Pharmacokinetics and anthelmintic efficacy of injectable eprinomectin in goats.

The objective of this study was to assess the pharmacokinetics and the anthelmintic efficacy of eprinomectin (Eprecis® 20mg/mL) following subcutaneous administration to goats. Forty non-lactating female Alpine goats aged between one and three years and weighing between 32.7 and 59.5kg, were randomly allocated to one of the following groups (8 animals per group): two groups were not infected and were treated at a dose of either 0.2 or 0.4mg/kg BW of eprinomectin, two groups were experimentally infected with nematodes and treated at 0.2 or 0.4mg/kg BW of eprinomectin similarly and one group was infected and left untreated (control). Infection consisted in a single and simultaneous administration of 5000 Haemonchus contortus and 12,500 Trichostrongylus colubriformis infective larvae. No local or general adverse reaction was visually observed whatever the dose rate. The maximal plasma concentrations (Cmax) were 20.68±12.85 vs 39.79±17.25µg/L and the plasma bioavailabilities (AUC) 83.45±34.75 vs 169.37±43.44µg*d/L for 0.2 vs 0.4mg/kg respectively. The efficacy against H. contortus and T. colubriformis was 97.8 and 98.7% at 0.2mg/kg and 98.4% and >99.9% at 0.4mg/kg respectively. The differences in worm burdens between the two dose rates were only significant for T. colubriformis. These results indicate that injectable eprinomectin is a potent anthelmintic against the two major gastrointestinal nematodes in goats. Additional information is needed regarding pharmacokinetics in lactating goats and milk residues.

Asymmetrical transmission of human herpesvirus 8 among spouses of patients with Kaposi sarcoma.

BACKGROUND: Among heterosexuals, the sexual transmission of human herpesvirus 8 (HHV8) has not been established. OBJECTIVES: To assess HHV8 seroprevalence in spouses of patients with classic and endemic Kaposi sarcoma (KS) and to suggest possible routes of transmission. METHODS: A case-control study was carried out in a teaching hospital among spouses of human immunodeficiency virus-negative patients with KS (cases - exposed subjects) and controls who did not have KS nor were related to patients with KS (nonexposed subjects). HHV8 seroprevalence in spouses of patients with KS was compared with HHV8 seroprevalence in controls matched for age, gender and place of birth. Other serology tests were compared between cases and controls. Among heterosexual couples, HHV8-seropositive and HHV8-seronegative spouses were compared for possible risk factors for virus transmission. RESULTS: HHV8 seroprevalence was significantly higher among spouses of patients with KS (13 of 22; 59%) than among matched controls (19 of 58; 33%; P = 0.043). Among heterosexual couples, five of five (100%) male spouses were HHV8 positive vs. six of 15 (40%) female spouses (P = 0.04). There was no significant difference between HHV8-seropositive and HHV8-seronegative spouses for all other factors screened for among heterosexual couples. CONCLUSIONS: Being a spouse of a patient with KS is a risk factor for HHV8 seropositivity. Our results suggest that female-to-male HHV8 transmission could be more efficient than male-to-female transmission among couples including a patient with KS. Transmission could involve distinctive behaviours, or currently unknown biological properties of HHV8.

Virologic and immunologic parameters that predict clinical response of AIDS-associated Kaposi's sarcoma to highly active antiretroviral therapy.

The purpose of the work was to assess the predictive value of biologic factors on the efficacy of highly active antiretroviral therapy alone or combined with chemotherapy on AIDS-associated Kaposi's sarcoma. Twenty-six AIDS-Kaposi's sarcoma patients who started therapy with protease inhibitors were investigated. No baseline chemotherapy was associated with less severe initial clinical status. Median follow-up was 652 d. The main outcome measures were as follows: best Kaposi's sarcoma clinical response; Kaposi's-sarcoma-associated herpesviral load in peripheral blood mononuclear cells using real-time quantitative polymerase chain reaction (non-detectable if less than 100 copies per microg); human immunodeficiency viral charge in plasma (non-detectable if less than 200 copies per ml); and CD4 lymphocyte count. Time to undetectable Kaposi's-sarcoma-associated herpesviral load, time to undetectable human immunodeficiency viral charge, and time to CD4 >or= 150 per microl were also recorded over time, from 2 mo measurements. Patients were staged according to the AIDS Clinical Trials Group-based tumor, immune, systemic staging system criteria. At baseline, Kaposi's sarcoma was progressive for 25 (96%) of the 26 enrolled patients. Complete or partial response to highly active antiretroviral therapy alone or combined with chemotherapy was achieved in 22 patients (85%). Median time to clinical response was estimated at 251 d. Clinical response was faster in patients without chemotherapy at baseline (p = 0.003) as well as in patients not previously treated with reverse transcriptase inhibitors (p = 0.0012). Using univariable analyses, predictive factors of clinical response were undetectable Kaposi's-sarcoma-associated herpesviremia (p = 0.013), undetectable human immunodeficiency viremia (p = 0.03), and relative variation of CD4 lymphocytes (p = 0.004). Using multivariable analysis, undetectable Kaposi's-sarcoma-associated herpesviremia (p = 0.009) and relative variation of CD4 (p = 0.005) were independently selected as having a predictive value for clinical response. Occurrence of nondetection of either Kaposi's-sarcoma-associated herpesvirus or human immunodeficiency virus was not associated with baseline CD4 value. Kaposi's-sarcoma-associated herpesvirus quantitative viral charge is an independent predictive factor of the efficacy of highly active antiretroviral therapy on AIDS-Kaposi's sarcoma. Our results support immune reconstitution as a mechanism of response of Kaposi's sarcoma to highly active antiretroviral therapy.

[Kaposi's disease and HHV8: a new model of virus-induced tumorigenesis]. / Maladie de Kaposi et HHV8: un nouveau modèle de tumorigenèse viro-induite?

Kaposi's sarcoma (KS) is a skin disease characterised by spindle cells proliferation and neovascularisation which, in 1994, was associated with a new Gammaherpesvirinae, named human herpesvirus 8 (HHV8). The HHV8 genome, containing more than 140 kilobases, includes genes encoding structural proteins and enzymes, and some homologues to cellular genes which could have been captured in host cells during viral evolution. Several HHV8 proteins interfere with the host cellular cycle either by inhibiting apoptosis or by positive regulation of the cell cycle (viral cyclin or v-cyclin, v-bcl-2, v-FLIP). HHV8 also contains potential oncogenes (v-IRF and v-GPCR, which promote angiogenesis, in particular the secretion of VEGF) as well as homologues of human cytokines and chemokines (v-IL6, v-MIP). HHV8 is clearly associated with KS, multicentric Castleman disease and primary effusion lymphoma. Most of the cells are infected by latent virus, resulting in persistent infection of the lesions. Only a minority of infected cells yield infectious viral particles, and their role in the development of KS and other associated diseases has not been clearly established. The molecular mechanisms and cofactors involved in the physiopathology of this infection have yet to be identified.

No evidence for a role of human herpesvirus type 8 in sarcoidosis: molecular and serological analysis.

The aim of this study was to analyse the association between human herpesvirus type 8 (HHV8) and sarcoidosis. Using nested polymerase chain reaction (PCR), we tested the presence of HHV8 DNA sequences in 13 skin specimens and peripheral blood mononuclear cells from eight patients suffering from sarcoidosis. We also looked for the presence of HHV8 antibodies in the sera of 28 patients with sarcoidosis using three techniques: two indirect immunofluorescence assays and an enzyme-linked immunosorbent assay with recombinant capsid protein fragment encoded by open-reading frame 65. HHV8 PCR analysis was negative while HHV8 serological studies showed an overall prevalence of 18% among patients suffering from sarcoidosis: 43% in patients from sub-Saharan Africa, 17% in patients from Northern Africa, 12.5% in patients from the French West Indies and 0% in French patients. In conclusion, our results do not indicate an association between HHV8 and sarcoidosis but reflect the seroepidemiology of this virus in different geographical regions.

Human herpes virus-8 and other risk factors for Kaposi's sarcoma in kidney transplant recipients. Groupe Cooperatif de Transplantation d' Ile de France (GCIF).

BACKGROUND: The exact reasons for the high incidence of Kaposi's sarcoma (KS) after kidney transplantation are still unknown. Immunosuppression is classically considered as the main risk factor, but the relative risk contributed by the patient's geographic origin and by human herpes virus (HHV)-8 infection still has to be determined. METHODS: We carried out a retrospective and a prospective study among kidney transplant recipients (TP) to identify the risk factors for posttransplantation KS. Each of 30 KS patients was matched with two controls to investigate the association with geographic origin, immunosuppressive regimen, HHV-8 antibodies before and after transplantation, and other infections. Among TP with new onset of KS, we prospectively evaluated HHV-8 serology and viremia in response to decreased immunosuppression. RESULTS: African and Middle East origins, past infection with hepatitis B, hemoglobin level <12 g/dl, lymphocyte count <750/mm3 at the time of diagnosis and initial use of polyclonal antilymphocyte sera were risk factors for KS. After multivariate analysis, origin in Africa or Middle East and use of antilymphocyte sera for induction remained as independent risk factors. Sixty-eight percent (17/25) of TP with HHV-8 antibodies before or after transplantation developed KS compared with 3% (1/33) of seronegative TP (P<0.00001). HHV-8 DNA was detectable in seven of nine peripheral blood mononuclear cells (PBMC) and in six of six KS lesions at diagnosis; it became negative in PBMC in three of five patients in parallel with tumor regression. CONCLUSION: African and Middle East geographic origins, HHV-8 infection before and after kidney transplantation, and initial use of polyclonal antilymphocyte sera were independent risk factors for KS. The presence of HHV-8 antibodies before or after transplantation was highly predictive of the emergence of posttransplantation KS and conferred a 28-fold increased risk of KS (odds ratio=28.4; 95% confidence interval: 4.9-279). Detection of HHV-8 DNA within PBMC and KS lesions seems related to tumor burden and evolution.

Clinical and biological impact of antiretroviral therapy with protease inhibitors on HIV-related Kaposi's sarcoma.

OBJECTIVE: To evaluate the clinical and biological impact of protease inhibitors on HIV-associated Kaposi's sarcoma. DESIGN AND SETTING: A cohort of 10 patients included prospectively from April 1996 to June 1997 were studied in one institutional centre after initiation of protease inhibitors. PATIENTS AND METHODS: All patients but one (stable disease) had progressive Kaposi's sarcoma. Three out of 10 patients had stopped specific chemotherapy for Kaposi's sarcoma for more than 4 weeks, three were still under chemotherapy, and four had never received specific treatment of Kaposi's sarcoma. Plasma HIV viral load, human herpesvirus (HHV)-8 viraemia in peripheral blood mononuclear cells (PBMC), and CD4 cell count were sequentially assessed from the beginning of therapy. For six patients, a semiquantitative evaluation of HHV-8 viral load in the Kaposi's sarcoma lesions was performed during treatment using polymerase chain reaction. RESULTS: After initiation of HIV triple therapy with protease inhibitors, we observed six complete responses, two partial responses, and two patients with progressive disease. All patients had undetectable plasma HIV viral load within 2 months of treatment. Undetectable HHV-8 viraemia in PBMC occurred in seven out of eight patients with partial or complete response and in none of the progressive patients. A decrease or negation of HHV-8 viral load in Kaposi's sarcoma lesions was observed in two complete responders. CONCLUSION: Our results suggest that antiviral therapy with protease inhibitors are clinically efficient in HIV-associated Kaposi's sarcoma and that there exists a correlation between clinical response and negation of HHV-8 viraemia.

Serological determination of hepatitis C virus genotype: comparison with a standardized genotyping assay.

In patients with chronic hepatitis C, determination of hepatitis C virus (HCV) genotype could be routinely run in the future to tailor treatment schedules. The suitabilities of two versions of a serological, so-called serotyping assay (Murex HCV Serotyping Assay version 1-3 [SA1-3] and Murex HCV Serotyping Assay version 1-6 [SA1-6]; Murex Diagnostics Ltd.), based on the detection of genotype-specific antibodies directed to epitopes encoded by the NS4 region of the genome, for the routine determination of HCV genotypes were studied. The results were compared with those of a molecular biology-based genotyping method (HCV Line Probe Assay [INNO-LiPA HCV]; Innogenetics S.A.), based on hybridization of PCR products onto genotype-specific probes designed in the 5' noncoding region of the genome, obtained with pretreatment serum samples from 88 patients with chronic hepatitis C eligible for interferon therapy. Definitive genotyping was performed by sequence analysis of three regions of the viral genome in all samples with discrepant typing results found among at least two of the three assays studied. In all instances, sequence analysis confirmed the result of the INNO-LiPA HCV test. The sensitivity of SA1-3 was 75% relative to the results obtained by the genotyping assay. The results were concordant with those of genotyping for 92% of the samples typeable by SA1-3. The sensitivity of SA1-6 was 89% relative to the results obtained by the genotyping assay. The results were concordant with those of genotyping for 94% of the samples typeable by SA1-6. Overall, SA1-6 had increased sensitivity relative to SA1-3 but remained less sensitive than the genotyping assay on the basis of PCR amplification of HCV RNA. Cross-reactivities between different HCV genotypes could be responsible for the mistyping of 8 (SA1-3) and 6% (SA1-6) of the samples. Subtyping of 1a and 1b is still not possible with the existing peptides, but discriminating between subtypes may not be necessary for routine use.

Significance of indeterminate third-generation hepatitis C virus recombinant immunoblot assay.

Indeterminate hepatitis C virus (HCV) third-generation recombinant immunoblot assay (RIBA3.0; Ortho Diagnostic Systems) patterns were arbitrarily defined by the manufacturer as the detection of only one antibody out of the four that were sought, namely, c100 (NS4 encoded), c22 (core encoded), c33c (NS3 encoded), and NS5 (NS5 encoded). The aims of the present study were (i) to determine the prevalence of indeterminate RIBA3.0 patterns in patients consecutively tested for anti-HCV antibodies in a university hospital; (ii) to evaluate the significance of these patterns in terms of viral replication, liver disease, and risk factors for HCV; and (iii) to get an insight into the mechanism underlying this peculiar immune response. Among 3,074 serum samples consecutively tested for anti-HCV antibodies, 588 were found to be positive by screening assays. Fifty-nine of them (10%) were RIBA3.0 indeterminate and were compared with 59 RIBA3.0-positive ones. Thirty-one RIBA3.0-indeterminate and 53 RIBA3.0-positive serum samples were HCV RNA positive by PCR (53 versus 90%; P < 10(-6). RIBA3.0-indeterminate and RIBA-3.0-positive patients with positive PCR results were not significantly different for the prevalence of risk factors for HCV infection and elevated serum alanine aminotransferase activities. Immunosuppression, attributable to coexisting human immunodeficiency virus infection, organ transplantation, or the administration of immunosuppressive drugs, was significantly more frequent in PCR-positive, RIBA3.0-indeterminate patients than in PCR-negative, RIBA3.0 indeterminate patients (P < 0.001) and PCR-positive patients with a positive RIBA3.0 result (P < 0.01). The distribution of HCV genotypes did not differ significantly between HCV RNA-positive patients with indeterminate or positive RIBA3.0 results. In conclusion, the prevalence of indeterminate RIBA3.0 patterns in virology laboratories is about 10%; in about half of these patients HCV replication is detected by PCR; the main factor responsible for indeterminate RIBA3.0 patterns could be immunosuppression, whereas HCV genotypes do not seem to play major role.

Interferon alfa therapy in patients with chronic hepatitis C and persistently normal aminotransferase activity.

BACKGROUND & AIMS: Chronic hepatitis C virus carriers may have repeatedly normal aminotransferase activity despite detectable viremia and histological hepatitis. The aim of this study was to determine the effect of interferon in this population. METHODS: Three million units of interferon alfa was administered 3 times weekly for 6 months in 10 patients with chronic hepatitis C virus infection, repeatedly normal alanine aminotransferase activity, and chronic hepatitis on liver biopsy. Serum hepatitis C virus RNA was detected by polymerase chain reaction and quantified by branched DNA before, at the end, and 1 year after treatment. A liver biopsy was performed 1 year after treatment withdrawal. RESULTS: At treatment withdrawal, hepatitis C virus RNA levels had significantly decreased, but RNA was still detectable by polymerase chain reaction in 8 of 10 patients. During the 1-year follow-up period, 6 of 9 patients had elevated aminotransferase activity on at least one occasion. One year after treatment withdrawal, RNA levels had returned to pretreatment values and no significant histological improvement was observed in the 7 patients who underwent liver biopsy. CONCLUSIONS: In patients with chronic hepatitis C and repeatedly normal aminotransferase activity, standard interferon therapy does not lead to sustained virological or histological responses despite a transient effect on hepatitis C virus replication.

Significance of anti-hepatitis C virus core IgM antibodies in patients with chronic hepatitis C.

Antihepatitis C virus (HCV) IgM antibodies were found in patients with both acute and chronic hepatitis C. The aims of the study were to determine the significance, in terms of liver disease and virological parameters, of anti-HCV core IgM antibodies in the serum of patients with chronic hepatitis C, and the possible relationship between the presence of these antibodies before treatment and biochemical and virological responses to interferon therapy. Sixty-one patients with chronic hepatitis C were studied. Tests for serum anti-HCV core IgM antibodies were carried out before treatment. The patients received 3 mega units of interferon alpha-2a subcutaneously thrice weekly for at least 3 months (6 months when alanine aminotransferase activity was normal at month 3). A biochemical response to interferon therapy was defined as normal alanine aminotransferase activity at the end of treatment (month 6: biochemical response) and 6 months later (month 12: sustained biochemical response). A sustained virological response was defined as serum HCV RNA negativity by a polymerase chain reaction-based detection method (PCR) in patients with normal alanine aminotransferase at month 12. Anti-HCV core IgM antibodies were detected in 28 of the 61 patients (46%). The prevalence of these antibodies was significantly higher in patients infected with HCV genotype 1 (including subtypes 1a and 1b) than in patients infected with other genotypes (including 2a and 3a) (57% vs. 17%; P < 0.01). No significant difference was found between IgM-positive and IgM-negative patients as regards the mean age, sex ratio, serum alanine aminotransferase and gamma-glutamyl transpeptidase activities, the prevalence of cirrhosis in liver biopsy specimens, detection of HCV RNA by PCR, and quantitation by branched DNA assay. At month 6 of interferon therapy, normal alanine aminotransferase activity was significantly more frequent in IgM-negative than in IgM-positive patients (52% vs. 21%, respectively; P < 0.02). At month 12, normal alanine aminotransferase activity and PCR negativity were significantly more frequent in IgM-negative than in IgM-positive patients (18% vs. 0%, P < 0.04). It is concluded that anti-HCV core IgM antibodies in serum are significantly more frequent in patients infected by HCV type 1 than by other types. This suggests that their overall prevalence in patients with chronic hepatitis C in industrialized countries, where HCV type 1 accounts for the majority of infections, would be of the order of 50%, that anti-HCV core IgM antibodies are not associated with characteristic features of liver disease, and that their presence before treatment is associated with a failure of interferon alpha therapy to clear the virus.

Relationship between hepatitis C virus genotypes and sources of infection in patients with chronic hepatitis C.

This study examined the relationships between hepatitis C virus (HCV) genotypes and the routes of HCV transmission in 101 patients with chronic hepatitis C. Patients who received blood transfusions (43%) and those with chronic hepatitis C of unknown cause (37%) had similar mean ages, age distribution, and HCV genotype distribution (1a, 19% vs. 14%; 1b, 52% vs. 54%; 3a, 10% vs. 9%; other, 19% vs. 23%). Intravenous drug users (IVDUs) were significantly younger and had a different genotype distribution (1a, 33%; 1b, 0; 3a, 63%; other, 5%; P < .001). Transmission of HCV 3a has been observed only over the past 20 years; other genotypes were transmitted up to 40 years ago. These results suggest that for 20 years there have been two independent ongoing hepatitis C epidemics. One affects persons who received blood transfusions or whose source of infection is unknown. These persons are older and are mainly infected by HCV 1b. The second type of infection occurs in IVDUs and infects younger persons, mainly with HCV 3a.

Influence of hepatitis C virus (HCV) genotypes on HCV recombinant immunoblot assay patterns.

Ninety-six patients with chronic hepatitis C were studied. A second-generation recombinant immunoblot assay detected anti-NS4 antibodies significantly more often in patients infected by hepatitis C virus genotype 1 than in patients infected by other types. By a third-generation recombinant immunoblot assay, the prevalences of the four antibodies measured did not differ according to the hepatitis C virus genotype.

[Cholelithiasis and ileo-ceco-cystoplasty (study of 39 patients)]. / Lithiase vésiculaire et iléo-caeco-cystoplastie (etude à propos de 39 patients).

Limited data have been reported concerning the gallstone complications of ileocaecocystoplasty. A retrospective study of gallbladder ultrasonography looking for gallstones was performed in 35 of 39 patients undergoing ileocaecocystoplasty between July 1987 and July 1991. The surgical technique used in 33 men and 2 women with a mean age of 64.5 years was that described by Thuroff (Mainz Pouch). Gallbladder ultrasonography was performed a fortnight before the operation and then regularly during follow-up, with a mean follow-up of 24 months. Eight patients, an average of 13 months after ileocaecocystoplasty, developed ultrasonographically detectable gallstones, whose prevalence in the male study population was four times higher than that of the general male population. This study raises the question of whether cholecystectomy should be performed routinely during ileocaecocystoplasty and further studies should be conducted to clarify this problem.