Hepatitis B virus depicts a high degree of conservation during the immune-tolerant phase in familiarly transmitted chronic hepatitis B infection: deep-sequencing and phylogenetic analysis.

When intrafamilial transmission of hepatitis B virus (HBV) occurs, a virus with the same characteristics interacts with diverse hosts' immune systems and may thus result in different mutations to escape immune pressure. In this study, the HBV genomic characterization was assessed longitudinally after intrafamilial transmission using nucleotide sequence data of phylogenetic and mutational analyses, including those obtained by deep-sequencing for the first time. Furthermore, HBeAg-anti-HBe profile and variability of HBV core-derived epitopes were also evaluated. Strong evidence was obtained from intrafamilial transmission of HBV genotype D1 by phylogenetic inferences. HBV isolates exhibited high degree (~99%) of genomic conservation for almost 20 years, when patients were persistently HBeAg positive with normal amino transferase levels. This identity remained high among immune-tolerant siblings. In contrast, it diminished significantly (P = 0.02) when the mother cleared HBeAg (immune clearance phase). By deep-sequencing, the quantitative analysis of the dynamics of basal core promoter (BCP) (A1762T, G1764A; A1766C; T1773C; 8-bp deletion; and other) and precore (G1896A) variants among HBV isolates from family members exhibited differences during the follow-up. However, only those from the mother showed amino acid variations at core protein that would impair their MHC-II binding. Hence, when intrafamilial transmission occurs, HBV was highly conserved under the immune-tolerant phase, but it exhibited mutations more frequently during the immune clearance phase. The analysis of the HBV BCP and precore mutants after intrafamilial HBV transmission contributes to a better understanding of how they evolve over time.

Molecular diagnosis of polycystic echinococcosis due to Echinococcus vogeli in a Paraguayan immigrant in Argentina.

Polycystic echinococcosis due to Echinococcus vogeli is a rare parasitic infection that occurs in rural areas of Central and South America. Only molecular identification performed on formalin-fixed paraffin-embedded liver tissue samples gave an unequivocal diagnosis of this disease in a Paraguayan immigrant in Argentina.

Genotypes B and C hepatocellular carcinoma-associated hepatitis B virus pre-S mutants: their detection among F1b and A2 - but not F4 - isolates from Argentina.

Prevalence rates of hepatocellular carcinoma (HCC)-associated hepatitis B virus (HBV) pre-S mutants among most genotypes are still lacking. In this study, viral (sub)genotypes of 70 Argentine nucleotide sequences (33 newly obtained) were determined by phylogenetic analysis, and the presence of such mutants was assessed in the American continent for the first time. Nucleotide substitutions of the pre-S2 start codon were observed in 10% of the HBV/A2 sequences. Ten per cent of the HBV/A2 and 12.5% of the HBV/F1b - but none of HBV/F4 - exhibited a deletion in the pre-S1/pre-S2 region. The contribution of these variants to liver cirrhosis (LC) and/or HCC development among HBV/F and HBV/A isolates deserves further prospective clinical studies.

Risk factors for acute pancreatitis in patients with migrating gallstones.

Stool screening for gallstones and ultrasound monitoring of diameter changes of the biliary and pancreatic duct were performed in 129 patients with choledocholithiasis. Gallstone migration was found in 44 patients, all of whom were operated on electively. At surgery, acute pancreatic lesions were found in 16 patients; in the remaining 28 there was no evidence of pancreatic inflammation. There were no significant differences among patients in both groups regarding sex, age, stone size, shape or number found in stools, interval between admission and migration, or the presence of a dilated pancreatic duct before migration. Pancreatic duct reflux, however, was significantly more frequent in cholangiograms of patients with acute pancreatitis, implying that a common channel may be a major factor relating to acute pancreatitis in patients with migrating gallstones.

Epidemiological surveillance of human hydatidosis by means of ultrasonography: its contribution to the evaluation of control programs.

Epidemiological surveillance for hydatidosis is based on initial determination and follow-up of the infestation rate in man, sheep and dogs. The use of ultrasonography (US) as a screening method has demonstrated its usefulness in detecting asymptomatic human carriers of abdominal hydatidosis. To evaluate the contribution of US to epidemiological surveillance within the framework of disease control programs, we performed 719 US studies in school children from 7 to 13 years of age, in rural areas of Río Negro, Argentina, subjected to canine deparasitization during 1984/1986 and 1997/1998. In the first period, 15/268 (5.6%), while in the second, 5/451 (1.1%) carriers were detected (P < 0.0004). The average diameter of the cysts was 4.71 cm in 1984/1986 and 2.14 in 1997/1998. US as a mass screening method allows evaluation of early changes in human prevalence rates, closely related to infestation rates in sheeps and dogs, thus providing a sensitive indicator of the evaluation of control programs.

[Human hydatidosis: field echography for the determination of groups at high risk in the evaluation of a control program]. / Hidatidosis humana: ecografía de campo para la determinación de grupos de alto riesgo en la evaluación de un programa de control.

The development of small ultrasonographic equipment has permitted to incorporate this technique to the methods of early detection of Human Hydatidosis. 689 echographies were carried out in this experience, and 5.51% of compatible images in the whole population, and 12.24% in groups of risk (those living together with the operated ones) were found. It was observed a significant decrease in the infection percentages in man in the whole population of areas under Control Program, using the echographic screening. In conclusion, echography might be incorporated to the epidemiologic surveillance system of human Hydatidosis.

[Propranolol and 5-isosorbide mononitrate in patients with cirrhosis: systemic and portal hemodynamic events]. / Propranolol y 5-mononitrato de isosorbide en pacientes con cirrosis: episodios hemodinámicos sistémicos y portales.

AIM: To study the acute variations in portal and systemic hemodynamics after propranolol and 5-isosorbide mononitrate (IMN) administration in cirrhotic patients. PATIENTS AND METHODS: Seventeen cirrhotic patients with portal hypertension were studied with catheterization and Doppler duplex Ultrasound Systemic hemodynamics. Hepatic venous pressure gradient (HVPG), portal blood flow and resistance were evaluated in baseline, after intravenous propranolol (0.15 mg/kg), and after 20 mg p.o. of IMN. Patients who showed a decrease > or = 20% and/or < 12 mm/hg in HVPG were considered responders. RESULTS: There were no significant differences in clinical or portal hemodynamic baseline data between responders and non-responders to the drugs. After propranolol administration cardiac index decreased (p < 0.05) and pulmonary capillary pressure increased (p < 0.0001). Six patients (35%) were responders; lack of response was associated with an insufficient decrease in portal blood flow or with an increase in portal resistance. After IMN administration cardiac index decreased (p < 0.05) with normalization of pulmonary capillary pressure (p < 0.05). Seven patients were responders to the addition of IMN (5 non-responders to propranolol) and showed a decrease in HVPG associated with a reduction in portal blood flow and resistance; in the remaining 10 patients HVPG did not decrease despite a reduction in portal blood flow, with an increase in portal resistance. CONCLUSIONS: Addition of IMN increased the number of responders and reduced portal blood flow with a variable effect in portal resistance.

Relationship between diversity of hepatitis C quasispecies and histological severity of liver disease.

The aim of this work was to assess if the diversity of hepatitis C virus (HCV) quasispecies is related to histological severity and duration of infection in a cohort of untreated patients with an estimated onset of the disease. A total of 27 patients with diagnosis of chronic liver disease and history of blood transfusion (n = 16) or intravenous drug use (IDU) (n = 11) were included. All were anti-HCV positive and had detectable serum HCV-RNA. The onset and the duration of the disease were estimated from the time of the transfusion or the first drug injection. Patients who consumed drugs for more than 2 years, or were coinfected with HBV or HIV were excluded. History of alcohol intake (> 80 g/day), ALT level and age at infection were recorded. Histological assessment of grading and staging was performed according to Knodell score. The quasispecies diversity was investigated by single strand conformation polymorphism (SSCP) targeted to HVR-E2 region and SSCP pattern was evaluated as a single or multiple bands. The number of quasispecies did not correlate with the estimated duration of the disease. Patients who acquired hepatitis C by blood transfusion did not differ in number of bands from patients who were IDU. There was no correlation between the heterogeneity of HCV quasispecies and age, serum ALT, Knodell score, HAI and fibrosis. In conclusion the quasispecies diversity of E2 had no correlation with grade and stage of chronic HCV infection and the presence of quasispecies was independent of the duration of the disease.

[The problem of hepatitis C virus infection in blood donors. Clinico-epidemiological, virological and histological evaluation]. / Problemática de la infección por el virus de la hepatitis C en donantes de sangre. Evaluación Clínico-epidemiologica, virologica e hestologica.

Our aim was to identify prospectively, with a case-control survey, risk factors for hepatitis C virus (HCV) infection in volunteer blood donors and to assess the histological features and their correlation with transaminase (ALT) level and viremia. In a Liver Unit of a referral-based University Hospital, 248 blood donors were evaluated for risk factors, according to definitive ELISA test and 132 were considered true positive. Of these, 132 anti-HCV(+) blood donors were age and sex-matched with the anti-HCV-negative group (n = 116). There was a high frequency in the anti-HCV(+) group of intravenous drug abuse (IVDA) (22%), history of major surgery (20.4%), tattooing (12.1%), non IVDA (17.4%), and multiple sexual partners or history of sexual transmitted diseases (25.7%). At least one risk factor was identified in 76.52% of the antiHCV(+) donors vs 34.4% in the anti-HCV (-) group (p = 0.000). A total of 71 patients accepted a liver biopsy; chronic liver disease was present in 85.9% (n = 61) (mean Knodell score 6.75). ALT was elevated in 69% (n = 49) and HCV RNA was detectable in 76% of patients. It can be concluded that in our study 76.5% of anti HCV positive blood donors showed at least one risk factor for HCV infection detected by a second highly efficient interview. Twenty two percent admitted to prior intravenous drug use although this disqualifies them for blood donation, but was not identified by the screening process. Most blood donors with anti HCV(+) had chronic hepatitis C regardless of their serum ALT levels. Normal ALT did not exclude liver disease.

Diagnosis of human hydatidosis. Predictive value of a rural ultrasonographic survey in an apparently healthy population.

The usefulness of ultrasonography (US) in the early diagnosis of hydatidosis, applied in large-scale surveys to populations lacking clinical symptoms of the disease, has been amply documented. However, the rate of false positive and negative results is poorly described. Due to this, the present paper is aimed to evaluate the sensitivity, specificity and predictive value of a conventional rural ultrasonographic survey in comparison with higher imaging complexity. Accordingly, during 1997 and 1998 a total of 1054 children from 7 to 14 years of age were evaluated by means of US, in the town of Ingeniero Jacobacci, Province of Rio Negro, Argentina, employing a portable device for population studies. All detected cases were referred to a high complexity center specialized in imaging diagnosis for their re-evaluation with US, CT scanning and X-rays. A control group comprising 3 children negative by US for each positive case in the mass screening survey was selected and reexamined by US and X-rays and CT scanning in doubtful situations. Twenty-seven asymptomatic carriers were referred with images compatible with hydatid cysts, while 66 were classified as disease free. At reexamination, 24 of those diagnosed as carriers and the totality of those classified as healthy were confirmed. On the basis of our results, a sensitivity of 100%, a specificity of 95.6% and a global test value of 96.7% were estimated.

Genomic and phylogenetic analysis of hepatitis C virus strains from Argentina.

HCV genomic characterization was performed by nucleotide sequence analysis (n=50) combined with restriction fragment length polymorphism (RFLP) of the 5' UTR region in 82 isolates corresponding to different Argentine groups. Genotype 1 was detected in 70.7% of the samples (58 out of 82), genotype 2 in 21.9% (18 of 82) and genotype 3 in the remaining 6 sera (7.3%). HCV 1b subtype contributed with 35.3% to the whole population studied (29 to 82) and was detected in 6 out of 21 sporadic cases. Besides their epidemiological significance, these results should be taken into account when future vaccines are considered on the basis of geographical HCV genotypic prevalence.

[Echography of non-lithiasic pathology of the gallbladder. Personal experience]. / Ecografía de la patología no litiásica de la vesicula biliar: experiencia personal.

Ultrasonography offers rich information of the gallbladders wall and its contents. Its high sensibility and specificity depends of the type of pathology. Its impossible to pretend an echographic histologic diagnosis but the sensibility of this method to reveal macroscopic alterations that correspond with anatomic macroscopic pathology is of sufficient hierarchy to induce a precocious surgical exploration. 85 cases of non lithiasis pathology of the gallbladder is presented. They were found in a revision of 2831 hepatobiliary echographic studies. A correlation with surgery and other methods with the ultrasonographic finding is made.

[Echographic specificity in the early diagnosis of human hydatidosis]. / Especificidad de la ecografía en el diagnóstico precoz de la hidatidosis humana.

Ultrasonography (US) were used for screening of hydatid disease in 1321 asymptomatic individuals fron Río Negro Province, Argentina. A total of 76 positive subjects were identified with possible hydatid disease. Surgery was performed in 30 patients, conforming in 28 of them sonographic diagnosis (93.34%).

[An echographic, serologic and radiologic register of human hydatidosis. Contributions to a control program]. / Catastro ecográfico, serológico y radiológico en hidatidosis humana. Aporte a un programa de control.

Between June and November 1984, 904 asymptomatic people from endemic areas of hydatidosis in Rio Negro, Argentina, were studied by means of ultrasonography (U.S.) and double diffusion for arc five (dd5). The population included 272 inhabitants from Pilcaniyeu, 55 patients from the Zonal Bariloche Hospital admitted for diseases other than hydatidosis and 577 recruits from different departments of the Rio Negro Province. A chest X-ray was performed in every recruits. 47 (5.20%) cases of hepatic hydatidosis were detected by U.S.; 11 (1.22%) were detected by dd5 (p 0.01); 2 (0.34%) were detected by chest X-ray. Due to the low sensitivity of dd5 a presumptive diagnosis of hydatidosis should be made in every patient proceeding from an endemic area with a liver cyst diagnosed by ultrasound, even if dd5 is negative. U.S. must be incorporated as an elective method associated with chest X-ray and dd5 in epidemiological yielding and monitoring of control programs for hydatidosis.

[Professional risk: hepatitis B. Vaccination strategies in a general hospital]. / Riesgo profesional: hepatitis B. Estrategias de vacunación en un hospital general.

The risk of contracting hepatitis B: (HBV) by health workers is widely accepted. In 1989 our Hepatology Service started a voluntary anti-HBV vaccination program, employing recombinant vaccine (SKF) by intramuscular route with a 0-1-6 month schedule after screening with antibody against the anti-core HBV antigen (AntiHBc Elisa Abbott). Initially, it was planned to monitor antibody titers against superficial antigen (Anti-HBs) 30 days after the last dose. An epidemiological form listing personal data, working area, profession, seniority, written consent for blood extraction and tentative acceptance of vaccination, was completed by 357 hospital staff members. After serological screening, only 184 (51%) workers agreed to receive vaccination. Given the paucity of volunteers, an attempt was made to explain this degree of reluctance by a randomized blind voluntary survey, to which 349 hospital staff members and 40 medical students replied. Questions were related to knowledge concerning vaccination in general, hepatitis and particularly hepatitis B, and specific anti-HBV vaccination. An appraisal of data gathered disclosed a considerable lack of information not only on the risk of HBV infection and its complications, but also on the existence of a suitable vaccine. Non-existent adverse effects of vaccination were mentioned, including AIDS (Acquired Immuno-Deficiency Syndrome), hepatitis and cirrhosis, among others. To overcome this obstacle, we held a two-day workshop on hepatitis B prevention and prophylaxis intended for medical and ancillary staff. After the meeting, which were attended by 221 members, 48 individuals, comprising 25 physicians and 23 nurses, spontaneously requested to be vaccinated.(ABSTRACT TRUNCATED AT 250 WORDS)

[Evaluation of propranolol response by catheterization and Doppler ultrasonography in patients with cirrhosis]. / Evaluación de la respuesta al propranolol por cateterismo y ecografía Doppler en pacientes con cirrosis.

AIMS: To analyze the changes in portal pressure, blood flow and resistance after propranolol administration, and to assess the predictive value of the variations of Doppler Duplex Ultrasonography (DDU) measurements according to the response of the hepatic venous pressure gradient (HVPG). PATIENTS & METHODS: 30 cirrhotic patients were studied. Assessment of systemic hemodynamics and HVPG were performed in baseline and after intravenous propranolol administration (0.15 mg/kg). Patients who showed a decrease > or = 20% &/or < 12 mm/Hg in HVPG were considered responders. The DDU study was performed in blind conditions, in baseline and after propranolol. Measurement of blood flow of the portal vein, splenic vein and femoral artery were performed. Portal resistance was calculated as HVPG/portal blood flow. RESULTS: All patients were beta blocked and 14 (47%) were responders. There were no significant differences in systemic or splachnic hemodynamic baseline data between responders and non responders. Femoral blood flow decreased in both groups. Splenic and portal blood flow decreased significantly only in responders. No significant difference was found in the variation of portal resistance between responders and non responders; when these changes were considered individually, a great variability was found in both groups. A decrease > or = 15% in splenic blood flow showed a positive predictive value of 88%, a lack of a similar decrease in portal blood flow showed a negative predictive value of 86%. CONCLUSIONS: The decrease in portal blood flow was the main factor in determining the response to propranolol.

[Pancreatic tumor: an unusual presentation of an occult breast carcinoma]. / Tumor de páncreas: presentación infrecuente de un carcinoma de mama oculto.

A case of cholestasis due to a synchronous pancreatic head metastasis from an occult lobular breast carcinoma is presented. The patient had a clinical and radiological picture compatible with a pancreatic head primary tumor with cholestasis and ascites. Ultrasonographically guided fine needle aspiration cytology demonstrated a metastatic breast lobular cancer (positive for cytokeratin AE1 and AE3, cytokeratin 7 and epithelial membrane antigen and negative for cytokeratin 20, CA 19.9, CA 125, CEA and estrogenic receptors). The same cytologic findings were observed in skin and subcutaneous armpit nodules. Clinical and radiological breast examination was unable to demonstrate any tumor in the breast. Pancreatic metastases are rare events and the majority of them are secondary to renal and lung cancer and rarely to breast cancer. In these latter cases, metastases are usually disclosed after a disease-free interval of months or years between primary tumor resection and recognition of the pancreatic tumor. Synchronous presentation is extremely rare. Metastases of epithelial origin are uncommon in pancreas and generally are first misdiagnosed as primary pancreatic cancer. Fine needle aspiration is a useful tool for the differential diagnosis in patients with widespread disease.

Prevalence of hepatitis C in health care workers investigated by 2nd generation enzyme-linked and line immunoassays.

The aim of this study was to evaluate the prevalence of antibodies to hepatitis C virus (anti-HCV) in health-care workers (HCW). Sera from 439 unselected HCW were assessed for anti-HCV by 2nd generation enzyme-linked immunoassay (ELISA) and anti-HBc by ELISA. Anti-HCV (+) sera were evaluated by line immunoassay (LIA) (LiaTeK, Organon). Anti-HCV proved positive by ELISA in 12 (2.73%) subjects, 6 of whom were reactive by LIA, one was indeterminate and 5 non reactive. The prevalence of anti-HCV confirmed by LIA was 1.59% (7 subjects). Positive anti-HCV results with an ELISA ratio greater than 3 were LIA reactive in 6/6 as compared with 5 LIA non reactive with an ELISA ratio less than 2, while in the indeterminate serum the ratio was 2.5. No differences in age, profession, seniority, history of hepatitis or transfusions were found between anti-HCV (+) and (-) subjects, but females predominate significantly. The areas of higher risk were hemodialysis, obstetrics, surgery and intensive care. Anti-HBc was (+) in 85.7% (6/7) of the anti-HCV (+) subjects. Follow-up of anti-HCV (+) subjects showed raised alaninoaminotransferase levels in 4 cases, while liver biopsies in 3 disclosed cirrhosis, chronic active hepatitis and chronic persistent hepatitis. The anti-HCV prevalence in HCW is low compared with other risk groups perhaps due to the peculiar epidemiological features of HCV. In low risk groups for HCV infection a positive ELISA result with a ratio lower than 3 should be confirmed by more specific tests.

HBV and HCV infection in kidney transplant recipients.

The risk of HBV and HCV liver infection in kidney graft recipients was evaluated in 35 patients. All were tested for anti-HBc, HBsAg, HBeAg, anti-HBs, Anti-HBe, anti-HCV (c-100-3 and c-100-3, c-22, 33-c), anti-HDV and anti-HIV by ELISA, and for HBV-DNA by hybridization. Liver biopsy, immunostaining for HBcAg and Knodell's hepatic inflammatory index were performed in 18. Mean time elapsing form transplant to inclusion was 20.7 months (range 1-108). HBsAg was the only marker searched for prior to transplant. Twenty six (74.2%) patients presented HBV and/or HCV markers, while 9 (25.8%) had none; 16 (45%) proved anti-HBc+, 6(17.1%) HBsAg+, (3 HBeAg+ and 3 anti-HBe+), 7 (20%) anti-HBs+ and 3 (8.5%) isolated anti-HBc. Anti-HCV (C-100-3) was positive in 9/32 (28.1%), while 2nd. generation anti-HCV was positive in 20/35 (57.1%) cases. No false positives for 1st. generation test were found. Both anti-HDV and anti-HIV were negative in all the sample. Raised aminotransferases were present in 13/30 (43.3%), 7 in anti-HCV+, one in HBsAg+ and 3 in HBsAg+/HCV+ cases, but normal in 17/30 (56.6%). History of Transfusion and Hemodialysis time showed no significant differences between anti-HCV+ and anti-HCV negative cases. Biopsy disclosed 10 chronic persistent hepatitis (CPH), one chronic active hepatitis (CAH) with cirrhosis, one inactive cirrhosis (Ci) 4 minimal lesions (MHL) and 2 normal. Seven CPH, 3 MHL. one normal and both cirrhosis cases proved anti-HCV+. HBsAg was positive in the single CAH, in 2 CPH and in one MHL.(ABSTRACT TRUNCATED AT 250 WORDS)