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Analysis of parapoxvirus genomes.

Eight stomatitis papulosa (SP), four orf and two milker's nodes (MN) virus isolates were compared by restriction enzyme analysis. Considerable genetic heterogeneity was found not only between isolates belonging to the three different taxonomic groups but also between members of the same group. This heterogeneity precludes classification of parapoxviruses simply by comparison of their DNA cleavage patterns. Restriction maps were therefore prepared for 12 parapoxvirus DNAs. Fragments from defined regions of the genome were then selected and used as probes for cross-hybridizations to all other parapoxvirus DNAs. DNA fragments derived from an internal region of the genome hybridized strongly to all parapoxvirus isolates examined. In contrast, cross-hybridization of the end region of the DNA molecule was observed only between members of the same virus group. Molecular hybridization as a means of classifying parapoxvirus isolates is discussed.

DNA Restriction Enzymes↗

Adventures with poxviruses of vertebrates.

Because they were the largest of all viruses and could be visualised with a light microscope, the poxviruses were the first viruses to be intensively studied in the laboratory. It was clear from an early date that they caused important diseases of humans and their domestic animals, such as smallpox, cowpox, camelpox, sheeppox, fowlpox and goatpox. This essay recounts some of the early history of their recognition and classification and then expands on aspects of research on poxviruses in which the author has been involved. Studies on the best-known genus, Orthopoxvirus, relate to the use of infectious ectromelia of mice as a model for smallpox, embracing both experimental epidemiology and pathogenesis, studies on the genetics of vaccinia virus and the problem of non-genetic reactivation (previously termed 'transformation') and the campaign for the global eradication of smallpox. The other group of poxviruses described here, the genus Leporipoxvirus, came to prominence when the myxoma virus was used for the biological control of Australian wild rabbits. This provided a unique natural experiment on the coevolution of a virus and its host. Future research will include further studies of the many immunomodulatory genes found in all poxviruses of vertebrates, since these provide clues about the workings of the immune system and how viruses have evolved to evade it. Some of the many recombinant poxvirus constructs currently being studied may come into use as vaccines or for immunocontraception. A field that warrants study but will probably remain neglected is the natural history of skunkpox, raccoonpox, taterapox, yabapox, tanapox and other little-known poxviruses. A dismal prospect is the possible use of smallpox virus for bioterrorism.

Animals↗

Strain-specific elements in influenza antigens.

Rabbit antisera were prepared against ten antigenically different influenza A strains. These sera were absorbed with one or more heterologous strains and in each case all the heterologous or crossing antibody was removed; the anti-bodies remaining after this treatment were specific for the immunizing strain or group of strains. On the basis of reactions with absorbed sera, the strains fell into seven groups. Absorbed specific antisera of these groups were used to test the HI titer against a large number of influenza A viruses. Most of the strains were inhibited by a single serum, a few were inhibited by none of the sera, and only one strain was inhibited by two antisera. The grouping of strains by this method was less equivocal than classifications based on previous tests. When more fully developed, this technique promises to be of interest and assistance in the study of influenza, especially from the epidemiological and prophylactic standpoints.

Animals↗

Disappearance of anti-p17 correlates with successful isolation of human immunodeficiency virus and deterioration in clinical status.

Anti-p17 was examined as a human immunodeficiency virus (HIV) marker predicting the onset of the acquired immunodeficiency syndrome (AIDS). Two different comparisons were done: (1) anti-p17 negativity and successful HIV isolation; and (2) anti-p17 negativity and clinical status, using the Centers for Disease Control classification. Anti-p17 negativity was not only significantly correlated with deterioration in clinical status (P less than 0.01), but also with successful HIV isolation (P less than 0.05). HIV isolation was affected by several drugs, e.g. zidovudine. However, the results of the antibody test were not affected. It is significant that anti-p17 can compensate for the defects of virus isolation.

Acquired Immunodeficiency Syndrome↗

Detection and genotyping of bovine diarrhea virus by reverse transcription-polymerase chain amplification of the 5' untranslated region.

A reverse-transcription polymerase chain reaction (RT-PCR) was developed to differentiate the bovine diarrhea virus (BVDV) from other pestiviruses, and to determine the genotype of the BVDV isolates. For this purpose, primer pairs were selected in the 5' untranslated region (5'UTR). The primers BE and B2 were located in highly conserved regions and were pestivirus-specific. Two primer pairs named B3B4 and B5B6 were specific of BVDV genotypes I and II, respectively. With this technique, an amplification product of the expected size was obtained with either the B3B4 or the B5B6 primer pairs for the 107 BVDV isolates tested but not for BDV or CSFV. For some isolates that were grouped in the genotype II, sequence analysis of the PCR fragments confirmed their classification into this genotype.

5' Untranslated Regions↗

The natural history of chronic hepatitis B infection.

Chronic hepatitis B infection is a global health problem that affects about 300 million people. Of these, 75% are Chinese. Most Chinese who become chronic carriers, contract the virus during the perinatal period. The natural history of these chronic hepatitis B carriers includes an initial immune tolerance phase, followed by immune clearance and an inactive hepatitis B non-replicative phase with the development of cirrhosis that may be complicated by hepatocellular carcinoma. The classification of hepato-cellular carcinoma has recently been revised. Based on immunohistochemical studies, it has been found that patients with hepatocellular carcinoma and biliary markers have a poorer survival than patients with hepatocellular carcinoma but who have negative biliary markers. Sometimes, a fourth phase, a hepatitis B envelope-negative hepatitis B virus replicative phase, reflecting the emergence of a pre-core mutant strain, may follow. Our improved understanding of the natural history of chronic hepatitis B infection has led to more effective approaches towards the control of this viral infection and its sequelae. Most importantly, immunisation against hepatitis B virus in the perinatal setting has been shown to prevent chronic infection.

Journal Article↗

Sequence of the structural proteins of tick-borne encephalitis virus (western subtype) and comparative analysis with other flaviviruses.

Tick-borne encephalitis (TBE) virus (Western subtype strain Neudoerfl) was cloned and the sequence of 2450 nucleotides of the 5'-terminal region of the genome was determined. By amino acid sequencing and sequence comparisons with other flaviviruses the amino-termini of the structural proteins and protein NS1 were localized. Sequence homologies with other flaviviruses were determined and corresponded well to the established serological classification system of the Flavivirus family. N-Glycosylation sites were found to be conserved to a large extent among members of the same serological subgroup, but not between members of different subgroups. Hydrophilicity plots and sequence comparisons revealed that the TBE virus capsid protein exhibited features distinct from all other flaviviruses. Additionally, the capsid protein coding region of another natural isolate of TBE virus (strain ZZ-9) was sequenced in order to analyze why the capsid protein of this strain exhibited a significantly faster migration rate on SDS-polyacrylamide gels than other TBE virus strains.

Amino Acid Sequence↗

The epidemiology of Hodgkin's disease.

Much of the epidemiological heterogeneity of HD incidence reflects the behaviour of the NS subtype, at least in the USA. Incidence variation across races (except Asians) and time periods is most marked in this subtype. In young adults with HD, there is compelling evidence for social class modification of risk consistent with an infectious aetiology; limited data suggest that this effect occurs within the NS subtype, but considerable evidence indicates that it does not primarily involve EBV infection. Findings from familial aggregation studies and HLA associations point to inherited susceptibility to this subtype. Despite little sex difference for NS in young adulthood in the latest incidence data, parity nevertheless appears to be protective against this subtype for women. Therefore, the greater increase for females than males in the incidence of young-adult NS in recent years may reflect the impact of population trends towards later childbearing and lower parity. This change, as well as the concomitant smaller family sizes and growing affluence, could explain part of the burgeoning incidence of NS in young adults in the USA. These observations suggest that NS in young adults constitutes a separate disease, probably of infectious origin. The incongruous occurrence of this subtype in older adults, and the presence of EBV in some NS cases, could reflect heterogeneity within NS, for example, representing features of the cellular phase of NS (Cozen et al, 1992; Medeiros and Greiner, 1995). For the non-NS subtypes, many of the factors that predict risk of NS may also be relevant. Patterns of social class determinants in children and older adults, the age groups at risk for MC, support involvement of an infectious precursor given intense exposure, and EBV is a likely candidate, based on its high prevalence in these groups. However, little aetiological research has been directed explicitly at the non-NS subtypes. Considerable effort has gone into exploring an infectious aetiology of HD. Recently, this line of investigation has moved beyond social class determinants to molecular epidemiological studies of EBV and, to a lesser degree, other potentially involved viruses. The roles of genetic susceptibility and sex hormones also represent promising areas for exploration, particularly in their possible interaction with infectious agents and other environmental factors. Ultimately, clearer epidemiological understanding of HD will be aided by more precise classification of this disease at the molecular level.

Adult↗

The relationship between upper respiratory infections and hospital admissions for asthma: a time-trend analysis.

We have shown that viruses are associated with 80 to 85% of asthma exacerbations in school-age children in the community. We hypothesize that viral infections are also associated with severe attacks of asthma precipitating hospital admissions. To investigate this, we conducted a time-trend analysis, comparing the seasonal patterns of respiratory infections and hospital admissions for asthma in adults and children. During a 1-yr study in the Southampton area of the United Kingdom, 108 school-age children monitored upper and lower respiratory symptoms and took peak expiratory flow rate (PEFR) recordings. From children reporting a symptomatic episode or a decrease in PEFR, samples were taken for detection of viruses and atypical bacteria. A total of 232 respiratory viruses and four atypical bacteria were detected. The half-monthly rates of upper respiratory infection were compared with the half-monthly rates for hospital admissions for asthma (International Classification of Diseases [ICD] code 493) for the same time period for the hospitals serving the areas from which the cohort of schoolchildren was drawn. The relationships of upper respiratory infections and hospital admissions for asthma with school attendance were studied. Strong correlations were found between the seasonal patterns of upper respiratory infections and hospital admissions for asthma (r = 0.72; p < 0.0001). This relationship was stronger for pediatric (r = 0.68; p < 0.0001) than for adult admissions (r = 0.53; p < 0.01). Upper respiratory infections and admissions for asthma were more frequent during periods of school attendance (87% of pediatric and 84% of total admissions), than during school holiday periods (p < 0.001). These relationships remained significant when allowance was made for linear trend and seasonal variation using multiple regression analysis (p < 0.01). Not surprisingly, school attendance, because it is a major factor in respiratory virus transmission, was found to be a major confounding variable in children. This study demonstrates that upper respiratory viral infections are strongly associated in time with hospital admissions for asthma in children and adults. Rhinoviruses were the major pathogen implicated, and the majority of viral infections and asthma admissions occurred during school attendance.

Adolescent↗

Genetic heterogeneity of bovine viral diarrhoea virus in Italy.

The genetic characteristics, of 38 field isolates of bovine viral diarrhoea virus (BVDV) collected in 1999 from sick or healthy and persistently infected cattle of dairy farms situated in northern Italy, were investigated. A partial 5'-untranslated region (5'-UTR) sequence of each isolate was determined and a phylogenetic analysis was performed. All the isolates were classified as belonging to the BVDV-1 genotype and could be assigned to different BVDV-1 groups, namely BVDV-1b (n = 20), BVDV-1d (n = 6) and BVDV-1e (n = 10). Two remaining isolates could be classified as BVDV-1f and BVDV-1h, respectively. These results provided evidence for genetic heterogeneity of BVDV in Italy, and contribute to a better knowledge of the circulation of BVDV strains, and to their classification.

5' Untranslated Regions↗

Immunity to Haemophilus influenzae type b polysaccharide capsule after vaccination with the complete series of oligosaccharide CRM197 conjugate vaccine in infants with human immunodeficiency virus infection.

We evaluated immunity to Haemophilus influenzae type b (Hib) in 18 human immunodeficiency virus-infected infants who were vaccinated with a complete series of Hib conjugate vaccine. Four months after the primary series, the geometric mean anticapsular antibody concentration in 11 children was 0.40 microgram/ml. There were no significant differences in CD4+ cell counts or in the Centers for Disease Control and Prevention disease classification according to the presence of immunity to Hib. Four months after the booster dose, the geometric mean anticapsular antibody concentration in the 18 children was 0.82 microgram/ml. Children with immunity were more likely than children lacking immunity to have higher CD4+ cell counts and mild human immunodeficiency virus-related disease. The majority of the anticapsular antibody concentrations were lower than in healthy children.

Antibodies, Bacterial↗

Pathogenesis and classification of primary central nervous system lymphoma: an update.

Primary central nervous system lymphoma has undergone a remarkable increase in incidence over the last decade, both in immunosuppressed and immunocompetent individuals. Its clinicopathologic evaluation requires knowledge of current hematopathologic systems for lymphoma classification, as well as specific understanding of unique central nervous system determinants. In immunocompromised individuals, the tumor has a constant association with Epstein-Barr virus. The form of Epstein-Barr virus in these tumors appears to be predominantly latent, however, the precise mechanism relating virus to tumor pathogenesis is still unclear. In immunocompetent patients, risk factors and oncogenic associations are completely unknown. Critical to the formation of these tumors is the trafficking of B-cells, whether pre- or post-transformation, across the blood-brain barrier. These B-cell migrations may require perturbations to the barrier that originate in or around neural parenchyma, such as localized injury or infection.

Blood-Brain Barrier↗

Beta 2-microglobulin and neopterin: predictive markers for human immunodeficiency virus type 1 infection in children?

The value of beta 2-microglobulin and neopterin concentrations in serum for early diagnosis of infants born to human immunodeficiency virus type 1 (HIV-1)-infected mothers was assessed. Concentrations of both markers were measured in serum samples from pediatric patients (Centers for Disease Control classifications P0, P1, and P2), as well as in age-matched normal subjects. Both beta 2-microglobulin and neopterin were significantly increased in HIV-1-infected symptomatic subjects (P2) compared to controls. Seventy-five percent of asymptomatic patients (P1) also had increased values. On the other hand, a significant overlap in concentrations of both markers in serum was found between controls and P0 patients. Thirty-eight percent of the P0 patients had values comparable to those of the P2 group. Persistently high concentrations of both markers in P0 patients may be indicative of HIV-1 infection.

Biomarkers↗

Outcome of untreated infection with Entamoeba histolytica in homosexual men with and without HIV antibody.

Among homosexual men the prevalence of infection with Entamoeba histolytica is high. To determine the clinical importance of this infection 55 homosexual men carrying the parasite were investigated in detail. No clinical, serological, or histological evidence of invasive amoebiasis was found in any of them. The patients were not treated and were followed up for 12 to 29 months (mean 21.6 months), during which period none developed symptoms that could be attributed to E histolytica. Spontaneous loss of the parasite occurred in 17 patients, some of whom later became reinfected. Sixteen patients had antibody to human immunodeficiency virus, and infection with E histolytica showed the same benign course in them as in the patients who did not have antibody. Throughout the study classification of the isolates of E histolytica consistently showed that they belonged only to non-pathogenic zymodemes. The findings provide further evidence that E histolytica in homosexual men is a commensal organism.

Animals↗

High prevalence of high risk human papillomavirus-capsid antibodies in human immunodeficiency virus-seropositive men: a serological study.

BACKGROUND: Serological study of human papillomavirus (HPV)-antibodies in order to estimate the HPV-prevalence as risk factor for the development of HPV-associated malignancies in human immunodeficiency virus (HIV)-positive men. METHODS: Sera from 168 HIV-positive men and 330 HIV-negative individuals (including 198 controls) were tested using a direct HPV-ELISA specific to HPV-6, -11, -16, -18, -31 and bovine PV-1 L1-virus-like particles. Serological results were correlated with the presence of HPV-associated lesions, the history of other sexually transmitted diseases (STD) and HIV classification groups. RESULTS: In HIV-negative men low risk HPV-antibodies were prevailing and associated with condylomatous warts (25.4%). Strikingly, HIV-positive men were more likely to have antibodies to the high-risk HPV types -16, -18, -31, and low risk antibodies were not increased in a comparable range. Even those HIV-positive heterosexual individuals without any HPV-associated lesions exhibited preferentially antibody responses to the oncogenic HPV-types (cumulative 31.1%). The highest antibody detection rate (88,8%) was observed within the subgroup of nine HIV-positive homosexual men with anogenital warts. Three HIV-positive patients had HPV-associated carcinomas, in all of them HPV-16 antibodies were detected. Drug use and mean CD4-cell counts on the day of serologic testing had no influence on HPV-IgG antibody prevalence, as had prior antiretroviral therapy or clinical category of HIV-disease. CONCLUSION: High risk HPV-antibodies in HIV-infected and homosexual men suggest a continuous exposure to HPV-proteins throughout the course of their HIV infection, reflecting the known increased risk for anogenital malignancies in these populations. The extensive increase of high risk antibodies (compared to low risk antibodies) in HIV-positive patients cannot be explained by differences in exposure history alone, but suggests defects of the immunological control of oncogenic HPV-types. HPV-serology is economic and can detect past or present HPV-infection, independently of an anatomical region. Therefore HPV-serology could help to better understand the natural history of anogenital HPV-infection in HIV-positive men in the era of antiretroviral therapy.

Antibodies, Viral↗

Regional blood flow in avian sarcoma virus (ASV)-induced brain tumors.

Regional blood flow (F) was measured in avian sarcoma virus (ASV)-induced brain tumors in rats. Blood flow was variable in individual as well as different tumors; tumor F did not correlate with histologic classification, tumor size, central versus peripheral tumor regions, intraparenchymal location, cell density, or specific cytologic characteristics. Low values of F did correlate with tumor necrosis and hydrocephalus; high values of F correlated with tumorous invasion or association with choroid plexus. Mean tumor F was not significantly different from that of the same anatomic, tumor-free brain region of the contralateral hemisphere, but F in brain tissue adjacent to the tumor was depressed significantly. Depression of F was observed in tumor-free cortex and corpus callosum, especially in the hemisphere ipsilateral to the main tumor mass and in those animals with hydrocephalus.

Animals↗

Distribution of HCV genotypes among different exposure categories in Brazil.

Hepatitis C virus (HCV) infection is widespread and responsible for more than 60% of chronic hepatitis cases. HCV presents a genetic variability which has led to viral classification into at least 6 genotypes and a series of subtypes. These variants present characteristic geographical distribution, but their association with different responses to treatment with interferon and severity of disease still remains controversial. The aim of this study was to investigate the patterns of distribution of HCV genotypes among different exposure categories in Brazil. Two hundred and fifty anti-HCV positive samples were submitted to HCV-RNA detection by RT-PCR and their genotype was determined by restriction fragment length polymorphism (RFLP) analysis. In addition, the genotype/subtype of 60 samples was also determined by a reverse hybridization assay. HCV 1 was the most prevalent (72.0%), followed by type 3 (25.3%), HCV 2 (2.0%) and HCV 4 (0.7%). The HCV genotype distribution varied among the different exposure categories, with HCV 1 being more frequent among blood donors, hemophiliacs and hemodialysis patients. A high frequency of HCV 3 was observed in cirrhotic patients, blood donors from the South of Brazil and injecting drug users (IDUs). The general distribution of the HCV genotype in Brazil is similar to that in other regions of the world.

Blood Donors↗

The epidemiology of non-Hodgkin's lymphoma: why the increased incidence?

The incidence of non-Hodgkin's lymphoma continues to increase in the United States. Improvements in diagnostic techniques, changes in disease classifications, and the increase in AIDS-related lymphomas account for only a small percentage of the increase, leaving most of the increased incidence unexplained. Viruses clearly play a major role in the pathogenesis of some subtypes of non-Hodgkin's lymphoma, especially Epstein-Barr virus and HTLV-1, and improvements in molecular diagnostic techniques may identify additional viruses that have a role in the pathogenesis of lymphomas. Immunodeficiencies, whether acquired, congenital, or iatrogenic, clearly predispose to the development of non-Hodgkin's lymphoma. Numerous environmental exposures have also been linked to an increased risk of non-Hodgkin's lymphoma and may account for a portion of the continuing increased incidence of the disease.

Acquired Immunodeficiency Syndrome↗