Structure-activity relationship studies of 4-substituted-2-guanidinothiazoles: reversible inhibitors of gastric (H+/K+)-ATPase

To investigate the incidence of child's HCV infection in our area, 637 children with different background, including 65 posttransfusion cases, 419 hepatitis patients (250 cases of acute hepatitis A, 156 cases of chronic hepatitis B and 13 cases of non-A, non-B hepatitis), 50 infantile hepatitis syndrome (IHS) infants and 103 healthy day-cared children were tested for serum anti-HCV antibody (EIA) and HCV RNA (nested PCR). It was found that posttransfusion children had significantly higher anti-HCV positive rate (30.8%) and HCV infection incidence (43.1%) than hepatitis patients (4.3% and 5.3%), IHS infants (6.0% and 8.0%) and day-cared children (2.9% and 2.9%). 25 of 33 cases with posttransfusion hepatitis (PTH) developed hepatitis C, which was the leading cause of PTH (75.8%) and NANB PTH (25/30, 83.3%). The incidence of HCV infection in NANBH patients was 23.1% (3/13) which was apparently higher than that in day-cared children (P < 0.02) and lower than that in PTH patients (P < 0.001), but not statistically different from that in AHA and CHB patients (P > 0.05). Mother-infant paired study in IHS group showed that 4 pairs of mother-infant had HCV infection, one boy aged 8 months and his mother were anti-HCV positive, and another 3 pairs possessed HCV RNA in sera. 3 of 103 healthy day-cared children were found to have inapparent HCV infection, who were anti-HCV and HCV RNA positive.     

Conjugation of 5-azido-3-oxapentyl glycosides with thiolated proteins through the use of thiophilic derivatives

Hepatitis C virus (HCV) is a leading cause of chronic liver disease. The prevalence in blood donors in the USA and West Europe is around 0.3%. However, there are few studies performed in East Europe. We have studied the prevalence of hepatitis C virus infection and risk factors associated with it in the Russian Republic of Daghestan. POPULATION AND METHODS: We included 10,682 volunteer blood donors, 267 commercial blood donors, 97 high risk patients (22 haemophiliacs, 41 in haemodialysis program and 34 parenteral drug addicts), and 87 patients with chronic liver disease (61 chronic hepatitis, 20 cirrhosis and 6 hepatocellular carcinoma). Antibodies against HCV were detected by second generation ELISA. RESULTS: 0.93% of volunteer blood donors were found to be HCV reactive. Factors with stronger association with seropositivity were previous blood transfusion and parenteral drug addiction. 66% of seropositive blood donors had an elevated level of ALT. Alcohol use correlated with more marked deterioration in liver function tests. 7.5% of commercial blood donors were seropositive. Prevalence in high risk patients and in patients with chronic liver disease was very high (50-80% and 40-50%, respectively). CONCLUSION: The Russian Republic of Daghestan has one of the lowest rates of HCV infection in East Europe. Commercial blood donors have a very high prevalence of HCV infection. The risk factors associated with HCV infection are similar to those found in other epidemiological studies.     

Detection of replicative form of HCV RNA in peripheral blood leukocytes and its clinical significance

Nested RT-PCR, done by using degenerated primer pair, was used to detect hepatitis C virus RNA (HCV RNA) in serum, plasma, liver and peripheral blood leukocytes (PBLC) of 30 patients with acute and chronic posttransfusion hepatitis C and 7 asymptomatic anti-HCV positive subjects. The results showed that the percentage of positive HCV RNA in PBLC, including both the plus and minus strands, in patients with chronic hepatitis C was significantly higher than that in acute hepatitis C and asymptomatic anti-HCV positive subjects (P < 0.05-0.001). All the 7 asymptomatic anti-HCV positive subjects did not have detectable minus strand of HCV RNA in their PBLC, serum or plasma. In 17 patients who had liver histologic examination, the positive rate of both strands of HCV RNA in PBLC of acute hepatitis (AH) was lower than that of chronic active hepatitis (CAH) (P < 0.05). Both strands of HCV RNA were detected in the liver of one AH and 6 CAH patients. The present data confirmed that PBLC of patients with hepatitis C were indeed infected by HCV. The longer the infection time, the more the chance of PBLC being infected by HCV. Patients with active liver disease (CAH) had usually higher positive rate of minus strands of HCV RNA in PBLC. In the serum and plasma of all the 37 cases, minus strand of HCV RNA was not detected and the positive rate of the plus strand of HCV RNA in their serum and plasma was similar. Futhermore, the positive rate of both plus and minus strands of HCV RNA in PBLC of 30 patients with chronic hepatitis C was also similar. It is suggested that HCV not only may infect PBLC, but also replicate in PBLC and that the occurrence of minus strand of HCV RNA is associated with activity of liver disease.     

Chronic hepatitis B and C in HIV-infected patients

BACKGROUND AND AIM: This retrospective study examined the prevalence of co-infections with hepatitis B virus (HBV) and hepatitis C virus (HCV) and the frequency of chronic hepatitis in HIV-infected patients with respect to both the different risk groups and the serological results. PATIENTS AND METHODS: All Zurich participants of the Swiss HIV Cohort Study were evaluated who had available results of hepatitis B and C serology and ALT. RESULTS: Of the total 279 patients, 52% belonged to the intravenous drug user, 34% to the homosexual, and 11% to the heterosexual risk category. Serologically, previously acquired infection with HBV alone could be demonstrated in 92 (33%), HCV alone in 9 (3%), and both HBV and HCV in 130 (47%) patients. Only 3% of patients with sexually acquired HIV infection had anti-HCV antibodies, whereas co-infection with HBV and HCV was present in 87% of intravenous drug users. Among the 222 patients with previous HBV contact, 25 (11%) had positive HBsAg and 91 (41%) had "anti-HBc alone", both assumed to represent active HBV infection. 66 (24%) of 279 patients had chronic hepatitis with ALT elevation lasting > or = 6 months. Chronic hepatitis was present in 46% of those with active HBV and HCV co-infection, in 36% of those with HCV infection alone and in 18% of those with active HBV infection alone (P < 0.001). Of the 66 cases of chronic hepatitis, 58 were associated with HCV infection, and only 2 cases had no serological signs of active HBV or HCV infection. CONCLUSION: In patients with sexually acquired HIV infection, HBV had frequently been co-transmitted. In contrast, almost all of those infected by means of intravenous drug use had a co-infection with both HBV and HCV. The latter seems to play the strongest role in the development of chronic hepatitis with persistent ALT elevation. A chronic ALT elevation was almost always associated with serologically active HBV or HCV infection.     

Prevalence of antibodies to hepatitis C virus among family members of patients with chronic hepatitis C

In this study, 108 family members of 40 chronically HCV-infected patients (19 post-transfusion and 21 sporadic), and 45 families of 16 anti-HCV-negative index cases (control group) were tested for anti-HCV antibodies. Anti-HCV antibodies were found in 16 (14.8%) families of anti-HCV-positive index cases (15% males and 14.6% females; p = NS), with no difference between families of index cases with post-transfusion and those with sporadic HCV infection. Out of the 16 anti-HCV positive family members, 12 (75%) had clinical and/or serological evidence of chronic liver damage. None of the control group subjects were anti-HCV-positive (p < 0.01). The rate of anti-HCV positivity was 34.4% among spouses, 14.3% among siblings, 16.7% among cohabitants and 2.3% among children; anti-HCV antibodies were not detected among parents. We found a positive correlation between the prevalence of anti-HCV antibodies among families and the severity of the HCV-related chronic liver damage of the index cases (p < 0.00005). In addition, to confirm that HCV infection and HCV-related chronic hepatitis may be transmitted intrafamiliarly, our findings also indicate that horizontal, especially sexual contact, is a more important route of HCV infection than vertical/perinatal transmission. Finally, the risk of acquiring HCV infection among families appears to be the highest when index cases are suffering from severe HCV-related chronic hepatitis.     

Astrocytic pathology in progressive supranuclear palsy: significance for neuropathological diagnosis

It has been shown that hepatitis C virus (HCV) infection is closely associated with mixed type cryoglobulinaemia. It is also known that HCV infection is rampant among chronic haemodialysis patients. We studied 531 renal failure patients on maintenance dialysis including 170 with positive HCV antibodies for cryoglobulinaemia, and its incidence was compared with controls which consisted of 242 chronic hepatitis C patients without renal failure and 183 healthy adults. Cryoglobulinaemia was present in 30.6% of dialysis patients with HCV infection, 10.8% of dialysis patients without HCV infection, 29.8% of patients with chronic hepatitis C without renal failure, and 0% of healthy adults. Among the 30 new renal failure patients who were started on dialysis within 6 months, four were positive for HCV antibodies, and one of them had cryoglobulinaemia; of the 26 HCV-negative patients, four (15%) were cryoglobulinaemic. The cryocrit values among dialysis patients were much lower than those of the control cases and other reports on non-dialysis cases. Patients with cryoglobulinaemia were generally younger compared with patients negative for this condition. There was no correlation between cryoglobulinaemia and past blood transfusion, underlying disease or length of dialysis. Cryoglobulinaemic patients seem to develop renal failure at relatively young ages and a considerable proportion of cryoglobulinaemic dialysis patients may have already had cryoglobulinaemia at the time of the start of haemodialysis. There was no indication that the presence of cryoglobulin in serum adversely affects the liver disease nor increases serum virus load in HCV-infected dialysis patients. Thus, it was concluded that although HCV infection has a certain role in the development of cryoglobulinaemia in dialysis patients, they develop cryoglobulinaemia less frequently and produce cryoglobulin to a lesser degree in the presence of HCV infection as compared with non-dialysis patients.     

The natural history of chronic hepatitis C in haemophiliacs

Most haemophiliacs treated with non-virally-inactivated clotting factor concentrates have been infected with hepatitis C virus (HCV). We have studied the natural history of chronic HCV infection by following all 138 HCV-positive patients from our centre for periods of up to 28 years. As well as the clinical and biochemical characteristics, we studied 116 liver samples from 63 patients obtained at elective biopsy (n = 103) or autopsy (n = 13). 36 (26%) of the patients were HIV positive, and three were chronic carriers of hepatitis B. Evidence of previous exposure to hepatitis A and B was found in 37.2% and 48.1% respectively. Raised transaminase levels were found in 82.6% of patients. 11 of 15 patients with normal transaminases tested by PCR for HCV RNA were positive, indicating that most patients, even in this group, have chronic hepatitis C infection. Cirrhosis was diagnosed by liver histology in 19 patients, and nine patients developed liver failure. The incidence of cirrhosis rose rapidly 15 years after HCV infection to 15.6 per 1000 person-years. Multivariate analysis showed that HIV status, length of time since HCV infection and age at HCV infection were independently associated with both the development of cirrhosis and liver failure. Two patients developed hepatocellular carcinoma: one of these was exposed only to a single batch of FVIII concentrate 11 years earlier. Chronic hepatitis C is increasingly recognized as a major cause for morbidity and mortality in haemophiliacs, especially those who are HIV positive and who were infected at an older age.     

The management of acute hepatitis C and needlestick accidents with hepatitis-C virus positive blood

Acute hepatitis C is rarely diagnosed, in part because of its usually subclinical course. Infection with the hepatitis C virus (HCV) has a high chronicity rate, 70-90%. The risk of infection after a needlestick accident with HCV positive blood is 3-10%. There are no efficacious preventive measures regarding HCV infection but treatment with the antiviral drug interferon alpha during the acute phase of the disease has shown to significantly reduce the risk of subsequent chronic infection. It is advised to evaluate individuals who were exposed to infected blood by a needlestick accident regarding HCV transmission, and to offer interferon treatment to them in case they become HCV positive, as demonstrated with a positive serum HCV-RNA test.     

Comparison of technetium-99m-MIBI and technetium-99m-tetrofosmin uptake by musculoskeletal sarcomas

The development of molecular biology techniques has enabled us to clone the genetic sequence of three new hepatitis viruses, namely, hepatitis C virus, hepatitis E virus and hepatitis G, or GBV-C virus in the last decade. Hepatitis C virus (HCV) is now known to account for the majority of the parenterally transmitted non-A, non-B hepatitis and hepatitis E virus (HEV) is the major cause of the epidemics of enterally transmitted non-A, non-B hepatitis in the less developed countries, mainly through contaminated water supplies. While 80% of those who were infected with HCV will become chronic carriers, there is no known chronic carrier state for HEV. To date, 6 major genotypes of HCV were identified, with various variants of genotype 6 described in Southeast Asia. In Singapore, the majority of our chronic hepatitis C patients are infected with genotype 1. The seroprevalence rate of HEV is 10% to 14% in Singapore. This probably reflects our suboptimal sanitary condition in the past rather than a reflection of a currently high HEV infection rate in our population. However, local cases of acute hepatitis E have been reported. As for the most recently cloned hepatitis G virus, its aetiologic role in the remaining cases of non-A, non-B hepatitis is still unclear. Some preliminary evidence suggests that it may be just an innocent bystander with limited pathogenicity and it certainly cannot account for all the remaining cases of non A-E hepatitis. Hence, the search for yet another hepatitis virus continues.     

Normal long axis function

Data from a surveillance system for type-specific acute viral hepatitis in Italy has been used to evaluate the risk of heterosexual transmission of hepatitis C virus (HCV) associated with sexual activity with multiple partners in subjects > or = 15 years of age. Hepatitis A cases were used as controls. During the period 1991-1996, 1,359 acute hepatitis C and 4,365 hepatitis A cases were recorded among subjects > or = 15 years of age. Intravenous drug use was the most frequent source of infection (35.9%) reported by HCV cases; two or more sexual partners during the 6 months before disease onset accounted for 34.9% of hepatitis C cases. Adjusting by multiple logistic regression analysis for the confounding effect of all risk factors considered (blood transfusion, intravenous drug use, surgical intervention, dental therapy, other parenteral exposure), and for age, sex, area of residence, and educational level of subjects, showed that having two or more sexual partners is an independent predictor of the likelihood of hepatitis C (OR=2.2; 95% CI=1.7-2.7). After excluding intravenous drug users and patients transfused with blood from analysis, the increase in the adjusted OR for the association between HCV and the number of sexual partners correlated with the increase in the number of sexual partners. The risk of hepatitis C was 2.0 times higher (95% CI=1.4-2.9) for subjects with two sexual partners and 2.8 times higher (95% CI=2.1-3.8) for subjects with three or more sexual partners, as compared to subjects with less than two sexual partners. These findings suggest that heterosexual transmission may play an important role in the spread of hepatitis C in Italy.     

Epidemiology of hepatitis C and G in sporadic and familial porphyria cutanea tarda

From 1995 to 1997, we prospectively evaluated the prevalence of hepatitis C virus (HCV) RNA in 124 patients with porphyria cutanea tarda (PCT) from Northern France (83 sporadic and 41 familial PCT). Serum samples were analyzed for ferritin, transaminases, HCV antibodies, and HCV RNA. In addition, genotyping of HCV and searches for HCV infection risk factors (blood transfusion, iv drug abuse, and surgical intervention) were performed. Twenty-six of 124 patients (21%; 95% CI: 13.9-28) were positive for serum HCV antibodies. All of them were also positive for HCV RNA. The prevalence of HCV infection was higher in the sporadic PCT group (26.5%, 22 out of 83) than in the familial PCT group (9.7%, 4 out of 41). Risk factors for hepatitis C infection were found to be significantly increased in the HCV-positive group when compared with the HCV-negative PCT group. In all HCV-positive patients with a risk factor, the suspected date of exposure to the virus always preceded the clinical onset of PCT. The HCV genotype pattern in PCT patients was similar to that observed in nonporphyric HCV patients in western European countries. Serum ferritin level was increased in both HCV-positive and HCV-negative porphyric patients. Transaminase levels were significantly higher in HCV-infected PCT patients. Sixty-seven out of 124 patients were retrospectively studied for hepatitis G virus (HGV) infection. Six of these 67 patients (8.9%; 95% CI: 2.1-15.8) were positive for HGV RNA. None of the six HGV-infected patients were positive for HCV RNA. The HGV-infected patients did not differ statistically from those without HGV infection with regard to age, ferritin, transaminase levels, and PCT treatment. These results support the view that sporadic cases of HGV infection may occur frequently. This study of a large cohort of HCV and PCT patients further documents an increasing gradient in HCV prevalence from northern to southern Europe, and shows that HCV infection acts as a triggering factor of PCT. Finally, the HGV prevalence found in the PCT patients was comparable with that found in French blood donors, suggesting that HGV is not a PCT triggering factor.     

Chronic hepatitis C virus infection after treatment for pediatric malignancy

Sera of 658 patients who had completed treatment for pediatric malignancy were analyzed by a second-generation enzyme-linked immunosorbent assay and recombinant immunoblot assay test to assess the prevalence of hepatitis C virus (HCV)-seropositivity. All HCV-seropositive patients underwent detailed clinical, laboratory, virologic, and histologic study to analyze the course of HCV infection. One hundred seventeen of the 658 patients (17.8%) were positive for HCV infection markers. Among the 117 anti-HCV+ patients, 41 (35%) were also positive for markers of hepatitis B virus infection with or without delta virus infection markers, 91 (77.8%) had previously received blood product transfusions, and 25 (21.4%) showed a normal alanine aminotransferase (ALT) level during the last 5-year follow-up (11 of them never had abnormal ALT levels). The remaining 92 patients showed ALT levels higher than the upper limit of normal range. Eighty-one of 117 (70%) anti-HCV+ patients were HCV-RNA+, with genotype 1b being present in most patients (54%). In univariate analysis, no risk factor for chronic liver disease was statistically significant. In this study, the prevalence of HCV infection was high in patients who were treated for a childhood malignancy. In about 20% of anti-HCV+ patients, routes other than blood transfusions are to be considered in the epidemiology of HCV infection. After a 14-year median follow-up, chronic liver disease of anti-HCV+ positive patients did not show progression to liver failure.     

Sutton''s disease

Acute and chronic liver diseases related to hepatitis viruses are the main indications for liver transplantation. The risk of viral reinfection after transplantation is the main limitating factor in these indications. HCV reinfection was demonstrated by demonstrating a sequence homology of the hypervariable region of HCV RNA in 2 patients before and after liver transplantation. HCV reinfection is almost constant, assessed by the persistence of HCV RNA in serum in 90% of cases. Acute lobular hepatitis appeared in 75% of patients at a median of 4 months post-transplantation with extremes between 23 days and 4 years. In our series, the 5 year actuarial rate of HCV acute hepatitis on the graft, chronic hepatitis and cirrhosis was 75%, 60% and 8%, respectively. HCV RNA level is dramatically increased after transplantation and seems to correlate with the occurrence of acute hepatitis on the graft. A positive relationship between genotype 1 b and prevalence and severity of HCV hepatitis on the graft have been suggested in European series. There is no demonstrated way to prevent HCV reinfection. The use of interferon for the treatment of HCV hepatitis on the graft was disappointing due to a poor antiviral effect and the occurrence of chronic rejection episodes in some patients. Promising results of the combination of interferon and ribavirin have been reported and need confirmation. The 5 year survival of patients transplanted for viral C cirrhosis in our Center is 78%. In conclusion, patients with endstage HCV cirrhosis are candidates for liver transplantation. Viral C reinfection is frequent, but medium term survival is good. However, longterm graft and patient survival remains unknown, and methods to prevent and treat HCV reinfection on the graft are needed.     

Chronic non-B, non-C hepatitis among blood donors assessed with HCV third generation tests and polymerase chain reaction

Forty five blood donors with increased serum aminotransferases levels had liver histologic assessment and were tested for antibodies to hepatitis C virus (anti-HCV) with second and third generation ELISAs and RIBAs, and for HCV RNA with polymerase chain reaction (PCR) in serum and liver tissue. Twenty-nine of these 45 donors (65%) had steatosis without chronic hepatitis. Sixteen (35%) had chronic hepatitis. Twelve had evidence of HCV infection. Four had no evidence of HCV infection demonstrable by ELISA, RIBA or PCR. These four patients had no known cause of chronic hepatitis and no risk factor for parenterally acquired viral infection was found in them. This observation supports the hypothesis that a non-B, non-C virus might be implicated in chronic hepatitis. However, this hypothesis remains to be demonstrated.     

Hepatitis C: Part I. Routine serologic testing and diagnosis

Hepatitis C, which is caused by the hepatitis C virus (HCV), is a major public health problem in the United States. HCV is most efficiently transmitted through large or repeated percutaneous exposures to blood. Most patients with acute HCV infection develop persistent infection, and 70 percent of patients develop chronic hepatitis. HCV-associated chronic liver disease results in 8,000 to 10,000 deaths per year, and the annual costs of acute and chronic hepatitis C exceed $600 million. An estimated 3.9 million Americans are currently infected with HCV, but most of these persons are asymptomatic and do not know they are infected. To identify them, primary health care professionals should obtain a history of high-risk practices associated with the transmission of HCV and other bloodborne pathogens from all patients. Routine testing is currently recommended only in patients who are most likely to be infected with HCV.     

Nested polymerase chain reaction (PCR) and multiple cloned antibody capture PCR for the examination of serum hepatitis B virus DNA in negative hepatitis B surface antigen patients suffering from liver diseases

In order to find out rapidly the causes of the liver diseases suffered by patients with negative hepatitis B surface antigen (HBsAg), nested polymerase chain reaction (PCR) and multiple cloned antibody capture PCR techniques were established to examine serum hepatitis B virus (HBV) DNA. By using both techniques along with the examination of hepatitis C virus (HCV) infection, the causes of chronic liver diseases with negative HBsAg were studied. It is found that nested-PCR can increase the sensitivity of single PCR more than 1,000 fold and multiple cloned antibody capture-PCR can detect concentration of HBV DNA as low as 0.1-0.01 pg/L. HBV DNA positive patients were found in 45.5%, 30.8%, 13.3% and 100% respectively of the patients suffering from liver cirhosis with negative HBsAg (group A, 22 cases), chronic hepatitis with negative HBsAg (group B, 13 cases), normal subjects with negative HBsAg and positive hepatitis B core antibody (HBcAb, group C, 30 cases) and liver cirhosis with positive HBsAg and negative HBeAg (group D, 12 cases). HBV DNA can be also found in the serum of HBsAb positive patients and subjects supposed to be healthy, 81.8% and 53.8% of the patients were infected with HBV and/or HCV in group A and group B respectively. All these results suggest that nested-PCR and multiple cloned antibody capture-PCR are rapid and highly sensitive methods for detection of serum HBV DNA. HBV infection is an important cause of chronic liver diseases in patients with negative HBsAg. The causes of most of the HBsAg-negative chronic liver diseases are related with infection of viruses. The clinical significance of serum HBsAb in naturally infected patients should be reconsidered.     

Altered expression of bladder mast cell growth factor receptor (c-kit) in interstitial cystitis

BACKGROUND/AIMS: Forty-two patients with the diagnosis of acute hepatitis C virus hepatitis were studied to investigate the relationship between hepatitis C virus genotype and progression to chronic infection. METHODS: The patients were followed for more than 1 year (mean age 29 years, male/female ratio 2.5). Intravenous drug use was documented in 15 cases, blood transfusion in four, surgical intervention, dental therapy or other parenteral exposure in 15, and unknown factors in the remaining eight. The evolution to chronicity was diagnosed on the basis of a persistent increase in transaminase levels, the presence of HCV-RNA and the histological pattern of chronic hepatitis. RESULTS: The majority of cases presented hepatitis C virus infection of subtype 1a (38.1%) or 1b (33.9%). Six cases showed the presence of genotype 3a (14.3%). Subtype 2c was observed in three out of four cases infected with genotype 2. No significant association was demonstrated with documented risk factors. The overall chronicity rate was 59.5%. This value increased to 92% in individuals infected with genotype 1b. By multivariate analysis the age-adjusted odds ratio for infection with genotype 1b as compared with all other genotypes was 14.4 (95% confidence interval; 1.52-137). Moreover, significant differences (p= 0.0002) were present in this group for histological activity index (8.7 as compared with 5-7). CONCLUSIONS: The results of this prospective study are consistent with an independent association between hepatitis C virus genotype 1b and a poor prognosis.