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Evaluation of nitric oxide (NO) levels in hepatitis C virus (HCV) infection: relationship to schistosomiasis and liver cirrhosis among Egyptian patients.

Nitric oxide (NO), a recently discovered free radical, is overproduced in liver cirrhosis. Hepatitis C virus (HCV) might increase NO levels via increased inducible NO synthase (iNOS). This work was carried out to study the effect of HCV-induced liver cirrhosis on NO levels among Egyptian patients. The study included 46 patients with liver cirrhosis, and 30 healthy individuals of matched age and sex. NO levels determined as the stable endproduct nitrate, showed a statistically significant increase among patients compared to the control group (P < 0.001). Furthermore, NO levels increased proportionally with the severity of liver cirrhosis as assessed by Child's classification (P < 0.05). Moreover, schistosomial infection enhanced NO levels in cirrhotic patients with HCV infection compared to non-bilharzial patients (P < 0.001). Polymerase chain reaction (PCR) and branched DNA assays were used for detection of HCV RNA positivity, and measurement of the virus load, respectively. Both showed a positive correlation with the NO levels (P < 0.001). At a nitrate cutoff value of 70 micromol/L, the sensitivity and specificity were 83.0% and 73.0%, respectively. Chi square analysis showed a significant correlation between ALT levels and both HCV RNA positivity by polymerase chain reaction (PCR) (P < 0.02), and virus load (P<0.05). Interestingly enough, there was a significant positive correlation between HCV RNA and schistosomal antibody titer as measured by hemaglutination inhibition assay (HAI) (P < 0.05). The data presented in this report indicated an association between NO levels and the development and progression of liver cirrhosis. Furthermore, the findings obtained from this study demonstrated that schistomiasis is an important risk factor involved in enhancement of NO levels and virus replication. The latter may aggravate liver cell injury and hence the development of cirrhosis.

Egypt↗

Interferon treatment of chronic hepatitis C in patients with hemophilia or von Willebrand's disease in Japan.

Seven patients with chronic hepatitis C, six hemophiliacs and a patient with von Willebrand's disease, were treated with interferon-alpha (IFN-alpha). Either 9 MU of recombinant IFN-alpha 2a or 3 MU of lymphoblastoid alpha-IFN was administered daily for 2 weeks and then three times a week for 22 weeks. Liver histology, hepatitis C virus (HCV) genotypes, and HCV-RNA levels in sera were investigated in all of the patients before IFN therapy was instituted. Liver histology was classified by the European classification. HCV genotyping conformed to the so-called Okamoto's classification. HCV-RNA levels in sera were quantitated by competitive polymerase chain reaction, using mutant RNA. Liver histology, HCV genotype, and serum HCV-RNA level (copies/ml) in each patient were: patient 1, chronic persistent hepatitis, type II, 3 x 10(3) respectively; patient 2, chronic active hepatitis (CAH) 2a, type III, 6 x 10(4); patient 3, CAH2a, type IV, 2 x 10(5); patient 4, CAH2b, type I, 2 x 10(7); patient 5, CAH2b, type II, 8 x 10(4); patient 6, CAH2b, type III, 7 x 10(6); and patient 7, CAH2b, type IV, 1 x 10(7). Sustained elimination of HCV was achieved in patient 3 and temporary elimination was achieved in patients 1 and 2. The other patients showed persistent HCV-RNA positivity in sera both during and after IFN treatment. Poor responsiveness to IFN was observed in patients with relatively progressive liver histology and high levels of HCV viremia.

Adult↗

Molecular diversity of HIV in Albania.

Little information is available on circulating human immunodeficiency virus (HIV) subtypes and resistance to antiretroviral drugs in Albania. To fill this gap, we studied 72 plasma samples from HIV-infected individuals from throughout the country. Subtype classification and genotypic resistance analysis were performed on the HIV pol gene region. The analysis was successfully performed on 66 (91.6%) plasma samples and showed that 43 (65.2%) strains were non-B subtypes (mostly subtype A, as determined by analysis of pol gene sequences). No major mutations in the protease gene were found, whereas analysis of the reverse transcriptase gene revealed a few major mutations associated with resistance. In conclusion, non-B subtypes are predominant in Albania, and the prevalence of resistance to antiretroviral drugs is still low.

Albania↗

Lymphoproliferative responses to mitogen and antigen in HIV-infected children.

We endeavored to study lymphoproliferative responses in children with human immunodeficiency virus (HIV) infection and to compare them with normal control children. Children were grouped according to age; 6-18 months and greater than 18 months, and according to CDC classification: asymptomatic (P1), mildly symptomatic (P2A), and advanced symptoms (P2D). Absolute CD4 and CD8 numbers were compared and found to be higher in the younger age groups. The children in P1 and P2A classes demonstrated an increase in CD8+ cells; only the children with AIDS showed a significant decrease in CD4+ cells. Lymphoproliferative responses to phytohemagglutinin A (PHA) were compared to tetanus toxoid. Only the children with acquired immunodeficiency syndrome (AIDS) (P2D) in the older group and only the symptomatic children (P2A and P2D) in the younger group showed a significant decrease in proliferative responses to PHA. All classes of infected children demonstrated a significant decrease in response to tetanus toxoid. We have been able to demonstrate a loss of antigen responsiveness which precedes the loss of mitogenic responsiveness. Furthermore, we have been able to demonstrate an age related increase in lymphoproliferative responses to both PHA and tetanus in HIV-infected and control children. Therefore, we conclude that children are particularly susceptible to the immunologic effects of HIV infection. Loss of lymphoproliferative responses to antigen occurs early in infected children and precedes the loss of CD4+ helper cells and of PHA responsiveness. This increased susceptibility to the immunopathogenesis of HIV infection is due, at least in part, to the relative immunodeficiency of infancy.

Age Factors↗

Beta 2-microglobulin and immunoglobulins are more useful markers of disease progression in HIV than neopterin and adenosine deaminase.

Reference change values of six biochemical quantities (beta 2-microglobulin, neopterin, adenosine deaminase and immunoglobulins IgA, IgG and IgM) have been established in asymptomatic human immunodeficiency virus (HIV)-infected patients following the method described by Harris and Yasaka in 1983. Patients included in the evaluation were classified as A1, A2 or A3 according to the classification of the Centers for Disease Control (CDC) (January 1993). All patients were followed-up quarterly, with a minimum of four samples each available for statistical analysis. The main objective of this paper was to study whether differences found to be greater than calculated reference change values could predict clinical or immunological worsening in patients' status. Retrospective analysis was made in asymptomatic patients (n = 256) included in an HIV infection protocol carried out in our hospital. Of these patients, 179 showed clinical or immunological worsening during the study period and 77 maintained their clinical and immunological status. Changes in beta 2-microglobulin showed the greatest sensitivity to detect clinical or immunological worsening (43.0%), whereas changes in adenosine deaminase showed the lowest (21.8%). Clinical or immunological worsening in 169 of the 179 patients was detected by one of the six biochemical quantities evaluated. Ten patients showed clinical or immunological worsening, although differences between measurements were lower than the reference change values calculated. Of 77 patients whose clinical state did not deteriorate, there was a change in biochemical analytes greater than the reference value calculated in 29 patients (a period of 12 months had elapsed since detection). In 48 patients, no increases greater than calculated reference change values were detected. The sensitivity obtained using the six analytes was 94.4% and the specificity was 62.3%.

Adenosine Deaminase↗

[Pneumocystis carinii is still a dangerous opportunist. The infection is continuously a threat to immunocompromised patients].

Despite recent decrease in the incidence of Pneumocystis carinii pneumonia (PCP) among patients infected with HIV (human immunodeficiency virus), PCP remains a threat to other categories of immunocompromised patients. The article provides an outline of recent, mainly molecular genetic, findings in P. carinii research, including its new classification as a primitive fungus, host specificity and verified de novo infection in HIV-infected subjects. As the pathogen still defies propagation in vitro, laboratory diagnosis is dependent on microscopic demonstration of the organism. Diagnostic specificity can be enhanced by generating specific PCR (polymerase chain reaction) products which can be sequenced for genotyping. Findings in animal studies and epidemiological observations (e.g., in outbreaks of PCP among immunocompromised hospital patients), suggest transmission of PCP infection to be airborne. Genetic methods have been used to study the mode of P. carinii transmission. Nucleic acids of the human form of P. carinii (P. carinii f. sp. hominis) have been detected in the air of hospital wards, indicating susceptible patients to be at risk. By contrast, findings obtained with the same methods in studies of person-to-person transmission of P. carinii among clustered cases of PCP in hospitals suggest infection to be environmentally acquired. Thus, many questions remain to be answered regarding the occurrence and transmission of P. carinii infection.

AIDS-Related Opportunistic Infections↗

Effect of granulocyte-macrophage colony-stimulating growth factor on interferon and tumor necrosis factor production in whole blood cell cultures of patients with acute myelogenous leukemia.

The effect of recombinat human granulocyte-macrophage colony-stimulating growth factor (rHuGM-CSF) treatment on in vitro interferon (IFN) and tumor necrosis factor (TNF) production in peripheral blood cells of 46 patients with acute myelogenous leukemia (AML) was examined. GM-CSF significantly enhanced virus-induced IFN-alpha production in blood cells (containing 68% of blasts) of 28 patients with M4-M5 AML according to the French-American-British (FAB) classification and also phytohemagglutinin (PHA)-induced IFN-gamma production in blood cells (containing 70% of blasts) of 18 patients with AML MO-M3 type. In control blood cells (25 healthy persons) GM-CSF enhanced PHA-induced IFN-gamma but did not influence IFN-alpha production. In the presence of GM-CSF, TNF-alpha titers induced with lipopolysaccharide were also higher in control blood cells but not in cells of patients with M0-M3 or M4-M5 type of AML. The significance of GM-CSF-enhanced IFN-alpha and IFN-gamma production in antimicrobial and anti-leukemic immune reactions which can develop during GM-CSF therapy is discussed.

Adolescent↗

[Cutaneous and oral manifestations in HIV-infected children and adults--169 cases].

UNLABELLED: The aim of the study was to evaluate cutaneous and oral manifestations in infected HIV patients. MATERIALS AND METHODS: We retrospectively analyzed 169 cases admitted in Infectiouse Disease Department of Iaşi in 2001-2002 period. RESULTS: Cutaneous and oral manifestations were: candidiasis (99 cases), herpes virus infectious (36 cases), scabies and straphylococcal/streptococcal skin disease (26 cases), prurigo nodularis, psoriasis and verruca vulgaris (9 cases). Children of 0-13 year old group was 75.73 percent. Classification of HIV infection was related with CD4 count for 166 cases. Twelve cases with oral pharyngitis candidiasis, scabies and streptococcal skin diseases was 2-3 recurrent episodes of manifestations. Etiotrop treatment was associated with HAART therapy. CONCLUSIONS: Cutaneous and oral manifestations are occurred frequently in HIV infected patients, with a various etiology, but the severity, persistence and its evolution did not evaluate.

AIDS-Related Opportunistic Infections↗

Taxonomy and evolutionary studies on lyssaviruses with special reference to Africa.

Currently, the Lyssavirus genus is composed of four serotypes and the ungrouped European bat lyssaviruses (EBL). Using previously described PCR amplification and direct sequencing methods, the N genes of 70 representative lyssaviruses collected from 40 different countries were examined. From the results, a phylogenetic tree with six branches (genotypes) was constructed: genotypes 1-4 correlated with the classification of serotypes 1-4 and in addition EBL 1 and EBL 2 were identified as two further genotypes (5 and 6). Analysis of the 51 rabies (genotype 1) viruses in the study identified nine distinct groups which could be correlated with their geographical origins. A comparison of the six genotypes with four principal Vesiculovirus serotypes showed that their relationship was closer than that of even the two most closely related Vesiculovirus serotypes. Antigenic site sequences of isolates from vaccine/treatment failures were similar to vaccine and field strains, suggesting that failures were not due to genetic variation at the N protein level.

Africa↗

Dilated cardiomyopathy--new insights into the pathogenesis.

There is increasing evidence for a viral aetiology in many patients with dilated cardiomyopathy, although the term viral heart disease is not currently recognized in the WHO/ISFC classification of the cardiomyopathies. The prognosis of dilated cardiomyopathy remains poor with a high morbidity and mortality. New insights into the pathogenesis of dilated cardiomyopathy and the effect of viruses on its prognosis, offer potential for future treatments to modify the natural history of the disease.

Cardiomyopathy, Dilated↗

[Brain stem encephalitis related with viral infection].

There is no definite classification of brainstem encephalitis, its etiology is not clear. Bickerstaff's brainstem encephalitis (BBE) was established by clinical diagnosis without etiology. Recently there are great progression to identify a virus by using immunological methods and PCR (polymerase chain reaction) methods. Sometimes the progression of MRI revealed subdiagnostic images of brainstem lesion. Nowadays we can investigate anti-ganglioside antibodies in some BBE patients but not all. High IgG anti-GQ1b antibody titers were present in some BBE sera samples but decreased with the clinical course of illness. A girl was suffered from BBE whose Cytomegarovirus-DNA was detected in the CSF by PCR. Serum anti-GQ1b antibody was positive. The neurological symptoms disappeared with decreasing of anti-GQ1b antibody. Immuno adsorption therapy is effective for BBE with anti-GQ1b antibody positive patients.

Adult↗

Absence of Epstein-Barr virus in anaplastic large cell lymphoma: a study of 64 cases classified according to World Health Organization criteria.

The frequency of Epstein-Barr virus (EBV) in anaplastic large cell lymphoma (ALCL) has been controversial. The interpretation of previous studies is complicated by the use of nonuniform EBV detection methods and the inclusion of cases of CD30-positive diffuse large B-cell lymphoma and so-called "ALCL, Hodgkin-like," as defined in the Revised European-American Lymphoma classification scheme. In the current World Health Organization (WHO) classification system, both of these tumors are excluded from the ALCL category. Also, recently developed antibodies (eg, the antibody specific for PAX-5/B-cell-specific activator protein [BSAP]) provide new, sensitive tools for identifying neoplasms of B-cell lineage that can morphologically resemble ALCL. In this study we evaluated 64 cases of ALCL of T- or null-cell lineage, defined according to the WHO classification system, for the presence of EBV. All tumors were negative for B-cell antigens, including PAX-5/BSAP and CD20 or CD79a. The study group included 27 (42%) anaplastic lymphoma kinase (ALK)-positive (18 T-cell and 9 null-cell) and 37 (58%) ALK-negative (30 T-cell and 7 null-cell) tumors analyzed by in situ hybridization for EBV-encoded RNA (EBER) or immunohistochemistry for EBV-latent membrane protein type 1. All 64 cases were negative for EBV. We conclude, based on the current definition of ALCL in the WHO classification, there is no role for EBV in ALCL arising in Western patients. We suggest that published reports of EBV in a small proportion of ALCL cases in Western patients can be explained by the inclusion of tumors no longer considered to be in the current classification of ALCL, such as CD30-positive anaplastic tumors of B-cell origin.

Anaplastic Lymphoma Kinase↗

Pyothorax-associated lymphoma: relationship with Epstein-Barr virus, human herpes virus-8 and body cavity-based high grade lymphomas.

Pyothorax-associated lymphoma (PAL) is a newly-described entity developing several decades after artificial pneumothorax treatment for pulmonary or pleural tuberculosis. It is known to be associated with Epstein-Barr virus (EBV) with constant expression of the two latent membrane proteins: latent membrane protein (LMP)-1 and EBV-associated nuclear antigen (EBNA)-2. We are reporting three new cases of PAL. All of the tumours were of B-cell lineage and classified as large-cell diffuse lymphomas according to the International Working Formulation for the Classification of Lymphomas. The EBV genome was detected in two of the cases with LMP-1 and EBNA-2 expression. No EBV could be detected in the third case suggesting that different mechanisms may be involved in the pathogenesis of the disease. Body cavity-based high grade lymphomas (BCBL) represent a new disease, developing mainly in human immunodeficiency virus (HIV) infected patients: the tumoural cells often contain both human herpes virus (HHV)-8 (or Kaposi's sarcoma herpes virus) and EBV genomes, suggesting that these viruses might co-operate in the pathogenesis of the disease. The pleural location and the association of EBV have led to speculation that PAL could also be related to HHV-8 infection. However, no HHV-8 genome could be detected in any of the 14 tested cases already reported in the literature nor in the two cases we studied (one EBV-positive and one EBV-negative), suggesting that PAL and BCBL are two different entities.

Aged↗

Aggressive Epstein-Barr virus-associated, CD8+, CD30+, CD56+, surface CD3-, natural killer (NK)-like cytotoxic T-cell lymphoma.

We report an unusual case of aggressive natural killer (NK)-like cytotoxic T-cell lymphoma in a previously healthy immunocompetent West African male. He presented with a fever of unknown origin, subsequently developed erythematous skin nodules, generalized lymphadenopathy, and hepatosplenomegaly, and then died of multiple organ failure. A skin nodule and lymph node biopsy showed an infiltrate of pleomorphic atypical medium and large lymphoid cells with extensive necrosis and prominent apoptosis. Peripheral blood and ascites also harbored these cells, with cytology revealing irregular nuclear folding and basophilic cytoplasm, and some with azurophilic cytoplasmic granules. Flow cytometry and immunohistochemistry demonstrated the expression of CD2, CD7, CD8, CD30, CD56, and cytoplasmic but not surface CD3. In situ hybridization demonstrated Epstein-Barr virus transcripts. A monoclonal T-cell receptor gamma chain gene rearrangement was detected by polymerase chain reaction. This is the first reported case of an NK-like T-cell lymphoma with these unusual features, making precise classification difficult. Some features suggest an NK1.1 or NKT lymphocyte origin. Because the earliest clinical manifestation was splenomegaly and abnormal liver function, the normal cellular counterpart may be a distinct subset of NK1.1 cells normally present in hepatosplenic sinusoids. This tumor disseminated early and pursued a fulminant clinical course, thus emphasizing the importance of early recognition and diagnosis.

Adult↗

A second group of hepatitis C viruses.

cDNA clone 11-7 was isolated by immunoscreening a cDNA library that was prepared from a pooled plasma of non-A non-B hepatitis (NANBH) patients using expression vector lambda gt11. This cDNA corresponds to known nucleotide positions 3983-4745 of the genome of hepatitis C virus (HCV). This clone was used as a probe for screening the HCV-related cDNAs in a cDNA library similarly prepared by using lambda gt10. As a result, six more cDNA clones were isolated and analyzed for their nucleotide sequences. The results strongly suggested that there are at least two groups of HCV, group I and group II. According to our classification, the prototype HCV and clone 11-7 belong to group I HCV, and their nucleotide and deduced amino acid sequences were diverged from those of group II HCV. Genetic variation observed in the nucleotide and the amino acid sequences between the two groups resembles that in the NS3 region of the genome between Japanese encephalitis virus and West Nile fever virus. Polypeptides produced in Escherichia coli carrying a clone 11-7 or a group II cDNA clone E reacted with antibodies in the blood of 12 or 4 out of 14 individual chronic NANBH patients, respectively. Our data clearly indicate the existence of a second group of HCV.

Amino Acid Sequence↗

Use of reverse transcriptase polymerase chain reaction for detection of vaccine contamination by avian leukosis virus.

A reverse transcriptase polymerase chain reaction (RT-PCR) for avian leukosis virus (ALV) was developed for the detection of contamination of vaccines produced in embryonated eggs and cell cultures derived from chicken. ALV is highly pathogenic and induces a wide spectrum of disease in infected animals. ALV can be divided into five subgroups (A-E). The envelope glycoprotein (env gp85) is the main antigen determinant and responsible for subgroup classification. Viral RNA of all subgroups (A-E) was isolated and amplified using three sets of primers. Subsequently, restriction endonuclease analysis confirmed the product identity and discriminated between subgroups. In specific pathogen free (SPF) eggs experimentally inoculated with ALV, viral RNA was found in allantoic fluids, as well as in vaccines spiked with different subgroups of ALV. No adventitious virus was detected in commercially available preparations. This system provides a rapid and specific in vitro method for the detection of ALV RNA as an extraneous agent and may be applied for quality control of avian vaccines.

Animals↗