Cryoglobulinaemic membranoproliferative glomerulonephritis and hepatitis C virus infection.

BACKGROUND/AIM: A striking correlation between mixed cryoglobulinaemia and chronic hepatitis C virus infection has recently been described. Since membrano-proliferative glomerulonephritis is a rare complication of mixed cryoglobulinaemia, this study was undertaken to determine the prevalence of Hepatitis C virus infection in membrano-proliferative glomerulonephritis. PATIENTS: Eighteen patients, selected among a group of 121 affected by mixed cryoglobulinaemia, with renal involvement were included in the present study. A group of 148 patients affected by renal disease of different aetiology and the general population (6,917 people) were used as control groups. METHODS: The presence of anti-hepatitis C virus antibodies was determined by a commercial kit. The hepatitis C virus genotype was determined according to Okamoto. All patients underwent kidney and bone marrow biopsy, while the hepatic biopsy was performed in those showing signs of chronic liver disease. RESULTS: In patients with renal involvement, the kidney biopsy showed the presence of membrano-proliferative glomerulonephritis Type I in all cases. Chronic liver disease was present in eleven patients (61%). All patients were positive for serum hepatitis C virus-RNA. Bone marrow biopsy was normal in five cases, while in the others paratrabecular foci of infiltration by small lymphocytes were present. In six of these, the massive bone marrow infiltration by lymphoplas-macytoid lymphocytes suggested the diagnosis of low grade non-Hodgkin's lymphoma. In the group of patients affected by other chronic renal disease, the prevalence of hepatitis C virus infection (3.1%) was not different from that of the general population (3.2%). CONCLUSIONS: Hepatitis C virus seems to be the aetiologic agent of mixed cryoglobulinaemia and, consequently, of membrano-proliferative glomerulonephritis.

Hepatitis C virus and immunoglobulin gene rearrangements: an early step in lymphomagenesis?

A clonal expansion of peripheral blood mononuclear cells committed to IgM cryoprecipitating rheumatoid factor production has been demonstrated in hepatitis C virus (HCV)-associated mixed cryoglobulinemia (MC). To determine the role of HCV in B cell gene rearrangements we studied a series of 57 HCV-infected patients with and without MC. Clonal Ig gene rearrangements of both RNA and DNA were detected in 10 of the 13 patients with type II MC, 1 patient had gene rearrangement of the DNA only, and 2 had polyclonal patterns. 2 of the 17 patients with type III MC showed clonal rearrangement of both RNA and DNA, in 6 only the DNA was rearranged clonally and in 9 the patterns were completely normal. 14 of 27 patients with cryocrit <1% or without cryoglobulins had clonal DNA rearrangements without any in the RNA. These results suggest that clonal lesions in the DNA are related to HCV infection and that these changes antedate the appearance of mixed cryoglobulinemia.

Systemic manifestations and liver disease in patients with chronic hepatitis C and type II or III mixed cryoglobulinaemia.

The aim of this study was to evaluate the prevalence of cryoglobulins in patients with chronic hepatitis B and C virus infection and to investigate the association of type II and type III mixed cryoglobulinaemia with systemic manifestations and liver disease stage and outcome in hepatitis C virus (HCV)-positive patients. We analysed the prevalence of cryoglobulinaemia in a cohort of patients with chronic liver disease and compared the systemic manifestations and liver involvement in HCV-positive patients with type II or type III mixed cryoglobulinaemia. The prevalence of serum cryoglobulins was significantly higher in HCV-positive patients than in hepatitis B surface antigen (HBsAg)-positive patients (55.4 vs 20.6%). In HCV-positive patients, stage of liver disease correlated with the prevalence of cryoglobulinaemia. Patients with type II cryoglobulins showed a significantly higher risk of cirrhosis and of extrahepatic manifestations while patients with type III cryoglobulins had a significantly higher prevalence of hepatocellular carcinoma. During follow-up the former had an odds ratio of 11.9 of death from extrahepatic complications while the latter had an odds ratio of 3.4 of dying from hepatic disease. Our study confirms the high frequency of mixed cryoglobulinaemia in patients with chronic hepatitis C virus infection. The presence and type of cryoglobulins seem to be associated with different clinical manifestations and outcome.

Interferon therapy in HCV-positive mixed cryoglobulinaemia: viral and host factors contributing to efficacy of the therapy.

BACKGROUND: In a previous paper, we reported on the short-term efficacy of alpha-interferon in the treatment of hepatitis C virus positive mixed cryoglobulinaemia. AIMS: We investigated the long-term effects of therapy in a larger group of patients, and the viral and host factors able to influence the response to treatment. METHODS: In 27 females and 15 males (mean age 54.8 +/- 9.1 years) affected by mixed cryoglobulinaemia, bone marrow biopsy and phenotyping of marrow cells were performed before treatment and at the end of follow-up. A liver biopsy was obtained from patients showing biochemical signs of chronic liver disease. The presence of hepatitis C virus was assessed by detection of serum anti-hepatitis C virus antibodies, and hepatitis C virus-RNA. The treatment schedule was 3 million units of recombinant interferon alpha-2b three times a week for one year. Follow-up lasted for 1 year after the end of treatment. The response was classified as follows: 1) Complete response: Disappearance of the cryocrit (or reduction of more than 50%) and of all clinical manifestations of the disease. 2) Partial response: Disappearance of all clinical signs of the disease, but reduction of cryocrit of less than 50%. 3) Minor response: Reduction of cryocrit of less than 20% associated with the disappearance of one or more (but not all) signs of vasculitis. RESULTS: Anti-hepatitis C virus antibodies were present in 41 (95%) patients, and hepatitis C virus-RNA was detectable in all cases. Before therapy, marrow histology showed a massive monomorphous infiltration by plasmacytoid lymphocytes indicating the presence of low-grade non-Hodgkin lymphoma in 7 cases (16.6%). After therapy, 13 (31%) patients achieved a complete response, 23 patients (55%) a partial response, and 6 patients (14%) a minor response. Seven of the responders and all patients showing partial or minor responses relapsed a few months after withdrawal of therapy. At the end of the follow-up, only 6 patients had obtained complete remission. Bone marrow examination showed that B-lymphocytic monoclonal infiltrate had disappeared in 3 long-term responders. No difference was found between responders and non-responders/relapsers in terms of age, sex, duration of the disease, severity of symptoms, liver function tests, rheumatoid factor or complement levels, while the lack of response was associated with the presence of genotype 1b, liver cirrhosis, and high cryoglobulin level. CONCLUSIONS: Mixed cryoglobulinaemia is associated with a high prevalence of B-cell lymphomas. Alpha-Interferon is an effective agent for the treatment of this disease and seems able to determine regression of the lymphoproliferative disorder. The hepatitis C virus genotype and cryoglobulin level are the most important predictive factors of response to therapy.

HCV and lymphoproliferative diseases.

METHODS: Genomic and replicative forms of HCV-RNA in B lymphocytes were detected by RT-PCR, and HCV genotyping was performed using universal and type-specific primers for the core region. Immunoglobulin gene rearrangement was detected by RT-PCR. RESULTS: The presence of genomic and replicative forms of HCV-RNA in the 5'NC region was investigated on total RNA extracted from subpopulations of PBMC. The frequency of HCV-RNA was higher in the B lymphocytes than in other PBMC. In two patients a larger sized band was present in the B lymphocytes and PMN; this band could represent either another form of HCV-RNA or a cross-reaction between cellular RNA and HCV primers. HCV-RNA detected using primers for the core region was negative in the patients examined. Immunoglobulin monoclonal gene rearrangement was present on the cDNA in all of the HCV and type II cryoglobulinemia positive samples except two; in contrast, it was absent in the HCV positive and cryoglobulinemia negative samples. The analysis of immunoglobulin monoclonal gene rearrangement on DNA showed the presence of new positive samples among the HCV positive, type II cryoglobulinemia negative patients, who had been negative when PCR was performed on cDNA. Denaturing sequencing gel showed clearer results than agarose gel. CONCLUSIONS: The early detection of immunoglobulin monoclonal gene rearrangement and expression is very important because it could provide evidence of the possible lymphoproliferative evolution of HCV infection. In addition, these investigations together with PCR product sequencing could show us the steps in the clonal selection of B lymphocytes towards malignant transformation, in which HCV plays a direct and/or indirect role.

Clinical and virological findings in mixed cryoglobulinaemia.

OBJECTIVES: As a close relationship has been established between mixed cryoglobulinaemia and hepatitis C virus (HCV) infection, the clinical, histological and virological findings of patients affected by mixed cryoglobulinaemia were determined. DESIGN: Hepatitis C virus infection was investigated by the presence of anti-HCV antibodies and PCR amplification of the 5' untranslated region (5' UTR), and the genotype of HCV was also determined according to Okamoto. A bone marrow biopsy was performed in all patients and liver and kidney biopsies when indicated. SUBJECTS: Eighty-two subjects affected by mixed cryoglobulinaemia were enrolled in this study. RESULTS: The prevalence of anti-HCV antibodies was high (83%); PCR amplification of the 5'UTR region was performed in 52 subjects and in 44 of them (85%) the results were positive. In the same subjects, the Core region amplification was positive in 46 cases (88%). A high prevalence of genotype II was found (54%). Chronic liver disease was present in 55 patients (67%). Bone marrow biopsies showed the presence of low-grade non-Hodgkin's lymphomas in 11 cases (13%). Membrano-proliferative glomerulonephritis was found in seven subjects (8%). CONCLUSIONS: Mixed cryoglobulinaemia is associated with HCV infection in the nearly all cases. Several HCV genotypes are involved in the pathogenesis of this disease. Mixed cryoglobulinaemia is associated with a high prevalence of chronic liver disease, low-grade non-Hodgkin's lymphomas and membrano-proliferative glomerulonephritis.

Effects of alpha-interferon and steroids on CD23 expression and release in B-cell chronic lymphocytic leukemia.

BACKGROUND: Since high CD23 expression and release have been reported in B-chronic lymphocytic leukemia (B-CLL), we investigated whether alpha-interferon or corticosteroids were able to modulate the expression and/or the release of this factor. METHODS: CD23 expression was determined with FITC-labelled anti-CD23 monoclonal antibody, and sCD23 release with a sandwich enzyme immunoassay. Twenty-one patients affected by B-CLL (stage A or B) were studied before and after three different treatment regimens (alpha-interferon, corticosteroids, alpha-interferon+corticosteroids). RESULTS: CD23 was highly expressed in the B-cells of all patients, and expression was not modified by any of the therapies, sCD23 release from leukemic cells was significantly greater (p < 0.00001) in untreated subjects than controls, and in vitro treatment with phorbol myristate acetate (PMA) led to a 10-fold increase (p < 0.0001) in sCD23 secretion. On the contrary, PMA did not increase sCD23 release in normal B cells. Treatment with corticosteroids (either alone or associated with alpha-interferon) reduced sCD23 secretion from leukemic cells, whereas alpha-interferon alone was not able to modify sCD23 release. CONCLUSIONS: Our data support the hypothesis that CD23 plays a role in the maintenance and progression of B-CLL and that the pharmacological modulation of this receptor/lymphokine could be useful in the therapy of B-CLL.

Infections in an Alpine environment: antibodies to hantaviruses, leptospira, rickettsiae, and Borrelia burgdorferi in defined Italian populations.

From 1987 to 1991, a seroepidemiologic survey for antibodies to hantaviruses, leptospira, rickettsiae, and Borrelia was conducted in selected Italian population groups. In the mountainous areas of northeastern Italy, the prevalence of antibody to hantaviruses, as detected by indirect immunofluorescent antibody (IFA) assay, was 7.1%, 4.8%, 4.3%, and 4% in 265 forestry workers, 82 rangers, 395 farmers, and 75 hunters, respectively. Among 299 Alpine soldiers, the prevalence was lower (0.7%). Of those with Hantaan antibody, the reactivity pattern using Hantaan, Puumala, and Fojnica viruses suggested a prevalence of antibody to Hantaan virus, with titers reaching levels of 128. The presence of leptospiral antibodies (by microagglutination test), which included the prevalence of antibodies to Leptospira icterohaemorrhagiae, L. bratislava, and L. saxkoening serotypes, was observed in 10-12% of the farmers and forestry workers in these Alpine mountain regions. Only a few sporadic clinical cases of leptospirosis have been reported from these regions. Antibodies to Borrelia burgdorferi (by IFA) were observed in 19% of the rangers and forestry workers, with lower values in farmers (10%) and hunters (8%). These data suggest the presence of a large number of asymptomatic infections with B. burgdorferi and the leptospires in the densely wooded areas of the Alpine Italian regions. Furthermore, the recent identification of a case of Hantaan acute nephropathy in a man living in the mountainous northeastern area of Italy confirms the presence of hantavirus in the Italian Alpine zones, especially those near the Slovenian border.

Seroprevalence of antibodies to hantaviruses and leptospires in selected Italian population groups.

A study of hantaviral and leptospiral antibodies in selected population groups was performed. Among high risk subjects in the Rome area, Hantaan antibody was found in mammalogists (10%) and dialysis patients (6%), while none of the trappers, oarsmen, river policemen and firemen studied tested positive for antibodies to hantaviruses. In occupationally-exposed subjects (farmers, rangers, lumbermen, hunters) from rural and densely forested areas of northern Italian regions, the prevalence of Hantaan antibody ranged from 3.3% to 8.8%. In the positive cases the comparative antibody titration using different hantaviruses showed a predominance of Hantaan virus (titer 1:128) compared to Puumala virus (titer 1:32); no reactivity was observed with Seoul or Prospect Hill viruses. In Rome, leptospiral antibodies were found in trappers (21%) and oarsmen (5%) at a titer of 1:50 or more, with a predominance for the L. icterohaemorrhagiae serotype (85%). In the Alpine areas the leptospiral antibody prevalence was 12% and L. icterohaemorrhagiae and L. bratislava were the predominant serotypes. The presence of hantavirus infections, suspected after the first epidemiological survey conducted in central Italy, is now supported by the new data obtained in northern Italian regions. Furthermore, the recent observation of one case of Hemorrhagic Fever with Renal Syndrome (HFRS) in the Udine area, not far from the Yugoslavian border, strongly confirms the presence of one or more hantaviruses in Italy.

Relación entre albúminas e inmunoglobulinas séricas y licuorales en hanseniano

Los resultados de mayor interés obtenidos sobre las muestras de LCD y suero de 7 casos de hanseniasis (H.D.) (2 borderline: B 3 lepromatosos: L de los cuales el 4 L grave y con déficit inmunitario; 2 tuberculoides: T), son: I.º) Los índices LCR/suero de la albúmina y de las IgG por tests cinético-nefelometría y de las IgG anti PGL-1 por ELISA, en cuanto separan los casos según sus tipos y forma mediante valores típicamente uniformes: los más bajos de albúmina para los L; los más bajos de IgG anti PGL-1 para los B; los medios de IgG para los T, siendo los B y L muy deformantes. II.º) Los index de LINK indican producción intratecal de IgG si superan el V.N. en nuestros casos los de IgG anti PGL-1 han resultado todos superiores y altos, sólo los B menos altos; los IgG todos inferiores; pero, hay que considerar que por la policlonía de la IgG, una parte de ellas podría resultar superior si fuese revelada aisladamente por su antígeno específico. III.º) Una mayor casuística con test de medidas homólogas para los dos de IgG podrá confirmar o no nuestros resultados, y también modificar nuestra de ducción por la cual los epítopes HLA-DR de los queratinocitos uniéndose con las IgG intratecales determinarían la respuesta al test lepromínico, es decir, negativa en los L por la falta de los HLA-DR; positiva la precoz en los B po la unión con la IgG; positiva la tardía y a menudo la precoz en los T por la unión con la IgG anti PGL-1 y las IgG. Otro caso de diagnosis incierta resulta positivo para la H.D. de tipo T, pero atípico o la debido a la asociación de otra enfermedad del S.N.C. como la esclerosis múltiple o la siringomielía.

Borreliosis risk after tick bite in north-eastern Italy.

Lyme Borreliosis is an infectious disease caused by the Borrelia burgdorferi spirochete transmitted to man by a tick bite. The aim of this study was to assess the predictive value of tick bites for the diagnosis of Borreliosis in a sample of 266 subjects residing in an endemic area of north-eastern Italy. In the serological diagnosis of Borreliosis, positive and negative predictive values of tick bites were found to be 24% and 88% respectively.

Risk factors for human immunodeficiency virus infection in 581 intravenous drug users, northeast Italy, 1984-1988. The AIDS and Related Syndromes Study Group.

Prevalence and risk factors for human immunodeficiency (HIV) virus infection were assessed in 581 heterosexual intravenous drug users (IVDUs), recruited from eight public drug assistance centres in the northeast of Italy, an area at low risk for AIDS. The overall seroprevalence for HIV antibody was rather high, 39% (95% confidence interval (CI): 35-43%). HIV prevalence had risen from 32% in 1984-1985 to 52% in 1987-1988. Thirty-three per cent of female and 41% of male IVDUs had antibodies to HIV (p = 0.07). Seropositivity rates showed a strong east-west geographical gradient (from 16% to 60%, p = 0.005): IVDUs living in the western part of the study area had a nearly sevenfold higher risk of infection (odds ratio (OR) = 6.9, 95% CI: 4.4-13.9) than those living in the eastern part. Sharing of drug injection equipment (OR = 3.6), duration of drug addiction (OR = 2.6), use of heroin in high-incidence cities (OR = 2.3), use of cocaine in addition to heroin (OR = 1.5) and history of prostitution (OR = 2.3) increased the risk of acquiring HIV infection. Over the study period, the ORs associated with area of residence decreased, whereas those associated with the use of heroin in high-incidence cities and with prostitution increased.

Fusarium peritonitis in peritoneal dialysis: report of two cases.

This report describes two cases of peritoneal dialysis-related peritonitis caused by Fusarium species which have been incriminated in a variety of disease conditions. Treatment with 5-Fluorocytosine was unsuccessful, but removal of the catheter resulted in a rapid resolution of the infections.

Evaluation of an immunoluminometric assay for the AFP determination.

We evaluated a two-site immunoluminometric assay (AFP LIA-mat byk) for the alpha-fetoprotein (AFP) determination. The assay, involving two monoclonal antibodies which recognize two different epitopes of the AFP, is rapid (approximately 4 h), with a wide working range (0-600 IU/ml) and a good sensitivity of detection. It presents a good reproducibility (CV less than 10% for the total working range). The regression equation for the AFP LIA-mat (Y) and the AFP bridge Serono (X) was Y = 1.175X-2.27 (n = 95, r = 0.996). It didn't present hook effect and, when we assessed the linearity by assaying a high concentration of AFP, it gave a linear response down to 3.5 IU/ml.

Ofloxacin: in vitro activity against recently isolated gram-negative bacterial strains.

The activity of ofloxacin was determined against 117 Enterobacteriaceae, 13 Acinetobacter var, anitratus, 124 Pseudomonas aeruginosa in comparison with other antibiotics. Its activity was very high: against Enterobacteriaceae the minimum inhibitory concentration (MIC)50 was 0.125 micrograms/ml, the MIC90 1 micrograms/ml, and the geometric mean (GM) was 0.4 micrograms/ml against Acinetobacter var. anitratus the MIC50 1 micrograms/ml, MIC90 4 micrograms/ml, GM 1.7 micrograms/ml. Unlike other authors we found that the activity of ofloxacin was influenced by the selection of P. aeruginosa resistant to carbenicillin and gentamicin.

Time-related shape control modifications during erythrocyte storage with additive solutions.

To obtain more detailed information on the reversibility of shape alterations in blood bank stored erythrocytes, we have studied shape recovery after chemical crenation and rheological properties in 8 PAGGS-sorbitol preserved erythrocyte concentrates during a five week storage period under blood bank conditions. Our results show that red cell capability to regain a normal discoid shape after chemical crenation decreases during storage but is not lost over a five week period. Moreover there is a significant but weak correlation between red cell ATP content and both shape recovery capability and viscosity. Our results confirm suspicious that red cell shape perturbations following blood bank storage are widely reversible. Two different mechanisms may be involved in reducing shape recovery capability during storage, namely an ATP-dependent mechanism and an energy-independent one. The energy dependent mechanism may be preserved by the previous addition of solutions which maintain higher energy levels during storage.

Critical evaluation of bioluminescence methods for quantifying bacterial adhesion to polystyrene.

A total of 134 strains of Escherichia coli which included 60 fecal and 74 urinary isolates, cultured in liquid and on solid media, were examined for adhesive properties using bioluminescence and haemagglutination methods. The study aimed to verify whether irrespective of the absence or presence of flagella, there is any relation between haemagglutination and bioluminescence test. Examining the results we failed to note any correlation between the two methods: strains bearing MS, MR or MS-MR adhesins adhered to polystyrene at random. Even though it is fast and easy to perform, bioluminescence is not an alternative to traditional methods to reveal MS adhesins.