Reactivation of latent herpes simplex virus infection of the autonomic nervous system by postganglionic neurectomy. Latent herpes simplex virus infection of the superior cervical autonomic ganglion was reactivated in vivo by postganglionic neurectomy. Two methods were used to demonstrate viral reactivation: (i) recovery of infectious herpes simplex virus in ganglion homogenates and (ii) acceleration of virus expression in ganglion explants in culture. Both the percentage of mice exhibiting reactivated ganglion infection and the viral titers detected in ganglia increased when neurectomized mice were treated with cyclophosphamide. Antithymocyte serum treatment prolonged the time course over which neurectomy-induced virus could be detected, but neither antithymocyte serum nor cyclophosphamide reactivated herpes simplex virus in the absence of neurectomy. These results demonstrate that postganlionic neurectomy provides a specific stimulus for herpes simplex virus reactivation and that cell-mediated immune defense are involved in the highly efficient elimination of reactivated virus from the ganglion in vivo.


Search for latent cytomegalovirus in renal allografts. Cytomegalovirus (CMV) infection is common after renal transplantation, and cytomegaloviruria occurs in about two-thirds of the recipients. These observations suggest that the allografts may be a site of latent infection with CMV and its reactivation may be the source of viruria. To investigate this possibility, 130 kidney specimens from 85 persons were cultured, and simultaneous explants were made of 63 of them from 50 people. No CMV was received from 33 normal kidney or cadaver donors or from 19 allograft recipients who had no evidence of posttransplantation infection with CMV. The experiment included 37 primary organ explants that yielded no evidence of latent virus. Among 33 allograft recipients with posttransplantation CMV infection, overt infectious virus was isolated from 6 of 57 allograft biopsies. All six positive specimens were from four patients, all of whom had viruria simultaneously. Organ explants from 20 of the recipients with demonstrated postoperative CMV infection, including 13 viruric patients, failed to unmask any latent CMV. Thus, allograft kidneys were infrequently infected with CMV (6%), even in patients with viruria (24%). The kidney parenchyma appears to be an uncommon site of latent CMV infection and may not be the usual source of virus in patients with viruria.


Pivmecillinam treatment of chronic urinary tract infection. Twenty-eight patients with chronic urinary tract infections were treated with 400 mg pivmecillinam orally three times daily for 10 to 15 days. The diagnosis was confirmed by a history of cystitis or cystopyelitis four to six times annually, microscopy of urine sediment, and growth of pathogens in urine specimens obtained by suprapubic bladder puncture. Three days, three and six weeks after completion of therapy the success rates were 24/28, 20/28 and 19/28 respectively. Pivmecillinam was well tolerated. Two patients developed nausea and vomiting. Other side-effects were not observed. Pivmecillinam is a useful drug in the treatment of chronic urinary tract infections.


Effect of immunization on the cyclic AMP level and 3H-thymidine incorporation in cultured lymphoid cells. The cyclic AMP level and the uptake of 3H-thymidine were studied in short-term suspension cultures of guinea pig lymphoid cells. Spleen cells from animals immunized 3 days previously with typhoid vaccine were shown to differ in three respects from the normal cells: (1) The level of cyclic AMP was higher. (2) The cyclic AMP increase following addition of isoproterenol or adenosine was abolished. (3) Isoproterenol (10(-5) to 10(-4) M) stimulated the uptake of 3H-thymidine in cells from immunized animals, while the uptake into normal cells was inhibited by these concentrations. A higher dose of isoproterenol (10(-3) M) inhibited 3H-thymidine uptake in both immunized and normal cells. The results reveal certain biochemical alterations in splenic lymphocytes after immunization. An altered 3H-thymidine uptake may be caused by the modified cyclic AMP metabolism in cells from immunized animals.


Establishment of paired tumor cells and autologous virus-transformed cell lines to define humoral immune responses in melanoma and sarcoma patients. Peripheral blood lymphocytes and skin fibroblasts from 12 cancer patients were infected with Epstein-Barr virus (EBV) or SV40 virus. The EBV-transformed lymphoblasts and SV40-transformed fibroblasts were grown as continuous cell lines and expressed the same histocompatibility antigens as tumor cell lines established from the same cancer patients. Sera from 350 melanoma and 195 sarcoma patients were tested for antibody reactive with membrane antigens on three of these tumor cell lines (two melanomas and one sarcoma) by immune adherence (IA) and indirect immunofluorescence (IMI) assays. Antibodies to HLA and other non-tumor-related antigens were completely removed from the most reactive sera by quantitative absorption with 4 x 10(7) lymphoblasts or 10(7) transformed fibroblasts autologous to the tumor target cells. These paired cell lines were used to monitor humoral immune responses in melanoma and sarcoma patients receiving allogeneic tumor cell vaccines.


The frequency of occurrence of feline leukaemia virus subgroups in cats. Feline leukaemia viruses (FeLV) were isolated from cats in Glasgow and New York with lymphosarcoma and from apparently healthy carrier cats. The subgroup composition of each isolate was determined. All isolates contained FeLV of subgroup A(FeLV-A) and a high proportion also contained subgroup B virus (FeLV-B). Virus of subgroup C (FeLV-C) was rare and occurred in association with FeLV-A and, in some isolates, with FeLV-B as well. The same pattern was observed in isolates from British and American cats. The frequency of FeLV subgroups was different in cats with lymphosarcoma and in healthy carrier cats. In cats with lymphosarcoma, 42% had FeLV-A and 58% had FeLV-AB; there was no obvious correlation between virus subgroup and type of disease. In FeLV-positive healthy cats, 65% had FeLV-A and 33% had FeLV-AB. FeLV-C was isolated only from cats with disease. The healthy carrier cats were from multiple-cat households (MCH). Two distinct types of FeLV-infected MCH were found: MCH-A in which the carrier cats yielded only FeLV-A, and MCH-AB in which cats with either FeLV-A or FeLV-AB were present. In MCH-AB half of the cats had FeLV-A and half had FeLV-AB. Overall, the proportion of cats in MCH which were viraemic with FeLV was 42%. However, there was a marked difference in the prevalence of carrier cats in each type of MCH: in MCH-A, 28% were FeLV-positive while in MCH-AB, 53% were viraemic.


Design of a photoaffinity label for the hormone binding site of neurophysin. The photolabile peptide, L-methionyl-L-tyrosyl-p-azido-L-phenylalaninamide, was synthesized by solution methods. This peptide, as well as the analogous species containing tritiated methionine, were found to bind reversibly and specifically, in the dark, to bovine neurophysin II. The dissociation constant, stoichiometry, and pH-dependence of this noncovalent interaction are typical of those properties for hormone (oxytocin) and hormone-like ligand binding to neurophysin II. Under photolytic conditions, methionyl-tyrosyl-p-azidophenylalaninamide causes irreversible inhibition of the noncovalent ligand binding activity of neurophysin II. This inactivation was achieved to the extent of about 90%. Both the dark and light (photolytic) interactions of the photolabile peptide with neurophysin II indicate its reaction at the hormone binding site of the protein and thus its potential use to identify amino acid residues at this site by covalent photoaffinity labelling.


Radical formation in single crystals of hypoxanthine.HCl.H2O, inosine, and the disodium salt of 5'-inosine-monophosphate. Radical formation in single crystals of hypoxanthine.HCl.H2O, inosine and Na2-5'-IMP.(7.5 H2O) by X-irradiation has been studied using electron-spin-resonance spectroscopy at 9.5 and 35 GHz. In all crystals both H-addition radicals at position C2 and C8 of the purine ring are found. The coupling constants of these two radicals are different and depend strongly on the protonation state of the base. INDO-calculations indicate that the C8-radical is protonated at O6. In Na2-5'-IMP OH-addition radicals at position C2 of the purine ring are formed. Electron adduct radicals are found in the neutral and the N7-protonated base after X-irradiation at 77 K. In Na2-5'-IMP no electron adduct is formed but a radical which probably is the cation. In hypoxanthine.HCl.H2O a radical could be observed after X-irradiation at 77 K, which results from addition of a Cl- to the nitrogen N1.


Sensory testing: a sensitive method in the follow-up of nerve involvement. Sensory testing (ST) as a parameter in assessing nerve damage and as a guide in anti-reaction treatment is discussed in comparison with established methods like voluntary muscle testing (VMT) and motor nerve conduction velocity (MCV). Sensory testing is shown to be a valuable addition to the existing tests and may even be used as a single parameter.


The action of ionizing radiation on protein. III. Radical formation in L-phenylalanine.HCl. The radicals produced by X-irradiation in L-phenylalanine.HCl crystals have been analysed by electron-spin-resonance (e.s.r.) spectroscopy. Four radicals have been identified: the radical resulting from electron capture by the carboxylic group, the radical resulting from deamination, an hydrogen addition radical to the benzene ring, and the radical resulting from hydrogen abstraction from the methylene group bonded to Cbeta. These identifications are supported by INDO calculations.


The interaction of cytochrome C with phospholipids. A pulse-radiolysis study. The reactions of the hydrated electron (eaq-), produced during pulse radiolysis, have been used to study the binding of phosphatidyl choline (PC), phosphatidyl serine (PS), phosphatidyl ethanolamine (PE), and phosphatidyl inositol (PI) vesicles with horse-heart cytochrome C. An interaction could only be detected between cytochrome C and either PS or PI. An apparent equivalence point in the binding was reached for both phospholipids at a molar ratio of phospholipid : protein of 6 : 1. At this point, the reactivity of (eaq-) towards the cytochrome C was very markedly reduced. Indeed, the rate of disappearance of (eaq-) under such conditions was the same as the rate of eaq- disappearance in triply-distilled water. The inclusion of cholesterol at a molar ratio of 1 : 1 within the phospholipid vesicles changed the stoichiometry of the interaction. Evidence that protonated epsilon-amino groups of lysine residues are involved in the interaction is presented. Possible models for the complexes formed are discussed.


The influence of protonation or alkylation of the phosphate group on the e.s.r. spectra and on the rate of phosphate elimination from 2-methoxyethyl phosphate 2-yl radicals. The e.s.r. spectra of 1-yl, 2-yl and 3'-yl methoxyethyl phosphate radicals derived from CH3OCH2CH2-OPO3H2 by hydrogen abstraction have been measured in aqueous solutions and the hyperfine constants determined. The coupling constants vary strongly with protonation or alkylation of the phosphate group. The 2-yl radicals eliminate phosphate. The rate-constants for the elimination (ke) have been estimated by e.s.r. measurements and by product studies as a function of pH using 60Co gamma-radiolysis. The ke values vary from approximately 0.3 s(-1) for the CH3OCHCH2OPO3--radical and approximately 10(3) s-1 for CH3OCHCH2OPO3H-, to approximately 3 X 10(6) S-1 for CH3OCHCH2OPO3H2. Alkylation of the phosphate group increases the elimination rate-constant to a similar extent as protonation. The results support a recent mechanism which described the OH-radical-induced single-strand breaks of DNA in aqueous solution starting from the C-4' radical of the sugar moiety. It is further concluded the C-4' radical of DNA eliminates the 3'-phosphate group faster than the 5'-phosphate group.


Battered women: a medical problem requiring detection. A one month study of the surgical and psychiatric service of a university hospital emergency room identified thirty-seven women coming for treatment as a result of physical abuse. Thirty-three of the thirty-seven women presented at the surgical services are represented 3.8 per cent of surgical admissions in the emergency room.  These women came from all ages, social and ethnic groups. They usually presented with contusions or other injuries to the head and neck. The majority would not accept psychiatric treatment but were willing to discuss the problem with the emergency room physician if encouraged. The preventive implications of these findings for the nonpsychiatric physician are discussed.


Invasive primary mucinous carcinoma of the skin. An unusual case of primary mucinous carcinoma of the skin has been reported. The tumor progressed very rapidly and recurred twice within a short period after adequate local excision. It had wide spread lymph node metastases and a fatal ending. The characteristic clinical and histologic differentiation from sweat gland carcinoma has been discussed.


Effects of dexamethasone on the morphogenesis of two mutants of Rous sarcoma virus. Japanese quail cells transformed by the replication-defective, Bryan high-titer strain of Rous sarcoma virus, BH RSV(-)Q, clone 3, revealed intracytoplasmic A-type particles after dexamethasone treatment. In the absence of dexamethasone, or when superinfected with a helper virus in the presence or absence of dexamethasone, no such particles were observed. Chick embryo cells (CEC) infected with a temperature-sensitive sarcoma virus mutant, LA 334, coordinately defective for transformation and viral replication, showed abnormal accumulation of viral core substances and aberrant budding at the non-permissive temperature (41 degrees). CEC infected with LA 334 and treated with dexamethasone at 41 degrees resulted in more extensive accumulation of abnormal budding without increased release of viral particles. Dexamethasone, however, did not lead to abnormal morphogenesis of virus under permissive conditions. The selective effects of dexamethasone on the morphogenesis of these two mutants of avian sarcoma virus are discussed.


Contrasting characteristics of Marek's disease herpesvirus isolated from chickens with and without avian leukosis virus infection. Marek's disease herpesvirus (MDHV) isolated from chickens free of naturally occurring avian leukosis virus (ALV) infection produced characteristic foci in both chicken embryo fibroblast (CEF) and chicken kidney cell (CKC) cultures. MDHV-A, which was extracted from the feather follicle epithelium of chickens naturally infected with ALV, did not induce cytologic changes in CEF cultures, but did cause focus formation in CK cultures. ALV was detected in MDHV-A, but not in MDHV preparations. MDHV-A (reconstructed in vivo) and MDHV were further distinguished from one another by inoculation of ALV-free LSI-SPF chickens. MDHV-A elicited a high incidence of early mortality which was not accompanied by the gross tumor spectrum characteristic of Marek's disease, although extensive histologic lesions were present. The differences between MDHV and MDHV-A were not as striking in another line of ALV-free chickens (SPAFAS). By contrast, among conventional chickens with naturally occurring ALV infection, neither MDHV-A nor MDHV caused appreciable early mortality although both were highly oncogenic (gross tumor development). These observations demonstrate that the presence of an oncornavirus (ALV), detected by radioimmune but not complement fixation assays, can influence the in vitro and in vivo characteristics of an oncogenic herpesvirus (MDHV). The observations recorded here resolve some of the inconsistencies reported in the literature. Thus, the apparent failure by some to find interactions between MDHV and oncornaviruses can be ascribed to the comparatively limited sensitivity of the complement fixation assay used to detect oncornaviruses (ALV). We have shown that the presence of oncornavirus detectable by radioimmune assay, but not by complement fixation, can influence the in vitro and in vivo responses of an oncogenic herpesvirus (MDHV). Our observations relating to viral interaction do not imply that MDHV required the presence of ALV to produce disease.


The polypeptides of hepatitis A virus.

The polypeptides of hepatitis A virus. Hepatitis A virus was purified from fecal specimens obtained from 3 patients with naturally acquired hepatitis A, by a process of differential centrifugation, chloroform extraction, column chromatography, and isopycnic ultracentrifugation. Analysis of purified virus by discontinuous SDS-PAGE revealed three major polypeptides with molecular weights of 34,000, 25,500, and 23,000 daltons. These polypeptides appear to be specific for hepatitis A virus and have similar molecular weights to three of the four major polypeptides reported for members of the genus Enterovirus within the family Picornaviridae.


Transabdominal lymphography after intraperitoneal injection of Rous virus in newborn rats. Newborn rats were inoculated in the peritoneal cavity with Rous sarcoma virus (Schmidt-Ruppin strain). They were studied lymphographically and histologically after various intervals of time. As a result of lesions of the lymph vessels and nodes, the lymphatic drainage of the peritoneal cavity was increasingly impaired. Pleural effusion developed as a result of leakage through the walls of the lymphatics. The passage through the lymph nodes was blocked and large lymph cysts arose in the mediastinum and retroperitoneally.


Rapid identification and subtyping of herpes simplex virus by complement-dependent cytotoxicity. A simple method for rapid identification and subtyping of herpes simplex virus from clinical specimens is presented. Inoculated cultures demonstrating cytopathic changes were treated with type-specific cytotoxic antisera against herpes simplex virus and complement. Lysis of cells was revealed by staining with trypan blue. The technique allowed identification and subtyping of herpes simplex virus within 2 h after the appearance of CPE.


Observation of togavirus-like particles in nonhuman primates. Particles resembling togaviruses were observed by electron microscopy in the urinary bladder lamina propria of one capuchin monkey and in placental syncytiotrophoblasts of one baboon. Type C viruses were observed in the baboon placenta, but not in the tissues of the capuchin monkey.


Autoradiographic detection of Epstein-Barr virus (EBV)-associated early antigen in a variety of EBV DNA-containing lymphoblastoid cell lines previously designated as nonproducers. By using autoradiography combined with 125I-labeled IgG prepared from human sera containing high anti-EA (early antigen) antibody titers, EA has been detected in a low fraction of cells in a variety of EBV DNA-containing cell lines previously designated as negative for this antigen. These positive cell lines are highly inducible for EA following treatment with iododeoxyuridine (IUDR) and also contain multiple copies of the virus genome on average. The correlation between IUDR inducibility and spontaneous expression of EA, in particular, suggests that the ability to express EA in both situations is interrelated.


Distributions of 137Cs, 201Tl, 203Hg, 203Pb and 57Co in a rat hepatoma model. Comparison with 67Ga. The distribution of carrier-free 203Pb-acetate, 203HgCl2, 57 CoCl2, 137CsCl and 201TlCl was investigated in rats bearing thigh-implanted Morris 7777 hepatomas. Viable and nonviable tumor tissue was collected in order to determine the relative affinities of the radiopharmaceuticals for these tissues. The animals were sacrificed at 4, 24, 48, 72 and 96 hrs following intravenous injection. Washout of the radioisotope from the viable tumor tissue was rapid, the maximum concentration being reached on or before 4 hrs following injection. In contrast, residual activity within the nonviable tumor tissue decreased much more slowly and in some cases even increased with time. Viable tumor-to-muscle and nonviable tumor-to-muscle ratios for 203Pb, 203Hg and 57Co were comparable to the analogous ratios reported for 67Ga. However, none of these isotopes approached 67Ga as a potential tumor imaging agent because the large ratios were the result of low muscle uptake rather than high tumor uptake. Blood clearance of 67Ga was faster than any of the five cations, while viable and nonviable tumor affinity for 67Ga was greater than for any of the radiopharmaceuticals studied.


Relationships between base-exchange in phospholipid metabolism and cyclic-AMP levels of synaptosomal membranes. The effect of the base-exchange reaction upon the production of cyclic-AMP at the level of rat brain synaptic membranes has been examined. The exchange with ethanolamine produces a significant decrease of the NaF-stimulated production of the cyclic nucleotide, whereas it increases the noradrenaline-induced production. With some exceptions, the exchange with L-serine produces opposite effects.


Polymeric forms of yeast nicotinamide-adenine dinucleotide-specific isocitrate dehydrogenase in the presence of various ligands. The molecular weight of NAD-specific isocitrate dehydrogenase, purified from baker's yeast, has been studied by molecular sieve chromatography. By elution of the enzyme from columns of Sepharose 6B with 0.05 M phosphate buffer, pH 7.6, a mol. wt of 151,000 was measured. Higher values of the mol. wt were measured in presence of the following ligands (mol. wt in parentheses): the substrate, isocitrate (224,000); the activators, citrate (203,000) and AMP (275,000); the inhibitor, NaCl (360,000). A mol. wt of 337,000 was measured when AMP, which antagonizes the inhibition by chloride, was present together with NaCl. The results indicate the absence of a correlation between the aggregation form of the enzyme in presence of the ligands and the effects of these ligands on the enzyme activity.


Cancer: the eighth plague--a suggestion of pathogenesis. The current treatment of cancer is based on the assumption that this disease is the result of autonomous clonal proliferation of aberrant cells which must be excised, irradiated or selectively poisoned to achieve a cure. The presumption that the malignant cell constitutes the disease is now challenged by a variety of clinical observations and experimental studies. Like the grasshopper, which is transformed into a locust under altered environmental conditions, the cancer cell may be the manifestation of a defect in homeostatic regulation of cellular proliferation. Identification of the specific regulatory defect could allow for a more rational and more effective treatment of cancer in man.


Effects of prophylactic treatment with the methanol extraction residue fraction of tubercle bacilli (MER) on the development of Rous sarcomas of chickens following challenge with the Rous sarcoma virus. Three-month-old chickens were treated with the methanol extraction residue fraction of tubercle bacilli (MER) under different conditions, and subsequently challenged with living Rous sarcoma virus. The birds developed progressively growing sarcomas following viral challenge. A substantial proportion of the hosts which had been pretreated with MER under optimal circumstances (38 to 58%) showed complete and long-lasting regression of the neoplasms, and the survival of many of the animals that did succumb to progressively growing tumors was prolonged. An absolute condition for prophylactic efficacy of MER treatment in this model system was injection of the agent into the same body area (wing) into which subsequent viral challenge was introduced. The quantity of MER employed and the timing of the prophylactic administration were also decisive variables.


Affinity chromatography of yeast nicotinamide-adenine dinucleotide-specific isocitrate dehydrogenase on immobilized nicotinamide-adenine dinucleotide. Effects of ligands. The method of affinity chromatography has been used for studying the effects of some ligands of yeast NAD-specific isocitrate dehydrogenase on the affinity of the enzyme for NAD+ immobilized on Sepharose 4B. In absence of ligands, the enzyme is eluted from NAD+-Sepharose columns by 0.1 M phosphate buffer, pH 7.6, in a highly purified form. The elution of enzyme is accelerated by NAD+ and, more effectively, by AMP; and retarded by isocitrate and citrate. The elution patterns show a rather irregular shape, probably due to the occurrence of aggregation processes of the enzyme protein.


Inert gas a-A differences: a direct reflection of V/Q distribution. A computer model was developed to study the relationship between ventilation-to-perfusion (V/Q) mismatch and the development of inert gas arterial-to-alveolar partial pressure differences (a-A differences). Increasing inhomogeneity of V/Q ratio is revealed directly as an increase in the a-A difference of each gas. The quantitative relationships between the Q vs. V/Q distribution and the fractional a-A difference solubility plot (a-A difference plot) were studied and described. These studies demonstrated that for log normally distributed V/Q ratios, the area under the a-A difference plot is linearly related to the log variance of the V/Q distribution and can be estimated directly from the values obtained from six gases. The maximum a-A difference occurs for a gas whose solubility is numerically equal to the mean V/Q. The effects of departure from log normality and multimodality are discussed. We conclude from these studies that quantitative information regarding the degree of inhomogeneity of V/Q for log normal distribution is available from direct calculations of inert gas retention and excretion data. Qualitative information is also available indicating the departure from log normality and the region toward which the distribution is skewed.


Longitudinal mixing in pulmonary airways--normal subjects respiring at a constant flow. We have measured the impulse response of helium and sulfur hexafluoride in the airways of five normal human subjects at a respiratory flow of 400 ml/s. The longitudinal mixing of the inert gases was characterized by the increased volume variance of the expired concentration response. This parameter was measured over the largest possible range of airway penetrations, 30-290 ml. Employing a symmetrical model of the airway geometry, we have computed the values of a mean mixing coefficient from the volume variance data. This mixing coefficient is largest in the large airways and decreases rapidly with increasing penetration; it may be as much as 4,000 times greater than the molecular diffusivity; and it is relatively independent of the inert gas tested, at least up to an airway penetration of 180 ml. These observations are consistent with several preivously proposed mixing mechanisms including axial streaming, turbulent dispersion, and mixing by geometric asymmetry. However, the latter observation appears to rule out the importance of laminar dispersion since mixing by this mechanism is inversely dependent on the molecular diffusivity.


Biochemical activities of the derivatives of dehydrodicaffeic acid dilactone. Activities of derivatives of dehydrodicaffeic acid dilactone (DDCAD) to inhibit catechol-O-methyltransferase (COMT), cyclic AMP phosphodiesterase (PDE) and DOPA decarboxylase (DDC) were examined. Among those tested, 2,6-bis-(5',6'-dibromo-4'-hydroxy-3'-methoxyphenyl)-3,7-dioxabicyclo-[3,3,0]-octane 4,8-dione was found to be the strongest inhibitor of both COMT and PDE. There were no derivatives which showed a stronger inhibition against DDC than the original compound, DDCAD.


Hypoglycemia: an overview.

Hypoglycemia: an overview. Hypoglycemia was not described as a human clinical syndrome until exogenous insulin was discovered. Much of our current knowledge of the symptoms of hypoglycemia derived from the reactions of schizophrenics to insulin coma therapy in the late 1920's. The diagnosis of hypoglycemia cannot be made simply on the basis of a blood glucose level since many asymptomatic persons have levels below 50 mg/100 ml. The diagnosis should be made only if symptoms occur at the glucose nadir and are relieved by the administration of glucose. The most common organic cause of hypoglycemia is functioning islet-cell tumor. By far the most prevalent type of hypoglycemia is idiopathic (function), a condition whose pathophysiology is not understood and which has given rise to a popular lay "mythology."


Hysterical symptoms masking brain stem glioma. To function effectively as primary care specialists, psychiatrists must remain ever alert to the possibility of organic disorders in patients who at first show only psychiatric symptoms. A case is presented in which hysterical overlay led to misdiagnosis in a 31 year woman, who dies of a diffuse medullary glioma 3 1/2 years after onset of "conversion" symptoms. The authors point out how the label "hysterical" clouds longitudinal objective diagnostic observations especially when initial clinical and laboratory data fail to support a definitive organic diagnosis.


Clinic versus private treatment of psychosis. Thirty patients with a hospital diagnosis of schizophrenia were randomly assigned upon discharge to either 1) a specialized aftercare clinic of 2) to private psychiatrists. At the end of six months, the clinic patients had spent significantly less time as hospital readmissions and were significantly more occupied (i.e. employed or in school) than the private patients. It seems possible that attendance at clinic provides group pressure to maintain drug compliance and may thus work to reduce rehospitalization. It is also likely that the network of clinic paramedical and volunteer staff, by its flexibility and availability, aids the patient's transition into employment.


Antibody and cell-mediated immunity to herpes simplex virus in psychotic depression. The incidence and antibody titers to herpes simplex virus (HSV) were found significantly higher in patients with psychotic depression as compared to normal controls. Furthermore, the cell-mediated immunity (CMI) to HSV in psychotic depression was similar to that observed after acute HSV infection or recurrence. The results suggest therefore an association between HSV infection and psychotic depression.


Micro determination of unsaturated disaccharide formed by the action of acidic glycosaminoglycan-endoeliminases. An application of the thiobarbituric acid method to the assay of D-gluco-4-enepyranosyluronic acid-containing disaccharides. An improved method is described for the micro determination of acidic glycosaminoglycans after digestion with chondroitinase-ABC and -AC. The determination is based on the color production of D-gluco-4-enepyranosyluronic acid-containing disaccharides produced by the action of chondroitinase-ABC and -AC (acidic glycosaminoglycans-endoeliminase) when the periodate-thiobarbituric acid method is applied to the alpha,beta-unsaturated disaccharides. Suitable conditions for the quantitative assay are described.


C-type cytochromes isloated from an extreme thermophile, Thermus thermophilus HB8. Two cytochromes of the C-type, c-554 and c-549, were isolated from the soluble fraction of an extreme thermophile, Thermus thermophilus HB8. Highly purified cytochrome c-554 had absorption maxima at 554, 522, and 417 nm in the reduced state, and at 410 nm in the oxidized state. The alpha-band of the reduced state resembled that of "split-alpha" cytochromes. The isoelectric point was at pH 4.9, and the molecular weight was about 29,000. Cytochrome c-549, partially purified, had absorption maxima a6 549,520, and 416 nm in the reduced form, and at 408 nm in the oxidized form. The molecular weight was about 25,000. Both were slowly auto-oxidizable, and did not combine with CO.


A single cleavage of Simian virus 40 (SV40) DNA by a site specific endonuclease from Thermus aquaticus, Taq I. A site specific endonuclease from Thermus aquaticus, Taq I, cleaves Simian virus 40 (SV40) DNA at a single site. The cleavage site was localized on the physical map by double digestions, using the previously characterized fragments produced by digestion with Hae II, Hae III, AluI, HhaI, HinfI, or BstI. The Taq I site is located at the position that is 56.5% of the unit length from the Eco RI site.


Presence of the sites for interacting with cyclic AMP and with catalytic subunit on small fragments of protein kinase regulatory subunit. During the purification of cyclic AMP binding proteins from rat liver, some smaller active fragments were obtained, possibly as the result of proteolysis. The binding proteins detected had approximate molecular weights of 50,000, 36,000, and 10,000. Each of these components bound cyclic [3H]AMP with high affinity (apparent dissociation constants ranging from 2 to 10 nM) and had a similar ability to inhibit the purified catalytic subunit of rat liver protein kinase. Cyclic AMP prevented this inhibition in each instance. These results suggest that the binding site for cyclic AMP and the site for interacting with catalytic subunit occur relatively close to one another on the regulatory subunit and can remain functional when a substantial fraction of the subunit is lost.


Phosphoglycolate phosphatase. Purification and properties. Phosphoglycolate phosphatase (EC 3.1.3.18) was purified 1500-fold from field-grown tobacco leaves by acetone fractionation, DEAE-cellulose and molecular sieve chromatography, and preparative polyacrylamide gel electrophoresis. Preparations were judged 90 to 95% homogeneous by chromatography on DEAE-cellulose, polyacrylamide gel electrophoresis, and by isoelectric focusing. The highest specific activity obtained was 468 mumol of phosphate released/min/mg of protein. The native protein has a molecular weight of 80,500 by Ferguson plot analysis and 86,300 by sedimentation velocity on sucrose density gradients. Sodium dodecyl sulfate-polyacrylamide gels gave a molecular weight of 20,700, indicating the P-glycolate phosphatase is a tetramer with identical or near identical subunits. The enzyme, freshly purified or in crude homogenates, had a pI of 3.8 to 3.9 pH units by isoelectric focusing. Phosphosphoglycolate phosphatase from spinach leaves has a molecular weight of 93,000 and, unlike the enzyme from tobacco leaves, it is extremely unstable after DEAE-cellulose chromatography and is inactivated by lipase (EC 3.1.1.3). The phosphatase from both plants was stabilized by the addition of citrate or isocitrate in the buffers. Ribose 5-phosphate is a competitive inhibitor of phosphoglycolate phosphatase at physiological concentration, while other phosphate esters of the photosynthetic carbon cycle were without effect.


Mechanism of phosphoglycolate phosphatase. Studies of hydrolysis and transphosphorylation, substrate analogs, and sulfhydryl inhibition. Enzymatic hydrolysis of phosphoglycolate proceeds through O-P bond cleavage as determined by reaction in H218O and analysis of the trimethylsilyl derivatives of the reaction products by mass spectrometry. No phosphate, hydroxyl, or carboxyl exchange occurred. End product inhibition was consistent with an ordered release of products, first the alcoholic product, glycolate, then phosphate. Analysis of the data indicated that the phosphate.enzyme complex dissociated very rapidly, and this was confirmed by use of alternative phosphomonoester substrates. Maximum velocity with these alternate substrates was found to be proportional to the pKa of of the corresponding alcoholic product, indicating the rate-limiting step in the reaction was protonation of the bridge oxygen. The use of substrate analogs further suggested that enzymatic specificity residues in exacting steric requirements for binding, and that large alkyl groups were excluded on this basis. Phosphoglycolate phosphatase catalyzed transphorylation to a wide range of acceptors and was inhibited at the active site by diisopropyl-fluorophosphate. The data suggest that the reaction sequence proceeds via a phosphoenzyme intermediate. N-Ethylmaleimide slowly inactivated the enzyme, the rate being greatly increased by P-glycolate, but not by magnesium or phosphate ions. The data suggest a conformational change is necessary to induce the transition state complex and phosphoenzyme formation. This may account for the phosphate acceptor specificity and is consistent with the failure to observe an enzyme-mediated H2O-phosphate oxygen exchange.


Interaction of cytochrome c with cytochrome c oxidase. Photoaffinity labeling of beef heart cytochrome c oxidase with arylazido-cytochrome c. Cytochrome c derivatives labeled with a 3-nitrophenylazido group at lysine 13, at lysine 22, or at both residues have been prepared. The interaction of the cytochrome c derivatives with beef heart cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1) in the presence of ultrviolet light results in formation of a covalent complex between cytochrome c and the oxidase. Using the lysine 22 derivative, the polypeptide composition of the oxidase is not modified, nor is its catalytic activity, whereas with the lysine 13 derivative, the gel electrophoretic pattern is altered and the catalytic activity of the complex diminished. The data are consisten with a specfic covalent interaction of the lysine 13 derivative of cytochrome c with the polypeptide of molecular weight 23,700 (Subunit II) of cytochrome c oxidase.


Cyclic nucleotide phosphodiesterase. Partial purification and characterization of a high affinity enzyme activity from human lung tissue. Part of the soluble cyclic nucleotide phosphodiesterase activity of crude human lung tissue can be attributed to a thermosensitive (37 degrees) enzyme with a high apparent affinity for both adenosine 3':5'-monophosphate (cyclic AMP) and guanosine 3':5'-monophosphate (cyclic GMP). The enzyme can be partially purified by DEAE-Sephadex chromatography. In the presence of 0.1 mM EDTA or ethylene glycol bis(beta-aminoethyl ether)N,N'-tetraacetic acid (EGTA), it is eluted from the column immediately before a cyclic GMP-specific phosphodiesterase, but in the presence of 0.2 mM Ca2+, the elution follows that of the cyclic GMP-specific enzyme. The two forms of the nonspecific phosphodiesterase activity are referred to as DEAD-Sephadex Fractions Ia and Ic, respectively. Their apparent molecular weights, recorded at gel filtration, vary with different preparations from 230,000 to 150,000. Occasionally, corresponding recordings for main peaks of activity also cluster round the values 120,000, 105,000, and 78,000. The enzymatic properties of Fractions Ia and Ic closely resemble each other. The enzyme activity is blocked by EDTA, partially inhibited in the presence of 1,10-phenanthroline, but only slightly affected by EGTA. The inhibitory effect of EDTA can be overcome by Mg2+ and Mn2+ and that of 1,10-phenanthroline, in part, by Zn2+; this cation in itself is inhibitory at millimolar concentrations. With submicromolar substrate concentrations, the activity of either fraction obeys linear kinetics displaying an apparent Km of approximately 0.4 micron for both substrates. Reciprocal inhibition experiments suggest that hydrolysis of both cyclic AMP and cyclic GMP is performed by the same active site. Examination of the activity using extended substrate concentration ranges indicates nonlinear kinetics; Hill plots of such data also show nonlinear curvature. The activity is inhibited by micromolar concentrations of inosine 3':5'-monophosphate (cyclic IMP), 3-isobutyl-1-methylxanthine, papervine, and some antiallergic agents. Theophylline and disodium cromoglycate are less potent inhibitors. Inhibition of activity by Lubrol PX follows a biphasic dose response curve. The activity of Fraction Ia can be enhanced 2- to 3-fold by a Ca2+-dependent activator prepared from lung tissue, whose action is counteracted by chlorpromazine, and by lysophosphatidylcholine. It is initially enhanced but subsequently decreased at exposure to trypsin. Fraction Ic is less prone to activation by these agents. The results indicate that the present activity represents an enzyme form that differs from three previously described phosphodiesterases of human lung tissue. It is apparently related to, but also shows distinct differences from the Ca2+-dependent enzyme(s) of brain and heart tissue.


The purine nucleotide cycle. Studies of ammonia production and interconversions of adenine and hypoxanthine nucleotides and nucleosides by rat brain in situ. Electrical shock treatment produces a rapid loss of high energy phosphates in rat brain. The [ATP]/[ADP] ratio decreases to one-third of its control value within 10 s. The ammonia content increases 3-fold during the first minute after starting the stimulus. The total adenine nucleotide plus adenosine content of brain decreases an equivalent amount of hypoxanthine-containing compounds appears. Adenosine, inosine, and hypoxanthine accumulate, and there is a transitory accumulation of adenylosuccinate. The contents of ATP and creatine phosphate, and the [ATP]/[ADP] ratio, are rapidly restored to control values, but other metabolite contents are restored more slowly. The transient rise in adenylosuccinate and IMP provides evidence that the ammonia production is due in part, and possibly in whole, to the operation of the purine nucleotide cycle.


Similarities in the membrane fluidity of 3T3 and SV101-3T3 cells and its relation to concanavalin A- and wheat germ agglutinin-induced agglutination. Intact, viable ultransformed 3T3 and transformed SV101-3T3 cells were labeled with fatty acid spin labels and with 2,2,6,6-tetramethylpiperidine-1-oxyl in order to measure the fluidity properties of membrane lipids. Both cell types were grown in regular calf serum and in a lipid-depleted serum supplemented with either oleate or elaidate. The temperature dependence of the spectra obtained revealed inflections that correlate with the temperature below which agglutination with concanavalin A is inhibited, and another inflection that correlates with the temperature below which agglutination with wheat germ agglutinin is inhibited, suggesting that (a) the lipid phase(s) in the vicinity of the receptor(s) for these two lectins differ, and (b) a fluid membrane in the vicinity of the lectin receptor(s) is necessary for agglutination with either concanavalin A or wheat germ agglutinin. Studies with a partially characterized plasma membrane fraction suggest that the plasma membrane fluidity parameters closely resemble those of the intact cell. 3T3 and SV101-3T3 cells show virtually identical fluidity profiles by all of the tests we have applied.


DNA-directed in vitro synthesis of Escherichia coli beta-isopropylmalate dehydrogenase. The in vitro synthesis of beta-isopropylmalate dehydrogenase (EC 1.1.1.85), an enzyme involved in leucine biosynthesis, has been obtained using as template DNA from the hybrid plasmid (pLC1) which contains the Escherichia coli leucine operon. Enzyme synthesis in vitro is stimulated about 2-fold by guanosine-5'-diphosphate-3'-diphosphate and inhibited about 60% by 2 X 10(-4) M L-leucine.


Active phosphate ion transport in plasma membrane vesicles isolated from mouse fibroblasts. Inorganic phosphate accumulated 8-fold in plasma membrane vesicles derived from simian virus 40-transformed 3T3 mouse fibroblasts when a NaCl gradient (external greater than internal) was artificially imposed across the membrane. Preincubation with Na+ or addition of monensin markedly reduced phosphate accumulation. Na+-stimulated phosphate transport was not affected by addition of either dicarboxylic acids, antimycin A, or ouabain and persisted after addition of proton ionophores. The coupling of phosphate transport to Na+ gradients was pH-dependent, with maximal stimulation by Na+ below pH 7. These findings suggest that monovalent phosphate anion moves across the plasma membrane in co-transport with sodium ion.


Aspartokinase I-homoserine dehydrogenase I of Escherichia coli K12. Concentration-dependent dissociation to dimers in the presence of L-threonine. The concentration-dependent association-dissociation equilibrium of the bifunctional enzyme aspartokinase I-homoserine dehydrogenase I of Escherichia coli K12 has been investigated at pH 7.6 in the presence of 10 mM L-threonine and 0.1 M KCl by equilibrium gel permeation monitored by a single-photon counting spectrophotometer. The results obtained are consistent with the existence of a dimer-tetramer equilibrium with the association constant of 2.6 X 10(7) M-1 (deltaG0 = -9.9 kcal/mol of dimer). The limiting partition cross-sections estimated by a three-parameter least squares minimization procedure indicate that the molecular radii of the dimer and tetramer are 53.8 A and 70 A, respectively. Both the dimeric and tetrameric forms of the enzyme possess dehydrogenase activity. Treatment of the enzyme with the chaotropic salts, potassium thiocyanate or potassium trichloroacetate, generates a monomeric form that is devoid of dehydrogenase activity. The catalytically inactive monomeric form of the enzyme has a molecular radius between 43 and 45.5 A and a molecular weight of approximately 80,000 as determined by small zone gel chromatography and sedimentation equilibrium studies.


The preparation and characterization of highly purified, enzymically active complex III from baker's yeast. A soluble enzymically active cytochrome b.c1 complex has been purified from baker's yeast mitochondria by a procedure involving solubilization in cholate, differential fractionation with ammonium sulfate, and ultracentrifugation. The resulting particle is free of both cytochrome c oxidase and succinate dehydrogenase activities. The complex contains cytochromes b and c1 in a ratio of 2:1 and quinone and iron-sulfur protein in amounts roughly stoichiometric with cytochrome c1. EPR spectroscopy has shown the iron-sulfur protein to be present mainly as the Rieske protein. EPR spectroscopy also shows a heterogeneity in the cytochrome b population with resonances appearing at g = 3.60 (cytochrome bK) and g = 3.76 (cytochrome bT). A third EPR resonance appearing in the region associated with low spin ferric hemes (g = 3.49) is assigned to cytochrome c1. Anaerobic titration of the complex with dithionite confirmed the heterogeneity in the cytochrome b population and demonstrated that the oxidation-reduction potential of the iron-sulfur protein is approximately 30 mV more positive than cytochrome c1. An intense EPR signal assigned to the coenzyme Q free radical appeared midway in the reductive titration; this signal disappeared toward the end of the titration. A conformational change in the iron-sulfur protein attendant on reduction of a low potential species was noted.


The electronic state of heme in cytochrome oxidase II. Oxidation-reduction potential interactions and heme iron spin state behavior observed in reductive titrations. Magnetic circular dichroism (MCD), electron paramagnetic resonance (EPR), and optical absorption spectroscopies have been used to monitor the concentrations of oxidized and reduced heme and copper during stoichiometric reductive titrations of purified beef heart cytochrome oxidase. The MCD data are deconvoluted to obtain the concentrations of reduced cytochromes a and a3 during the titrations; analysis of the EPR spectra provides complementary data on the concentrations of the EPR-detectable species. For the native enzyme in the absence of exogenous ligands, cytochromes a and a3 are reduced to approximately the same extent at all points in the titration. The reduction of the EPR-detectable copper, on the other hand, initially lags the reduction of the two cytochromes but in the final stages of the titration is completely reduced prior to either cytochrome a or a3. These non-Nernstian titration results are interpreted to indicate that the primary mode of heme-heme interaction in cytochrome oxidase involves shifts in oxidation-reduction potential for each of the two cytochromes such that a change in oxidation state for one of the hemes lowers the oxidation-reduction potential of the second heme by approximately 135 mV. In these titrations high spin species are detected which account for 0.25 spin/oxidase maximally. Evidence is presented to indicate that at least some of these signals can be attributed to cytochrome a3+ which has undergone a low-spin to high-spin state transition in the course of the titration. In the presence of carbon monoxide the oxidation-reduction properties of cytochromes a and a3 are markedly altered. The a32+. CO complex is fully formed prior to reduction of either cytochrome a3+ or the EPR-detectable copper. The g = 3 EPR signal attributed to cytochrome a3+ decreases as the MCD intensity of cytochrome a2+ increases; no significant high-spin intensity is observed at any intermediate stage of reduction. We interpret these Nernstian titration results to indicate that in the presence of ligands the oxidation-reduction potential of cytochrome a relative to cytochrome a3 is determined by the oxidation-reduction state of the stabilized cytochrome a3 ligand complex; if ligand binding occurs to reduced cytochrome a3 then cytochrome a titrates with a lower potential; cytochrome a titrates with a higher potential if oxidized cytochrome a3 is stabilized by ligand binding.


Molecular symmetry of fructose-1,6-diphosphatase by X-ray diffraction analysis. Large single crystals of rabbit liver fructose-1,6-diphosphatase suitable for a high resolution structure analysis have been grown from polyethylene glycol. The space group of these crystals is I222 with a = 75 A, b = 81 A, and c = 132 A and there are 2 tetrameric molecules in the unit cell. These crystals have one protein subunit as the crystallographic asymmetric unit and establish point group symmetry 222 as the molecular symmetry.


Insulin regulation of Leydig cell luteinizing hormone receptors. The effect of experimentally induced diabetes on the luteinizing hormone-human chorionic gonadotropin receptors in rat testes was examined. Pancreatectomy and streptozotocin diabetes the gonadotropin receptors and the administration of insulin restored the binding capacity to normal values. No differences in the association constant as well as the sedimentation coefficients of the luteinizing hormone-human chorionic gonadotropin receptor complexes were observed between normal and diabetic animals.


The soluble "high potential" type iron-sulfur protein from mitochondria is aconitase. Properties of soluble high potential type iron-sulfur protein (HiPIP) from beef heart mitochondria were compared to those of aconitase from pig heart. The two proteins when purified to homogeneity by the criteria of sodium dodecyl sulfate (SDS)-polyacrylamide electrophoresis show identical light absorption characteristics. EPR signals of the HiPIP type centered at g = 2.01 when oxidized, isoelectric points at pH 8.5 to 8.6, are inseparable by SDS-polyacrylamide electrophoresis, and exhibit aconitase activity when activated by reducing agents in the presence of ferrous iron. The requirement for activation goes parallel to the intensity of the signal from the oxidized iron-sulfur cluster, i.e. the cluster is reduced in the active enzyme. We conclude that the soluble mitochondrial HiPIP is identical with aconitase. The relationships of iron to labile sulfide, molecular weight and unpaired spins in the EPR signal, and implications of our findings for the role of iron in aconitase are discussed.


Cytidine 3':5'-monophosphate phosphodiesterase in mammalian tissues. Occurrence and biological involvement. A phosphodiesterase activity that preferentially hydrolyzed cytidine 3':5'-monophosphate was partially purified from rat liver extract. The enzyme was best activated by Fe2+ (5 to 10 mM). Mn2+ and Mg2+ were less effective, whereas Zn2+, Co2+, and Ca2+ were ineffective. It exhibited kinetics typical of a high Km phosphodiesterase, with a Km for cycli CMP of 2.4 mM. The enzyme, inhibited by theophylline and 1-methyl-3-isobutyl xanthine to much less extents than cyclic AMP and cyclic GMP phosphodiesterases, was found in all rat tissues examined, with highest levels seen in the liver, kidney, and intestine, and lowest levels found in the skeletal muscle, cerebellum, aorta, and blood cells. The enzyme levels in the regenerating liver were found to be about 40% lower than the control liver of rats; they were also 3 to 10 times lower in the fetal liver, lung, and heart than the corresponding adult tissues of guinea pigs. These findings suggest that depressed cyclic CMP phosphodiesterase may be in part related to cell proliferation, in line with reports that the regenerating liver has higher levels of cyclic CMP (Bloch, A. (1975) Adv. Cycli Nucleotide Res. 5, 331-338) and cytidylate cyclase (Cech, S. Y., and Ignarro, L.J. (1977) Science 198, 1063-1065).


Demonstration, in leukemia L-1210 cells, of a phosphodiesterase acting on 3':5'-cyclic CMP but not on 3':5'-cyclic AMP or 3':5'-cyclic GMP. cCMP-specific phosphodiesterase activity was demonstrated in the 80 to 100% ammonium sulfate fraction obtained from disrupted leukemia L-1210 cells. The activity was linear with time (up to 60 min), was a function of protein concentration, and was markedly stimulated by Mg2+ and by ammonium sulfate. Under identical assay conditions, no significant hydrolysis of cAMP or cGMP was observed, although these cyclic nucleotides served as substrates for phosphodiesterase(s) present in all the fractions obtained by less than 80% ammonium sulfate saturation. This is the first demonstration of a cCMP-specific phosphodiesterase.


The indispensability of phospholipid and ubiquinone in mitochondrial electron transfer from succinate to cytochrome c. The indispensability of phospholipid and ubiquinone (Q) in mitochondrial electron transfer was studied by depleting phospholipid and Q in succinate-cytochrome c reductase and then replenishing the depleted enzyme. More than 90% of phospholipid and Q was removed by repeated ammonium sulfate-cholate fractionation. The depleted succinate-cytochrome c reductase showed no enzymatic activity for succinate leads to c or QH2 leads to c and yet retained most of the succinate leads to Q activity. All enzymatic activity was restored upon the addition of Q and phospholipid. Restoration required the addition of Q prior to the addition of phospholipid. Reversing the addition sequence or addition of a mixture of phospholipid and Q resulted only in a small restoration of activities. The conditions for restoration are given in detail. Removal of phospholipid from succinate-cytochrome c reductase resulted in reduction of cytochrome c1 in the absence of exogenous electron donor. Replenishing the preparation with phospholipid brought about the reoxidation of cytochrome c1 in the absence of electron acceptor or oxygen.


Partial purification and properties of the Epstein-Barr virus-associated nuclear antigen. The Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA) was purified 85-fold from a nuclear pellet derived from an EBV-transformed B lyphoblastoid cell line by a five-step procedure consisting of preparation of extract, heating at 80 degrees C in phosphate buffer, ammonium sulfate precipitation, preparative ultracentrifugation, and affinity chromatography on double-stranded DNA-cellulose. The purified complement fixing antigen specifically blocked the anticomplement immunofluorescence assay for EBNA. Several properties indicate a close association of EBNA with chromatin, viz. 1) precipitation of antigenic activity by phosphate buffer and subsequent thermal fractionation; 2) partial sensitivity of antigenic activity to DNase (but not to RNase) and restoration of activity by addition of calf thymus DNA; and 3) specific binding of EBNA to double-stranded DNA-cellulose. Other properties of EBNA, including its unusual heat stability, are described.


Immune adherence hemagglutination test applied to the study of herpes simplex and varicella-zoster virus infections. The immune adherence hemagglutination (IAHA) test has been used successfully to detect antibody to herpes simplex (HS) virus and varicella-zoster (V-Z) virus. Comparative studies between the complement-fixation (CF) test and the IAHA test revealed that, in most cases, the IAHA test was more sensitive than the CF test. Furthermore, diagnosis on the basis of a fourfold change in antibody titer was made more rapidly by the IAHA test. The IAHA test was found to be a very simple and practical technique requiring only a few hours for completion compared with the conventional CF test which required up to 24 h. In addition, both sera and cerebrospinal fluids could be tested in very low dilutions in the IAHA test, so that very low levels of antibody could be detected. Also, the IAHA test detected antibody to V-Z virus more frequently than did the CF test in adults with a history of varicella occurring 9 to 30 years prior to sampling. The level of cross-reaction between HS and V-Z viruses was examined by both the CF test and the IAHA test, and no major differences between the two techniques were found.


Enzyme-linked immunosorbent assay for detection of hepatitis A antigen in stool and antibody to hepatitis A antigen in sera: comparison with solid-phase radioimmunoassay, immune electron microscopy, and immune adherence hemagglutination assay. Previously described techniques for detection of hepatitis A antigen (HA Ag) and antibody (anti-HA) have required purified HA Ag and expensive equipment. Herein is described an enzyme-linked immunosorbent assay (ELISA) for specific detection of HA Ag in human stool filtrates and of anti-HA in sera by using selected HA Ag-containing human stool filtrates as the antigen source. Because human stools often react nonspecifically in serological tests for HA Ag, blocking with preexposure and hyperimmune anti-HA sera from a chimpanzee inoculated with hepatitis A virus was used to confirm specific detection of HA Ag. The sensitivity of ELISA was found to be comparable to that of solid-phase radioimmunoassay (SPRIA) and immune electron microscopy (IEM). Of 37 acute-phase stools collected from nine patients, 16 were positive for HA Ag by ELISA. In 13 of these, HA Ag particles were found by IEM, and an additional 3 stools negative by ELISA contained HA Ag particles by IEM. Eight control stools were negative by both ELISA and IEM. Anti-HA was measured in sera by demonstrating its ability to block binding of the enzyme conjugate to HA Ag in a stool without detectable nonspecificity. This test (blocking ELISA) was as sensitive and specific as blocking SPIRA, IEM, and immune adherence hemagglutination and, like SPRIA and IEM, detected early-developing antibody. The ELISA is simple to perform and requires only a minimum of equipment. It is useful for screening stools for HA Ag and for monitoring HA Ag during purification, as well as for detecting early and late anti-HA in sera.


Hepatocellular carcinoma and hepatic cirrhosis in the west of Scotland: a 25-year necropsy review. A two-fold increase in the incidence of hepatocellular carcinoma in the west of Scotland is reported on the basis of a 25-year retrospective necropsy review (313 cases). This increase is not accompanied by a corresponding increase in the incidence of hepatic cirrhosis. The relationship between hepatocellular carcinoma and hepatic cirrhosis is discussed in the light of these findings.


Routine diagnosis of human rotaviruses in stools. Electron microscopy, immune electron microscopy and complement fixation as methods of detecting rotavirus in the stools of young children with gastroenteritis were compared in a blind study during the winter of 1975-6. Complement fixation was the simplest to perform, was as sensitive as the other two, and allowed a quantitative measurement of viral excretion. Absorption of faecal extracts with fetal calf serum usually removed the anticomplementary activity of faecal extracts.


Laboratory diagnosis of EB virus infection in some cases presenting as hepatitis. In 1975 and 1976, 7580 serum specimens were tested for HBsAg by the passive haemagglutination test, Hepanosticon. Serum from 38 people gave a positive result after absorption with the absorbent provided, but the presence of HBsAg could not be confirmed by other tests. Tests for current infection with EB virus, namely, the presence of heterophil antibody and EB virus specific IgM, were performed and were positive in 11 of the 38. In six of these the clinical picture was of hepatitis rather than infectious mononucleosis.


Changes in peripheral nerve function with long-term hemofiltration treatment. The changes in the peripheral nerve function of three patients with chronic renal failure who were treated over a period of 1-1/2 to 2 years by hemofiltration have been analyzed in the form of a longitudinal study by quantitative measurements of vibratory perception threshold and nerve conduction velocity. Changes in vibratory perception threshold were measured in six patients both before and after treatment. Despite increased values of creatinine and BUN, an improvement in the peripheral nerve function of all patients undergoing hemofiltration could be observed, although vibratory perception and nerve conduction velocity did not return to normal. All of the vibratory perception threshold measurements made directly after hemofiltration showed an improved vibratory perception in comparison to the original values. A comparison of the measurement methods showed a good correlation and clearly indicated the advantage of vibratory perception threshold measurements as a means for the routine diagnosis of nephrogenic polyneuropathy.


The glucagonoma syndrome.

The glucagonoma syndrome. The glucagonoma syndrome is characterized by necrolytic migratory erythema, glossitis, ungual dystrophy, diabetes mellitus, anemia, weight loss, elevated plasma glucagon levels and an alpha-cell glucagon-secreting neoplasm of the pancreas. We are reporting a case of this syndrome in a middle-aged woman, in whom the first complaints and signs were cutaneous. The recognition of the distinctive skin manifestations of the syndrome led to early diagnosis and treatment of the underlying malignant pancreatic tumor.


Microspectrophotometric quantitation of the diaminobenzidine reaction for histochemical demonstration of cytochrome oxidase activity. Polyacrylamide models in which an extract of cattle heart mitochondria was incorporated, as well as cryostat sections of tongue muscle and epithelium, were used to set up the conditions under which the histochemical reaction for the demonstration of cytochrome oxidase can be quantitated. Using diaminobenzidine in a concentration of 5.5 mM, cytochrome C in a fixed concentration of 76 micron and keeping the incubation medium away from direct light action, enzyme activity can be evaluated by means of direct microphotometry on tissue sections. Each biologic model requires previous individual determination of the measurement limits. These limits can be readily established by using a small chamber for the incubation medium, which can be placed in the microphotometer, allowing the reaction rate to be following using a single section.


Bluetongue studies with sentinel cattle in Kenya. Bluetongue antibody of 19 different serological types was found in a group of sentinel cattle near Nairobi. A continuous challenge by these 19 strains occurred each year in this location. The sero-conversion rates were shown to vary from year to year and rainfall had no obvious effect upon the conversion rates. The 19 different strains of virus were also shown to be active at five widely scattered sites in different parts of the country.


Scanning electron microscopic appearance of viral-antigen-coated polystyrene balls. A radioimmunoassay (RIA) was recently developed for the detection of antiviral IgG and IgM class-specific antibodies using antigen-coated polystyrene balls as the RIA solid-phase. In this communication the attachment and distribution of herpes simplex virus (HSV) capsid and envelope antigens and rubella viruses on the surface of the balls was examined by scanning electron microscopy (SEM). In SEM the surface of the untreated 'clear frosted' polystyrene balls appeared very uneven with innumerable pits and grooves. The viral particles were haphazardly distributed both in the grooves and on the exposed surface of the balls. The strength of adsorption of the viral antigens onto the balls seemed to be remarkably resistant to outside mechanical forces. HSV antigens frequently appeared in clusters, whereas rubella viruses were mostly found as single particles.


Epinephrine activation of pig skin adenylate cyclase in vivo and subsequent refractoriness to activation. Epinephrine injected intradermally activated pig skin adenylate cyclase and increased the epidermal cyclic AMP level in vivo. This biphasic response reached a maximum in 5 min and gradually decreased thereafter. The simultaneous injection of a cyclic AMP phosphodiesterase inhibitor, isobutyl methyl xanthin (IBMX) potentiated the increase. The simultaneous injection of a specific beta-adrenergic receptor inhibitor, propranolol, inhibited this accumulation of cyclic AMP. After the first activation by epinephrine in vivo, there was a marked refractoriness of the skin (epidermal) adenylate cyclase to subsequent epinephrine stimulation vivo and in vitro. This refractoriness was specific for catecholamine stimulation as responses to histamine were normal. Recovery from refractoriness started at 48 hr and was completed at 1 week after the injection of epinephrine.


Experimental infection of Tupaia belangeri (tree shrews) with herpes simplex virus types 1 and 2. The susceptibility of Tupaia belangeri (tree shrews, which are primitive prosimian primates) to infection with herpes simplex virus (HSV) and the pathogenesis of HSV in these animals were investigated. Juvenile (28--45 days old) and adult (150 days old) animals were inoculated intravenously, intraperitoneally, or subcutaneously with HSV type 1 or 2 (25--10(5) plaque-forming units per animal). Clinical illness usually appeared in juvenile animals on the second day after inoculation, and the animals died between two and 14 days after inoculation. High titers of infectious HSV were recovered from liver and spleen. The histopathologic examination always showed severe liver changes with numerous necrotic areas. The morphologic events in the liver were designated as herpetic hepatitis. The next most common morphologic findings were encephalitis and fibrosis in the spleen. These results demonstrate the high pathogenicity of HSV types 1 and 2 in juvenile T. belangeri. In contrast, adult animals did not develop acute clinical disease and survived the HSV infection.


Rotavirus in travelers' diarrhea: study of an adult student population in Mexico. The role of rotavirus in adult diarrhea was evaluated in 165 students attending a Mexican university. Students were divided into three groups: newly arrived summer students from the United Sttes, regular students from the United States, and Mexican and Venezuelan students. Ninety-one students with diarrhea and 74 corresponding, matched, asymptomatic control students were included in the study. The frequency of rotavirus in stools was determined by electron microscopy with use of the pseudoreplica technique. Twenty-five percent of those who were ill and 12% of the controls had rotavirus in their stools. A significantly (P less than 0.05) greater number of newly arrived United States summer students with diarrhea had rotavirus in their stool than did matched controls (26% vs. 3%). There was no significant difference in rate of recovery of bacterial pathogens from rotavirus-positive and rotavirus-negative stools (52% vs. 53%) from students with diarrhea. Although significantly more rotavirus was identified from ill American summer students than from controls, the role of rotavirus as a cause of diarrhea in these students could not be established in all cases since bacterial pathogens were also commonly found in stool.


Hepatitis A virus infection: new insights from seroepidemiologic studies. The prevalence of exposure to hepatitis A virus (HAV) increases with increasing age; decreases with increasing socioeconomic class; increases with increasing serologic evidence of prior hepatitis B virus (HBV) exposure but is much more common than HBV exposure; is independent of sex and race; varies in different parts of the world as a function of hygienic, developmental, and unrecognized geographic factors; and is not affected by immune deficiency or immaturity. Transmission of type A hepatitis is enhanced by poor personal hygiene such as that seen in institutions for the mentally retarded. On the other hand, there is no increased exposure to HAV among homosexuals, who have frequent and intimate contact with multiple sexual partners; among hemodialysis patients and staff; or among multiply transfused individuals, all of whom are at significantly increased risk of exposure to HBV. No epidemiologic evidence has confirmed the existence of viremic or intestinal carriers of HAV, and the virus is rarely, if ever, spread by parenteral mechanisms. Finally, HAV appears to play no role in chronic liver disease and a very minor role in fulminant hepatitis; however, HAV is responsible for a sizable proportion (approximately 20%--40%) of sporadic hepatitis among urban adults.


Measurment of rhesus monkey (Macaca mulatta) apolipoprotein B in serum by radioimmunoassay: comparison of immunoreactivities of rhesus and human low density lipoproteins. A sensitive and specific double antibody radio-immunoassay for the major apolipoprotein (apoB) of rhesus (Macaca mulatta) serum very low density lipoprotein (VLDL) and low density lipoprotein (LDL) is described. The anti-serum was raised to LDL (d 1.030-1.040 g/ml) and the LDL(2) (d 1.020-1.050 g/ml) was labeled with (125)I by the chloramine-T or iodine monochloride method. The assay, which was sensitive to 0.02-0.5 micro g of LDL(2), had an inter-assay coefficient of variation of 4.5%. This assay was successfully used to measure apoB in the whole serum and low density lipoproteins of control monkeys maintained on a standard Purina monkey chow (PMC) diet and of three groups of monkeys fed atherogenic diets: an "average American diet," a 25% peanut oil and 2% cholesterol-supplemented PMC diet, and a 25% coconut oil and 2% cholesterol-supplemented PMC diet. The control monkeys (n = 13) had a serum cholesterol of 146 +/- 28 mg/dl and an apoB of 50 +/- 18 mg/dl. In the monkeys maintained on the atherogenic diets the serum apoB was elevated: 103 +/- 28 mg/dl (American), 102 +/- 35 mg/dl (peanut oil), and 312 +/- 88 mg/dl (coconut oil). The values for serum total cholesterol were 333 +/- 65 mg/dl (American), 606 +/- 212 mg/dl (peanut oil), and 864 +/- 233 mg/dl (coconut oil) and were elevated relative to controls (P < 0.001). For each of the diets, total serum cholesterol correlated with serum apoB (P < 0.001). The slopes of the regression lines of serum apoB vs. cholesterol for the monkeys on the PMC, American, and coconut oil diets were similar (m = 0.531, 0.401, and 0.359, respectively), but differed from that of monkeys on the peanut oil diet (m = 0.121). The immunoreactivities of rhesus and human LDL were compared using specific antisera raised against these antigens. In homologous assay systems, monkey and human LDL exhibited unique immunological determinants. The same results were obtained with the delipidated preparations of the two LDLs using antisera raised against either monkey or human apoB. Crossover studies using a heterologous tracer with each anti-serum resulted in the selection of a specific population of antibodies directed against antigenic sites shared by these two LDL species.


Effect of temperature and plasma on the exchange of apolipoproteins and phospholipids between rat plasma very low and high density lipoproteins. The effect of temperature and plasma on the exchange of apoprotein C and phospholipids between VLDL and HDL was studied using 125I-labeled and 32P-labeled VLDL. Temperature affected the exchange of apoproteins and phospholipids similarly, and both were enhanced by increasing the temperature of incubation to 20 degrees C and higher. The exchange of apoC was almost complete within 5 minutes of incubation and was not influenced by the addition of plasma to the incubation mixture. The exchange of phospholipids occurred much more slowly and was enhanced 5- to 10-fold in the presence of plasma. These results indicate that the exchange of both phospholipids and apoC is dependent on the temperature of the incubation, but that they exchange independently, at least in part. It is suggested that the exchange of apoC occurs through dissociation of the apoprotein from the lipoprotein surface to the water phase, whereas the exchange of phospholipids is mediated mainly by a carrier molecule present in plasma and absent in lipoproteins.


Chylomicron apoprotein localization within rat intestinal epithelium: studies of normal and impaired lipid absorption. Monospecific antisera were produced to two chylomicron apoproteins (apoB, apoA-I) and utilized for indirect immunofluorescent localization of these apoproteins within rat intestinal epithelium during normal and impaired lipid absorption. Isolated intestinal epithelial cells prepared after different periods of lipid absorption from in situ intestinal segments revealed a rapid increase in fluorescence for both apoproteins that filled the entire apical portion of the cell. Prolonged lipid absorption for as long as 5 hr demonstrated sustained immunofluorescence and gave no indication of a depletion of the intestinal mucosa for either apoprotein during normal lipid absorption. [(3)H]Leucine incorporation into mesenteric lymph chylomicron apoproteins showed a linear decrease in specific activity of total chylomicron protein as well as apoB over 4 hr of a continuous lipid infusion indicating sustained active apoprotein synthesis during prolonged lipid absorption. Acetoxycloheximide, a potent inhibitor of protein synthesis, was employed to determine the dynamics of chylomicron apoproteins during an experimental condition of impaired lipid absorption. In animals with inhibited protein synthesis, fluorescence for both apoproteins was present early in the course of lipid absorption; however, at 60 min after the onset of lipid absorption, fluorescence for both apoproteins was absent. Fluorescence for both apoproteins returned during the recovery of protein synthesis. The present studies have confirmed previous results that localized two chylomicron apoproteins within intestinal epithelial cells. The present studies extend these observations and disclose a rapid and sustained synthesis of these apoproteins during prolonged chylomicron formation. During an experimental condition of impaired protein synthesis there was a marked reduction in the mucosal content of both apoA-I and apoB. These results are the first demonstration of impaired mucosal apoprotein synthesis during an experimental model of impaired lipid absorption.


Pregnant mare serum gonadotrophin and follicle-stimulating hormone stimulation of cyclic AMP production in rat seminiferous tubule cells. Pregnant mare serum gonadotrophin (PMSG) was found to stimulate the production of cyclic AMP in isolated rat seminiferous tubule cells. The PMSG was equipotent with highly purified ovine FSH. The subunits of PMSG were inactive in this system but recombination resulted in partial restoration of activity, and the combination of PMSG alpha-subunit with FSH beta-subunit was even more active. Desialation of PMSG, but not of ovine FSH, resulted in a reduction of slope and potency of this response, suggesting that sialic acid may be important for the interaction of PMSG and the Sertoli cell. Further tests of the responsiveness of this cell preparation demonstrated activity with several species of mammalian FSH molecules, but not with FSH molecules obtained from non-mammalian species.


Effect of progesterone on insulin secretion in the rat. Implants of progesterone resulted in an increased amount of insulin in plasma in response to intravenous administration of glucose in the rat. Isolated islets of Langerhans from progesterone-treated animals showed a greater maximum secretory response to glucose than islets from control animals but their sensitivity to low concentrations of glucose was unchanged. Theophylline increased glucose-induced secretion of insulin to a greater extent in progesterone-treated than in control rats and also produced a greater increase in the concentration of cyclic AMP in isolated islets from hormone-treated animals. These results suggest that the effect of progesterone on insulin secretion may be mediated by a change in cyclic AMP levels in the beta cell. The possible role of progesterone in increasing the secretion of insulin in pregnancy is discussed.


Genetic studies of autoimmunity and retrovirus expression in crosses of New Zealand black mice I. Xenotropic virus. The relationship between expression of xenotropic virus and the development of autoimmunization was studied in the progeny of crosses between New Zealand Black (NZB) and SWR mice. The (F1 X SWR) and F2 progeny segregated into three phenotypes: high-virus, low-virus, and virus-negative; F1 and (F1 X NZB) progeny were always high-virus. Autoantibodies, immune deposit nephritis and lymphomas developed in the progeny of these crosses. The virological phenotype of the animal could be dissociated from the presence of either autoantibodies or nephritis. For example, mice that expressed titers of virus as high as the NZB parent failed to develop signs of autoimmunization, even up to 24 mo of age. By contrast, some (F1 X SWR) and F2 mice that expressed low titers of virus developed autoimmune disease. Furthermore, a proportion of virus-negative mice produced autoantibodies and were found to have typical immune deposit nephritis. No viral antigens could be detected in the renal lesions of such virus-negative animals. By contrast with the dissociation between expression of virus and occurrence of nephritis, the presence of antibodies to DNA correlated with the development of renal lesions. We conclude that the genes that determine the expression of infectious xenotropic virus in NZB mice segregate independently from those that are involved in the autoimmune disease of these animals.


Genetic studies of autoimmunity and retrovirus expression in crosses of New Zealand black mice. II. The viral envelope glycoprotein gp70. The retroviral envelope glycoprotein, gp70 was measured in the serum of New Zealand Black (NZB) and SWR mice and the progeny of their crosses. The serum gp70 values segregated to "NZB-like" and "SWR-like" levels in these mice. A complex mechanism determined the inheritance of NZB-like serum gp70 levels. We found that the factors determining the expression of this retroviral protein were independent of the genes (Nzv-1 and Nzv-2) that determined the expression of infectious xenotropic virus. Autoimmune disease, including immune deposit nephritis could be dissociated from the degree of expression of serum gp70. By contrast, presence of circulating immune complexes and anti-DNA antibody did correlate with the development of nephritis in these crosses. A significant correlation was found between high grade expression of serum gp70 and the presence of lymphomas in these mice.


Interferon action III. The rate of primary transcription of vesicular stomatitis virus is inhibited by interferon action. Transcription of vesicular stomatitis virus (VSV) in Vero cells was confined to the synthesis of parentally-derived mRNA (primary transcription) by the use of cycloheximide and/or a ts mutant, G41(IV), at a non-permissive temperature (40 degrees C). More transcripts accumulated in the presence of cycloheximide than in its absence. This so-called "cycloheximide effect" results from higher rates of virus transcription sustained for longer periods of time. The rate of VSV transcription initially increases linearly for 1 to 2 h after injection. Interferon reduces this rate (congruent to fourfold with 50 units/ml interferon) irrespective of the presence or absence of cycloheximide. The VSV mRNA transcripts synthesized in mock- or interferon-treated cells were equal in size and had an equivalent half-life of 17 h at 40 degrees C. It seems likely that once transcription is initiated in interferon-treated cells, it is completed successfully. Since interferon reduces the rate of early VSV primary transcript synthesis to below that achieved in the presence of cycloheximide, we conclude that interferon has an effect on transcription beyond that attributable solely to protein synthesis inhibition. We postulate that interferon decreases the probabiltiy of initiating virus transcription. Virus mRNA escaping this facet of interferon action may then encounter other facets such as post-transcriptional modification and/or inhibition of translation. However, the mandatory sequence of primary transcription leads to primary translation for negative-strand viruses like VSV dictates that the overall inhibitory effect of interferon on translation would derive in part from this prior inhibition of transcription. Thus, to apply the term "primary effect" to one particular facet of interferon action may not always be meaningful.


Isolation of thioguanine resistant variants of K-BALB cells non-inducible for type C viruses by 5-iododeoxyuridine. Cones of thioguanine resistant K-BALB mouse cells wereisolated which were inducible for endogenous type C virus synthesis by cycloheximide and dexamethsone, but not 5-iododeoxyuridine. A comparison of the number of foci formed on NRK and SC-I cells suggested that the xenotropic virus was suppressed. The variants were not defective in the incorporation of thymidine or iododeoxyuridine or deficient in thymidine kinase, but were deficient in hypoxanthine-guanine phosphoribosyltransferase and the incorporation of hypoxanthine into nucleic acid. Because these cells are blocked at some point in the expression of endogenous virus, they may prove useful in establishing the steps involved in chemical activation of virus synthesis.


Reduction of intercellular adhesiveness of chick heart cells by herpes simplex viruses 1 and 2. Infection with herpes simplex viruses type 1 or 2 prevented the aggregation of 7-day-old chick heart cells into smooth, spheroidal, spontaneously beating aggregates. Virus infection also caused a loosening of peripheral cells in aggregates formed from initially uninfected cells. Measurements of rate of attachment of labelled single heart cells to a monolayer of like cells (homotypic), to HEp-2 cells (heterotypic), or to plastic substrata (nonspecific adhesion) indicated that virus infection caused a significant but differential loss of homotypic and nonspecific adhesiveness, but no alteration in heterotypic attachment rates. These observations indicate that those cells surface changes induced by viruses which are related to cell adhesion can be quantified by techniques measuring attachment rates.


Epstein-Barr (EB) virus genome-containing, EB nuclear antigen-negative B-lymphocyte populations in blood in acute infectious mononucleosis. Experiments have been performed to identify the type and size of cell infected by EB virus in the blood of acute infectious mononucleosis (IM) patients, and to investigate the nature of the infection. Virus-infected cells, recognized by their ability to give rise to lymphoblastoid cell lines when co-cultivated with foetal lymphocytes, were shown to be restricted to the B-lymphocyte population. Samples of this population from each of eight IM patients were found to be negative for EB nuclear antigen (EBNA) staining. Thereafter, fractions of IM B-lymphocytes prepared on the basis of cell size were assayed either by co-cultivation, for the incidence of virus-infected cells, or by immunofluorescence staining for the presence of cells expressing EBNA. The great majority of virus-infected cells were found in the fractions of normal sized B-lymphocytes and yet these fractions were unequivocally EBNA-negative B-cell populations in IM blood is discussed in terms of the type of infection established by EB virus in the circulation of IM patients.


Mitogen induction of murine C-type viruses. IV. Effects of lipoprotein E. coli, pokeweed mitogen and dextran sulphate. Lipoprotein E. coli, a B-cell mitogen, is identified as a new agent inducing the release of endogenous C-type virus from mouse spleen cells. Like lipopolysaccharide, a previously identified inducer, this compound has a synergistic effect with 5-bromo-2'-deoxyuridine. Induced virus has the characteristic density as well as morphology of C-type viruses. Budding viruses are detected on cultured BALB/c cells by electron microscopy 2 to 4 days following culturing in the presence of lipoprotein. Pokeweed mitogen, a compound mitogenic for T- and B-cells was negative when tested for virus induction, both alone and in combination with 5-bromo-2'-deoxyuridine. Dextran sulphate, another B-cell mitogen, was negative for induction as well. However, when, combined with lipopolysaccharide, it enhanced the virus release induced by this mitogen. In contrast, no additive effects were observed either by combining dextran sulphate with other virus amplifying mitogens or by combinations of mitogens which both have virus-inducing ability. This finding is discussed with respect to B-cell differentiation.


Aggregation of enterovirus small plaque variants and polioviruses under low ionic strength conditions. Virion aggregation in low ionic conditions was observed with small plaque variants of Coxsackievirus type B3 and Echovirus types 4 and 11 by sedimentation and filtration methods. Inclusion of salts or DEAE-dextran into the media prevented or reversed virion aggregation. The effect of pH on aggregate formation in low ionic strength solutions was also investigated with various strains of poliovirus. Type I Sabin strain formed aggregates even at high pH, while Mahoney strains did so only below pH 6.5. Type 2 virus, Sabin and MEF1 strains, and type 3 virus, Sabin, Saukett and Suwa strains, showed an intermediate behaviour between the two type 1 strains, except MEF1-LB strain, a clone obtained from MEF1 strain under acidic overlay, which showed little tendency to aggregate. These results were compared with the degree of the d character of the strains. Besides the effect of inhibiting virion aggregation, the inclusion of DEAE-dextran into a sucrose gradient slowed the sedimentation of some of the viruses in low ionic strength solutions.


Involvement of microtubules in cytopathic effects of animal viruses: early proteins of adenovirus and herpesvirus inhibit formation of microtubular paracrystals in HeLa-S3 cells. In order to examine the involvement of microtubules in the virus-induced cytopathic effect (c.p.e.), the effect of virus infection on the formation of microtubular paracrystals (PC) induced by 10 microgram/ml of vinblastine sulphate in HeLa-S3 cells was examined by phase-contrast microscopy. In poliovirus-infected cells, c.p.e. (cell rounding) and the inhibition of PC formation proceeded in parallel, starting 4 h post-infection. In Sendai virus-infected cells, however, PC formation was not inhibited even 24 h post-infection when most infected cells clearly showed c.p.e. (syncytial formation). In adenovirus-infected cells, the inhibition of PC formation was observed 9 h before the appearance of c.p.e. Cytosine arabinoside (ara C) did not block the inhibition of PC formation in infected cells, but blocked the appearance of late c.p.e. (nuclear alteration). Cycloheximide blocked both the inhibition of PC formation and the induction of late c.p.e. These results suggest that an early protein synthesized de novo by adenovirus is required for direct or indirect inhibition of the microtubular PC formation. Furthermore, on ultraviolet (u.v.) inactivation of adenovirus both activities (induction of early c.p.e. shown by shrinkage of cytoplasm, and inhibition of PC formation) followed the same inactivation curve and were inactivated at a slower rate than viral infectivity and the activity leading to late c.p.e. The u.v. light sensitive target responsible for the induction of early c.p.e. and the inhibition of PC formation is about 20% of that for infectivity and is in accord with the genome size of the early functioning virus genes. In herpes simplex virus (HSV)-infected cells, the inhibition of PC formation, the appearance of c.p.e. (cell rounding and disappearance of nucleoli) and the synthesis of V antigen proceeded in parallel. These three functions of HSV were not blocked in infected cells even when the de novo synthesis of virus DNA was inhibited by ara C or phosphonoacetic acid (PAA), whereas these three functions were blocked by cycloheximide, suggesting that a protein coded by the input virus genome early after infection inhibits the microtubular PC formation and is responsible for c.p.e. From the u.v. inactivation curve of HSV, it was confirmed that only one-tenth of virus genome was responsible for both activities (induction of c.p.e. and inhibition of PC formation).


Isoelectric points of polypeptides of standard poliovirus particles of different serological types and of empty capsids and dense particles of poliovirus type 1. The isoelectric points of polypeptides of standard and dense poliovirus particles and of empty capsids have been determined by isoelectric focusing in urea and by two-dimensional analysis. Comparing virus strains belonging to the three serological types of poliovirus, differences in the pI of some, but not all of the structural polypeptides are found. The pI of polypeptides of dense particles and of empty capsids are identical with those of standard particles. Polypeptide VPo present in empty capsids has a pI between those of VP4 and VP2.


Cytomegalovirus as a frequent cause of Guillain-Barré syndrome. In 10 patients with Guillain-Barré syndrome a preceeding cytomegalovirus infection could be demonstrated by virus-specific IgM antibodies that were present in high titers in 9 of the 10 patients in the first serum specimen. The IgM antibodies to cytomegalovirus were detected by a sensitive "double" indirect immunofluorescence technique. In most of our cases (8 of 10) the complement-fixing antibody titers had already reached high levels on admission into a hospital, and significant titer changes were not observed. Cytomegalovirus was isolated from the urine of five patients.


A study of the prevalence of rotavirus infection in children with gastroenteritis admitted to an infectious diseases hospital. In a 12 month survey of infants and children with gastroenteritis admitted to Fairfield Hospital, Melbourne, rotavirus was found in approximately 42% of patients. This virus was detected more often during the winter months, particularly in children aged between 12 months and 3 years. Detection of rotavirus by electron microscopy was found to be more sensitive than by counterimmunoelectrophoresis. Routine bacterial and viral studies revealed that bacterial pathogens and common enteric viruses were associated with relatively few cases of gastroenteritis. There is little doubt that rotavirus is the most important aetiological agent of acute gastroenteritis in yvirus is the most important aetiological agent of acute gastroenteritis in young children in Melbourne.


Studies of an outbreak of acute hepatitis A: II. Antibody changes to cytomegalovirus and herpersvirus. The acute and convalescent sera from 14 schoolchildren with acute hepatitis A were tested for antibody changes to 70 viral antigens. Marked decreases were noted in the levels of antibody to cytomegalovirus in 5 of the 14 children and in the levels of antibody to herpesvirus type 1 in 3. No such changes were noted in 9 sex- and age-matched healthy control children from the same classes.


Seroepidemiological investigation of patients and family contacts in an epidemic of hepatitis A. Serial blood and faecal samples were collected from patients and family contacts during an outbreak of hepatitis A in a village and tested by a solid-phase competitive type radioimmunoassay for hepatitis A antigen and hepatitis A antibody. The amount and duration of excretion of hepatitis A antigen was correlated with the severity of the illness. In 2 severe clinical cases, hepatitis A antigen was demonstrated in faecal extracts 11 days before the onset of jaundice and continuing for 10 days thereafter, with maximum shedding during the late incubation period. Faecal antigen was demonstrated in low concentrations for only 2 days in a patient with mild disease and in a person with subclinical infection. There was an inverse correlation between the incidence of infection and prevalence of hepatitis A antibody and age. Of 24 infections, 19 (79%) occurred in persons in the age group 0 to 20 years, a group in which only 6% of individuals had pre-existing antibody. Hepatitis A antibody was present in the serum of 3 persons in low titres of 1:20 to 1:40 on the day jaundice developed. The antibody titres increased very rapidly during the following 2 weeks of illness and slowly during the following months, reaching titres of 1:900 to 1:3500. In a separate study, a mean antibody titre of 1:591 was found in 13 patients, 12 years after clinical hepatitis A with jaundice.


Incidence and distribution of herpes simplex virus types 1 and 2 from genital lesions in college women. During a 9-month period, 9,772 women were treated at the student health center's gynecology clinic. Herpes simplex virus was isolated from 30 of 57 patients clinically diagnosed as suffering from a herpetic or herpetic-like genial infection for a virological incidence rate of 0.31%. Using virus plaque diameter in chick embryo cells and heat stability of viral thymidine kinase, 37% of the isolates were classified as herpes simplex virus type 1 and 63% were classified as herpes simplex virus type 2.


Immunotherapy of viral infections with transfer factor. It has been reported that dialysable leucocyte extract preparations, thought to contain transfer factor, may be used therapeutically for the treatment of a variety of immunodeficiency syndromes. Clinical and laboratory studies have suggested that such preparations, in addition to transferring specific cellular immunity may also contain non-specific adjuvant activities. Attempts at immunotherapy of viral infections are described against a background of current research on the biological and biochemical properties of leucocyte dialysates.


Multiple buoyant densities of hepatitis A virus in cesium chloride gradients. Hepatitis A virus (HAV) recovered from stools of human cases of hepatitis A and from stools of chimpanzees experimentally infected with HAV was shown to possess multiple buoyant densities in CsCl gradients. The greatest proportion of HAV was most frequently found at a buoyant density of 1.32-1.34 g/cm3, however, large proportions of HAV were also frequently found at higher densities, including 1.36-1.37, 1.40-1.42, and 1.45-1.48 g/cm3. These findings are consistent with the notion that HAV may be a parvovirus.


Picorna- and togavirus infection of cells detected by gas chromatography. Viral infection of cells causes chemical and metabolic changes, which can be detected by gas chromatography (GC) of ether extracts of supernatant fluids and cell homogenates before any significant damage to the cells is observable microscopically. The characteristic and specific GC patterns obtained from BHK-21 and Vero cell cultures infected with encephalomyocarditis, polio, echoviruses, and a togavirus make it possible to distinguish between these infecting viruses. The appearance of 1 or 2 compounds, represented by GC peaks with TR values of 302 and 677 seconds seems to be specific for these viruses. Other peaks found in the supernatant media 1-2 hours after infection probably represent cell constituents leaking into the medium as a result of damage to the cell membrane by the invading virus.


Peripheral neuropathy and solitary plasmacytoma. Three patients with peripheral neuropathy and a solitary plasmacytoma are presented, and the literature is reviewed. It is suggested that middle-aged men with an obscure progressive sensorimotor neuropathy, a raised CSF protein, and otherwise negative investigations should have a full skeletal survey since irradiation of a plasmacytoma may lead to a considerable improvement in the associated neurological disability.


Neuromuscular transmission in human single motor units. A method, multiple point stimulation, has been reported to isolate 5-20 single hypothenar or thenar motor units for investigation. This method is attractive for testing neuromuscular transmission because the required stimulus intensities are much less than for supramaximal nerve stimulation, and movement artefact is less of a problem. In this investigation of controls, the changes in hypothenar and thenar motor unit surface voltage and latency of single muscle fibre action potentials belonging to the motor unit have been measured in response to trains of stimuli delivered to the motor nerve. In healthy motor units, increases in the surface peak-to-peak voltage and corresponding reductions in the peak-to-peak duration occurred, the changes being maximum at the shorter stimulus intervals and accompanied by increased synchronisation of motor unit muscle fibre action potential discharges. The investigations of the responses for healthy single motor units to repetitive stimulation provide the basis for investigations of neuromuscular transmission at the level of the motor unit in diseases of neuromuscular transmission.


Neuromuscular transmission in myasthenic single motor units. In patients with myasthenia gravis neuromuscular transmission has been tested in individual hypothenar and thenar motor units using trains of near threshold electrical stimuli delivered to the motor nerve. The most important observations included: (1) the proportion of motor units with pathological decrements varied from 0 to 90%, (2) the decrements in surface voltage were frequently much more normal in individual motor units than in the corresponding maximum compound potentials evoked by supramaximal nerve stimulation, and (3) the most abnormal decrements were observed in motor units at the low end of the surface voltage range. These observations can be interpreted to suggest important neurogenic factors in the pathogenesis of myasthenia gravis. An attractive alternative would be to suggest that the small motor units have the most abnormal neuromuscular transmission because they normally have a lower margin for safe neuromuscular transmission and, as a consequence, fail first in diseases of neuromuscular transmission.