Prevalence of TT virus infection in Italian dialysis patients.

OBJECTIVE: A novel DNA virus, transfusion-transmitted virus (TTV) is identified from the serum of a patient from acute hepatitis non-A-E. Few reports have been published on patients with renal disease. Patients on dialysis were at high risk of blood-borne viral infections and little is known about the prevalence of this virus in dialysis patients. In order to verify the prevalence of infection from TTV in these patients, we have examined the incidence of TTV in Italian patients with dialysis. METHODS: Serum samples of 85 patients and 65 healthy individuals were examined. In order to evidence the presence of the TTV virus, a method of the seminested polymerase chain reaction (PCR) with TTV-specific primers derives from the N22 region deIl'ORF-1 of the virus has been used and products were analyzed by agarose-gel electrophoresis. All serum samples were also analyzed to markers HBV and HCV. RESULTS: The prevalence of TTV DNA in dialysis patients [35/85 (41.7%)] was significantly higher than in healthy population [7/65 (10.7%)]. Among TTV positive dialysis patients, HCV coinfection was present in six cases. The positivity rate for TTV-DNA tends to increase with age. CONCLUSION: Transfusion-transmitted virus had a high prevalence in Italian-dialysis patients. In our study the virus did not have an important clinical effect on patients; but remains the possibility that it may aggravate liver disease caused by HCV. However, the question of whether TTV infection might have a possible effect on dialysis patients requires further investigation in larger groups.

Synthesis and in vitro selective anti-Helicobacter pylori activity of pyrazoline derivatives.

In order to develop new anti-Helicobacter pylori agents, a series of N1-substituted 3,5-diphenyl pyrazolines P1-P13 was prepared and evaluated for their antibacterial activity. All synthesized compounds showed little or no activity against different species of Gram-positive and Gram-negative bacteria of clinical relevance and against various strains of pathogenic fungi. The same derivatives exhibited a significant degree of activity against a range of H. pylori strains, including those resistant to the reference compound metronidazole. Among the prepared compounds those with an N1-acetyl group and a 4-methoxy substituent in the 5-phenyl ring showed the best activity against H. pylori metronidazole resistant strains in the 1-4 microg/mL MIC range.

Low-dose intramuscular hepatitis B immune globulin and lamivudine for long-term prophylaxis of hepatitis B recurrence after liver transplantation.

The combination of lamivudine and hepatitis B immunoglobulins (HBIg) to prevent recurrence of HBV hepatitis has significantly improved the survival of patients transplanted for HBV-related end-stage liver disease. Generally, HBIg are administered intravenously. We evaluated the efficacy, tolerability, and cost savings of long-term intramuscular HBIg and lamivudine in 28 patients (23 men and 5 women), who received liver transplants for acute or chronic HBV-related liver disease. Twelve patients started lamivudine before and 16 at the time of liver transplantation. HBIg were administered intravenously during the first week (50 to 70,000 IU) and intramuscularly thereafter (1200 IU every 3 to 6 weeks) to maintain an HbsAb titer >100 IU/L. Mean follow-up was 20 +/- 13 months. Only one patient experienced HBV recurrence (9 months after transplantation). This patient had failed to follow the scheduled prophylaxis. Cumulative survival at 3 years was 83%. Intramuscular HBIg were well tolerated in all cases. Cost analysis comparing intramuscular vs intravenous HBIg administration showed that 39,490 Euros were saved per patient per year. These preliminary results show that low-dose intramuscular HBIg and lamivudine are efficacious and cost-effective for long-term prophylaxis of hepatitis B recurrence after liver transplantation.

Occult hepatitis C virus infection in type II mixed cryoglobulinaemia.

Mixed cryoglobulinaemia, when not secondary to other well-defined immunological disorders, is commonly associated with hepatitis C virus (HCV) infection. However, a minority of cases lack evidence of HCV infection and are, therefore, defined as 'true essential' mixed cryoglobulinaemias. We thoroughly investigated three such patients to determine the aetiology of this disorder. Antibodies to HCV (anti-HCV) and HCV RNA, detected by sensitive enzyme-linked immunosorbent and polymerase chain reaction assays in serum and in concentrated cryoglobulins, were repeatedly negative in the three patients. Despite the lack of evidence for HCV infection, two of them were still treated with interferon alpha-2a assuming unrecognized viral infection. Both patients demonstrated excellent clinical and laboratory responses, but cryoglobulinaemia relapsed after the withdrawal of therapy. At the time of relapse, HCV RNA genomic sequences were detected for the first time in the cryoprecipitates of both patients. In the third case, HCV RNA was demonstrated for the first time during a flare of cryoglobulinaemia coincident with varicella infection. In all three patients anti-HCV antibodies remained negative throughout follow-up. We conclude that some apparently 'essential' forms of mixed cryoglobulinaemia can be caused by occult HCV infection. Interferon therapy can be taken into consideration in such HCV-negative cases.

Use of oligoprobes on amplified DNA in the diagnosis of bacterial meningitis.

An approach based on the 16 S rDNA polymerase chain reaction (16S PCR) and oligoprobe hybridization was applied to 77 cerebrospinal fluid samples submitted to the clinical microbiology laboratory for culture. Broad-range 16S rDNA primers were selected in conserved regions of the gene. Oligoprobes specific for Neisseria meningitidis, Haemophilus influenzae, Streptococcus spp., and Mycobacterium tuberculosis were selected in specific variable regions of the amplified 600 base pairs (bp) in the 16S rDNA. None of the oligoprobes cross hybridized with DNA from the other bacteria or from common contaminants. There were no false-negative results in culture-positive cerebrospinal fluid samples. Ten cases of meningitis caused by bacteria other than the four probes were not identified by any of the four probes. In culture-negative cerebrospinal fluid samples with some abnormal chemical parameters, there were 14 amplicons -- one of Haemophilus influenzae, three of Streptococcus spp., six of Mycobacterium tuberculosis, and four not identified -- while in normal cerebrospinal fluid samples there were no amplicons.

Diagnostic value of anti-hepatitis C virus (HCV) core immunoglobulin M in recurrence of HCV infection after orthotopic liver transplantation.

The significance of anti-hepatitis C virus (HCV) core immunoglobulin M (IgM) and its relationship with genotypes, alanine aminotransferase abnormality, and histological data were studied for 18 patients who had undergone orthotopic liver transplantation due to HCV-related end-stage disease. During follow-up, IgM response seemed to be associated with the recurrence of HCV infection but did not correlate with abnormal alanine aminotransferase levels and histological data. In addition, the results of this study indicated that the detection of HCV RNA is critical for diagnosis of reinfection in liver transplantation.

Recurrence of hepatitis C virus infection after orthotopic liver transplantation: role of genotypes.

In this study, we evaluated the correlation between alanine aminotrasferase levels and hepatitis C virus genotypes in liver transplant patients. We studied 18 patients who had undergone orthotopic liver transplantation because of end-stage cirrhosis (n = 9) or hepatocellular carcinoma (n = 9) hepatitis C virus related. Serum HCV-RNA testing was performed monthly on all the 18 series of serum samples from the first week after liver transplant until the end of the follow up, this period ranging from 1 to 39 months. After liver transplantation, serum HCV-RNA was detected in 14 patients (78%). Of the 8 patients infected with subtype 1b. 1 remained asymptomatic, 2 developed acute liver failure and 5 developed chronic hepatitis. In patients infected with types 1a (Choo et al., 1989), 2a (Choo et al., 1989), with a mixed infection 1b/3 (Kuo et al., 1989) or with an undetermined genotype, significant laboratory abnormalities were not observed. Recurrence of hepatitis C virus infection after liver transplantation is common, and recurrent hepatitis occurs in 50% of cases. Genotype 1b appears to be associated with a higher rate of recurrent hepatitis, compared to other genotypes.

Hepatitis C virus in patients with porphyria cutanea tarda: relationship to HCV-genotypes.

Porphyria cutanea tarda (PCT) is a rare metabolic disorder characterized by an abnormal porphyrin metabolism and typical cutaneous lesions. Recently a strong association between PCT and hepatitis C virus (HCV) has been proposed. Studies in south Europe have shown high prevalence (53 to 91%) of HCV markers in patients with PCT. We studied HCV genotypes in 72 subjects: 40 with PCT and 32 patients with chronic liver disease. A high rate of HCV-RNA positive PCT patients (84%) was observed, reflecting an active HCV replication, the genotypes study showed a prevalence of genotype 1b in PCT patients (61.2%). These findings implicate HCV in the aetiology of PCT-associated liver disease suggesting that hepatitis C serological and virological testing could be indicated in all patients with PCT.

Different reactivity against HCV proteins (RIBA 3) according to progression of liver damage.

BACKGROUND: Aim of the study was to assess the correlation between clinical stage of HCV-related liver disease and viraemia to immune response to different viral antigens. METHODS: We considered 1330 patients with HCV chronic infection followed up from 6 months up to 6 years divided into two groups according to RIBA 3 (Abbott) response: Group I, 1231 patients with positivity for at least two bands (83 subjects with asymptomatic infection, 941 with chronic hepatitis, 201 with cirrhosis and 6 with HCC); Group II, 99 patients with positivity at only one band (45 with asymptomatic infection, 53 with chronic hepatitis and 1 cirrhotic). RESULTS: We noticed a major percentage of positive patients for at least three bands in more severe clinical forms (90% of chronic hepatitis or cirrhosis versus 60% of asymptomatics, p < 0.005, chi 2 test). Moreover we noticed a percentage increase of positivity for antibodies anti-c100 and anti-NS5 with the progression of liver damage, statistically significant differences between asymptomatics and patients with chronic forms. We also observed that viraemia is related neither to clinical stage nor to different reactivity to RIBA 3, albeit viraemia is usually detected more frequently among patients with liver damage, but unrelated to different reactivities. CONCLUSIONS: Our results show a clear correlation between number of reactivities towards HCV proteins and progression of liver damage, pointing out that immune response plays a direct role in the long-term outcome of HCV infection.

IgM anti-hepatitis C virus in patients with chronic non-A, non-B hepatitis and their relationship to viral replication.

Patients with hepatitis C virus (HCV) infection may have different patterns of antibody response to various structural and non-structural viral antigens. We have correlated the serological patterns to the clinical features of chronic infection and to viral replication in 68 HCV-Ab-positive patients with chronic liver disease at different stages (19 with cirrhosis-hepatocellular carcinoma, 38 with chronic active hepatitis and 11 with chronic persistent hepatitis). Serum samples from each patient were assayed for HCV-IgM by enzyme immunoassay and for HCV-RNA by the polymerase chain reaction using primer sets derived from the 5'-non-coding region. The prevalence of HCV-IgM was high (54 patients (79.4%)) and the study showed a good correlation between high values of anti-HCV-IgM and the presence of HCV-RNA in serum, since HCV-RNA was detected in 35 of the 54 IgM-positive patients (64.8%) and notably in 19 of the 20 subjects with high levels of specific IgM. Conversely, all the 35 sera containing HCV-RNA were also reactive for HCV-IgM, while none of the HCV-IgM-negative sera was HCV-RNA reactive. Positivity rates for both HCV-RNA and IgM anti-HCV were higher in the more advanced stages of disease; thus, the clinical pattern of HCV chronic hepatitis seems to be strictly related to the serological pattern and the presence of HCV-RNA.

Evaluation of indeterminate sera for hepatitis C virus by assays using synthetic peptides.

The development of new immunodiagnostic systems to study antibodies anti-HCV based on the use of synthetic peptides are potentially very important in the evaluation of indeterminate samples. We have compared two immunodiagnostic tests, the 3-RIBA and LIA-HCV test system, to examine the samples rated as indeterminate by the 2-RIBA test. The results showed that the specificity of the new tests has improved, in fact 29/40 of the indeterminate samples studied became positive with both tests. However, the 3-RIBA test showed higher sensitivity, since two samples negative by LIA-HCV, were positive with this system.

Detection of antibodies to hepatitis C virus in dialysis patients.

Dialysis patients are at risk for infection by a variety of blood-borne agents transmitted within dialysis units. The development of the hepatitis C virus (HCV) screening test prompted many studies on the prevalence of anti-HCV among dialysis patients. The authors have evaluated the prevalence of anti-HCV in 405 hemodialysis patients both by Elisa screening and 4-RIBA test system with a follow-up of two years. The study showed a difference in the incidence of antibodies to HCV by year. In 1990, 15.2% were positive with an increase to 20.8% in 1991. There was an increase of 5% in dialysis patients and only 1.9% in the personnel working in the dialysis ward. Another control group of volunteers did not show any positivity. In addition, the correlations of the antibodies against HCV with markers of hepatitis B virus (HBV) and history of transfusion were evaluated. These findings suggest that the patients found to be positive should be dialyzed on separate machines and special precautions must be undertaken to reduce the risk of transmission.

Effect of cyclosporin A on natural killer cells' response during viral infections.

In the this study the modification of lymphocyte subsets (T3, T4, T8) and Natural Killer (NK) cells in organ transplanted patients treated with Cyclosporin A (CyA) in the course of viral infection, have been analyzed. Different subsets have been studied with the monoclonal antibody method and infective processes have been verified by serological data of seroconversion. Our study has shown that CyA at the adopted doses does not alter NK response to viral infection; in fact, in patients with seroconversion, higher NK values and lower OKT4/OKT8 ratio values have been found with respect to patients who did not show any viral infection serologic data. Furthermore an increased incidence of reject crisis has been observed in patients with seroconversion.

[Sero-epidemiological results in subjects receiving a renal transplant]. / Indagine sieroepidemiologica in soggetti sottoposti a trapianto renale.

In this study a sero-epidemiological investigation on 127 renal allograft recipients was examined. In these patients, treated with Cyclosporine A or conventional drugs, antibody response to various antigens (Cytomegalovirus, Herpes simplex, Varicellae/Zoster and Mycoplasma pneumoniae) was examined. The data were compared to the healthy population and dialyzed subjects.

In vitro susceptibility tests with cefoperazone compared with cefotaxime, cefoxitin, cefuroxime, cephaloridine and gentamicin.

Minimum inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs) and sensitivity disc tests of cefoperazone, cefotaxime, cefoxitin, cefuroxime, cephaloridine and gentamicin were determined from 300 Gram-positive and -negative isolates from kidney transplant recipients. Cefoperazone inhibited 50% of Pseudomonas aeruginosa strains at a concentration of 6.25 micrograms/ml. Only gentamicin had similar activity but if we consider the sources of our strains and the nephrotoxicity of the compound, its use is not suitable. Moreover, cefoperazone was able to inhibit from 50% to 90% of the other strains at variable concentrations between 0.39 micrograms/ml and 3.12 micrograms/ml.