HBV mutations and their clinical significance.

PURPOSE: The launch of the vaccine against HBV in 1983 significantly reduced the number of HBV infections in the world. But there still remain a large group of people with chronic hepatitis B who were infected before beginning of vaccination programs or in whom the vaccine was - for various reasons - ineffective. Current therapy of HBV infection based on PEG-IFN α-2a or nucleotide/nucleoside analogues does not guarantee sustained virologic response in the large majority of chronically infected persons. Treatment with some nucleoside analogues is associated with mutations and subsequent selection of resistant strains resulting with therapeutic failure, risk of cross-resistance to other drugs and finally selection of mutants with oncogenic properties.

Treatment of chronic hepatitis C in a patient with Fanconi anaemia.

Fanconi anaemia is a rare autosomal recessive disorder with progressive bone marrow failure and predisposition to malignancy. We report a case of a 26-year-old female patient with Fanconi anaemia and severe chronic active hepatitis C virus infection. Her past medical history included treatment with multiple blood transfusions and bone marrow transplantation at the age of 13. The decision to treat the infection was taken, and history of hematologic disease contributed to the introduction of therapy with leukocyte interferon-α n3 and ribavirin combined with a granulocyte - colony stimulating factor. The treatment was well tolerated and after 48 weeks a reduction of the viral load and alanine aminotransferase activity were achieved. No adverse effects on bone marrow functioning were noted.

Concentrations of ssDNA in liver tissue and its correlation with sFas and sFasL in serum of patients infected with HBV, HCV, HCV and HIV.

PURPOSE: The concentration of nucleic acids that undergo apoptosis (ssDNA) determines the actual activity of programmed cell death. ssDNA concentrations in liver tissue of patients with chronic HBV, HCV and HCV and HIV infections were assessed. The concentration of this nucleic acid was analyzed in relation to the concentrations of serous apoptosis indicators, sFas and sFasL receptor proteins, the activity of inflammatory processes and fibrosis in liver tissue as well as HBV, HCV and HIV viraemia. PATIENTS: The study included 153 patients: 48 chronic HBV infected, 86 chronic HCV infected and 19 HCV and HIV infected. PATIENTS AND METHODS: The concentrations of HBV-DNA, HCV-RNA and HIV-RNA were determined by use of RT-PCR method. CD3+, CD4+ and CD8+ lymphocytes count were detected in HIV infected patients' blood by use of a flow cytometer. The concentration of ssDNA was determined by use of monoclonal antibodies and ELISA tests. The concentrations of sFas and sFasL in serum were determined by use of an immunoenzymatic method (ELISA). RESULTS: The concentration of ssDNA in liver tissue of both HCV and HBV infected patients was higher in comparison to those co-infected with HCV and HIV (1332 x 10(-6) g/mg, +/-664 x 10(-6); vs 1508 x 10(-6) microg/mg, +/-810 x 10(-6); vs 886 x 10(-6) microg/mg, +/- 388 x 10(-6); p < 0.004). No correlation between ssDNA concentration and HBV and HCV viraemia was observed. In patients infected with HCV genotype 3, the concentration of ssDNA was 1343 x 10(-6) microg/mg, +/-700 x 10(-6), comparable from patients infected with genotype 1, 296 x 10(-6) microg/mg, +/- 615 x 10(-6). The highest concentration of ssDNA in liver tissue was detected in HBV infected patients with low inflammatory activity (1645 x 10(-6) microg/mg, +/-987) and low fibrosis (1606 x 10(-6) microg/mg, +/- 876 x 10(-6). Mild inflammatory changes and low fibrosis were observed in all HCV and HIV infected patients. No correlation between ssDNA concentration in liver tissue and HIV viraemia (r = 0.03; p = 0.90), HCV, CD8+ and CD4+ count (r = -11; p = 0.66) was observed. The concentration of ssDNA among HCV and HIV infected patients correlated with the concentration of sFas in serum (r = 0.52; p < 0.02). CONCLUSIONS: HCV, HBV and HIV viraemias do not correlate with ssDNA concentration in liver tissue. In patients with HCV and HIV infections, CD4+ and CD3+ counts do not correlate with the concentration of ssDNA in liver tissue. HIV infection seems to inhibit apoptosis processes in liver tissue of HCV and HIV co-infected patients. In the case of HCV and HIV infections, the concentration of sFas in serum correlates with the concentration of ssDNA in liver tissue.

Levels of HBV-DNA, sFas and sFasL among healthy HBsAg carriers in period of three years.

PURPOSE: The object of the study was the usefulness of sFas and sFasL concentration in the prognosis of disease development in healthy HBsAg carriers. PATIENTS: 34 healthy HBsAg carriers were examined over a three-years period. MATERIAL AND METHODS: HBV-DNA was extracted using the Gene Elute Mammalian Genomic DNA Miniprep Kit (Sigma, USA). HBV-DNA concentration and YMDD mutations were measured by RT-PCR based on TaqMan Universal Master Mix (Applied Biosystems, USA). HBeAg and anti-HBe in serum were detected by MEIA method (ABBOTT, Germany). The concentration of sFas and sFasL in serum was estimated by ELISA method (Bender MedSystems, Austria). RESULTS: Within three year observation period the number of carriers with absent HBV-DNA increased from 19% to 33%. HBV-DNA above 105 copies/ml, which was detected in 63% of carriers, decreased to 11% (p < 0.05). After 3 years, a reduction of HBV-DNA levels was observed in 89% of carriers (p < 0.05). The occurrence of sFasL decreased from 56% to 48%. sFasL correlated with HBV-DNA (p < 0.05). The concentration of sFas decreased (p < 0.01). Chronic hepatitis B developed in 11% of men carriers, and 11% eliminated HBeAg, anti-HBe and HBV-DNA. YMDD mutant was not detected in any of the HBsAg carriers. CONCLUSIONS: High concentration of sFasL in serum may suggest the development of chronic hepatitis and it seems that sFasL detection is never a good prognostic factor.

HBV-DNA and sFas, sFasL concentrations in serum of healthy HBsAg carriers.

PURPOSE: Increased HBV-DNA concentration is a prognostic factor of disease progression in chronic hepatitis B patients. Moreover, active hepatic inflammation during HBV replication influences apoptosis intensification. The aim of this study was to estimate occurrence of HBV replication among carriers of HBsAg. Furthermore, we analysed the correlation between HBV replication and HBeAg or anti-HBe presence as well as known apoptosis indicators--sFas and sFasL concentration. MATERIAL AND METHODS: The study included 34 HBV infected patients, aged 20-43 yrs defined as HBsAg healthy carriers. HBV-DNA was extracted from patients' serum using two different DNA isolation kits: the QIAamp DNA Mini Kit (QIAGEN Ltd, USA) and the Gene Elute Mammalian Genomic DNA Miniprep Kit (Sigma, USA). HBV-DNA concentration in serum was measured by RT-PCR based on TaqMan Universal Master Mix (Applied Biosystems). The detection limit of this system was as few as 10 HBV-DNA copies/mL of serum. HBV-DNA concentration was calculated from a linear standard curve obtained between 10 and 10(8) DNA copies/reaction. HBeAg and anti-HBe in serum were detected by MEIA method (ABBOTT, Germany). The concentration of sFas and sFasL in serum was-estimated by ELISA method (Bender MedSystems, Austria). RESULTS: HBV active replication was detected in 79% HBsAg carriers. The HBV-DNA levels exceeding 10(5) copies/mL were observed in 64% patients. Among HBsAg carriers presenting HBeAg, HBV replication occurred more often and was more intensify than in HBsAg carriers presenting anti-HBe antibodies. The sFasL occurrence in serum of 56% HBsAg carriers shows an active apoptosis, independent from ALT and AST activity within normal ranges.

The incidence of HLA DRB1*0103 in ulcerative colitis patients in north-eastern Poland.

BACKGROUND/AIMS: Susceptibility to inflammatory bowel disease is partially genetically determined and the HLA (human leukocyte antigen) alloantigens and genes located in the HLA region have been studied over the course of many years as the candidate genes responsible for ulcerative colitis. Improvements in molecular genotyping have allowed disease association with HLA to be narrowed down to specific subtypes. For class II antigens, increasing phenotype frequency of DRB1*0103, DRB1*1502 is observed and positive correlation to disease susceptibility is proposed. We investigated the incidence of HLA DRB1*0103 in ulcerative colitis patients in North-Eastern Poland and possible association with overall disease susceptibility and clinical course of the disease. METHODOLOGY: 41 patients and 45 healthy control blood donors were included in this study. All subjects were Polish. RESULTS: The incidence of HLA DRB1*0103 was low (2.44%), but was associated with fulminant course of the disease (pancolitis with megacolon toxicum). None of the ethnically matched healthy control blood donors possessed the HLA DRB1*0103 allele (0.00%). CONCLUSIONS: The results gained in the presented study confirm, that in the Polish population HLA DRB1*0103 allele is uncommon and it would not be a useful marker of disease susceptibility.

Search for hepatitis delta virus (HDV) infection in hepatitis C patients in north-eastern Poland. Comparison with anti-HDV prevalence in chronic hepatitis B.

Hepatitis C virus and hepatitis D virus have been shown to suppress HBsAg synthesis. Thus it is possible that HDV infection occurs despite the lack of detectable HBsAg. The aim of our study was to (a) determine the prevalence of HDV infection in patients with chronic hepatitis C (b) compare it with the prevalence of HDV infection in HBsAg positive patients with hepatitis B. The study group consisted of 51 chronic hepatitis C patients, 30 HIV infected drug addicts (27 of them were also positive for anti-HCV) and 102 hepatitis B patients. The participants were tested for anti-HDV, anti-HCV and HBsAg. All anti-HCV positive patients were negative for anti-HDV. Four individuals with anti-HDV belonged to hepatitis B group and constituted 3.9% of all HBsAg positive subjects. We conclude that (a) there is currently no evidence of HDV infection among HCV infected patients in our region (b) hepatitis delta infection is rare in north-eastern Poland.

The levels of IL-1beta, IL-4 and IL-6 in the serum and the liver tissue of chronic HCV-infected patients.

The pro-inflammatory interleukines play a major role in the progress of chronic hepatitis C. Among the patients with chronic hepatitis C virus (HCV) infection, the morphology of the liver was assessed and the levels of serum and liver-tissue IL-1beta, IL-4 and IL-6 were determined. The levels of the cytokines were related to the liver-tissue changes. RNA-HCV was measured by the RT-PCR method. Cytokine levels of the serum and liver tissue were measured by the Quantikine High Sensitivity test. The levels of serum IL-1beta, IL-4 and IL-6 (0.221, 0.104 and 1.393 pg/ml) in all HCV patients were higher in comparison with healthy adults (0.188, 0.025 and 0.600 pg/ml). The levels of liver tissue IL-1beta, IL-4 and IL-6 (4291.3, p<0.05; 1624.6, p<0.05; 1158.7 pg/g protein) in all HCV patients were higher compared with patients with liver cirrhosis without HBV or HCV infection (2319.9, 553.6 and 756.2 pg/g protein). Patients with HCV infection demonstrated a significant correlation between serum and liver-tissue levels of IL-1beta (Pearson: 0.61, p<0.05) and IL-4 (Pearson: 0.51). The level of serum IL-6 in patients with moderate chronic active hepatitis was higher when compared with patients with mild chronic persistent hepatitis. Among the patients with mild chronic persistent hepatitis, the levels of liver-tissue IL-6 were higher compared with those with moderate chronic active hepatitis. There was no correlation between histology changes and the levels of serum and liver-tissue IL-1beta and IL-4.

[Plasmapheresis in medical practice]. / Zastosowanie plazmaferezy w praktyce lekarskiej.

Therapeutic plasmapheresis means the removing of patient's plasma with simultaneous administration of blood products. The expected therapeutic effect is connected with mechanical removal of certain compounds present in plasma: abnormal cellular metabolites, antigens, immunoglobulins and immunoglobulin complexes. The procedure activates immune system. Therapeutic plasmapheresis has been used in the management of different diseases. However, some complications associated with this therapy have been noted and therefore critical evaluation of this therapeutic method is necessary.

[Chronic viral hepatitis C in patient with Hand-Schüller-Christian disease]. / Przewlekle wirusowe zapalenie watroby typu C u pacjenta z choroba Handa-Schüllera-Christiana.

Chronic hepatitis C often occurs among patients with different diseases and may influence their natural history. From the other hand coexisting diseases may modify the hepatitis C infection. We present a case of a male patient with the histiocytosis X, earlier called Hand-Schüller-Christian disease with hepatitis C virus infection. The mild course of the chronic hepatitis C, not requiring anti-viral therapy, may be the result of immunomodulation relevant to Hand-Schüller-Christian disease.

[Activation of acute phase proteins in patients with chronic hepatitis C treated with interferon-alpha 2a]. / Aktywnosc bialek ostrej fazy u chorych na przewlekle zapalenie watroby typu C leczonych interferonem alpha 2a.

The course of chronic viral hepatitis C (CVH C) depends in large degree from on non-specific immunological response. Interferon-alpha 2a (IFN-alpha 2a) affects immunological responses including acute phase proteins. The aim of the study was to determine whether chronic hepatitis C affects acute phase proteins and whether treatment with interferon-alpha restores disturbed acute phase proteins concentrations. Concentrations of acute phase proteins were compared with aminotransferases activity, a standard indicator of hepatocyte damage. We studied twenty patients (from 21 to 53 years old) with CHV C treated with interferon-alpha 2a (6 million units, three subcutaneous injections a week). CRP, fibrinogen, C3 and C4 components of complement and albumin serum concentrations were determined before treatment, after 6 and 36 doses of interferon (after 2 and 12 weeks of therapy). Alanine aminotransferase activity was checked at the same time. No abnormalities in CRP, fibrinogen and albumin levels were observed before and during the treatment. It is regarded that low concentrations of C3 and C4 can lead to cryoglobulinemia, vasculitis, and polineuropathy. Both C3 and C4 were decreased among patients with CVH C. During therapy with interferon; levels of both proteins increased and achieved normal levels, especially among patients with effectiveness therapy of IFN-alpha.

[Liver biopsy: the mandatory test in liver diagnosis in adults]. / Biopsja watroby--niezbedny element w diagnostyce chorób watroby u doroslych.

The aim of the study was clinical and morphological analysis patients with liver biopsy. In this work technique of percutaneous liver biopsy, indications, contraindications and complications were presented. Among 547 liver biopsy 56% patients was HCV infected and a 14% patient was HBV infected. Microscopic examination was performed the similar percentage the stage of inflammation and fibrosis. Among patients without viral infection (HBV and HCV) the most finding was steatosis and liver cirrhosis. Agreement of clinical diagnosis with results of microscopic investigations with reference to of inflammable changes carried out 90%, the less agreement with steatosis, cholestasis and hemosiderosis. In 30% patients after biopsy were pains in place executed of intervention and right arm for 6 hours. 0.5% of patients was sub capsular bleeding after intervention. We no observed dangerous complications after liver percutaneous biopsy. Liver biopsy is very important investigation in liver diagnosis, comparatively safety if contraindications are complains.

[The joint changes and presence of HLA-B27 antigen in patients with ulcerative colitis]. / Zmiany stawowe oraz wystepowanie antygenu HLA-B27 u chorych z wrzodziejacym zapaleniem jelita grubego.

The extra-bowel manifestation can be found among patients with ulcerative colitis (UC). The most common are the joint changes. A group of 18 patients with ulcerative colitis was investigated. The physical and radiological signs of joint inflammation and presence of HLA-B27 antigen in blood were examined. Appearance of HLA-B27 antigen among 28% of patients with ulcerative colitis can support involvement of autoimmunologic mechanism in joint inflammation. The joint changes (swelling of small joints of hand, presence of nodes, sacroileitis, and particularly spondyloarthritis) were observed in 50% of patients.

The levels of IL-1 nu beta, IL-2, IL-4, IL-6 and IFN-gamma among patients with chronic hepatitis type C treated with IFN-alpha.

Interferon regulates the activity of other cytokines. Alterations in non-specific immune response, particularly cytokine level changes result in therapeutic difficulties in patients with chronic hepatitis C. Serum levels of IL-1 beta, IL-2, IL-4, IL-6 and IFN-gamma were measured in patients with chronic hepatitis C before the interferon therapy, in the early phase of the treatment (after the 6th dose of the drug), and in the later stable phase of the treatment (after the 36th dose of the drug). Concentrations of cytokines during the interferon therapy were compared to the changes in alanine aminotransferase activity. The IL-1 beta, IL-2, IL-4 and IL-6 serum concentrations were initially elevated in all patients. The increase in IL-1 beta level after the 6th dose and the decrease after 36th dose of IFN appeared to be an indicator of a good response to treatment. High levels of IL-6 before the IFN therapy followed by a decrease after the 6th dose and high levels of IL-4 followed by a decrease after 36th IFN dose also indicated good response. High levels of IL-2, stable throughout the treatment suggested a poor response. No changes in the IFN-gamma values before or during the treatment were observed.

[Exposure to toxocariasis and tularemia among workers at Bialowieza National Park]. / Zagrozenia tokosokaroza i tularemia pracowników Bialowieskiego Parku Narodowego.

Everyday contact of the workers of Bialowieza sanctuary with animals may create a risk of infection by microorganisms attacking animals. The reported study was undertaken for carrying out an epidemiological analysis of tularemia and toxocarosis in this group of workers. Tularaemia infection was not found in then. In 12% of these workers antibodies to Toxocara canis were found, and fire workers had articular borreliosis. Antibodies to taxocara were disclosed in 10% of the Bialowieza population not employed in the sanctuary and 30% of the population of the village Kruklanka in the Pisz Forests. The workers of the Bialowieza Sanctuary are not exposed to Toxocara canis infection more than the remaining population of Bialowieza and the population in the Kruklanka village in a region far from Bialowieza.

[Practical use of anti-HB immunoglobulins]. / Praktyczne zastosowanie immunoglobuliny anty HBs.

Viral hepatitis B remains a significant problem among iatrogenic infections. The development of transplantology met a barrier in the form of active hepatitis B. Wide but still inadequate prophylaxis against viral hepatitis B and the necessity of protection of patients after organ transplantation against secondary HBV infection require establishing indications for specific, passive immunotherapy.

[The neuropsychiatric side effects of the interferon therapy in patients with chronic hepatitis B and C]. / Neurologiczno-psychiatryczne objawy niepozadane leczenia interferonem u chorych na przewlekle zapalenia watroby typu B i C.

Interferons are usage in therapy patients with chronic viral hepatitis (ch). Independently apart from therapy effects, lots of side effects are observed. The neuropsychiatrical side effects are most often and maybe hinder of interferon therapy. Among patients with ch B and C were estimated efficiency, type and frequency of side effects after interferon therapy. Especially symptoms disorders of central nervous system were observed between all side effects. The 67 patients: 20 with ch B, 45 with ch C and 2 with ch B retherapy by interferon were estimated. The lymphoblastoid interferon: alpha n1, Wellferon (Wellcome/Great Britain) and recombinant interferon alpha 2b, Roferon (Roche/Switzerland) and Intron-A (Schering-Plough/USA) were administrated. The efficiencies of treatment were observed after 6 month from finished interferon therapy. Elimination of polymerase HBV-DNA, antigens: HBs, HBe and antibody HBcIgM and normalization of aminotransferase activity in serum, were evaluated in patients with ch B. Elimination of HCV-RNA and normalization of aminotransferase activity in serum was evaluated in patients with ch C. Among neuropsychiatrical side effects was study. The frequently of irritability, fatigue neurosis, disorders of psychomotion activity, vegetative symptoms, somatic symptoms and disorder of higher mind processes. Grow excitability of sensorimotor, affective, irritability of vegetative system, were examined among irritability symptoms. Deficiency activity of polymerase HBV-DNA was observed in 40% patients with ch B. Deficiency of RNA-HCV in serum was observed in 31% patients with ch C. The flu-likely syndrome was observed at the beginning interferon therapy in 85% patients. Among patients treated with interferon were observed side effects: hair loss (29% patients), disorder of vision (29%), thrombocytopenia (< 50,000 mm3) (29%), leukopenia (< 2,000 mm3) (16%). The neurasthenia was detected in 60% patients with ch B and 50% patients with ch C. Neurasthenia most often manifested by irritable and quickly exhaustion of strength. 53% patients with ch B and 42% patients with ch C manifested disorders of psychomotion activity. Disorders of higher mind processes were in 50% patients with ch B and 33% with ch C. The neurological side effects were not influence on modification therapy. The flu-likely syndrome was observed at the beginning interferon therapy in majority patients treat with interferon. The neuropsychiatrical disorders are frequently watching of interferon therapy in patients with ch B and C.

Efficacy of interferon therapy on patients with HCV infection in dependence from the time of treatment.

Among 45 patients with chronic hepatitis C the efficiency of interferon alpha 2b treatment was evaluated. The efficiency of interferon therapy was determined after 6 months of treatment in the group of 28 patients (group I) treated for 6 months and 17 patients (group II) treated for 12 months. The side effects were investigated with respect to such a factor as time of treatment. 11 (39%) patients from group I had eliminated HCV RNA and 14 (50%) patients had normalized AlAT levels in the serum. Five (29%) patients from group II eliminated HCV RNA and 8 (47%) patients had normalized AlAT levels in the serum. The flu-like syndrome, thrombocytopenia, vision disorder, depression and somnolence were most often the side effects observed in treated patients. Proportionally to time of treatment vision disorder, depression and somnolence increased.

Plasmapheresis in the treatment of tropical malaria.

A case of tropical malaria imported from Kenya was described. Plasmodium falciparum (an etiological agent of this disease) is often drug resistant, therefore pharmacological treatment of the patients was supplemented with plasmapheresis. Full recovery has been achieved. It is suggested that plasmapheresis may be recommended for a complex therapy of severe forms of tropical malaria.

[Use of plasmapheresis in patients with AIDS and peripheral polyneuropathy]. / Zastosowanie plazmaferezy u chorych na AIDS z polineuropatiami obwodowymi.

The plasmapheresis is procedure removal of plasma and simultaneously, together with plasma, toxins, incorrect metabolites, antigens and immunological complexes. In patients with HIV infection often appearance periphery polyneuropathy. This is connected with autoantibody action on the damage of neurone. The aim of this study was evaluation effectiveness of plasmapheresis in treatment patients with AIDS and polyneuropathy. Seven patients with AIDS--C3 and polyneuropathy coexistence were treatment by plasmapheresis. The symptoms of polyneuropathy decrease in all patients. Were don't influence on immunological system and side effect performance plasmapheresis.