Dynamics of hepatitis B virus quasispecies heterogeneity in association with nucleos(t)ide analogue treatment determined by MALDI-TOF MS.

Minor drug-resistant variants may preexist in every subject infected with hepatitis B virus (HBV). However, understanding the dynamic of genotypic evolution within the HBV population requires accurately following allele frequencies through time. We used MALDI-TOF MS (matrix-assisted laser desorption-ionization time-of-flight mass spectrometry) for localization and quantitative allele frequency detection to investigate preexisting HBV quasispecies and the genotypic evolution of drug-resistant variants during nucleos(t)ide analogue therapy. We found a significant difference between the genotypic evolution of drug-resistant variants depending on response to treatment.

Coexistence of HFE and rare UGT1A1 genes mutations in patients with iron overload related liver injury.

This report describes two patients hospitalised in Hepatology Unit, Infectious Diseases Department Medical University of Gdansk because of liver damage discovered in family doctor's practice. Hereditary hemochromatosis was diagnosed in both cases. Diagnosis was established basing on medical records review, and biochemical, molecular and liver specimen tests. The analysis of polymorphism of UGT1A1 gene was done in these cases because those patients were a part of the larger study on prevalence of UGT1A1 gene mutations in patients with hereditary hemochromatosis. We discovered rare variant forms of UGT1A1 gene coexisting with HFE gene mutations.

Hepatitis delta virus infection in chronically HBV-infected patients from northern Poland.

The objective of this study was to estimate the presence of hepatitis delta virus RNA in chronically HBV-infected patients from northern Poland. Three out of 63 studied samples (4.8%) were positive in a qualitative test for total antibodies to HDV antigen. Five samples (7.9%) turned out to be HDV-RNA-positive by RT-PCR, four of them were sequenced in the region of L-HDAg, and phylogenetic analysis was performed. All four examined samples belonged to genotype I. Two RNA-positive/anti-HD-negative samples possessed a few uncommon nucleotide substitution sites within the L-HDAg sequence, which could suggest unique variants in the Polish population of HDV-infected patients.

Value of magnetic resonance cholangiography (MRC) in primary sclerosing cholangitis (PSC).

BACKGROUND: Magnetic resonance cholangiography is a relatively new method, hence it has not gained a proper status yet in the diagnostic algorithm in patients with bile duct diseases. Many authors believe that MRC is an effective method for the visualisation of bile ducts. The aim of the study is to evaluate MRC usefulness in patients with PSC. MATERIAL AND METHODS: The study material consisted of 4 patients with PSC (3 men and 1 woman) aged 23-50 years. Patients were subjected to liver MRI in transverse plane, and T1 and T2-weighted images were obtained. Additionally, magnetic resonance cholangiography was performed with Gyroscan 0.5 T unit and Magnevist contrast medium. RESULTS: The patients manifested cholestasis and hepatitis of different duration (range: 2-14 years). The biochemical analyses revealed FA ranging from 563 to 1114 IU/l, GGTP 157-1270 IU/L, AlAT 56-295 IU/L, AspAT 80-190 IU/L. Three out of four patients displayed significant increase in the level of ANCA antibodies. Histopathological investigation of the liver, conducted in 3 out of 4 patients, showed cholangitis et pericholangitis. ERCP was attempted in all the patients. In one case, irregular, overlapping and narrowed bile ducts were found, while in two cases it was impossible to fill intrahepatic ducts. ERCP was not performed due to technical obstacles encountered in one patient. All the subjects underwent MRC. In three cases, the segmental irregular dilatation of intrahepatic ducts was observed, while in one case intrahepatic ducts were poorly marked. The defect in the filling of initial segment of common bile duct was diagnosed in one patient. Another patient had a lesion adjacent to common bile duct--low signal intensity in T1 and the intensification after the administration of contrast medium indicate the presence of expansive process. CONCLUSIONS: 1. MRC is an accurate examination enabling the visualisation of all bile ducts in their natural size. 2. Simultaneous MRI of the liver allows for the diagnosis of focal lesions. 3. MRC may be used in those cases, where ERCP cannot be performed for technical reasons or when it is objected by the patient.

Autoimmune reactions in HBV and HCV.

BACKGROUND: Hepatotropic viruses HBV and HCV are capable of triggering autoimmune reactions (AIH). The aim of the study was the assessment of clinical course of AIH in patients with HBV and HCV, and the revision of treatment methods employed in these cases. MATERIAL AND METHODS: Among 120 patients with AIH and those qualified for antiviral therapy, 21 (17.5%) subjects were selected, including 16 with HCV (7 men and 9 women) and 5 with HBV (including 2 men and 3 women). AIH diagnosis was based on international criteria taking into account biochemical tests, autoantibodies and morphological picture of the liver. HBV infection was confirmed with the determination of HBV markers and HCV--with the presence of antiHCV and HCV RNA. RESULTS: The duration of infection at AIH diagnosis was difficult to assess in patients with HCV. AIH was diagnosed when patients were qualified for antiviral treatment. Three patients displayed high anti-LKM1, 8--ANA and SMA, two pts--ANA, two pts--SMA, one pt--ANA + pANCA and one--SMA + pANCA. Nine patients did not show hyperproteinaemia, and hypergammaglobulinaemia was not observed in 2 patients. Six patients suffered from other immunity disorders--thrombocytopenia, vasculitis, arthritis, visceral lupus erythematosus. The diagnosis of chronic hepatitis was confirmed by morphological examinations in 15 patients. Ten subjects received adrenocortical hormones, 3 patients were treated with adrenocortical hormones and azathioprine, 4 received interferon and 3 received no treatment. All patients with HBV proved HBeAg(-). In these patients, AIH symptoms developed 5-18 years after the diagnosis of HBV infection. Liver biopsy confirmed the diagnosis in 4 patients. SMA was observed in 2 subjects, ANA and SMA--in the remaining patients. All the subjects manifested typical biochemical changes as well as high IgG values. Extrahepatic exponents of immune process were observed in 3 patients. Three subjects were treated with Encorton (Prednisone), while 2 patients received Encorton and Azathioprine. CONCLUSIONS: Patients with HBV and HCV infections may manifest the features of AIH in the course of the disease, which requires careful attention while selecting treatment.

Correlation between liver damage and degree of gastric mucose colonization by Helicobacter pylori in subjects with parenchymatous liver damage.

BACKGROUND: Helicobacter pylori (H. pylori) is an important factor responsible for chronic inflammatory conditions of the gastric mucosa. It has been demonstrated in numerous animal studies that some Helicobacter species may cause parenchymatous liver damage. The aim of the study was to investigate whether there is any correlation between the incidence of parenchymatous liver damage, and the incidence and degree of colonization of the gastric mucosa by H. pylori. MATERIAL AND METHODS: The study was carried out in the group of 30 patients (14 females, 16 males) whose mean age was 37 years, hospitalized because of parenchymatous liver damage without clinical symptoms of cirrhosis. All the patients had gastroscopy and urease tests performed, and mucosal biopsies were taken for immunomorphological investigations. The patients were divided into groups, group I comprising those with positive, and group II with negative urease test results. RESULTS: Positive urease tests were obtained in 26/30 patients (group I), 18/26 of whom demonstrated macroscopic changes of the gastric mucosa visible in gastroscopy. Group II with negative urease test results comprised 4/30 patients, 2/4 of whom had detectable changes in the gastric mucosa. The presence of H. pylori antigens was demonstrated by gastric mucosa immunomorphology in all 30 patients. The degree of invasion of H. pylori was visualized by immunofluorescence, which allowed to differentiate deep mucosal invasion of H. pylori (bacterial antigens present in lymph follicles and at the base of muciferous glands) observed in group I in 14/26 and in group II in 1/4 cases and superficial invasion (epithelium and mucosal surface) observed in group I in 12/26, in group II in 3/4. CONCLUSIONS: The obtained results may suggest more frequent H. pylori infections in subjects with parenchymatous liver damage than in the population without liver damage. Immunofluorescence seems to be a highly sensitive method allowing for detection of even small degrees of gastric mucosa colonization by H. pylori.