Krzysztof Piotr Bielawski

Publications of Krzysztof Piotr Bielawski

• Iron overload and HFE gene mutations in Polish patients with liver cirrhosis.

  Authors: Katarzyna Sikorska, Tomasz Romanowski, Piotr Stalke, Ewa Iżycka-Świeszewska, Krzysztof Piotr Bielawski

  Hepatobiliary & pancreatic diseases international : HBPD INT. 06/2011; 10(3):270-5.

  Increased liver iron stores may contribute to the progression of liver injury and fibrosis, and are associated with a higher risk of hepatocellular carcinoma development. Pre-transplant symptoms ofIncreased liver iron stores may contribute to the progression of liver injury and fibrosis, and are associated with a higher risk of hepatocellular carcinoma development. Pre-transplant symptoms of iron overload in patients with liver cirrhosis are associated with higher risk of infectious and malignant complications in liver transplant recipients. HFE gene mutations may be involved in the pathogenesis of liver iron overload and influence the progression of chronic liver diseases of different origins. This study was designed to determine the prevalence of iron overload in relation to HFE gene mutations among Polish patients with liver cirrhosis. Sixty-one patients with liver cirrhosis included in the study were compared with a control group of 42 consecutive patients subjected to liver biopsy because of chronic liver diseases. Liver function tests and serum iron markers were assessed in both groups. All patients were screened for HFE mutations (C282Y, H63D, S65C). Thirty-six of 61 patients from the study group and all controls had liver biopsy performed with semiquantitative assessment of iron deposits in hepatocytes. The biochemical markers of iron overload and iron deposits in the liver were detected with a higher frequency (70% and 47% respectively) in patients with liver cirrhosis. There were no differences in the prevalence of all HFE mutations in both groups. In patients with a diagnosis of hepatocellular carcinoma, no significant associations with iron disorders and HFE gene mutations were found. Iron disorders were detected in patients with liver cirrhosis frequently but without significant association with HFE gene mutations. Only the homozygous C282Y mutation seems to occur more frequently in the selected population of patients with liver cirrhosis. As elevated biochemical iron indices accompanied liver iron deposits more frequently in liver cirrhosis compared to controls with chronic liver disease, there is a need for more extensive studies searching for the possible influence of non-HFE iron homeostasis regulators and their modulation on the course of chronic liver disease and liver cirrhosis.

• MRSA distribution and epidemiological procedures evaluation at two hospitals in Northern Poland.

  Authors: Krystyna Paszko, Ewa Michnowska, Julianna Kurlenda, Mariusz Grinholc, Joanna Nakonieczna, Krzysztof Piotr Bielawski

  GMS Krankenhaushygiene interdisziplinär. 01/2011; 6(1):Doc19.

  In the present study we have analyzed the impact of modified MRSA screening of carriers and patients on epidemiological situation of MRSA during 2008-2010, comparing two regional hospitals withIn the present study we have analyzed the impact of modified MRSA screening of carriers and patients on epidemiological situation of MRSA during 2008-2010, comparing two regional hospitals with similar bed numbers and similar ward profiles in Northern Poland. In 2008 the proportion of MRSA to all S. aureus isolates was 14.4% resp. 6.0%, in 2009 8.3% resp. 4.7% and in 2010 6.5% in both hospitals. Independent of the different prevention and intervention strategy in both hospitals the different MRSA incidence seems to be due to regional epidemic settings.

• [The influence of hepatitis B virus polymorphism on the progression of chronic liver disease].

  Authors: Magda Rybicka, Piotr Stalke, Urszula Charmuszko, Krzysztof Piotr Bielawski

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2011; 65:244-54.

  Hepatitis B virus (HBV) infection is one of the major human health problems worldwide. It is estimated that chronic HBV infection affects more than 350 million people globally. It is one of theHepatitis B virus (HBV) infection is one of the major human health problems worldwide. It is estimated that chronic HBV infection affects more than 350 million people globally. It is one of the leading causes of liver cirrhosis and hepatocellular carcinoma. High genetic variability is a characteristic feature of HBV as the viral polymerase lacks proofreading activity. The nucleotide substitution rate for HBV is 10-fold higher than for other DNA viruses. Genetic variations of HBV influence the clinical outcome of HBV infection. There are eight genotypes of hepatitis B virus (A-H) that have a distinct geographical distribution. There is clinical significance of HBV genotype in terms of disease activity, risk of progression to cirrhosis, the development of hepatocellular carcinoma and response to antiviral treatments. Moreover, polymorphism in HBV viral polymerase influences the development of HBV mutants resistant to nucleotide analogue treatment that is a consequence of treatment failure.

• Host response to the presence of Helicobacter spp. DNA in the liver of patients with chronic liver diseases.

  Authors: Magda Rybicka, Joanna Nakonieczna, Piotr Stalke, Krzysztof Piotr Bielawski

  Polish journal of microbiology / Polskie Towarzystwo Mikrobiologów = The Polish Society of Microbiologists. 01/2011; 60(2):175-8.

  Literature data indicate an association between the presence of Helicobacter spp. in the liver and the development of hepatocellular carcinoma (HCC). However, the role of H. pylori infections inLiterature data indicate an association between the presence of Helicobacter spp. in the liver and the development of hepatocellular carcinoma (HCC). However, the role of H. pylori infections in chronic liver diseases (CLD) remains controversial. The aim of this study was to detect Helicobacter spp. DNA in patients with CLD, and to investigate the host response to the presence of the bacterium in the liver. Helicobacter spp. DNA was detected in 59% samples. H.pylori was the most prevalent species (94%). We estimated the expression level of IL-1 and IL-8 genes. The presence of Helicobacter spp. did not have a significant effect on the gene expression of IL-8 and IL-1.

• Detection of Helicobacter rodentium-like DNA in the liver tissue of patients with chronic liver diseases by polymerase chain reaction-denaturing gradient gel electrophoresis and DNA sequence analysis.

  Authors: Joanna Nakonieczna, Piotr Stalke, Waleed Abu Al-Soud, Torkel Wadström, Krzysztof Piotr Bielawski

  Diagnostic microbiology and infectious disease. 11/2010; 68(3):201-7.

  Many Helicobacter spp. were isolated from the stomach, intestinal tract, and liver of different animals and humans. The association between Helicobacter spp. and hepatobiliary diseases, includingMany Helicobacter spp. were isolated from the stomach, intestinal tract, and liver of different animals and humans. The association between Helicobacter spp. and hepatobiliary diseases, including hepatocellular carcinoma, was thoroughly examined, indicating a potential role of the bacteria in the progression toward cancer. In our work, we screened 97 liver biopsies from patients with chronic liver diseases for the presence of Helicobacter spp. DNA. With the use of genus-specific polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and subsequent sequencing, we found that the majority of Helicobacter spp. DNA detected was similar to Helicobacter rodentium DNA (71%). The DNA of other detected Helicobacter spp. was similar to Helicobacter pylori DNA. This is the first indication of H. rodentium-like DNA presence in human liver tissue. We also conclude that PCR-DGGE is a useful screening method for assigning species designation and heterogeneity.

• The role of iron overload and HFE gene mutations in the era of pegylated interferon and ribavirin treatment of chronic hepatitis C.

  Authors: Katarzyna Sikorska, Piotr Stalke, Ewa Izycka-Swieszewska, Tomasz Romanowski, Krzysztof Piotr Bielawski

  Medical science monitor : international medical journal of experimental and clinical research. 02/2010; 16(3):CR137-143.

  Iron overload observed in chronic hepatitis C (CHC) has been suggested to be one of the negative prognostic factors influencing liver disease progression and failure of treatment with recombinantIron overload observed in chronic hepatitis C (CHC) has been suggested to be one of the negative prognostic factors influencing liver disease progression and failure of treatment with recombinant interferon in monotherapy or in combination with ribavirin. The aim of this study was to assess occurrence of iron overload in relation to polymorphism of the HFE and the influence of both these factors on efficacy of antiviral treatment with pegylated interferon and ribavirin in patients with CHC. Liver function tests, serum indices of iron metabolism, and HFE mutations were assayed in 152 patients with CHC from Poland. Histopathological examination of the liver biopsy specimen was performed in 138 patients. Sixty-one patients were treated with pegylated interferon alfa-2 and ribavirin. The comparative analysis was performed in 2 groups of patients: those with and those without elevated serum indices of iron metabolism. Increased biochemical iron metabolism parameters correlated with older age, higher ALT activity, more advanced liver fibrosis and treatment failure. Iron deposits in liver specimens were not accompanied by exacerbation of necro-inflammatory activity and advanced fibrosis. Elevated biochemical values of iron metabolism parameters and presence of hepatic iron deposits correlated positively with C282Y mutations. The lack of sustained viral response after treatment with pegylated interferon and ribavirin was observed more frequently in carriers of HFE mutations. Iron overload was frequently detected in patients with CHC, and was associated only with C282Y alleles. Biochemical markers of iron overload and HFE gene mutations were negative prognostic factors of antiviral treatment.

• [Helicobacter spp. infections in chronic liver damage].

  Authors: Magda Rybicka, Piotr Stalke, Krzysztof Piotr Bielawski, Joanna Nakonieczna

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2010; 64:386-95.

  Liver is a key organ responsible for organism’s homeostasis. A proper function of this organ is crucial for detoxification of metabolic products and regulation of metabolic processes ofLiver is a key organ responsible for organism’s homeostasis. A proper function of this organ is crucial for detoxification of metabolic products and regulation of metabolic processes of macromolecules (proteins, lipids, carbohydrates). The most important infectious factors, leading to liver damage, are primary hepatotropic viruses, particularly those causing chronic inflammation of the organ (HBV, HCV, HDV), which may subsequently cause cirrhosis and/or primary hepatocellular carcinoma. There has been a growing interest in Helicobacter spp. liver infections as a potential factor promoting injury of the organ towards hepatocellular carcinoma. The association between hepatocellular carcinoma and the presence of Helicobacters in the liver has been well documented in animal models (Helicobacter hepaticus versus liver cancer in mice). Some reports also indicate similar association in humans, where the presence of Helicobacter antigens in patients with liver cancer is detected more often in comparison to healthy or chronically infected population. Although the molecular mechanisms underlying such a phenomenon are not well known, the knowledge on this subject has considerably increased during recent years. The review presents data on the association between the presence of Helicobacter spp. in the liver and injuries of the organ, as well as the role that is played by the bacteria in chronic liver diseases.

• UGT1A1 gene polymorphism as a potential factor inducing iron overload in the pathogenesis of type 1 hereditary hemochromatosis.

  Authors: Tomasz Romanowski, Katarzyna Sikorska, Krzysztof Piotr Bielawski

  Hepatology research : the official journal of the Japan Society of Hepatology. 01/2009;

  Aim Hereditary hemochromatosis is a common genetic disorder characterized by iron overload and subsequent organ damage. It is caused in most cases by HFE gene mutations which penetrance can beAim Hereditary hemochromatosis is a common genetic disorder characterized by iron overload and subsequent organ damage. It is caused in most cases by HFE gene mutations which penetrance can be affected by many factors. The aim of this study was to establish the role of UGT1A1 gene polymorphism and serum bilirubin concentration in the pathogenesis of hereditary hemochromatosis. Methods Biochemical, histopathological and genetic data indicating iron excess and serum total bilirubin concentration were determined in 32 patients with the type 1 hereditary hemochromatosis. Fluorescent molecular probes assays were used for genotyping of UGT1A1*28 and UGT1A1*60 mutations in these individuals. Results High incidence and a significant correlation of UGT1A1 gene mutations with increased serum bilirubin level and lower grades of liver tissue inflammatory activity were observed in study participants. UGT1A1*28 and UGT1A1*60 mutations were strongly linked together. Two of the subjects presented very rare genotypes of UGT1A1 gene: (TA)(5/7) and c.-64G>C heterozygotes. Conclusions UGT1A1 gene polymorphism and as its consequence of high serum bilirubin level may promote iron accumulation in hemochromatosis patients by reducing the activity of inflammation. We proposed a possible mechanism of this interaction.

• GUS and PMM1 as suitable reference genes for gene expression analysis in the liver tissue of patients with chronic hepatitis.

  Authors: Tomasz Romanowski, Katarzyna Sikorska, Krzysztof Piotr Bielawski

  Medical science monitor : international medical journal of experimental and clinical research. 07/2008; 14(7):BR147-152.

  Background: The proper application of quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) for the relative quantification of a target gene in gene profiling studiesBackground: The proper application of quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) for the relative quantification of a target gene in gene profiling studies requires reference genes to normalize sample variations. Stable housekeeping genes for this purpose have never been investigated in the liver tissue of patients with chronic hepatitis. Material/Methods: Expression profiles of six functionally distinct housekeeping genes (ACTB, CYCC, GUS, HPRT1, PMM1, POLR2L) were examined by RT-PCR in liver specimens from 12 individuals with chronic hepatitis C or B. Two software programs, geNorm and NormFinder, were used to assess the expression stability of the studied genes. Results: Crossing-point values of the candidate reference genes were recorded between 22 and 28. In three groups of patients (all patients, HCV patients, HBV patients) both programs identified GUS as the most stably expressed housekeeping gene (stability values: 0.275-0.360 and 0.095-0.107 determined by geNorm and NormFinder, respectively), followed by PMM1 (0.275-0.360 and 0.168-0.227), and POLR2L (0.347-0.397 and 0.319-0.388). Conclusions: The genes GUS and PMM1 are recommended for normalization purposes in gene expression studies of liver tissue from patients with chronic hepatitis C or B. Using these genes in combination will ensure very reliable results.

• Determination of lamivudine-resistant variants of hepatitis B virus by denaturing gradient gel electrophoresis: a novel approach to monitoring drug resistance.

  Authors: Krzysztof Piotr Bielawski, Waleed Abu Al-Soud, Piotr Stalke, Urszula Charmuszko, Torkel Wadstrom

  Medical science monitor : international medical journal of experimental and clinical research. 06/2008; 14(5):CR281-285.

  BACKGROUND: A DGGE-based assay for hepatitis B virus (HBV) drug-resistance monitoring was designed and checked for feasibility. It detects mutations within the YMDD motif related to lamivudineBACKGROUND: A DGGE-based assay for hepatitis B virus (HBV) drug-resistance monitoring was designed and checked for feasibility. It detects mutations within the YMDD motif related to lamivudine resistance. MATERIAL/METHODS: The YMDD motif of HBV polymerase was amplified by the set of primers designed in this study. DGGE analysis of the amplicons was performed on 9% polyacrylamide gels containing a 20-40% gradient of urea plus formamide and electrophoresis was performed. DNA sequencing was performed using a standard protocol. RESULTS: Based on the DGGE pattern of previously sequenced HBV variants, a reference ladder consisting of bands was constructed within and near the YMDD motif of HBV with excellent resolution. The genotypes of all the fragments included in the ladder were confirmed by sequencing after DGGE analysis. The flexibility of DGGE was demonstrated by the ability to add more bands to the migration ladder when new variants were discovered during the analysis of patient specimens. Clinical samples from HBV-infected patients were also used to demonstrate the performance of this approach. CONCLUSIONS: This preliminary feasibility study of HBV drug-resistance monitoring by means of DGGE shows the potential advantage of this approach for low-cost screening for viral drug resistance in clinical settings. The presented example can be extended to detect other mutations related to drug resistance in the HBV genome as well as other viruses.

• [Molecular markers of micrometastasis in the blood of hepatocellular carcinoma patients]

  Authors: Anna Stokowska, Piotr Stalke, Krzysztof Piotr Bielawski

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2007; 61:310-9.

  The frequent occurrence of metastases from the primary tumor present a major therapeutic problem in hepatocellular cancer because of the hematogenous spread of cancer cells. Standard imagingThe frequent occurrence of metastases from the primary tumor present a major therapeutic problem in hepatocellular cancer because of the hematogenous spread of cancer cells. Standard imaging diagnostics methods do not allow for an early detection of relapse of the disease, as opposed to the analysis of molecular cell markers, especially mRNA-based methods, in peripheral blood samples by RT-PCR. Analysis of alphafetoprotein expression is the "gold standard" in the diagnostics of HCC at the protein and mRNA level because of its specificity for liver cancer. However, working out an analysis protocol is problematic and the utility of the marker in monitoring a patient's status in the perioperative period remains controversial. New HCC markers are being searched for among the highly liver-specific ones and those shared between various cancers types. Markers associated with freely circulating nucleic acids are also studied and quantitative assays are used. An assay for several markers simultaneously could give satisfactory results.

• [Housekeeping genes as a reference in quantitative real-time RT-PCR]

  Authors: Tomasz Romanowski, Aleksandra Markiewicz, Natalia Bednarz, Krzysztof Piotr Bielawski

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2007; 61:500-10.

  The determination of gene expression levels in normal tissue is necessary for the analysis and interpretation of results of gene profiling studies in pathological samples. With the real-time reverseThe determination of gene expression levels in normal tissue is necessary for the analysis and interpretation of results of gene profiling studies in pathological samples. With the real-time reverse transcription-PCR technique, which enables one to detect the amplification rate during the process, assessment of the amount of gene transcript is fast and accurate. The most important problem in this type of analysis is the variability in the amount of genetic material between samples, caused mostly by changes in the efficiency of mRNA isolation and reverse transcription. Therefore, a reference gene to normalize sample variations is required. Quantification of the mRNA of the target and the reference gene in the sample ensures that the changes in transcript levels will influence both genes equally. To be used as a reference, a gene should show stable, unregulated expression in the analyzed sample type. Housekeeping genes (HKGs) fulfill this criterion and they are used for normalization purposes in most expression studies. However, transcript levels of HKGs can vary between different types of tissue (normal and pathological samples) and under different treatment conditions (drugs and chemicals). The aim of this study was to show the differences and the factors which can influence housekeeping gene expressions.

• Genetic variability of hepatitis B virus isolates in Poland.

  Authors: Krzysztof Piotr Bielawski, Urszula Charmuszko, Aleksandra Dybikowska, Piotr Stalke, Anna Jadwiga Podhajska

  Virus genes. 09/2006; 33(1):77-86.

  There is very limited knowledge about the genetic variability of HBV strains circulating in the population of Polish chronically infected HBV patients. The aim of this study was to analyse theThere is very limited knowledge about the genetic variability of HBV strains circulating in the population of Polish chronically infected HBV patients. The aim of this study was to analyse the phylogenetic relatedness and polymorphism in some functional domains of HBV genome among chronically infected patients from northern Poland. Fifty-one serum samples were included to analysis of HBV genomes due to the viral load sufficient for DNA preparation and sequencing. The sequences of the rt polymerase/S and preC/BCP regions of those isolates were analysed, compared to genome sequences of different variants of HBV from GenBank database and genetic relatedness of Polish genotypes to known reference strains was estimated. A phylogenetic tree of 41 analysed genotype A isolates as well as 8 genotype D strains was constructed showing relationship to know reference strains. Two isolates, initially classified as genotype F turned to be related to genotype H, newly described genotype deriving from genotype F, a very rare genotype in Europe. HBV genotypes' distribution pattern in Poland and phylogenetic relatedness seems to be different from our Eastern neighbours. Due to the fact that Poland is still ethnically uniform country, it is interesting to explore molecular epidemiology of HBV infections in our population.

• [Molecular basis of hereditary hemochromatosis]

  Authors: Tomasz Romanowski, Katarzyna Sikorska, Krzysztof Piotr Bielawski

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2006; 60:217-26.

  Hereditary hemochromatosis (HH) is a genetic metabolic disease characterized by increased intestinal iron absorption and progressive iron loading in the cells of various organs. Human body ironHereditary hemochromatosis (HH) is a genetic metabolic disease characterized by increased intestinal iron absorption and progressive iron loading in the cells of various organs. Human body iron homeostasis involves a number of complicated processes, some of which are not identified yet. Genetic analysis of patients affected by HH recently led to the discovery of many novel proteins and mechanisms that can influence the uptake, transport, storage, and excretion of iron. It also showed that hemochromatosis is a very complex disease and that the type of mutation can influence its clinical manifestation. This review presents the current knowledge about the mechanisms of iron metabolism and describes the types of hereditary hemochromatosis and the mutations which induce the disease.

• [Hereditary hemochromatosis: the most frequent inherited human disease]

  Authors: Katarzyna Sikorska, Krzysztof Piotr Bielawski, Tomasz Romanowski, Piotr Stalke

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2006; 60:667-76.

  Hereditary hemochromatosis is now recognized as a very common inherited disease of the Caucasian population. It is defined as a disorder of unique clinicopathology caused by mutations of genes thatHereditary hemochromatosis is now recognized as a very common inherited disease of the Caucasian population. It is defined as a disorder of unique clinicopathology caused by mutations of genes that control iron metabolism. Inappropriately increased intestinal iron absorption and accelerated recycling of iron by macrophages lead to progressive body iron accumulation and the generation of oxidative stress in tissues. This results in significant cellular damage, induction of inflammation, and fibrosis. Liver cirrhosis, hepatocellular carcinoma, diabetes mellitus, and cardiac insufficiency are diagnosed in the advanced phase of this disease. The natural course is modified by environmental factors and personal predisposition. Three forms of hemochromatosis with the pathophysiology of iron overload are described. Among them the classical form, juvenile hemochromatosis with severe course and circulatory insufficiency, and ferroportin disease are presented. Properly directed diagnostics makes early treatment protecting against disease progression and multiorgan insufficiency possible.

• Molecular epidemiology of chronic hepatitis B virus infection in northern Poland.

  Authors: Krzysztof Piotr Bielawski, Piotr Stalke

  Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology. 01/2006; 34 Suppl 1:S63-9.

  Hepatitis B virus (HBV) infection is a global health problem, with more than 350 million people chronically infected worldwide. The chronic HBV infection in Poland is also an essential medical andHepatitis B virus (HBV) infection is a global health problem, with more than 350 million people chronically infected worldwide. The chronic HBV infection in Poland is also an essential medical and social problem. Starting from 1993, a steady decline of the incidence of HBV has been observed, reaching the estimated rate of 4.5 per 100 000 in 2004. Nothing is known about the genetic variability of HBV in Poland, the occurrence and spreading of genetic variants and mutants of hepatitis B virus in the population of Polish patients during the course of the disease and in relation to antiviral treatment. It is very interesting to study the molecular epidemiology of the Polish population regarding hepatitis B virus infection as Poland is still ethnically a uniform country, with no more than 3-4% of ethnic minorities. The first results regarding distribution of HBV genotypes and serotypes in northern Poland have been published by our group in 2003 and 2004. This work was part of a scientific project supported by the Fifth Framework Programme initiative of the European Union, entitled "Emerging variants of hepatitis B virus: new tools for epidemiological survey, diagnosis of infection, and monitoring of drug resistance". In the course of the project more than 200 hepatitis B infected patients from the northern part of Poland have been enrolled, diagnosed and - if the viral load of HBV was suitable for analysis - genotyped by sequencing of the HBV pol/S gene fragment. This review presents the main characteristics and some interesting aspects of the studied cohort of chronically infected patients from northern Poland as well as the molecular epidemiology.

• HFE gene mutations in Polish patients with disturbances of iron metabolism: an initial assessment.

  Authors: Katarzyna Sikorska, Krzysztof Piotr Bielawski, Piotr Stalke, Elzbieta Anna Lakomy, Zofia Michalska, Krystyna Witczak-Malinowska, Tomasz Romanowski

  International journal of molecular medicine. 12/2005; 16(6):1151-6.

  Hereditary hemochromatosis is one of the most frequent genetic disorders in Europeans, but its prevalence in Poland is still unknown. The aim of the study was an initial assessment of the prevalenceHereditary hemochromatosis is one of the most frequent genetic disorders in Europeans, but its prevalence in Poland is still unknown. The aim of the study was an initial assessment of the prevalence of C282Y and H62D HFE gene mutations and their influence on the course of chronic hepatitis C. Forty-one patients were admitted to the Department of Infectious Diseases, Medical University of Gdansk in 2000-2004 because of chronic liver diseases with accompanying disturbances in iron metabolism. Genetic tests for the C282Y and H63D mutations were performed by PCR and restriction fragment length polymorphism (PCR-RFLP) analysis. The HFE gene mutations were confirmed in 24 of 41 (59%) cases with symptoms of chronic liver disease and iron overload, significantly more frequently in HCV-negative patients (12/14 vs. 12/27; chi2=8.28; p=0.05). The C282Y and H63D HFE gene mutations were detected in 16 of 41 (39%) and 9 of 41 (22%) cases, respectively. HCV-negative patients were C282Y carriers significantly more frequently than HCV-positive patients [9/14 vs. 2/27 C282Y homozygotes; 2/14 vs. 3/27 C282Y heterozygotes (p<0.0001)]. The carrier state of the H63D HFE gene mutation was not significantly more frequent in HCV-positive than HCV-negative patients. HCV infection seems to be a negative predictive marker of HFE gene mutations in patients with iron overload. The relationship of H63D HFE gene mutations with chronic hepatitis C and the possible influence of HCV infection on iron metabolism needs further analysis.

• Distribution of HBV genotypes and mutants among hepatitis B infected patients from northern Poland.

  Authors: Krzysztof Piotr Bielawski, Aleksandra Dybikowska, Urszula Lisowska-Charmuszko, Piotr Stalke, Katarzyna Gregorowicz, Hanna Trocha, Anna Podhajska

  International journal of molecular medicine. 09/2004; 14(2):301-4.

  Hepatitis B virus (HBV) infection is one of the major global epidemiological problems. The aim of our study was to determine the distribution of HBV genotypes in Poland since the data concerning theHepatitis B virus (HBV) infection is one of the major global epidemiological problems. The aim of our study was to determine the distribution of HBV genotypes in Poland since the data concerning the spread of HBV viruses in the central-eastern region of Europe is still very limited. HBV DNA was extracted from 58 serum samples. To quantify the level of HBV DNA the Roche Amplicor HBV Monitor Assay was used. To genotype and assign HBV subtypes DNA sequencing methods were performed. The HBV virus from 43 serum samples from hepatitis B infected patients was genotype A (74.1%), 12 cases had genotype D (20.7%), and 3 had the rare in Europe genotype F (5.2%). Prediction of HBV serological subtypes based on HBsAg sequencing showed almost 100% occurrence of subtype adw2 in the group of genotype A samples, three different subtypes in genotype D (ayw2, ayw3, and ayw4), and equal distribution of subtype adw4q- in all 3 cases of genotype F, also the most prevalent subtype in the Amerindians. Our results coincide with the general European HBV prevalence. However, HBV genotype F, which is not a common genotype in European countries, was detected and so was relatively high occurrence of genotype D, which may reflect historical and ethnical migration events in Poland in the past.

• Could iron deposits in hepatocytes serve as a prognostic marker of HFE gene mutations?

  Authors: Katarzyna Sikorska, Piotr Stalke, Kazimierz Jaskiewicz, Tomasz Romanowski, Krzysztof Piotr Bielawski

  Hepato-gastroenterology. 55(84):1024-8.

  BACKGROUND/AIMS: The diagnosis of hereditary hemochromatosis (HH) is based on qualitative measurement of tissue iron concentration and genetic tests. The aim of this study was to evaluate theBACKGROUND/AIMS: The diagnosis of hereditary hemochromatosis (HH) is based on qualitative measurement of tissue iron concentration and genetic tests. The aim of this study was to evaluate the correlation between the presence of iron deposits in the liver and the HFE gene mutations in patients with chronic liver diseases (CLD). METHODOLOGY: The 182 patients, age range 18-71 years, were hospitalized in Gdansk because of CLD. The C282Y, H63D and S65C HFE mutations were screened by PCR-RFLP analysis. Liver function tests, serological examinations for viral hepatitis, serum iron and ferritin concentration and semiquantitative assessment of liver iron were done in all subjects. Patients were divided into Group A without iron deposits in the liver, and Group B with deposits. The most frequent etiology of CLD was chronic hepatitis C. RESULTS: Biochemical parameters indicating iron storage and ALT activity were significantly higher in Group B. Either typical for diagnosis HH homozygotes C282Y/C282Y and combined heterozygotes C282Y/H63D or carriers of other HFE gene mutations were found significantly more frequently in Group B. CONCLUSIONS: The finding of iron deposits in routinely obtained liver specimen correlates with occurrence of the different HFE gene mutations.