Clinical evaluation of EBV serology in American patients with nasopharyngeal carcinoma. There is now extensive immunological, biological and biochemical evidence to support a possible etiological relationship between EBV and NPC in patients from different geographical locations. Besides providing information on the question of etiology, the results from immunological investigations suggest that antibodies to some of the EBV-associated antigens might also be of clinical importance in the diagnosis and prognosis of NPC. To determine the possible clinical application of EBV serology to American NPC, sera from patients seen at the Mayo Clinic and the National Institutes of Health were examined for antibodies to EBV-associated antigens in an effort to identify those parameters which most reliably distinguish NPC from other types of cancer. The results show that high antibody titres to EBV-induced EA and the presence of antibody to EBV antigens in the IgA immunoglobulin fraction were the two most specific discriminating parameters, although neither was infallible. These findings are discussed in relation to future studies that are needed in order to determine the potential clinical value of EBV serology to the diagnosis and prognosis of NPC.


Differences in EBV antibody titres of patients with nasopharyngeal carcinoma originating from high, intermediate and low incidence areas. In order to assess the differences in serological reactivities of NPC patients from high, intermediate and low incidence areas, 288 NPC sera from Hong Kong, Singapore, Tunis, East Africa, Paris and Los Angeles, together with sera from patients with ear, nose and throat tumours other than NPC and with those from normal individuals from the same areas, were tested 'blind' with the same batches of antigen. Important differences (up to 3-fold) in the GMTs of antibodies directed against VCA, EA and EBNA were observed between different ethnic groups. Although the stages of the disease varied somewhat between groups, the main cause of the variations appeared to lie in the socio-economic environment of the patients. Apart from these variations, and regardless of geographical or ethnic origins, NPC was consistently found to be associated with an active infection (or reactivation) by Epstein-Barr virus, which was not the case for patients with other tumours or for normal individuals from the same areas or of the same ethnic groups.


Cell-mediated immunity, Epstein-Barr virus and nasopharyngeal carcinoma. Cellular immunity is apparently important in host defenses against cancer. A growing body of information on CMI to EBV is here reviewed, providing a strong basis for studies of such reactivity in NPC. Present work suggests that CMI responses to virus- and tumour-associated antigens may have implications for etiology, diagnosis and treatment.


General immunological status of nasopharyngeal carcinoma patients in Singapore. Newly diagnosed NPC patients were found to have impaired general T-cell functions, as determined in vivo by the Mantoux test and in vitro by the PHA response assay. Treated remission patients were as hyporesponsiveness as newly diagnosed patients. PHA hyporesponsiveness was associated with the HLA profile of A2-B Sin 2 and with high antibody titres to EBV-related antigens, in particular to the early antigen. Impaired responses in the Mantoux or PHA tests were associated with poor survival. The impaired response could be partially or totally restored by in vitro treatment of the hyporesponsive lymphocytes with levamisole.


Autoantibodies (cold lymphocytotoxins, antiactin antibodies and antinuclear factors) in nasopharyngeal carcinoma patients. LTA, SMA and ANA were found at higher frequencies in sera from NPC patients than in those from matched controls. The frequency and GMTs of LTA-positive sera varied with the origin of the patient (Chinese greater than North African greater than Caucasian, thus paralleling the risk for NPC in each ethnic group) and the stage of the disease (stage IV greater than stage I). A positive correlation was found between LTA and anti-EBV titres with regard to anti-VCA and anti-EBNA antibodies. SMA were specific for actin, and their titres did not correlate with anti-EBV (VCA, EBNA and EA) titres. Although there was no evidence for an abnormal immunogenicity of the self components recognized by the three autoantibodies, their independent augmentations did not favour the alternative hypothesis of a common central mechanism for their production. The origin and the biological significance of these autoantibodies remains, therefore, to be explained.


In vivo cell-mediated immunity in Chinese patients with nasopharyngeal carcinoma. Delayed hypersensitivity to antigens derived from four EBV-related (KHLY 28, Raji, F265 and NC37) and one EBV-unrelated (Molt) T-lymphoid cell lines and to standard antigens (Trichophyton, Candida albicans and streptokinase/streptodornase) was measured in 104 NPC patients and 24 patients with other cancers. Of the NPC patients with non-disseminated disease, 55/86 (63.2%) had a positive skin reaction to HKLY 28 extract, compared with only 1/18 (5.6%) OC patients with non-disseminated disease. The frequencies of these NPC patients with positive skin reactions to the other cell-line extracts were significantly lower (1.7-5.4%). From the preliminary results of a longitudinal study, skin testing with this particular crude membrane extract from HKLY 28 cells appears to be of little practical value in monitoring the clinical evolution of NPC, although a significant association between clinical evolution and certain patterns of skin reactivity response to the extract was found in about two-thirds of the cases analysed.


Use of immunological studies in evaluating the clinical course of nasopharyngeal carcinoma. The prognosis of a NPC patient is influenced by many factors, including the therapeutic method used and the clinical stage and histological characteristics of the tumour. The immune response of the peripheral lymphocytes was examined in 27 patients with NPC, and it was concluded that this response accurately indicates the clinical course of the diseases as compared with clinical stage of disease, histological type of tumour and anti-VCA-antibody titre.


Prolonged prostaglandin E1 stimulation of cyclic AMP production in transformed and normal WI-38 fibroblasts. Long-term (48-hr) incubations of either the fibroblast strain WI-38 or its SV40-transformed counterpart, WI-38-VA13-2RA, in growth medium containing 1 micron prostaglandin E1 (PGE1) resulted in a sustained production and release of cyclic AMP from the cells into the medium. Despite the steady production, intracellular levels of the nucleotide decreased, reaching steady-state values within 4 hr of the initial exposure to PGE1. These values were maintained for the remainder of the 48-hr experimental period. The steady-state levels of intracellular cyclic AMP were higher than those observed in unstimulated cells, and cyclic AMP-dependent protein phosphokinase was in a highly activated state as compared to controls. Under these conditions little change in the growth or morphology of either the normal or transformed cells was observed. In contrast, inhibition of growth, apparent cell death, and unusual morphological changes were observed in both normal and transformed cells when high concentrations of either PGE1 (10 micron) or the phosphodiesterase inhibitor 1-methyl, 3-isobutylxanthine (0.5 mM to 2 mM) were used, which was indicative of toxic effects of the drugs. It was concluded that cyclic AMP-mediated activation of protein phosphokinase does not completely inhibit growth in WI-38 cells or restore normal growth and morphology to the SV40-transformed cells.


Cellular immune response to cytomegalovirus infection after renal transplantation. A prospective study of 15 patients who received renal transplants defined the effect of renal transplantation on the cellular immune response to cytomegalovirus infection. Of 15 patients, 14 developed cytomegalovirus infection, usually in the first 2 months after transplantation, and all infections were accompanied by a normal humoral immune response. After the initiation of immunosuppressive therapy and transplantation, there was a general depression of lymphocyte transformation, as reflected in the response to phytohemagglutinin, accompanied by a specific defect in cellular immunity, as indicated by lymphocyte transformation to cytomegalovirus antigen. Eleven patients had cellular immunity to cytomegalovirus before transplantation, and all of these became negative in the first month after transplantation. In subsequent months, only 6 of the 14 study patients with cytomegalovirus infection developed specific cellular immune responses to cytomegalovirus. This occurred most often in patients who had severe febrile illnesses in association with infection. The specific cellular immune response which developed in the posttransplant period did not persist in three of the patients. This study demonstrates the dissociation of the humoral and cellular immune response to cytomegalovirus infection in renal transplant patients and indicates the importance of the loss of cellular immunity in the appearance of infection. Previously infected patients lost their cell-mediated immunity and had reactivation infections despite the presence of serum antibody.


Penetration of cultured mouse fibroblasts (L cells) by Rickettsia prowazeki. The association of Rickettsia prowazeki with L cells was examined by using a novel radioactive assay in which [alpha-(32)P]ATP-labeled rickettsiae were incubated with L-cell monolayers. Rickettsial association with the monolayer involved adherence and internalization steps that could be experimentally distinguished. Since R. prowazeki but not L cells possess an ATP-ADP obligate exchange transport system, addition of excess unlabeled ATP resulted in exchange of the labeled ATP from external, adherent rickettsiae but not from internalized rickettsiae. Rickettsial association was temperature dependent and was a linear function of both time and concentration. More than 90% of the biologically active rickettsiae associated with L cells was internalized. Rickettsial internalization required active participation of both rickettsiae and L cells; inactivation of either greatly reduced internalization. Rickettsial adherence to poisoned L cells was a saturable function of time and concentration. Adherence showed less temperature dependence than did internalization, but like rickettsial internalization, the extent of adherence was extremely low at 0 degrees C. The rate and extent of adherence by inactivated and native rickettsiae to inactivated L cells were similar. Although inactive rickettsiae adhered to active and inactive L cells to a similar extent, inactive rickettsiae were internalized poorly by active L cells. These data form the basis for the hypothesis that R. prowazeki are internalized by the host cell through a process of "induced phagocytosis" and that inactivated rickettsiae adhere to the host cell differently from native rickettsiae, failing to trigger the endocytosis mechanism.


Protection against Marek's disease-derived tumor transplants by the nononcogenic SB-1 strain of Marek's disease virus. A series of experiments was conducted to study the in vivo protection against Marek's disease-derived tumor transplants by the nononcogenic SB-1 strain of Marek's disease virus. Intact, embryonally bursectomized (Bx), thymectomized (Tx), or cyclophosphamide (Cy)-treated chickens of four genetic lines were vaccinated with live or inactivated SB-1. JMV, a non-virus-producing transplant, and GA/Tr-1 and MDT-198, two virus-producing transplants were used for challenge. Optimal protection against JMV was present 7 days postvaccination, but there was significant protection even when SB-1 and JMV were administered together. Protection was abolished by an increase in the number of tumor cells used for challenge or by combined Tx and Cy treatment. Inactivated SB-1-infected cells were unable to induce protection against JMV challenge. Protection was also present against challenge with GA/Tr-1, but not against MDT-198, except in vaccinated, Bx chickens. It was concluded that protection against JMV was T-cell dependent and required the induction of neo-antigens not present in an inactivated SB-1 cellular preparation. The absence of protection in intact chickens against MDT-198 could not be explained.


Effect of Escherichia coli heat-stable enterotoxin on cyclic GMP levels in mouse intestine. Partially purified heat-stable enterotoxin obtained from Escherichia coli strain F11/P155 caused an accumulation of cyclic GMP in the intestines of 8-day-old mice.


Biological properties of equine herpesvirus type 1 DNA: transfectivity and transforming capacity. DNA extracted from purified virions of equine herpesvirus type 1 (EHV-1) was examined for its transfectivity and transforming ability. The infectivity of the herpesvirus DNA was demonstrated by addition of calcium phosphate-DNA  coprecipitates to monolayers of permissive horse cells, with resultant plaque formation. The efficiency of transfection (50 to 100 plaque-forming units/microgram of DNA) was reduced by treatment of the viral DNA with deoxyribonuclease or sonication but not with Pronase or antivirus neutralizing serum. When nonpermissive mouse 3T3 Cells lacking the enzyme thymidine kinase (TK-) were transfected with intact EHV-1 DNA, clones of cells transformed to the TK+ phenotype were isolated in selective HAT medium (hypoxanthine, aminopterin, thymidine), which prevents growth of the TK- parental phenotype. The efficiency of transformation ranged from one to five transformants per microgram of EHV-1 DNA. The TK activity of the biochemically transformed cells was characterized by biochemical, electrophoretic, and immunological techniques. By these criteria, the TK activity was identical to the EHV-1 TK and different from the host wild-type enzyme. In contrast to the parental TK+ 3T3 cells, the EHV-1-transformed TK+ cells were unable to grow in the presence of arabinosylthymine, a drug selectively phosphorylated by herpesvirus TKs. These results indicate that stable transfer of EHV-1 genes into nonpermissive cells can be achieved with purified viral DNA.


In vitro evaluation of opsonic and cellular granulocyte function by luminol-dependent chemiluminescence: utility in patients with severe neutropenia and cellular deficiency states. Actively phagocytizing polymorphonuclear leukocytes (PMN) emit light or chemiluminescence (CL) which has been shown to be linked to the oxidative activity of the PMN. The measurement of CL has been demonstrated to be a useful tool for the in vitro assessment of intracellular and opsonophagocytic function of PMN. We have increased the sensitivity of the CL measurement by the addition of luminol to the in vitro reaction of PMN, bacteria, and serum. The presence of luminol, which can be oxidized to emit light, amplifies the detection of CL and PMN cellular activity. This amplification effectively reduces the number of PMN that are necessary for assessment of PMN function from 1 x 10(7) to as low as 2 x 10(4) PMN/assay and permits the evaluation of PMN function in severely neutropenic patients (100 PMN/mm3) in whom cellular PMN function has been heretofore extremely difficult to assess by other methodology. When this luminol-dependent CL method was used, three of eight neutropenic leukemic patients with gram-negative septicemia were found to have deficient opsonic activity and/or increased or depressed cellular oxidative activity. Because the initial slope of CL is dependent on the amount of serum and heat-labile factors, this method can also be used effectively as a simple technique for the analysis of specific rates of opsonophagocytosis of various microorganisms. Additionally, this method can detect the cellular PMN abnormalities of chronic granulomatous disease and myeloperoxidase deficiency. The luminol-dependent CL method is a simple, sensitive, reproducible technique that provides useful information about PMN metabolic activity, particularly in studies in which the number of PMN is limited.


Enhancement of avian sarcoma virus-induced tumor growth after pretreatment with BCG. Pretreatment of chickens with BCG 1 week before inoculation with avian sarcoma virus had a stimulatory effect on virus-induced tumor growth in 15 of 17 cases tested. This outcome was dependent upon the injection of both BCG and the oncogenic agent into the same wing web. Injection of BCG into the wing web contralateral to that used for virus inoculation or injection of a subcellular fraction of BCG into the same wing web did not affect tumor development. Administration of BCG also gave rise to heightened levels of tumor-associated, cell-mediated immunity, both on the part of chickens which received a subsequent injection of oncogenic virus as well as normal, BCG-inoculated controls.


Comparison of various macrophage-inhibitory agents on vaginal and systemic herpes simplex virus type 2 infections. Pretreatment of mice intraperitoneally with silica, trypan blue, or dextran sulfate to inhibit macrophage function markedly increased the lethality of a systemic intravenous infection with herpes simplex virus type 2, but did not affect the lethality or local virus growth after vaginal infection of mice with herpes simplex virus type 2. Agents which inhibit macrophage function by different mechanisms decreased host resistance to herpes simplex virus type 2, but the effects of macrophage-inhibitory agents may vary according to the route of virus infection.


Comparative studies of five heat-labile toxic products of Escherichia coli. Five heat-labile, partially purified toxic products of Escherichia coli were distinguished by isoelectric focusing, molecular weight, and neutralization with homologous and heterologous antisera. Only two affected the morphology of Y-1 cells, induced fluid accumulation in rabbit ileal loops, and stimulated production of cyclic AMP in Vero cells; these two did not cross-neutralize and only one showed cross-neutralization with cholera antitoxin. The remaining three products were cytotoxic for Vero cells.


Induction of enhanced resistance against encephalomyocarditis virus infection of mice by nonviable Mycobacterium tuberculosis: mechanisms of protection. Nonviable Mycobacterium tuberculosis strain Jamaica suspended in oil-droplet emulsions was used to enhance resistance of mice against encephalomyocarditis virus (EMCV). The mycobacteria-injected mice were significantly resistant to 50,000 50% lethal doses of EMCV. Similar concentrations of virus in plasma of normal and mycobacteria-injected mice from 1 to 120 min after injection of EMCV showed that resistance was not a result of rapid elimination of virus from the circulation. Furthermore, survival of viremic mice indicated protective mechanisms were operative after EMCV had escaped primary surveillance. Resistance did not appear to be associated with the mouse major histocompatibility gene complex. The spleen was intimately associated with protection, and the thymus was nonessential for enhanced resistance to EMCV. Protection was significantly diminished by cyclophosphamide injected intraperitoneally from 3 days before to the day of virus challenge. Finally, silica given intraperitoneally 24 h before virus completely abrogated resistance of mycobacteria-injected mice to EMCV. These results suggest that macrophages functioning independently of T-lymphocytes are important effector cells in resistance to EMCV of mice injected with nonviable mycobacteria.


Suppression of in vitro growth of virulent and avirulent herpes simplex viruses by cell-mediated immune mechanisms, antibody, and interferon. A rounding cell-forming--GC strain, which is a variant of a syncytial giant cell-forming herpes simplex virus (+GC Miyama strain), was highly attenuated for Swiss, BALB/c nu/nu, and nu/+ mice, whereas +GC was highly virulent to all the mice tested. +GC and -GC were antigenically indistinguishable from each other by cross-neutralization and cross-immunization. Immunosuppression induced by cyclophosphamide converted the nonlethal -GC infection of mice into a fatal infection. -GC replication in tissue culture was more effectively suppressed by spleen cells immunized with either +GC or -GC than was the +GC replication. -GC replication was also inhibited more effectively by antibody or the antibody-dependent cell-mediated system than was the +GC replication. -GC is highly sensitive to mouse interferon, but +GC was relatively resistant. These findings indicate that attenuation of this avirulent -GC strain may be due to a high susceptibility of its replication to humoral and cell-mediated defense factors. The probable roles of each defense factor in recovery from the infection with virulent and attenuated herpes simplex virus are also discussed.


Characterization of mouse peritoneal exudate and associated leukocyte adherence inhibitory activity after intraperitoneal injection of either Bordetella pertussis or Corynebacterium parvum vaccines. Bordetella pertussis and Corynebacterium parvum are commonly used immunopotentiating agents. To explore the inflammatory environment induced by these agents, the peritoneal exudate response in mice following intraperitoneal injection of B. pertussis (PV) and C. parvum (CV) vaccines was investigated. The PV-induced exudate isolated by lavage was characterized by an early neutrophil influx followed by enhanced accumulation of mononuclear cells and fluid protein. The CV exudate was principally mononuclear in nature and displayed fewer numbers of cells and less fluid protein. Both vaccines also enhanced the leukocyte adherence inhibitory activity (LAIA) of peritoneal fluid as measured in vitro. The development of exudate LAIA was T lymphocyte independent. A similar LAIA was demonstrated in nonimmune mouse plasma and serum. Exudate fluid and serum LAIA were heat stable and trypsin sensitive. These studies suggest that significant differences exist in the composition of the local tissue environment following PV and CV injection and that exudate LAIA is serum derived. Further studies in this direction should result in a better understanding of the ways in which inflammatory cells and fluid substances affect lymphocyte-macrophage interaction subsequent to adjuvant administration.


Immunoglobulin G to virus-specific early antigens in congenital, primary, and reactivated human cytomegalovirus infections. Immunoglobulin G antibody to human cytomegalovirus (CMV)-specific early antigens (EA-Ab) was determined by the immunoperoxidase antibody technique in several cases of congenital, primary, and reactivated CMV infections. Mothers of congenitally infected infants and a group of leukemic children and pregnant women were also studied. In 11 cases of congenital infection, CMV EA-Ab was always associated with CMV excretion whether immunoglobulin M antibody was present or not. Nine mothers of congenitally infected infants had CMV EA-Ab for several months after delivery, but association with CMV elimination was not established when urine and/or saliva were tested for virus isolation. In all nine cases of primary CMV infection, CMV EA-Ab was present, and in five its detection was associated with CMV isolation. In one case, disappearance of EA-Ab occurred when virus excretion ceased. In five cases of reactivated CMV infections, a consistent association between CMV EA-Ab and virus isolation was found. Six of 31 leukemia children had CMV EA-Ab, and virus was isolated from 3 of these. Four of 28 pregnant women showed EA-Ab in their serum, but tests for isolation were not done. These data suggest that CMV EA-Ab is not a marker of a current primary CMV infection, as previously reported, but a marker of an active CMV replication which can take place in primary as well as in congenital and reactivated CMV infections.


Characterization of anemia induced by avian osteopetrosis virus. Chickens infected intravenously at 8 days after hatching with an avian osteopetrosis virus developed a severe, progressive anemia in the absence of osteopetrosis. The anemia was characterized as a pancytopenia, in which erythrocytes, granulocytes, and thrombocytes decreased concomitantly. Serum bilirubin levels were normal, whereas erythrocytes from infected chickens demonstrated a slightly elevated osmotic fragility. A negative Coombs test indicated that there was no evidence for erythrocyte-bound antibody. Erythrocytes from infected animals had slightly decreased 51Cr-labeled erythrocyte survival time when compared with normal. Examination of marrow histological preparations, together with ferrokinetic studies with 59Fe, indicated that marrow failure occurred during the acute phase of the anemia. Circulating virus was present during the development and acute phases of the anemia, but disappeared during the recovery phase of the disease. Neutralizing antibody appeared after the disappearance of circulating virus. It is concluded that virus infection induced both marrow failure (aplastic crisis) and decreased erythrocyte survival.


Effect of erythromycin and amoxycillin on Bordetella pertussis in the nasopharynx. The effectiveness of oral erythromycin and amoxycillin in eradicating Bordetella pertussis from the nasopharynx was compared. Erythromycin in a dosage of 40--50 mg/kg/day was significantly more effective than amoxycillin in a dosage of 25--30 mg/kg/day. The organism did not disappear in three cases receiving a lower dosage of erythromycin. As antibiotic treatment does not affect the clinical course of fully-developed whooping cough, erythromycin is indicated primarily when particularly susceptible individuals are threatened by exposure. In such cases erythromycin should be given as soon as whooping cough is suspected.


Tourist hepatitis, antibodies to hepatitis A virus and immune serum globulin prophylaxis. Immune serum globulin prophylaxis for foreign travellers should be reserved for those at high risk having no immunity against hepatitis A. In this study from Sweden the incidence of tourist hepatitis in different age groups was correlated to the prevalence of antibodies to hepatitis A virus (anti-HAV) in the population. 58% of individuals over 50 years had anti-HAV and travellers in these ages seldom experienced "tourist hepatitis". In younger age groups a low prevalence of anti-HAV (11%) was combined with a higher incidence of tourist hepatitis.


Decrease in incidence of hepatitis A infections in Germany. The hypothesis of a decline in the incidence of hepatitis A infections in Germany in recent decades was confirmed by determining the prevalence of hepatitis A antibody (anti-HAV) in sera collected in 1965 and in 1975 under the same conditions in North Germany. The prevalence of anti-HAV correlated with the year of birth and was independent of the time of serum sampling. The force of infection fell from 0.04 in 1945 to 0.005 in 1965 as judged from a catalytic epidemic model with a sigmoidal decrease.


Identification of murine endogenous xenotropic retrovirus in cultured multicellular tumour spheroids from nude-mouse-passaged nasopharyngeal carcinoma. After nude-mouse-passage, a carcinoma of the nasopharynx was found to contain a few scattered C-type retrovirus particles. Culture of this nude-mouse-grown material over solid agar allowed the human tumour cells to grow as multicellular spheroids which increased in number by budding and could be subcultured for up to 6 months. Within the spheroids the human tumour cells expressed increased retrovirus replication and large numbers of C-type particles were observed. Treatment with a halogenated pyrimidine further enhanced the virus production. This tissue culture system provided sufficient virus production for the identification of the retrovirus as an endogenous xenotropic murine leukaemia virus and not a human agent.


Burkitt's lymphoma in Iraq. Clinical and pathological study of forty-seven patients. The clinical and pathological features of 47 patients with Burkitt's lymphoma seen at the University of Baghdad Teaching Hospital over the years 1969-1977 are described. There were 30 males and 17 females with a mean age of 7.5 years. All patients presented with advanced disease; 44 were in Stage III and 3 in Stage IV, (1 with central nervous system involvement; 1 with bone-marrow involvement; 1 presented as acute leukaemia). The abdomen, with or without other sites, was involved in 42 patients. Thirty-four patients died within 18 months, 11 in the immediate post-operative period. Cyclophosphamide in variable dosages was administered to 29 patients with a good response in 15. The 3 patients with Stage IV disease died with poor response to therapy. Thus, in Iraq, a non-endemic area for the disease, Burkitt's lymphoma seems to have somewhat different clinical and pathological features from the endemic (African), or the rare, sporadic disease reported elsewhere.


14q+ marker chromosome in an EBV-genome-negative lymph node without signs of malignancy in a patient with EBV-genome-positive nasopharyngeal carcinoma. In a patient with an EBV-genome-positive nasopharyngeal carcinoma, an EBV-genome-negative inguinal lymph node without histological evidence of malignant lymphoma or metastatic carcinoma growth was found to contain a 14q+ marker chromosome, identified as an 8;14 translocation, in all cells analyzed. This observation indicates that chromosome aberrations may precede histological signs of malignancy. The possible implication of this finding in relation to the postulated role of the 14q+ marker and lymphoma development is discussed.


Correlated expression of a B-lymphocyte-specific glycoprotein (gp27/35) and the EBV receptor/C3 receptor complex in sublines from the same Burkitt lymphoma. A series of virus-producer and non-producer sublines, derived from the Burkitt lymphoma line Jijoye and its P3HR-1 clone, were previously found to differ in the expression of EBV receptors and, in parallel, C3 receptors. The differences could be related to an "internal selection" caused by the cytopathic P3HR-1 virus variant, favouring the growth of receptor-negative cells. We have now analyzed the same lines for the expression of a B-lymphocyte-specific glycoprotein (gp27/35). A close parallellism was found between the expression of the EBV receptor-complement receptor complex and the glycoprotein, measured by quantitative absorption. The results favour a relationship between gp27/35 and EBV/C3 receptor expression.


Cell-density-dependence for growth in agarose of two human lymphoma lines and its decrease after Epstein-Barr virus conversion. We have compared the growth in agarose medium of two EBV-negative Burkitt lymphoma lines, BJAB and Ramos, and that of their EBV-converted derivatives. Optimal conditions for growth in agarose medium are described. The original EBV-negative lines can grow in agarose to a high efficiency, provided a critical level of cell concentration is attained. Below this level, there is no growth at all. EBV-converted sublines form colonies at a much lower cell concentration, and their maximal plating efficiency is higher. These differences may be related to a super-transformation state induced in these cells by EBV.


Biochemical identification of primate lymphoid cell-surface glycoproteins. We have employed the galactose oxidase-tritiated sodium borohydride labelling method to examine the surface glycoproteins of cotton-topped marmoset and other primate cell lines either established from tumors or transformed in vitro by different lymphotropic herpesviruses. The labelled surface glycoproteins were separated on acrylamide gels in the presence of sodium dodecyl sulfate (SDS) and analyzed by fluorography. Our results indicate that (1) lymphocytes of the same class from different primate species are similar but can be distinguished; (2) T and B lymphocytes of the same species can be differentiated; (3) cotton-topped marmoset lymphocytes of the same class show marked similarities regardless of tumor or in vitro origin or virus used for transformation; (4) three cell lines established from different EBV-induced tumors of the same marmoset show essentially the same labelling pattern, supporting the hypothesis that they originated from a single clone.


Malignant fibrous histiocytoma of the temporomandiblar joint. Report of a case. A case of malignant fibrous histiocytoma of the temporomandibular joint in a 21-year-old Japanese female was presented. The patient died of the tumor with local aggressive growth but without any organ metastasis, despite the intensive treatment including radical operation of the tumor, irradiation, and chemotherapy. This case suggested that radical curative operation was the only method of the treatment for this tumor. The clinicopathological features of the tumor are discussed, making reference to the malignant fibrous histiocytoma.


An amphetamine model of manic depressive illness. Many features of manic-depressive illness can be mimicked in man by the use and withdrawal of amphetamines. In higher doses these drugs induce a syndrome virtually indistinguishable from paranoid schizophrenia. In this paper, some of the biochemical and physiological effects of the amphetamines are examined with the hope of clarifying the nature of the biological changes in these two major functional psychoses. The actions of the amphetamines are shown to reveal the operation of specific homeostatic or adaptive nervous mechanisms. These appear likely also to operate in adaptation to psychological stress. Evidence is presented that these mechanisms are malfunctioning in manic-depressive and acute schizophrenic states and that this accounts for many of the clinical features of these conditions.


Phagocytosis in leprosy. 2. Production of superoxide by circulating blood leukocytes from lepromatous patients. The ability of polymorphonuclear leukocytes from lepromatous patients to produce superoxide (O-2) anion while phagocytizing latex particles was studied. The results were compared with those obtained in a group composed of normal individuals. No differences were found between lepromatous and normal groups. No differences were found either when the comparison was made between patients showing any form of leprosy reaction and patients without leprosy complications at the time of the study. Together with other results previously reported, our findings suggest that PMN from lepromatous patients are not different from PMN from healthy individuals with respect to their ability to generate and show the metabolic oxidative changes (as measured by the NBT- and the O-2-tests) induced by in vitro phagocytosis of latex particles. These metabolic activities remain normal even when PMN may show alterations at other levels of the phagocytic process.


Circulating immune complexes in African mothers and their newborns. Circulating immune complexes were measured using the Clq-binding test in 185 African mothers and their newborns at delivery. Results were compared to non-pregnant African women, healthy European blood donors and to a group of European mothers and their newborns. Clq-binding activity was increased in African mothers and in non-pregnant African women. Circulating immune complexes rarely occurred in cord blood even when mothers showed a high Clq-binding activity. The levels of immune complexes were unrelated to malaria. Complement components Clq and C3 were lower in the newborns than in the mothers.


A study of 204 consecutive deliveries in Abidjan--Anthropometric data of newborns, mothers and placentas. Anthropometric measurements of newborns, mothers and placentas were taken in 204 consecutive deliveries at Adjamé Maternity Hospital in Abidjan. Aclinical assessment of gestational age was made. Low mean birthweight can be attributed to a higher incidence of pre-term newborns in the present sample. Incidence of small for gestational age and large for gestational age babies is similar to that found in developed countries. Detailed analysis of correlations between maternal, placental and neonatal variables show that birthweight is dependent on nutritional status of the mothers and on size of the placenta. Young primiparae are shown to become pregnant before reaching full growth. Their weight and weight for height are significantly lower than for multiparae. The main group at risk for impaired foetal growth, prematurity and low birthweight are the children born to primiparous mothers.


Maternal anaemia in Abidjan--Its influence on placenta and newborns. In a random population of 198 pregnant women, detailed haematological determinations were performed at delivery. The prevalence of anaemia (haemoglobin less than 10 g/100 ml) is 20%. Iron deficiency is even more frequent. Malaria is shown to play an important role. Maternal anaemia is also correlated with nutritional status. Primiparae are shown to be at higher risk of anaemia. The effect of maternal anaemia on anthropometric and haematological variables of the newborn are discussed. Mothers with sickle-cell or HbC trait are not at special risk of anaemia.


Malaria at delivery in Abidjan.

Malaria at delivery in Abidjan. In 198 consecutive deliveries in a region holoendemic for malaria, 39.4% of the mothers had malarial parasitaemia. Cord blood parasitaemia was found in 43 cases (55% of infected mothers). The incidence of malaria decreased with increasing parity possibly related to age. Malaria is partly responsible of the low birthweights and of the genesis of pre-term and small for date babies. A correlation between nutritional status and malarial infection was shown. According to IgM concentrations in cord blood, 2.5% of newborns had an intrauterine infection of unknown origin.


A year of deliveries at the Adjamé maternity hospital in Abidjan (Ivory Coast). This study based on hospital records is an analysis of 7433 deliveries in Adjamé Maternity Hospital, Abidjan, Ivory Coast. The incidence of low birthweight babies is 14.1%. Mean birthweight and height were shown to increase significantly from parity 1 to 9. Lowest mean birthweight and highest incidence of low birthweight babies were observed in primiparae. Incidence of stillbirths and abortions according to parity are described and an estimate of infant mortality rate has been attempted.


Haematological data of African newborns and their mothers in Abidjan. The analysis of cord blood samples of 198 African newborns and their mothers confirmed the high prevalence of iron deficiency. A hypochromia and microcytosis of red cells was found in 40% of the pregnant women at term. In the majority of newborns the haemoglobin concentration was below 16 g/100 ml. Hb S was found in 11%, Hb C in 9% and alpha-thalassaemia in 2% of the population examined. Haemoglobin, packed cell volume and mean cell volume of erythrocytes were lower than results of newborns from developed countries. Foetal sex had no influence on haematological variables.


Enzyme histochemical properties of kitten osteoclasts in bone imprint preparations. The enzyme histochemical characteristics of osteoclasts in imprints of the metaphyseal regions of femurs, from male kittens aged approximately 18 weeks, were investigated. A selected number of enzymes representative of a variety of metabolic pathways were studied. The enzyme profile, time for the first appearance of detectable reaction product, intensity of the reactions, and localization of the reaction products were noted. Osteoclasts are rich in enzymes, and metabolic pathways are well developed in respect of the utilization of the reduced coenzymes NADP and NADPH, succinic, malic, lactic, and isocitric acids, beta-hydroxybutyrate and glucose-6-phosphate, the reactions being mediated by the diaphorases and dehydrogenases. The activities of acid and neutral phosphatases, non-specific esterases, and leucine naphthylamidase were high in these cells. However, little or no activity was demonstrated in respect of glutamate and alpha-glycerophosphate dehydrogenases or of aryl sulphatase, glucose-6-phosphatase and alkaline phosphatase.


Ultrastructural localization of glycogen in the granulocytes of normal rabbit bone marrow. The glycogen of rabbit granulocytes has been studied in glutaraldehyde and osmium tetroxide fixed bone marrow by the periodic acid-thiocarbohydrazide-silver proteinate procedure (PA-TCH-SP). The PA-TCH-SP procedure involved the staining of intracytoplasmic glycogen more densely than the routine lead citrate staining. The PA-TCH-SP procedure demonstrated the intracytoplasmic glycogen in all three kinds of granulocytes. Though a sequence of intensity was observed in each stage of cell maturation, intracytoplasmic glycogen increased generally in accordance with cell maturation in the granulocytes. Functional significance of the glycogen in the granulocytes was discussed in relation to its staining. A very weak reaction in the granules of the granulocytes was described in relation to their contents.


Histochemical studies of some myocardial oxido-reductive enzymes after experimental beta-adrenergic blockade with propranolol. Histochemical studies of some myocardial oxido--reductive enzymes after a beta--adrenergic blockade with propranolol have been carried out. Succinate, isocitrate and glucose-6-phosphate dehydrogenases did not indicate any changes in activity, whereas the changes in reaction intensities concerning NADH and NADPH tetrazole reductases and glutamate dehydrogenase have rather a transitory and reversible character. Only lactate dehydrogenase showed an increase in the enzymatic activity which speaks for an increase in the glycolysis process in the heart muscle. In the light of our own presented research results we assume that the experimental beta--adrenergic blockade of the heart muscle in rats does not evoke more important enzymatic changes which are noticeable in histochemical microscope examination.


Waking and ventilatory responses to laryngeal stimulation in sleeping dogs. We studied waking and ventilatory responses to laryngeal stimulation during sleep in three dogs. The dogs breathed through an endotracheal tube inserted caudally into the trachea through a tracheostomy. Laryngeal stimulation was produced either by inflating a small balloon that was positioned in the rostral tracheal segment, or by squirting water onto the larynx through a catheter inserted through the tracheostomy. Airflow was measured with a pneumotachograph, and sleep state was determined by behavioral, electroencephalographic, and electromyographic criteria. We found that the degree of laryngeal stimulation required to produce arousal and coughing was higher in rapid-eye-movement (REM) sleep than in slow-wave sleep (SWS). Stimuli that failed to cause arousal from SWS often produced a single expiratory effort, or brief apnea (1--2 s) and bradycardia. In contrast, during REM sleep subarousal stimuli often resulted in prolonged apnea (greater than 10 s) and marked bradycardia. We conclude that during REM sleep arousal responses to laryngeal stimulation are depressed, but ventilatory and cardiac responses are intact.


O2 uptake and developed tension during and after fatigue, curare block, and ischemia. The purpose of this study was to investigate the effect of changes in developed tension on the ratio between O2 uptake and isometric developed tension in the in situ dog gastrocnemius-plantaris muscle group. O2 uptake and isometric developed tension of the muscle were measured during contractions at 1 twitch/s before and after fatigue with both single and twin impulses (6.5 V, 0.2-ms duration; the twins were separated by 10--20 ms). Twin impulses prior to fatigue raised developed tension to two times the tension developed with single impulses. Fatigue was obtained by stimulation at 10--20 impulses/s for 30--40 min. Twin impulses after fatigue returned developed tension to the level of single impulses before fatigue. O2 uptake and developed tension were also measured during the slower development of fatigue produced by continuous stimulation (3, 4, 5, and 6 impulses/s) as well as during "fatigue" induced by partial neuromuscular block with curare or ischemia. In all cases, there was no change in the O2 uptake per unit of tension developed, indicating a constant coupling between O2 uptake and developed tension.


Effect of exercise on collateral development in dogs with normal coronary arteries. To determine the effects of chronic exercise on the coronary collateral circulation of dogs with normal coronary arteries, 1-yr-old purebred beagles were divided into sedentary control and exercising groups. The latter were trained to run on a treadmill. A lower maximal heart rate during a standardized exercise test protocol after a 10- to 12-week training period and a higher gastrocnemius cytochrome oxidase activity in the runners attested to the presence of cardiovascular and skeletal muscle training effects. However, left ventricular weights, left ventricle/body weight ratios, myocardial myofibrillar and myosin ATPases, and hemodynamics were similar in sedentary and exercising dogs except for a significantly higher resting cardiac output in the runners. After occlusion of the left anterior descending coronary artery, both collateral conductance (retrograde flow/aortic pressure) and collateral flow measured with microspheres tended to be lower in the trained dogs, but differences were not significant. The endocardial/epicardial flow ratio in the ischemic area after coronary occlusion did not distinguish between exercisers and controls. Thus treadmill running in the dog with normal coronary arteries produced a training effect, but had no effect on coronary collateral vessels.


Triiodothyronine (T3) concentration and therapy in autistic children. The clinical and biochemical status of thyroid function of patients with an autistic syndrome was investigated. The study consisted of 13 patients between the ages of 7 and 21 years. There was no clinical evidence for hypothyroidism in any patient, and T3, T4, and TSH concentrations were within the normal range. Two patients who had retarded bone ages were treated with triiodothyronine for 6 months. Hyperthyroidism developed when T3 levels exceeded physiologic concentrations in these patients. The concept that the clinical response to triiodothyronine in autistic patients results from correction of thyroid dysfunction is not supported by these findings.


Mode of inhibition of globin synthesis by m7G5'p and m7G5'ppp. The mode of inhibition of rabbit globin synthesis by m7G5'p and m7G5'ppp ("cap analogs") was studied using the rabbit reticulocyte lysate system. The rate of globin synthesis was measured at various concentrations of both f[35S]Met-tRNAf and the cap analogs. The cap analogs were found to inhibit competitively the incorporation of f[35S]Met into hot trichloroacetic acid-insoluble material. Nascent chains prelabelled with f[35S]Met were released at various concentrations of m7G5'ppp. The release of nascent chains was not inhibited by m7G5'p (Suzuki, H. (1976) FEBS Lett. 72, 309) and m7G5'ppp, and it is therefore concluded that the cap analogs inhibit a step of initiation of globin synthesis. Under conditions such that the elongation of nascent chains was inhibited by sparsomycin, the formation of an 80S/fMet-tRNAf was inhibited by the cap analogs, but not that of a 40S/fMet-tRNAf complex. These data suggest that a factor which is required for the binding of globin mRNA with 40S/fMet-tRNAf complex forms an inactive complex with the cap analogs, so that the cap analogs inhibit globin synthesis.


Collagenolytic activities of cultured human malignant melanoma cells. Three human malignant melanomas were cultured in pure populations and one tumor was cloned into melanotic and amelanotic cell lines. In the homogenates of these cultured cells, specific collagenase activities were demonstrated by isotope release from 14C-labeled collagen, disc electrophoresis, and specific cleavage of collagen molecules as demonstrated in the segment long spacing form. No significant collagenase activity was observed in the culture media. Interestingly, early cultures had a high collagenase activity in the cells and as they were successively subcultured, the activity diminished. Cysteine completely inhibited the degradation of tropocollagen as determined by disc electrophoresis and EDTA partially inhibited the degradation. It is concluded that human malignant melanoma cells produce a specific collagenase in vitro which can be extracted in early culture directly from the homogenate.


Acyl-Coenzyme A synthetase and fatty acid oxidation in rat liver peroxisomes. Rat liver peroxisomes oxidized palmitate in the presence of ATP, CoA and NAD+, and the rate of palmitate oxidation exceeded that of palmitoyl-CoA oxidation. Acyl-CoA synthetase [acid: CoA ligase (AMP-forming); EC 6.2.1.3] was found in peroxisomes. The substrate specificity of the peroxisomal synthetase towards fatty acids with various carbon chain lengths was similar to that of the microsomal enzyme. The peroxisomal synthetase activity toward palmitate (40--100 nmol/min per mg protein) was higher than the rate of palmitate oxidation by the peroxisomal system (0.7--1.7 nmol/min per mg protein). The data show that peroxisomes activate long chain fatty acids and oxidize their acyl-CoA derivatives.


Primary avian tendon cells in culture. An improved system for understanding malignant transformation. Primary avian tendon (PAT) cells which maintain their differentiated state in culture are rapidly transformed by Rous sarcoma virus. By criteria of morphology, increased rate of 2-deoxyglucose uptake, and loss of density dependent growth control, PAT cells transform as well as their less differentiated counterpart, chick embryo fibroblasts. In addition, the percentage of collagen produced by PAT cells drops on transformation by an order of magnitude, from 23 to 2.5%, but is unaffected by viral replication of a transformation-defective mutant. The responsiveness of normal and transformed PAT cells to various environmental factors changes dramatically upon transformation. Normal PAT cells respond to the presence of ascorbate and high cell density by raising the level of collagen synthesis from 5 to 23%. Transformed PAT cells are totally unresponsive. These and previously reported results lead us to postulate that the break-down in the normal regulatory mechanisms used by the cell to maintain the differentiated state is related to or is responsible for the onset of malignant transformation.


Transport and phosphorylation of hexoses in normal and Rous sarcoma virus-transformed chick embryo fibroblasts. Effects of transformation by Rous sarcoma virus on sugar uptake and activity and the subcellular distribution of hexokinase isozymes in chick embryo fibroblasts were examined. Transformation caused a several-fold increase in the maximum velocity for uptake of 2-deoxyglucose without a significant change in Km. Cytochalasin B (CB), was used to differentiate between the effects of transformation on facilitated diffusion and the nonsaturable (CB-insensitive) mode. Transformation was found to stimulate 2-deoxyglucose transport by both mechanisms, but the increase in transport by the CB-insensitive mode was greater. Transformation enhances the activity of hexokinase, the enhancement being confined to the particulate fraction of the enzyme. Heat-inactivation and electrophoretic mobility studies showed that although hexokinase Type I is the major form in both normal and transformed fibroblasts, there is a significant increase in the proportion of the Type II isozyme in the transformed cells.


Measurements of metabolites during cAMP oscillations of Dictyostelium discoideum. During aggregation the cellular slime mold Dictyostelium discoideum synthesizes and releases pulses of cAMP about every five minutes. Current models proposed to explain this phenomenon postulate that oscillating levels of some key intracellular metabolite control the oscillatory synthesis of cAMP. We have assayed the levels of likely candidates for this metabolite during a cAMP oscillation, but have found them to remain constant. Compounds measured include ATP, GTP, glucose-1-phosphate, glucose-6-phosphate, isocitrate, alpha-ketoglutarate, amino acids, and other aminated metabolites. On the basis of this negative data, as well as results described elsewhere (Geller and Brenner, '78), we question whether the proposed models are correct, and discuss several alternatives.


Synergistic hemolysis phenomenon shown by an alpha-toxin-producing Clostridium perfingens and streptococcal CAMP factor in presumptive streptococcal grouping. A new phenomenon of synergistic hemolysis by Clostridium perfringens alpha-toxin and the streptococcal CAMP factor on human and guinea pig erythrocytes is described. A possible mode of action of the CAMP factors is suggested. On human blood agar all of the tested isolates of group B streptococci gave an arrowhead-shaped zone of hemolysis; 74% of group A gave a crescent-shaped lytic zone, whereas all isolates of groups C and G and the remaining 26% of group A streptococci gave a bullet-shaped lytic zone. By comparison, in the CAMP test incubated aerobically and anaerobically, 70 and 91%, respectively, of streptococci other than group B gave positive, arrowhead-shaped lytic zones. If all intermediate positive reactions in the CAMP tests were read as negative after aerobic incubation, only 89% of group B streptococci would be properly identified. The synergistic hemolysis phenomenon, using an alpha-toxin-producing C. perfringens and human blood agar, provided a reliable test for presumptive identification of group B streptococci, with promising potential to differentiate in the same test group A streptococci from other groups.


Method for evaluating broth culture media: application to Haemophilus. A method was devised to test the growth-promoting ability of a broth medium. The "dilute to extinction" method determines the inoculum required to develop heavy turbidity in a broth with overnight incubation. A statistical method using Poisson distribution was used to show that a single Haemophilus cell can develop heavy turbidity in an optimal broth. The dilute to extinction method was used to evaluate the shelf life of stored media, to titrate the growth factor requirements of Haemophilus, and to evaluate the use of purified hemin and nicotinamide adenine dinucleotide in a broth medium for the growth of Haemophilus. Of the media tested, the most suitable formulation was Mueller-Hinton broth supplemented with 10 microgram of hemin and 10 microgram of nicotinamide adenine dinucleotide per ml. The dilute to extinction method appears to be especially useful in the development of broth media for fastidious organisms. The method could also be used to assure the quality of other broth media which are required to support the growth of small inocula in the clinical or research laboratory.


Antibody assays for varicella-zoster virus: comparison of enzyme immunoassay with neutralization, immune adherence hemagglutination, and complement fixation. An enzyme immunossay (EIA) was adapted for detection of antibody to varicella-zoster virus, and its sensitivity and specificity were compared with those of neutralization, immune adherence hemagglutination (IAHA), and complement fixation tests. Test sera showed little nonspecific reactivity in the EIA system, and valid results could usually be obtained at serum dilutions as low as 1:8. Demonstration of the presence or absence of varicella-zoster viral antibody by EIA showed 94% correlation with results obtained in neutralization tests, but EIA titers were 2- to 16-fold higher than neutralizing antibody titers. Results by IAHA showed 87% correlation with those obtained by neutralization. No false positive IAHA results were seen, but a number of false negative IAHA results were seen at the 1:8 serum dilution, particularly in older individuals. With increasing age (>40 years), and presumably increased time from varicella infection, neutralizing antibody levels generally declined to 1:8 or 1:16, EIA levels fell to 1:128 or 1:256, and IAHA and complement fixation antibody titers were usually <1:8 or 1:8. EIA and IAHA were as reliable as the neutralization and complement fixation tests for serodiagnosis of varicella and zoster infections. All tests demonstrated heterotypic varicella-zoster antibody titer rises in selected patients with initial herpes simplex virus infections, but fewer heterotypic responses were seen by EIA than by the other methods. EIA offers a rapid, sensitive, and specific method for varicella-zoster antibody assay that is applicable to use in a clinical setting.


Comparison of pseudorabies virus inactivated by bromo-ethylene-imine, 60Co irradiation, and acridine dye in immune assay systems. Pseudorabies virus infections among animals, especially swine, have become prevalent in the United States in the past few years. The disease in swine is now economically important. Test systems and antigens are being developed for use in control and disease suppression efforts. Pseudorabies virus was inactivated by three methods: chemically, with bromo-ethylene-imine; physically, with 60Co irradiation; and chemically and physically, with 3,9-diaminoacridine dye followed by exposure to white visible light. The antigenicities of the preparations were determined in the presence of specific antibody in immunodiffusion tests and through immunoelectrophoresis. The latter technique permitted quantitation of either antigen or antibody. In the electrophoretic patterns, the antigenic mass in bromo-ethylene-imine preparations was estimated to be 42 mg/ml, the same as in the untreated control material. After 60Co irradiation, 22 mg/ml was present, in comparison with 50 mg/ml in the untreated control antigen. In contrast, 67 mg/ml was present in the acridine dye-light-treated preparation, in comparison with 58 mg/ml in the untreated control material. A possible explanation for the acridine dye-light-treated preparation values is that photodynamic inactivation interferes with viral maturation during the replicative cycle within cells, with a resulting production of a greater amount of antigen, at least some of which is in the form of defective particles.


Trophoblastic disease with coexistent fetus: a sonographic and clinical spectrum. The sonographic and clinical spectrum of coexistent live fetus and trophoblastic disease is illustrated and discussed. Two patients with a single placenta with trophoblastic change and one patient with two placentas, one normal and one with a hydatidiform mole, are described. The clinical and sonographic abnormalities in all three patients were characteristic. In the two patients with single placenta, the histologic findings were minimal. The clinical and sonographic presentations of coexistent mole and fetus are described and etiology is discussed. The implication of the lack of histologic confirmation of hydatidiform mole suggests a spectrum of findings in trophoblastic disease.


Ultrasonographic demonstration of inferior vena caval invasion. Whenever a liver neoplasm is seen ultrasonographically, an effort should be made to visualize the veins within the liver as well as the inferior vena cava for tumor extension. Valuable information can be obtained by the demonstration of this finding which may radically alter the further work-up and treatment of the patient.


Synaptic fine structure and the termination of corticospinal fibers in the lateral basal region of the cat spinal cord. The lateral basal region (LBR) of the spinal cord gray matter (Rexed's laminae IV-VII) by physiologic and anatomic criteria is the major terminal zone for the corticospinal (CS) tract in the cat. The neurons in this area are medium-sized with abundant spines on their dendrites. Axon terminals on the dendrites and somata of these neurons form synapses easily classified as asymmetric with spheroid vesicles and symmetric with flattened vesicles. There are rare exceptions to this. In a systematic count of terminals, 82% have spheroid and 18% flattened vesicles. The majority of all terminals are on dendrites (84.9%) and a minority on somata (14.1%). Less than 1% are axoaxonic. Degeneration of the corticospinal tract was produced by transecting one hemisphere of our experimental cats. Its termination in the lower cervical cord was studied for 17 hours to 7 days after surgery. Vesicle depletion and clumping and dense polymembranous inclusions were the most common forms of degeneration. Filamentous proliferation in the terminals was also prominent; dark degeneration, however, was infrequent. The percent of degenerating CS terminals in the LBR was the following: 17 hours - 2.2%, 36 hours - 4.19%, 2 days - 10.3%, 3 days - 8.4%, 4 days - 6.9%, 7 days - 10.25%; 84.8% of degenerating CS terminals were axodendritic and 15.2% axosomatic.


Membrane-limited granules in sinus hair gelatinous membrane in mice. Electron dense membrane limited granules were observed in basal cells of the outer root sheath and in connective tissue sheath of the anagen sinus hair of mice. These granules were oval or round in configuration, and were 150--180 nm in size. They were considered to represent structures related to collagenolytic activity to accelerate elongation of anagen hair.


Orogranulocyte peroxidase activity as a measure of inflammatory periodontal disease. Benzidine dihydrochloride in the presence of dimethylsulfoxide reacts with myeloperoxidase contained within the granules of polymorphonuclear leukocytes obtained from the oral cavity. A blue meriquinoid oxidation product is formed in the presence of hydrogen peroxide, which can be extracted from the granules with a combination of methyl alcohol and dimethylsulfoxide. A stable yellow-brown pigment is formed which can be measured spectrophotometrically. The values are expressed in optical density units and correlate with the Orogranulocytic Migratory Rate units obtained by other means. Consequently, the spectrophotometric method is an additional non-subjective technique available for the evaluation, monitoring, and indexing the inflammatory periodontal disease.


Angiolymphoid hyperplasia with eosinophilia: report of a case and a review of the literature. A case of angiolymphoid hyperplasia with eosinophilia mimicking multiple cylindromas (turban tumor) is presented in detailed clinical and histologic features, and the literature on the condition is reviewed.


The effects of the referral process on hospital in-patients. Decisions relating to the preparation for hospital discharge of medical patients can take place at any time after admission to hospital. Referrals to the hospital departments of social work, occupational therapy and physiotherapy, and to community nursing, health visiting and social services are a key factor in this preparation. The referral process is complex and multifaceted and can be analyzed in terms of the questions of who initiates the referral, for what reasons, and at what point in the patient's career. The variant views of role of those professionals involved in the referral process and their perceptions of the situation affect what subsequently takes place and can be examined in relation to each other and to the referral process itself. The complexity may be increased further by the intervention of a patient's relatives. Referrals are made on the basis of a social diagnosis, the sources of information for which are diverse, with the result that the referral process is unstandardized and unpredictable.


Paget's disease and the nervous system.

Paget's disease and the nervous system. Paget's disease usually is found in patients past the age of 40. Early presenting symptoms include headache, deafness, tinnitus, and pain due to radicular compression. The diagnosis is confirmed by radiographic features and elevated levels of serum alkaline phosphatase and urinary hydroxyproline. Bony overgrowth results in pressure on nearby soft tissues such as the brain, spinal cord, and certain peripheral nerves. The abnormally soft quality of the calvarial bone permits distortion by the weight of the brain. Dorsal inclination of the plane of the foramen magnum and the projection of the odontoid process into the posterior fossa lead to stretching of the brain stem over the odontoid process and the ventral margin of the foramen. Obstructive hydrocephalus may result. Sarcoma of the crainial vault may develop in cases of Paget's disease. Once cervicomedullary or spinal compression has occurred, surgical decompression may be necessary. Three drugs--calcitonin, disodium etidronate, and mithramycin--have been used with some benefit in the treatment of Paget's disease.


A review of psychophysiological research with hyperkinetic children. A number of studies have appeared which focus on the issue of whether hyperactive children are psychophysiologically different from normal children and what impact the stimulant drugs may have on psychophysiological characteristics of these children. The present paper reviews these studies under the type of measures that were employed. In general, the results suggest that hyperactive children are probably not under- or overaroused in their resting levels of autonomic functions, although some children may display resting cortical underarousal. However, the findings of studies on the impact of stimulation on autonomic or central functions intimate that some hyperactive children are probably underreactive to environmental stimulation, or are "underarousable." Indeed, where differences between hyperactive and normal children are found in such evoked-response studies, they are consistently in this direction of "underarousability." Results for the effects of stimulant drugs suggest that these drugs energize or increase the "arousal" of these children and enhance the impact of stimulation on the nervous system. The implications of these results for current theories of hyperactivity and for future research are discussed.


Cytochemical demonstration of sodium, potassium-adenosine triphosphatase by a hemepeptide derivative of ouabain. A cytochemical probe for the ultrastructural localization of the NaKATPase was devised, which utilizes the biological affinity of the noncompetitive inhibitor, ouabain (ouab) to which was coupled a hemepeptide (H11P) which possesses peroxidatic activity. The conjugate, ouab-H11P, had an apparent Ki of approximately 8 x 10(-7) M. When reacted with fixed tissue from the salt gland of osmotically stressed ducklings, the NaKATPase was localized to the basal and lateral infoldings of the plasma membranes of secretory epithelial cells. Reaction product consisted of fine textured deposits distributed in focal patches on the outer aspects of the membrane. Apical membranes were negative, as were intracellular membrane components. Preincubation of tissue with unlabeled ouabain or binding of ouab-H11P in the presence of 10 mM K+, no ATP and no Mg++, resulted in the absence or diminution of reaction product.


Quantitative introduction of a given macromolecule into cells by fusion with erythrocyte ghosts using a fluorescence activated cell sorter. FITC-conjugated bovine serum albumin (FITC-BSA) molecules were quantitatively introduced into human erythrocyte ghosts by gradual hemolysis. When the ghosts and L cells were fused with UV-inactivated HVJ (Sendai virus), FITC-BSA was introduced into the cytoplasm of the L cells and fluorescence could be observed inthe cells with a fluorescence microscope. A mixture of L cells and ghosts was introduced into a fluorescence activated cell sorter (FACS), which could separate the mononuclear cells on the basis of their light-scattering profile. Four distinct populations of mononuclear cells were found by fluorescence analysis. These populations were separated from the cell mixture and found to correspond to cells fused with one, two and three ghosts and unfused cells. After separation, the cells from each population could form colonies in culture. As a given macromolecule can be quantitatively introduced into erythrocyte ghosts with the FITC-BSA, after fusion of these ghosts with cells, this sorting method is useful for separating cells containing a definite number of macromolecules.


The relative effectiveness of commonly used disinfectants in inactivation of coxsackievirus B5. Coxsackievirus B5 in the presence of fetal calf serum was exposed to six commonly used disinfectants for times of 10, 20 and 30 s. At the end of exposure times skim milk neutralized the disinfectant activity, with residual virus assayed by the plaque technique. The six disinfectants considered were Javex, sodium hydroxide, ethanol, Wescodyne, One Stroke Ves-Phene and Sonacide. Although 95% (v/v) ethanol was significantly more virucidal than dilutions of the other five disinfectants tested causing a 10(6) reduction in 20 s, it may not be practical to use in many instances. Next to 95% (v/v) ethanol, 1/75 (800 parts/10(6) Javex,  0.25% (w/v) sodium hydroxide and 1/200 Wescodyne were the most effective virucides. These disinfectants were equal in effectiveness causing a 10(5) reduction of coxsackievirus B5 in 30 s. Of these three disinfectants Javex is the most practical to use since sodium hydoroxide is caustic and Wescodyne is selective in its virucidal action. Undiluted Sonacide was a less effective virucide causing a less than 10-fold reduction of coxsackievirus B5 in 30 s. A 1/50 dilution of One Stroke Ves-Phene was the least effective virucide tested since it did not significantly inactivate coxsackievirus B5 in 30 s.


Inherited enzyme variation among JAX strains of domestic rabbits. Twelve proteins were assayed for electrophoretic polymorphism in inbred and partially inbred JAX strains of the domestic rabbit. Four enzymes (mannosephosphate isomerase, soluble and mitochondrial malic enzyme, and methemoglobin reductase) were determined to be genetically polymorphic.


Excretion of foot-and-mouth disease virus in oesophageal-pharyngeal fluid and milk of cattle after intranasal infection. The virus growth in the pharyngeal area and the virus excretion in milk of susceptible and vaccinated dairy cows after intranasal instillation of foot-and-mouth disease (FMD) virus type O1 were examined. Ten vaccinated cows were purchased through a market. Of these, nine had delivered their first calf. The cows were inoculated 2-9 months after having received the last dose of vaccine. All vaccinated cows resisted the intranasal challenge. The virus multiplied in the pharyngeal area but, compared with two susceptible controls, to a limited extent. No clear relation was found between virus growth and the titre of circulating neutralizing antibody at the time of challenge. Virus was first detected in milk samples of the susceptible cows when generalized FMD lesions had developed on day four; the excretion lasted for 3-4 days. Up to 19 days after inoculation untreated milk of the vaccinated cows was examined for the presence of infectious FMD virus. Samples were inoculated onto cell cultures, fed to susceptible pigs and calves and injected intramuscularly and/or intradermolingually into susceptible steers. No infectious FMD virus could be detected, either in cell cultures or in susceptible animals. The animals did not develop neutralizing antibody against FMD virus and were subsequently shown to be fully susceptible to challenge. The results are discussed with particular reference to current problems regarding the export of milk products from countries where vaccination against FMD is practised to countries free of the disease.


Immunopathogenicity and oncogenicity of murine leukaemia virus. IV. Antinuclear antibody response and tumour induction in B10.A recombinant mice. The levels of murine leukaemia virus (MuLV) proteins p30 and gp70, antinuclear antibody (ANA), anti-soluble nuclear protein, anti-single-stranded DNA, anti-double-stranded DNA and anti-histone antibodies were measured in B10.A and B10.A recombinant mice neonatally infected with MuLV-Scripps (Lerner et al., 1972). The incidence and latency of lymphoma and the incidence of glomerulonephritis were also determined. The mice studied could be divided into high-responder and low-responder groups. Most of the high ANA antibody could be attributed to anti-histone antibody. High response was associated with the H-2b haplotype and recombinant strains on the B10 background which were identical at the I-A subregion derived from the H-2b parental stock. In contrast, low ANA response was associated with the I-A subregion derived from the H-2k haplotype. In all groups of virus-inoculated animals, most animals developed serum elevations of p30 and gp70 and at least 72% of the inoculated animals developed lymphomas. High serum p30 levels correlated inversely with latency and directly with gp70 values. From 4 to 28% of the animals in each virus-inoculated group had histological evidence of glomerulonephritis, although no clear genetic basis could be ascribed to the incidence of glomerulonephritis, serum p30 or gp70 levels, or latency of lymphoma development.


Evidence that endotoxin is the cyclic 3':5'-GMP--promoting factor in erythropoietin preparations. Since Ep preparations are contaminated with endotoxin, the possibility that the latter might be the factor in crude Ep which increases cGMP levels in rat fetal liver cells was examined. Endotoxin produced a striking elevation of cGMP in rat fetal liver cells without affecting cAMP levels or heme synthesis. Absorption with Limulus lysate of more than 99% of the endotoxin in a crude Ep preparation caused a parallel decrease in the cGMP-promoting activity without reduction of heme synthetic potency. It is concluded that endotoxin is the component of crude Ep which increases cGMP levels in rat fetal liver. The precise role of elevated cGMP in the action of endotoxin on cells and the universality of this effect remain to be determined.


Origin and transport of the A-I and arginine-rich apolipoproteins in mesenteric lymph of rats. Transport of apolipoprotein A-I and argininerich apolipoprotein in mesenteric lymph was examined in rats given constant intraduodenal infusions of saline, glucose in saline, or emulsified fat. Lymph flow in all groups was constant from 5 to 50 hr after beginning the infusions. Lymphatic transport of triglycerides was about 20-fold greater and transport of apoprotein A-I was about twofold greater in fat-infused rats than in the other two groups. In each group transport of apoprotein A-I bore a significant positive relationship to transport of triglycerides. Lymphatic transport of the arginine-rich apoprotein was only 6-12% of that of apoprotein A-I and was more closely related to lymphatic transport of total protein than to that of triglycerides. In fat-infused rats given [(3)H]lysine intraduodenally, about two-thirds of the (3)H in the chylomicron proteins was in apoprotein A-I and only about 1% was in the arginine-rich apoprotein. Estimated specific activity of chylomicron proteins was highest for apoprotein A-I and apoprotein A-IV, and lowest for the arginine-rich apoprotein and proteins of low molecular weight (mainly C apoproteins). In fat-infused rats given constant intravenous infusions of radioiodinated high density lipoproteins from blood plasma, the specific activity of apoprotein A-I in lymph chylomicrons was only about 5% of that of apoprotein A-I in blood high density lipoproteins, indicating that more than 90% of the apoprotein A-I in chylomicrons was synthesized in the intestine. From these and other data it is concluded that both the intestine and liver are significant sources of apoprotein A-I whereas only the liver synthesizes significant amounts of the arginine-rich apoprotein.


Purification of polyphosphoinositides by chromatography on immobilized neomycin. The binding of polyphosphoinositides (phosphatidylinositol phosphate and phosphatidylinositol bisphosphate) to the antibiotic neomycin is utilized for the purification of these lipids. Neomycin is reductively coupled to reactive glass beads (Glycophase-CPG) and serves as the stationary phase in column chromatography. A total lipid extract is prepared from tissues with chloroform-methanol-KC1 or chloroform-methanol-HC1 and washed once with acidified methanol-water. After the addition of an equal volume of methanolic 200 mM ammonium acetate, the extract is directly applied to the column. All lipids but the polyphosphoinositides are removed from the column by rinsing with 150 mM ammonium acetate in chloroform-methanol-water. Increasing the salt concentration to 600 mM elutes phosphatidylinositol phosphate. While further increases in ionic strength are not sufficient for a quantitative removal of phosphatidylinositol bisphosphate, the lipid is completely eluted by the addition of either ammonia or HC1 to the solvent. The column can be recycled and used repeatedly.


Endogenously labeled low density lipoprotein triglyceride and apoprotein B kinetics. The kinetics of endogenously labeled low density lipoprotein (LDL) triglycerides (TG) and apoprotein B (apoB) have been studied in four normal and in four hyperlipemic subjects using double tracers. Analysis of the data suggests that most LDL triglycerides turn over about 10 times faster than apoB (0.003/min vs. 0.0003/min) and that about 10% of the LDL particles contain most of the TG found with LDL. It is not possible to determine from the analysis whether each new LDL particle arrives with the excess TG or whether only a subpopulation of particles contains most of the TG. The kinetic analysis further suggests that triglyceride-rich LDL particles do not exchange with an extraplasma compartment as most LDL particles do, and thus, they behave more like very low density lipoprotein particles. A compartmental model accounting for both the LDL-TG and LDL-apoB kinetics is proposed.


Male--female variability in the dietary cholesterol-induced hyperlipoproteinemia of cynomolgus monkeys (Macaca fascicularis). The characteristics of dietary cholesterol-induced hyperlipoproteinemia were studied in adult Macaca fascicularis to determine if significant male--female differences could be identified. The plasma lipoproteins from individual animals were separated by agarose column chromatography and the resulting size populations of lipoproteins were chemically characterized. The distribution of constituents among the lipoprotein classes was determined. The lipoproteins that increased most significantly in response to dietary cholesterol were the low density lipoproteins (LDL) which were those of region III of the column elution profile. The LDL mass concentrations (in mg/dl of plasma) were not different between males and females. However, these lipoproteins showed a characteristic difference between males and females in response to dietary cholesterol. The primary response in the males was an increased LDL molecular weight. In females, the primary response was an increased number of LDL particles (measured as mumolar concentration). Thus, at the same LDL mass concentration, males had significantly larger LDL particles and yet fewer of them. Analysis of the chemical composition data showed that size differences between LDL of males and females was due primarily to the incorporation of more cholesteryl ester into the LDL particles from male animals; the content per LDL particle of free cholesterol, phospholipid, and protein showed the same proportionality to size for both sexes. Another significant difference between lipoproteins of males and females was in the extent to which the plasma HDL (identified in these studies as the lipoproteins of region IV of the elution profile) decreased in response to dietary cholesterol. Chemical composition differences were also noted. The HDL particles of males and of cholesterol-fed animals contained more cholesteryl ester than did HDL of females and control animals, respectively.


Bronchogenic carcinoma metastasizing to the orbit. A case report. A case of an orbital metastasis from a bronchogenic neoplasm is described. The initial clinical presentation was of unilateral facial pain and paraesthesia shortly followed by increasing left-sided external ophthalmoplegia and proptosis and the development of a slight swelling of the left temporal fossa. The diagnosis was established by a biopsy of the temporal mass. The patient died two months after the diagnosis was established. The significance of facial pain and paraesthesia and the literature pertaining to orbital cellulitis and orbital metastases is discussed.


Diurnal variations in the influence of the ultimobranchial glands on calcium homeostasis in the frog (Rana pipiens). The effect of the light-darkness cycle on the efficiency of the ultimobranchial and parathyroid glands in altering duodenal calcium transport and plasma and urinary concentrations of calcium was examined in the adult male frog (Rana pipiens). Frogs were unfed, but were allowed access to 0.05 M-CaCl2 in the surrounding medium after ultimobranchialectomy or parathyroidectomy. Calcium transport, as assayed by the everted gut-sac technique, was increased in ultimobranchialectomized frogs at sunrise, concomitant with acute hypercalcaemia and hypercalciuria. An opposite but chronic response was observed in parathyroidectomized frogs with intact ultimobranchial glands. The maximum response observed at sunrise occurred when the concentration of calcium in the plasma of control frogs was decreasing; the minimum response, which occurred 6 h after sunrise, was coincident with a diurnal peak in the concentration of calcium. Vitamin D3 (500 microgram/frog) enhanced calcium transport in ultimobranchialectomized frogs, which resulted in chronic hypercalcaemia and hypercalciuria. The results suggest that diurnal variations in the plasma concentration of calcium do not initiate ultimobranchial activity, but are a response to endocrine control synchronized with the transition from darkness to light.


Hypothalamic corticotrophin releasing factor activity in dogs with pituitary-dependent hyperadrenocorticism. Hypothalamic corticotrophin releasing factor (CRF) activity was determined in five dogs with spontaneous hyperadrenocorticism and in three control animals (one untreated, one treated with high doses of ACTH for 2 months and one treated with high doses of cortisone for 2 months). Hypothalamic CRF activity was low or undetectable in four dogs with Cushing's syndrome due to an adrenocortical tumour. The results are compatible with a pituitary origin for pituitary-dependent hyperadrenocorticism in the dog but are not conclusive; direct information about the rates of hypothalamic CRF secretion is required.


The regulation of the calcium sensitivity of the contractile system in mammalian cardiac muscle. Treatment of rat ventricular cells with 10 mM EGTA makes the sarcolemma highly permeable to small ions and molecules without removing its restriction of the diffusion of larger molecules or inactivating all of its enzymatic functions. These hyperpermeable cardiac cells have been used to study the regulation of the range of concentration of Ca over which activation of the contractile proteins occurs (Ca sensitivity). The Ca sensitivity can varied from three- to sixfold without any significant alteration in the general shape of the relation between force and Ca concentrations. Although cyclic nucleotides in concentrations of 10(-9) to 10(-5) M do not influence Ca sensitivity, in the presence of a phosphodiesterase inhibitor, cGMP increases and cAMP decreases Ca sensitivity. Treatment of the hyperpermeable cells with a nonionic detergent raises Ca sensitivity as does removal of the phosphate donor by complete substitution of CTP for ATP. These data indicate that Ca sensitivity is probably modulated by a cAMP-dependent phosphorylation that decreases Ca sensitivity. The sarcolemma is required for this reaction to take place. The effect of this reaction is antagonized by a cGMP-dependent reaction occurring inside the cell. Studies involving the perfusion of the heart with and without epinephrine before the exposure to EGTA indicate that epinephrine can regulate this system of control of Ca sensitivity. The functional considerations of this regulatory system are discussed.


The effect of pneumonia induced in mice with Mycoplasma pulmonis on resistance to subsequent bacterial infection and the effect of a respiratory infection with Sendai virus on the resistance of mice to Mycoplasma pulmonis. The effect of pneumonia induced by Mycoplasma pulmonis in mice on the resistance of the lung to additional bacterial infection was examined. The effect of pneumonia induced by Sendai virus on the resistance of mice to M. pulmonis was also investigated and compared with the effect of Sendai virus on resistance to Staphylococcus aureus. Sendai virus infection decreased subsequent resistance to M. pulmonis in proportion to the virus dose. Decreased resistance to subsequent S. aureus and M. pulmonis infection was greatest at about the same time after inoculation of virus and was related to virus-induced lesions. Besides affecting the resistance of mice to subsequent mycoplasma infection, Sendai virus could enhance an existing mycoplasma infection. Pneumonia induced by M. pulmonis did not decrease resistance to subsequent bacterial infection. The mechanism whereby Sendai virus decreases host resistance is therefore similar for bacteria and mycoplasmas, but pneumonia induced by mycoplasmas does not have the same effect.


Ultrastructural development of guinea pig cytomegalovirus in cultured guinea pig embryo cells. The ultrastructural development of guinea pig cytomegalovirus (GPCMV) in guinea pig embryo cells was studied using electron microscopy. Tubular structures were found in nuclei of virus infected cells, followed by the appearance of intranuclear inclusions containing virus nucleocapsids. While some nucleocapsids were enveloped at the inner nuclear membrane, others were released into the cytoplasm where they were associated with, or within, dense matrix which was subsequently enveloped by cytoplasmic membranes to form enveloped dense virions. Dense bodies without virus capsids were formed in the cytoplasm and enveloped in a similar manner. An involvement of the nuclear pores in the release of unenveloped virus capsids from the nucleus to the cytoplasm was postulated. Evidence that the enveloped dense virions and dense bodies shared common envelope antigen(s) was obtained by immunoelectron microscopy. The similarities and differences in the ultrastructural development of GPCMV and other cytomegaloviruses are discussed.


Pseudotypes of vesicular stomatitis virus and Pichinde virus. Super-infection of Pichinde virus-infected cells with vesicular stomatitis virus (VSV) resulted in the production of pseudotype virus which was not neutralized by antiserum to VSV but which was neutralized by antiserum to Pichinde virus. Analysis of pseudotype virus production in relation to the kinetics of replication of Pichinde virus demonstrated that pseudotype virus production occurred when super-infection with VSV was initiated 8 h or more after infecting the cells with Pichinde virus. The quantities of pseudotype virus produced correlated with the quantities of Pichinde virus antigen detected on the surface of the cells both during acute infection and in cells chronically infected with Pichinde virus. The observations indicate that pseudotype of VSV and Pichinde virus are readily formed and that the formation of pseudotype virus may be used to examine the Pichinde virus antigens expressed on the surface of infected cells.


Characterization of human TK- cell lines transformed to a TK+ phenotype by herpes simplex virus type 2 DNA. Human TK- cells carrying the HSV-2 TK gene as a result of transformation with virus DNA express a TK activity of virus origin and maintain the TK+ phenotype when grown in HAT medium. Under non-selective conditions, however, reversion to a TK- phenotype occurs with a significant frequency characteristic of each transformed line. Once reversion has occurred the TK- phenotype appears to be stable, since only very rare instances of TK- to TK+ reversion have been observed. TK- revertants were susceptible to re-transformation by virus DNA, but no reactivation of a silent virus TK gene could be obtained by superinfecting them with a TK- virus mutant. The data presented are consistent with the hypothesis that acquisition of the TK- phenotype is brought about by loss of the virus sequences coding for TK.


Murine cytomegalovirus-induced protein synthesis. Murine cytomegalovirus (MCMV)-induced protein synthesis in mouse embryo fibroblast (MEF) cells was studied using polyacrylamide gradient SDS gel electrophoresis and autoradiography. Synthesis of at least 14 virus induced proteins (VIPs) was consistently detected in a lytic cycle. They were designated VIPs 132, 118, 99, 98, 88, 81, 76, 74, 58, 56, 51, 38, 36 and 33 on the basis of their mol. wt. Judging from the pattern of the rate of protein synthesis, VIPs can be classified into three groups: group A VIPs were synthesized actively for a brief period of time and then their synthesis was no longer detectable. This group included two major VIPs, 98 and 88 and three minor VIPs, 58, 56 and 38. Group B VIPs 81, 74, 36 and 33 were similar to group A except that, following a brief period of active synthesis, a low level of synthesis continued during the entire lytic cycle. Group C VIPs 132, 118, 99, 76 and 51 were synthesized at low steady levels at all times after initiation and seemed to accumulate slowly. According to temporal sequences of initiation of VIP synthesis, these proteins can also be divided into three groups: immediate early, early and late VIPs. The synthesis of the immediate early VIPs 132, 98, 88, 81, 76, 74 and 38 was initiated immediately after virus infection. The early VIPs included 58, 56, 51, 36 and 33 and their synthesis was initiated from 1 to 3 h post-infection. VIPs 118, 99 and several minor VIPs were first synthesized during 12 to 13 h post-infection which corresponded to the time of initiation of virus DNA synthesis and they are classified as late VIPs. Cycloheximide reversal experiments indicated that the initiation of synthesis of early VIPs must be preceded by the synthesis of immediate early VIPs. In the presence of actinomycin D, the immediate early VIPs (0 to 1 h post-infection) were not synthesized indicating that immediate early VIPs are translated from virus mRNA synthesized after virus infection.


Varicella-zoster virus immunizes patas monkeys against simian varicella-like disease. To define further the antigenic relationship between human varicella-zoster virus and herpesviruses which produce varicella-like disease in certain simian species, patas monkeys were inoculated with varicella-zoster virus and then challenged with Delta herpesvirus, which uniformly produces severe, clinically apparent disease in susceptible animals. Protection against Delta herpesvirus was conferred both by hyperimmunization with varicella-zoster virus and by a single immunization with a cell-free preparation of varicella-zoster virus. Although the immunological relationship between the human and simian varicella viruses is not completely reciprocal, these studies confirmed that antigens which induce immunity are shared by the human and simian viruses. No clinical symptoms were seen in monkeys inoculated with varicella-zoster virus, but the rapid and marked antibody responses to the virus suggested that subclinical infection had occurred. In contrast, a chimpanzee inoculated with one of the same varicella-zoster virus preparations produced only low levels of antibody.


Concentration of human cytomegalovirus from large volumes of tissue culture fluids. Three methods of pelleting, ultracentrifugation (95000 g for 60 min), precipitation with polyethylene glycol 6000 (5% v/v), and with ammonium sulphate (38% w/v), were used to concentrate human cytomegalovirus (CMV) from tissue culture fluids. Maximum recovery of infectious virus particles was obtained with the polyethylene glycol (PEG) method. The precipitating activity of PEG 6000 and PEG 20000 was then compared at different concentrations. The best results were obtained with PEG 6000 at a final concentration of 5% (v/v). Changes in pH or salt concentration, treatment of the concentrates with Pronase and long periods of time at 4 degrees C significantly reduced the number of biologically active CMV particles recovered by PEG precipitation.


The kinetics of synthesis of infectious unintegrated virus DNA in rous sarcoma virus infected chicken cells. Infectious virus nucleic acids were extracted from the Hirt supernatant of chicken embryo fibroblasts infected for different times with the Schmidt-Ruppin strain of Rous sarcoma virus (RSV). Infectious DNA and DNA-RNA hybrid molecules could be recovered from 6 h after infection in experiments using 1.5 X 10(9) infected cells. Only small amounts of infectious virus DNA could be purified 6 h after infection whereas at 24 h approximately one infectious DNA molecule could be recovered for each input virus infectious unit. At 24 h, both infectious supercoiled and non-supercoiled molecules were found. The specific infectivity of the supercoiled fraction was less than that of the non-supercoiled fraction. Infectious supercoiled DNA could be recovered from 16 h after infection. Evidence is presented that both forms of unintegrated virus DNA may rest unintegrated for at least 8 days in the cell, though chronically infected cells were shown to contain less than one unintegrated molecule per 10(2) to 10(3) cells.


Rapid purification of lymphocytic choriomeningitis virus by density gradient centrifugation in colloidal silica. Lymphocytic choriomeningitis virus was purified from cell culture fluid by density gradient centrifugation in colloidal silica. The specific infectivity increased 5000-fold and the recovery of infectivity was about 10%.


Detection of polyoma virus DNA in PML-brain tissue by (in situ) hybridization. The human papova (JC) virus was extracted from brain of a patient with progressive multifocal leukoencephalopathy. A single band of virus was obtained at a density of 1.345 g/ml CsCl.JC virus DNA was purified and a highly specific cRNA was generated in vitro. In situ hybridization with JC virus cRNA and autoradiography on sections of the same brain revealed silver grains over oligodendrocytes, astrocytes and possible vascular endothelial cells, indicating the presence of JC virus DNA in these different cell classes.


Prevalence of antibody to the Norwalk agent by a newly developed immune adherence hemagglutination assay. An immune adherence hemagglutination assay (IAHA) for the detection of antibody to the Norwalk agent of acute epidemic nonbacterial gastroenteritis was developed using as antigen virus purified from stool from an experimentally infected volunteer. The assay was sensitive and specific and was efficient for detecting Norwalk antibody seroresponses. The prevalence of Norwalk antibody in various groups in the United States was studied. Antibody to the Norwalk agent was acquired gradually, beginning slowly in childhood and accelerating in the adult period so that by the fifth decade 50% possessed antibody. This pattern of antibody acquisition contrasted sharply with that for the human rotavirus of infantile gastroenteritis. Rotavirus antibody was acquired during early childhood by almost all individuals in the pediatric groups studied. Antibody to the Norwalk agent was also found in rural Bangladesh; in a small prevalence survey of 39 children and adults 21% possessed Norwalk IAHA antibody, whereas 95% possessed antibody to the human rotavirus.


Prevalence of antibody to the hepatitis A virus in a large institution for the mentally retarded. The prevalence of antibody to hepatitis A virus (anti-HAV) was determined in 854 people living in a large institution for the mentally retarded in Melbourne. Altogether 638 (74.4%) of the subjects were found to have specific antibody detectable by solid-phase radioimmunoassay (SPRIA). No difference in antibody prevalence was observed between males and females; however, the prevalence was higher among subjects with Down syndrome (84.1%) than those of other forms of mental retardation (72.8%). In both groups there was an increase in prevalence of antibody with increasing age and duration of institutionalization. When compared with the open community, patients in this institution have a higher prevalence of antibody and appear to acquire their infections at an earlier age.


Diagnosis of human coronavirus infection by immunofluorescence: method and application to respiratory disease in hospitalized children. Rabbit antisera were prepared against coronavirus strains 229E and OC43 and used successfully to detect viral antigen in epithelial cells shed from the nasopharynx of symptomatic volunteers who had received coronavirus inocula three to four days before. The same serologic reagents were applied to nasopharyngeal secretion cells obtained from 106 infants and children hospitalized with respiratory tract disease and apparently not infected with conventional respiratory viruses. No coronavirus infections were detected by this method. It appears that coronavirus OC43 or 229E infections were not common in children in Tyneside hospitals during the period of study. However, fluorescence is a useful method for detection of coronavirus infections in symptomatic human subjects.


Infection of human amnion cells with cytomegalovirus. Human cytomegalovirus (CMV), known to replicate in vitro in human fibroblastic cells, was found to replicate in epithelial human amnion (HA) cells. Large syncytia formed in these cells after infection with CMV; inclusion bodies were observed in the nuclei, and CMV antigens were demonstrated in both the cytoplasm and the nucleus by indirect immunofluorescence techniques. The synthesis of virus DNA was also detected, and the production of infectious virus was followed. The titers were lower (from 10(4) to 6 X 10(5) using different isolates of CMV) than those obtained in human embryo fibroblast (HEF) cells, and the replication cycle was slower in HA than in HEF cells.


Developments in viral hepatitis.

Developments in viral hepatitis. Viral hepatitis is a major public health problem occurring endemically in all parts of the world. The general term viral hepatitis refers to infections caused by hepatitis virus type A, type B and a more recently identified infection referred to as "non-A: non-B" hepatitis. These clinically and pathologically similar forms of hepatitis have been studied intensively following the discovery of a specific antigen, Australia antigen, one of the markers of infection with hepatitis B virus.