleep stages were markedly decreased by the same dose of p-CPA in the O.B. lesioned rats. In both sham and O.B. lesioned groups, the slow wave sleep and REM sleep stages decreased from 24 to 32 hours after 400 mg/kg of p-CPA and the percentage of decrease in the slow wave sleep stage was much larger with 400 mg/kg of p-CPA than with 200 mg/kg and 400 mg/kg of p-CPA. In the O.B. lesioned rats, the insomnia produced by 200 mg/kg and 400 mg/kg of p-CPA disappeared with 5-HTP (5 mg/kg). On the other hand, the insomnia produced by 200 mg/kg of p-CPA did not recur with 5-HTP in the sham lesioned rats, but with 400 mg/kg there was a recurrence. These results suggest that the enhanced effect of p-CPA and 5-HTP followed by p-CPA in the O.B. lesioned rats is due to changes in the sensitivity of the serotonergic system in the brain.


Potentiation of the NGF-mediated nerve fiber outgrowth by ginsenoside Rb1 in organ cultures of chicken dorsal root ganglia. The fiber outgrowth induced by ACh, dibutyryl cyclic AMP and dibutyryl cyclic GMP in explanted chick embryonic dorsal root ganglia differed distinctly from that be nerve growth factor (NGF) and submandibular gland extract of adult male mice. Ginsenoside Rb1 potentiated the effects of NGF and submandibular extract at concentrations of 3 and 30 muM, but did not potentiate the effects of ACh, dibutyryl cyclic AMP and dibutyryl cyclic GMP. NGF-antibody inhibited the effects of NGF, but not the effects of ACh, dibutyryl cyclic AMP and dibutyryl cyclic GMP, Concanavalin A and KCL did not promote fiber outgrowth.


The integrated trauma service concept.

The integrated trauma service concept. An integrated trauma service, through coordination of pre-existing manpower and resources, insures the availability of medical care to patients suffering unforeseen or critical physical injuries and their sequelae. Provided an examination of the overall emergency medical service capabilities of the community dictates the need for a trauma service, the first step is to secure a surgeon interested in the care of the trauma patient. Cooperation of physician colleagues is gained by a policy of mandatory consultation whenever injuries involve specialty areas. Trauma service involvement begins with prioritization, resuscitation, and assessment of the patient in the emergency department and extends through definitive care, management of injury sequelae and rehabilitation. The "isolated injury" is defined by the trauma service and the appropriate specialty service notified. Injuries involving two or more body systems dictate admission to the trauma service except for the patient with an unstable cervical spine injury or deteriorating neurologic status. Under such circumstances, a neurosurgical admission is instituted and the trauma service consults. When the trauma service is operational, educational programs, data collection and retrieval, and follow-up care are enhanced.


Progressive necrotizing external otitis: treatment with ticarcillin and tobramycin. The history of necrotizing external otitis, its diagnosis and management are reviewed. A case history is presented of a patient who was diagnosed as having progressive necrotizing external otitis with facial paralysis. In spite of standard medical treatment and aggressive surgical management, the disease process continued with progressive involvement of Cranial nerves IX and X. The Pseudomonas aeruginosa bacteria developed an increased minimal inhibitory concentration (MIC) to carbenicillin and gentamicin by requiring near toxic blood levels to be effective. Investigational ticarcillin (alpha-carboxy-3-thienylmethylpenicillin) and tobramycin were used successfully in resolving the infection.


Viral particles in nasopharyngeal carcinoma. Lymphoepithelioma of the nasopharynx was identified in a 17-year-old Chinese male. The Epstein-Barr virus titer was 1:3200. Tissue from the tumor mass was prepared for electron microscopy. Particles consistent with the morphological characteristics of herpes virus were found within the cytoplasm of malignant epithelial cells. This finding is discussed in relation to other data suggesting an oncogenic role of herpes virus in human malignancy.


The in vivo incorporation of [32P]-labeled orthophosphate into pyrophosphatidic acid and other phospholipids of Cryptococcus neoformans through cell growth. Cryptococcus neoformans was cultured in a liquid medium containing [32P]-orthophosphate and harvested at various stages of cell growth. An aliquot of the [32P]-labeled cells was transferred to a nonradioactive medium, and the culture was continued again for some hours. The [32P]-radioactivity composition and the phosphorus composition of individual phospholipids relative to the total phospholipid through the incubation periods were estimated. Although levels of major phospholipids remained constant throughout the cell growth, the distribution pattern of the [32P]-radioactivity of individual phospholipids changed remarkably along with the progress of cell growth. The changing patterns of the specific radioactivities of individual phospholipids through the growth phase demonstrated that phosphatidic acid was one of the most active metabolites in phospholipids and that pyrophosphatidic acid was also metabolically active.


[Structure and function of virus-induced antigens in cultured cells infected with Merek's disease and turkey herpes viruses. II. Isolation of intracellular antigens from infected cells (author's transl)]. Virus-induced antigens were isolated from MDV and HVT-infected cells by salt extraction with 3M KCl and purified by Con.A chromatography and isoelectric focusing. Electrophoretic analysis of 35S-methionine, 3H-fucose and 32P-orthophosphate labeled antigens revealed 7 different polypeptides, two of them containing labeled carbohydrates, and one phospholipid component. The isolated virus-induced antigens from infected cells were identified as serologically active membrane complexes carrying common antigenic determinants of MDV and HVT. They were able to bind to virus-neutralizing immunglobulins as shown by antibody binding tests. Summarizing the presented analytic and serologic findings it was proposed to classify MDV and HVT as different serotypes of a common Marek's disease virus group.


Hepatitis A-virus particles in stools of patients from a natural hepatitis outbreak in Germany. During a hepatitis outbreak in Southern Germany 27 nm particles were visualized by immune electron microscopy in stools of two patients. These particles were sereologically identical or similar to hepatitis A-virus particles identified in the USA. The buoyant density of these particles was 1.34 g/cm3 as shown by cesium chloride density centrifugation. The particles were first observed in small numbers in a stool obtained 11 days, and in large numbers in stools obtained 6 and 7 days before the onset of jaundice. Few particles were seen on the day of the onset of jaundice and none thereafter. In both patients a sereoconversion to hepatitis A-virus as judged by immune electron microscopy could be demonstrated.


Preparation in undenatured form of the main protein bound to heterogeneous nuclear RNA in liver and hepatoma cells. Particles carrying heterogeneous nuclear RNA (30 S-particles) were prepared from rat liver and Zajdela hepatoma ascites cell nuclei after ultrasonic disruption. The ribonucleoprotein structures were disintegrated in the presence of 100mM spermidine. Using chromatography on Sepharose-polyadenylate a protein component has been obtained which possessed high affinity for heterogeneous nuclear RNA, polyuridylate and polyadenylate, and double-stranded DNA. This protein was the main species of the ribonucleoprotein studied; it showed bands with molcular weights of 37000 and 40000 respectively in SDS gel electrophoresis. The RNA-binding proteins isolated from liver and hepatoma had identical molecular weights and the same affinity for Sepharose-polyadenylate used in the isolation.


Coding capacity of poly(A)+mRNA isolated from mengovirus-infected Ehrlich ascites tumor cells. Total poly (A)+mRNA was isolated from mengovirus-infected Ehrlich ascites tumor cells at various times postinfection and quantitated in a cell-free system derived from uninfected ascites cells. Basic proteins were separated from acidic proteins by carboxymethyl cellulose chromatography. At the end of the infectious cycle, 8h postinfection, the cellular contents of most mRNAs coding for basic ribosomal proteins are still between 70 and 90 percent of those measured at the beginning of infection or in uninfected cells. On the basis of this result, the rapid shutoff of host protein synthesis after mengovirus infection of Ehrlich ascites tumor cells cannot be the consequence of the inactivation of host template RNA.


New directions in prostaglandin research and their possible implications for immunologic and nonimmunologic inflammation. New information on prostaglandins and related structures derived from arachidonic acid is accumulating rapidly. Of particular interest is evidence for the existence of labile, highly biologically active, nonprostanoate derivatives. A role for the classic prostaglandins in immunologic inflammation is already strongly suspected, and it seems likely that the newly recognized arachidonic acid metabolites may prove to be even more important.


A protective effect of caffeine on drosophila larvae treated with methyl methanesulfonate. Lethality induced in larval populations of Drosophila melanogaster was recorded after treatment with (1) caffeine, (2) MMS or (3) caffeine plus MMS. The mixture of caffeine plus MMS was less toxic than expected from the effects observed after treatment with either substance individually. It is postulated that in the combined treatment the caffeine, by inhibiting semiconservative DNA replication, allows for some additional time for repair of alkylated DNA by a repair pathway which is not sensitive to caffeine, possibly excision repair.


[Study of intracellular enzymes in the genus Schizosaccharomyces. Sistematic implications]. In the genus Schizosaccharomyces intracellular osidases and nitrite and nitrate reductases are revealed; particularly all the species possessing invertase, alpha-glucosidase and alpha-galactosidase. These characters underline the homogeneity on the genus. On the basis of osidases, nitrite and nitrate reductases results, 2 groups can be distinguished in this genus.


Immunodiffusion test in avian encephalomyelitis. I. Standardization of procedure and detection of antigen in infected chickens and embryos. The double immunodiffusion technique was applied to avian encephalomyelitis virus (AEV). Agar gel medium containing such a high concentration of NaCl as 15% was more preferable for highly diluted quantities of reactants than any other NaCl-containing medium. A single precipitin line appeared on the 1st to 7th days of diffusion at room temperature. The specificity of reaction between AEV antigen and homologous immune chicken serum has been demonstrated by no cross reaction between heterologous viruses and specific absorption by homologous virus. The antigen was produced in the brain, viscera, eyeball, whole body and yolk sac of chick embryos inoculated via yolk sac, as well as in the thigh muscles of chicks subcutaneously inoculated at 2 days of age. Antigenicity was detectable in 50% emulsion of these organs with a virus titer more than 10(5.0) per 0.1 g of tissue weight.