High prevalence of genotype 4 among hepatitis C virus-infected intravenous drug users in north-eastern Poland.

The aim of this study was to describe the distribution of hepatitis C genotypes among intravenous drug users in north-eastern Poland. The study group included intravenous drug users recruited at a drug treatment center and a clinic for HIV-infected patients. HCV infection was confirmed by qualitative nested RT-PCR to test for the presence of HCV RNA. Genotypes were determined by 5'UTR sequencing and comparing the results with known genotype sequences. Among 111 HCV-infected and HCV-RNA-positive intravenous drug users, the most prevalent genotypes were 1 (38.7%), 3 (37.8%), and 4 (23.4%). Most infections with genotype 4 (88.5%) were found among HCV-HIV-coinfected drug users. The study demonstrated a high prevalence of genotype 4 (23.4%) among HCV-infected Polish drug users.

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.

Transforming growth factor beta1 and prostaglandin E2 concentrations are associated with bone formation markers in ulcerative colitis patients.

Osteoporosis is a significant complication of a multifactoral etiology associated with inflammatory bowel disease. The aim of study was to evaluate the relationships between bone mineral density as well as bone turnover markers and inflammatory activity modulators (i.e., PGE2 and TGFbeta1) in ulcerative colitis (UC). Twenty-one active ulcerative colitis subjects and 14 healthy individuals were included into the study. We observed no significant differences in serum concentrations of osteoprotegerin and osteocalcin, as well as bone mineral density between UC patients and healthy individuals. Plasma concentrations of PGE2, TGFbeta1 and TNF-alpha were significantly higher in UC patients than in controls. Serum osteocalcin demonstrated a positive correlation with both serum PGE2 and plasma TGFbeta1. Moreover there was significant correlation between osteoprotegerin and TGFbeta1 as well as serum TNF-alpha concentrations. In conclusion a positive association between PGE2 and TGFbeta1 and bone formation markers-osteoprotegerin and osteocalcin, as well as a comparable BMD in UC patients and healthy individuals was shown. Our results may indicate that increase of PGE2 as well as TGFbeta1 concentrations may play a protective role against bone loss in ulcerative colitis patients.

Medical procedures and the risk of iatrogenic hepatitis C infection: case-controlled study in north-eastern Poland.

Many hepatitis C virus (HCV)-infected individuals do not have any obvious risk factors and one of the putative sources of infection may be inadvertent exposure to infected blood or body fluids in the clinical setting. The aim of this study was to assess the role of medical procedures in transmission of hepatitis C in north-eastern Poland. In total, 194 patients with chronic hepatitis C were eligible for the study. The control group consisted of 275 age- and sex-matched individuals. Patients with a history of intravenous drug use were excluded. On multivariate analysis, transfusions [odds ratio (OR) = 3.7, 95% confidence interval (CI) 2.2-6.3], minor surgery (OR = 3.2, 95% CI 1.5-6.7) and dental care (OR = 2.3, 95% CI = 1.4-4.0) were independently associated with HCV infection. We conclude that apart from transfusion, minor medical procedures and dental care may carry a significant risk of hepatitis C infection. Improvements in basic hygiene routines and strict adherence to universal precautions may be essential to prevent iatrogenic transmission of the infection.

Metabolic disturbances in liver 1H MR spectroscopy in HIV and HCV co-infected patients as a potential marker of hepatocyte activation.

PURPOSE: To evaluate proton magnetic resonance spectroscopy (1H MRS) features in order to assess hepatocellular activation in chronic hepatitis C and human immunodeficiency virus/hepatitis C (HIV/HCV) co-infected patients. MATERIAL AND METHODS: Liver in vivo 1H MR spectra were obtained in 14 patients with hepatitis C virus infection (HCV), 20 HIV/HCV co-infected individuals, and 24 healthy volunteers. Resonances of lipids, glutamine/glutamate (Glx), phosphomonoesters (PME), glycogen/glucose (Glc) were assessed and metabolite ratios to total lipids (TL) were calculated. RESULTS: A significant increase in Glx/TL and PME/TL was observed in the HCV group as compared to healthy individuals. Patients with HIV and HCV co-infection had a further increase of all metabolite ratios. Changes in metabolite ratios were due to both the increase in particular metabolite contents and to the decrease in lipid levels. HIV/HCV-infected patients treated with highly active anti-retroviral therapy (HAART) showed elevated PME and Glx levels and significantly decreased TL compared to patients not undergoing anti-retroviral treatment. CONCLUSIONS: Our findings suggest clinical usefulness of liver 1H MR spectroscopy in detecting even slight disturbances in liver metabolism.

Serum hyaluronic acid during lamivudine treatment in chronic hepatitis B.

PURPOSE: We address the question whether lamivudine treatment modulates serum concentrations of hyaluronic acid and whether the pre-treatment HA level can give the information about the presumptive result of treatment and whether HA level evaluation can be useful in monitoring the antiviral therapy in chronic hepatitis B. MATERIAL AND METHODS: Forty-nine patients, 31 man, aged 40 +/- 2.5 years were treated with 100 mg lamivudine per day for 48 weeks. Serum hyaluronic acid level was determined using enzyme-linked binding protein commercial assay (Corgenix Inc., USA). RESULTS: The mean HA pre-treatment levels were higher than among controls (69.6 +/- 11.6 ng/ml vs 36.5 +/- 7.6 ng/ml, mean +/- SEM) and correlated with AST activity, p = 0.002; GGT activity, p = 0.006; ALP activity p < 0.001; prothrombin time, p = 0.01; and peripheral blood platelets count, p = 0.001 but did not correlated with ALT activity. The pre-treatment HA concentration correlated also with interlobular necroinflammatory activity score, p = 0.049 and with fibrosis score, p = 0.026, according to Scheuer classification. The mean HA levels decreased gradually during lamivudine treatment, up to levels lover than among controls (26.3 +/- 5.7 ng/ml). There were not significant differences in pre-treatment levels observed between patients neither with HBs seroconversion versus those without it, nor between patients with HBe seroconversion versus those without it and among patients with normalization of ALT activity versus ones without it. CONCLUSIONS: Serum hyaluronic acid level decreases during lamivudine treatment both in patients with HBeAg seroconversion and without it; serum hyaluronic acid pretherapy levels correlate with necroinflammatory lobular activity score and with liver fibrosis score; serum hyaluronic acid is of no predictive value for lamivudine therapy response; serum hyaluronan may be valuable complementary marker in chronically HBV infected patients.

MR and 1H MR spectroscopy of the brain in patients with liver cirrhosis and early stages of hepatic encephalopathy.

BACKGROUND/AIMS: Hepatic encephalopathy is a serious problem in patients with liver cirrhosis and precise pathophysiological mechanisms responsible for encephalopathy are not fully understood. Magnetic resonance imaging and magnetic resonance spectroscopy can be used to detect specific morphological and metabolic abnormalities in the brain even in patients with early stages of hepatic encephalopathy. METHODOLOGY: Twenty patients with liver cirrhosis and 14 patients with grade I-II hepatic encephalopathy were studied with magnetic resonance and proton magnetic resonance spectroscopy. Localized magnetic resonance spectra were acquired in the parietal gray/white matter regions and basal ganglia. Control group consisted of 20 healthy volunteers. RESULTS: Frequency and degree of brain atrophy and bilateral signal hyperintensities in globus pallidus were similar in groups with liver cirrhosis and with encephalopathy. Decreased myoinositol, choline and increased glutamine levels were noted in both groups whereas N-acetylaspartate levels were unchanged. The statistically significant differences between cirrhotic and encephalopathic groups were observed only in myoinositol/creatine ratio in basal ganglia. There were no significant differences in metabolic concentrations between parietal and basal ganglia regions. CONCLUSIONS: Metabolic brain alterations occur earlier than clinical evidence of hepatic encephalopathy but there is no correlation between presence of symptoms encephalopathy and magnetic resonance and magnetic resonance spectroscopy findings.

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.

Cerebral MR spectroscopy in neurologically asymptomatic HIV-infected patients.

PURPOSE: To detect early metabolic changes in the brain of neurologically asymptomatic HIV-infected patients with normal MR imaging and to find the correlation between 1H MR results and immune status. MATERIAL AND METHODS: Twenty neurologically asymptomatic HIV seropositive patients underwent MR imaging and single-voxel 1H MR spectroscopy (MRS) using a PRESS sequence. For all patients, the signals from N-acetyl-aspartate (NAA), choline-containing compounds (Cho), creatine-phosphocreatine (Cr) and myoinositol (mI) were compared with 32 healthy volunteers as metabolite ratios and metabolite areas to non-suppressed water area ratios. RESULTS: In HIV patients, the NAA/Cho ratio was significantly lower ( p < 0.01), but there were no changes in NAA/Cr ratio. A statistically significant reduction in NAA/H2O and Cr/H2O (both p < 0.05) was observed. For the immune status there was a statistically significant correlation (r=0.47, p<0.05) between CD4 counts and NAA/H2O ratio. A significant increase in Cho/Cr ( p<0.001) and mI/Cr ( p<0.01) ratios in HIV patients was found, but Cho/H2O and mI/H2O concentrations were non-significantly increased. CONCLUSION: These results indicate that neuronal loss and gliosis in HIV-infected patients may be associated with impairment of energy metabolism. The spectral changes found suggest that 1H MRS can be used for early detection of brain damage induced by HIV.

Immunological response in HIV-positive patients vaccinated against tick-borne encephalitis.

BACKGROUND: Vaccination against infectious diseases is a recommended preventive measure in patients with impaired immunity. The aim of the study was to estimate the immunogenicity of the vaccine against tick-borne encephalitis (TBE) in HIV-infected patients and to determine its safety for this group. PATIENTS AND METHODS: The TBE vaccine FSME-IMMUN-inject was tested on 29 HIV-positive patients and the response compared to that of 40 healthy controls. The vaccination protocol for the HIV-positive group was modified by the addition of a fourth dose according to a 0/1/2/9-month schedule. RESULTS: No serious adverse reactions were observed in patients with deficient immunity. A better response was obtained in HIV-infected patients with CD4 counts >or= 500/microl (55% of the patients had levels of IgG antibody > 126 VIEU/microl) than in those with CD4 counts of 200-499/microl (40%). However, the difference did not reach significance. 85% of healthy controls achieved protective antibody titers after a full course of vaccination. CONCLUSION: The correlation between post-vaccine seroconversion and CD4 lymphocyte count showed that the FSME-IMMUN-inject vaccine can be considered to be a CD4 cell-independent vaccine. The examinations carried out 1 year after the completed vaccination protocol showed maintenance of the anti-TBE response acquired after the third vaccination in healthy subjects and in HIV-infected patients.

[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.

Effect of ulcerative colitis activity on plasma concentration of transforming growth factor beta1.

Enhanced expression of transforming growth factor-beta1 (TGF-beta1) demonstrated in human colonic mucosa of patients with ulcerative colitis (UC), indicates its possible significance in the pathogenesis of this disease. The aim of this study was to evaluate plasma TGF-beta1 concentration in patients with different degrees of colonic mucosal injury, as a possible indicator of ulcerative colitis activity. TGF-beta1 concentration was measured with an enzyme immunoassay (EIA) in plasma of 45 patients with endoscopically confirmed UC. Values observed in UC patients (40.5+/-15.9 ng/ml) were significantly higher than in healthy people (18.3+/-11.6 ng/ml) and higher than in patients with irritable colon syndrome (ICS), (20.5+/-13.6 ng/ml). The highest plasma TGF-beta1 (58.6+/-112.1 ng/ml) was in patients with the severe UC course. TGF-beta1 level analysed in all UC patients revealed significant positive correlation with scored degree of mucosal injury (r=0.396;P<0.01). Among other possible laboratory markers of the disease activity, only C-reactive protein concentration demonstrated significant correlation. Enhanced production of TGF-beta1 can be related to inflammation activity. Measurement of plasma TGF-beta1 may be considered as a biomarker of the disease activity.

Activation of blood platelets in chronic hepatitis and liver cirrhosis P-selectin expression on blood platelets and secretory activity of beta-thromboglobulin and platelet factor-4.

BACKGROUND/AIMS: Blood platelets are cells that quite often undergo damage in chronic liver diseases. Endotoxemia and hyperkinetic circulation influence platelets in an active manner. The role of platelets in the development of hepatitis and liver fibrosis is speculative. The aim of the study was to determine the influence of chronic liver diseases on platelets morphologic parameters, their secretory activity and P-selectin expression. METHODOLOGY: The examination was completed in the group of 29 patients with chronic hepatitis and 27 with liver cirrhosis of postinflammatory etiology (HBV, HCV). Liver biopsies were carried out in all patients. Thirty-two healthy individuals were the control group. Platelets morphological parameters (number, volume, platelet crit, micro- and macrothrombocyte fraction) were estimated. beta-thromboglobulin concentration and platelet factor 4 in blood serum as well as P-selectin expression on resting platelets and after thrombin activation were also examined. RESULTS: Number, volume, and platelet crit decreased with the advancement of a liver disease. Megathrombocyte fraction increased inversely with the severity of liver damage. The concentration of beta-thromboglobulin and platelet factor 4 alpha-granule contents in blood serum was higher 2- and 7-times, respectively than in healthy controls. P-selectin expression on resting platelets was considerably higher. After stimulation with thrombin, P-selectin expression was equal (chronic hepatitis) or higher (liver cirrhosis) than in the control. CONCLUSIONS: There are changes of platelet morphological parameters, with accompanying megathrombocyte fraction increase that occur in chronic liver diseases. Thrombocytes in chronic liver diseases and liver cirrhosis are more activated. Platelet sensitivity to stimuli in these ailments is higher (liver cirrhosis) than in the healthy controls.

[The role of transforming growth factors beta in pathogenesis of ulcerative colitis]. / Rola transformujacych czynników wzrostu beta w patogenezie wrzodziejacego zapalenia jelita grubego.

Transforming growth factor beta is the protein playing a principal role in the intercellular signalling. The most important functions are: control of cellular growth, differentiation and migration. Moreover it stimulates synthesis of extracellular matrix proteins, modulates immune response and is responsible for angiogenesis, wound formation and tissue reconstruction. All these activities are involved in the development or healing of inflammatory bowel diseases. The role of transforming growth factors beta in the pathogenesis of ulcerative colitis are discussed in this paper.

[Central pontine myelinolysis in an eighteen year old woman]. / Mielinoliza centralna mostu i jader podstawy mózgu u osiemnastoletniej kobiety.

Central pontine myelinolysis is sporadically diagnosed disease. The disease has fulminant course and fatal prognosis. Myelinolysis is known for 40 years, but its cause remain still unclear and therapy is not established. A case of 18 year old woman hospitalized in the Department of Infectious Disease Medical Academy of Bialystoku is an example of such a difficulties.

[New imaging diagnostics of liver echinococcosis]. / Przydatnosc nowoczesnych technik obrazowania w diagnostyce bablowicy watroby.

Cystic and alveolar echinococcosis, parasitic diseases, caused in humans by the larval stage of tapeworm Echinococcus granulosus and E. multilocularis are rare diseases in Poland. We have diagnosed 71 patients with cystic echinococcosis and 10 patients with alveolar echinococcosis during last 12 years. Echinococcosis still create significant diagnostic difficulties, particularly alveolar one. Progress and actually available radiological imaging techniques, which can be applied in the cystic and alveolar echinococcosis diagnostics, are presented. Ultrasound appearance, CT, MRI images are described. Own experience with Proton Magnetic Resonans Spectroscopy in two patients with recurrent alveolar echinococcosis are presented.

Detection of Trichinella spiralis antigens in urine of men and animals.

The practical inability to diagnose Trichinella spiralis antibodies in man before day 20 post infection (dpi) has stimulated interest in the development of immunodiagnostic test to detect circulating antigens. Our previous experience showed that soon after infection immune complexes as well as uncomplexed parasite antigens in sera of infected rats could be detected. To diagnose the presence of antigen in urine, double sandwich-capture ELISA was applied using a peroxidase-conjugated rabbit immunoglobulin to T. spiralis larval antigens. The plates were coated with metabolic (AES) or somatic (AS) larval antigens. Mice were infected with 500 T. spiralis larvae. The urine samples from experimentally infected mice taken from 1 to 41 dpi. and the urine samples from patients of the Clinical Hospital in Bialystok taken from 3 to 120 dpi were examined. Before testing, the urine samples were heated for 6 min. at 100 degrees C and centrifuged for 6 min. at 5000 g, supernatants were used in ELISA. The presence of T. spiralis antigens in mice urine samples was detected between 6-26 days post infection (dpi) using double sandwich-capture ELISA. All samples taken later were negative as samples taken from uninfected mice. 3 from 9 human urine samples taken 3-10 dpi were positive, the remaining samples taken 3-10 and 10-30 dpi showed values near to "cut-off". In both mice and human urine samples the higher level of antigens was detected in ELISA when somatic larval antigen was used. The T. spiralis antigens were present in urine of infected men and mice in the first phase of infection.

[Nephrolithiasis in a HIV infected patient treated with indinavir]. / Kamica nerkowa u zakazonego HIV leczonego indinavinem.

Protease inhibitors together with reverse transcriptase inhibitors are used in antiretroviral treatment. Indinavir, precipitating in renal tubules, quite often can cause nephrolithiasis. The case of a HIV infected patient with renal colic probably caused by indinavir, diagnostic and therapeutic options in such cases are described in the paper.