The effects of long-term melatonin treatment on plasma liver enzymes levels and plasma concentrations of lipids and melatonin in patients with nonalcoholic steatohepatitis: a pilot study.

The present study represents the follow-up of our initial observations designed to investigate whether in patients with nonalcoholic steatohepatitis (NASH) the beneficial effect of 12-week course of melatonin (MT) on liver enzymes could be maintained with prolonged period of treatment and to analyze whether biochemical treatment responses could be sustainable after melatonin discontinuation. Forty two patients with histologically proven NASH (30 treated with melatonin 2x5 mg daily, 12 controls receiving placebo) enrolled to our previous 3-month study agreed to take part of subsequent 12 weeks treatment followed by 12-week follow-up period. Enrolled patients had biochemical determinations every six weeks during the melatonin treatment period and again after 12 weeks of follow-up. Significant reduction in median alanine aminotransferase (ALT) levels between baseline and week 18, week 24 and follow-up was observed in both MT-treated and control group: 43% and 31%, 42% and 33%, 32% and 31%. Aspartate aminotransferase (AST) and gamma-glutamyl transpeptidase (GGT) levels decrease significantly only in MT-treated group. In MT-treated group mean percentage change in AST level below baseline at week 18, at week 24 and at follow-up was 45%, 33% (p<0.05) and 8% (ns), respectively. The evolution of GGT levels was as follows: the mean percentage reduction in GGT below baseline level at week 18, 24 and follow-up was: 48%, 52% and 38% (p<0.05), respectively. In both MT-treated and control group plasma cholesterol, triglicerydes and glucose concentrations as well as plasma alkaline phosphatase persisted within normal values during the prolonged study period. Plasma concentration of melatonin (pg/ml) in MT-treated group averaged 7.5±3.5 at baseline and increased to 52.5±17.5 at 24th week. The results of our study demonstrating beneficial effect of melatonin on liver enzymes in patients with NASH would seem to encourage further controlled trials of melatonin given over a longer period of time with liver histology as end point.

The pilot study of 3-month course of melatonin treatment of patients with nonalcoholic steatohepatitis: effect on plasma levels of liver enzymes, lipids and melatonin.

The mechanism by which nonalcoholic fatty liver disease (NAFLD) progresses into nonalcoholic steatohepatitis (NASH) is unknown, however, the major process is oxidative stress with increased production of reactive oxygen species and excessive inflammatory cytokine generation. To date, there are no effective treatments for NASH and the published data with treatment using antioxidants are not satisfactory. Melatonin (MT), the potent endogenous antioxidant secreted in circadian rhythm by pinealocytes and in large amounts in the digestive system, was reported to improve oxidative status and to exert beneficial effects in NASH pathology in experimental animals, but no study attempted to determine the possible effectiveness of MT in humans with NASH. In this study, 42 patients (12 placebo controls and 30 MT-treated) with histological evidence (liver biopsy) of NASH and no history of alcohol abuse, were included. The treatment group took melatonin (2x5 mg/daily orally), while controls were treated with placebo. At baseline no significant differences between the groups were found for age, body mass index (BMI) as well as for plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), and concentrations of cholesterol, triglycerides (TG), glucose and MT. During the study period plasma ALT level and cholesterol concentration decreased significantly in both MT-treated and control groups, however AST and GGT levels decreased significantly only in MT-treated groups. Median value of AST level at baseline was 76.5 (64.2-114.2) IU/L and its percentage decrease at 4, 8 and 12 week was 20, 36 and 38%, resp. Baseline GGT median level was 113 (75.7-210.7) IU/L and its mean percentage decrease at week 4, 8 and 12 was 46, 48 and 47%, resp. Plasma ALP levels did not change significantly during MT treatment. Median value of plasma concentrations of MT (pg/mL) in MT-treated group rose from 7.5 (5.0-14.25) at baseline to 35.5(18.8-110.0), 43.5(17.0-102.5) and 49.5(18.0-99.5) at the end of 4, 8 and 12 week of treatment, respectively. Plasma levels of TG and glucose as well as BMI in controls and MT-treated patients were not significantly different from baseline. This study demonstrates for the first time in humans that three months treatment with MT significantly improves plasma liver enzymes in patients with NASH without causing any side-effect. Plasma MT levels during the whole period of MT treatment persisted above that at baseline. Our findings show that treatment with MT significantly improves plasma liver enzymes in NASH patients, but larger cohort trials and longer treatment with MT are required before this indole could be included into the spectrum of the NASH treatment.

Helicobacter pylori in liver diseases.

At the present stage of knowledge, the participation of the Helicobacter bacteria in the pathology of liver and the bile tract in humans has not been univocally documented. However, apparent are the premises so as to go on performing the examinations under discussion since the said participation cannot be excluded. If the more direct evidence of the etiologic role of the Helicobacter in the pathology of liver were available, it would create the chances for the more effective treatment of patients than the case has been so far. Cancer commonly derives from the chronic inflammation and infection and in case of hepatocellular carcinoma (HCC), may arise either from local liver derived progenitor cells (LPCs) or bone marrow originated stem cells (BMSCs) and future studies should disclose the role of either type of cells and of inflammatory factors such as generated by Helicobacter infection in the liver pathophysiology.

Sympathovagal balance fluctuates during colonoscopy.

BACKGROUND AND STUDY AIM: Colonoscopy is a common gastroenterological procedure for investigation of the bowel. The main side effects of colonoscopy are pain during investigation, cardiovascular complications and very rarely even death. The aim of this study was to compare the continuous fluctuation of heart rate variability (HRV) components during colonoscopy under normal conditions, analgesia/sedation, and total intravenous anesthesia. PATIENTS AND METHODS: 37 consecutive patients (aged 35 - 65), were randomly allocated to three groups: no sedation (control group 1); analgesia/sedation (group 2); and total intravenous anesthesia (group 3). Holter electrocardiography and subsequent frequency domain analysis were undertaken. The low-frequency (LF, 0.04 - 0.15 Hz) and the high-frequency (HF, 0.15 - 0.40 Hz) components were estimated using spectral analysis in the usual way. Normalized units (nu) were calculated from the following equations: LFnu = LF/(LF + HF), and HFnu = HF/(LF + HF). RESULTS: Groups 2 and 3 were found to have a significantly lower HFnu and higher LFnu than group 1 essentially throughout the procedure. A one-way analysis of variance and t-test confirmed that the differences were significant when the colonoscope reached the splenic flexure as were the LF/HF balances at the splenic and hepatic flexures and the cecum. The percentage change in LF/HF was also analyzed, and it was found that in group 3 the mean change was over 136 % when the colonoscope reached the sigmoid flexure, which was significantly higher than in the other two groups. CONCLUSION: Most changes in HRV components occurred during colonoscopy of the left side of the bowel. Analgesia/sedation and total intravenous anesthesia increased HRV by increasing the LF component.

Progastrin and its products from patients with chronic viral hepatitis and liver cirrhosis.

BACKGROUND: Gastrin and its precursor, progastrin, are synthesized in the stomach, particularly when infected with Helicobacter pylori, and they are metabolized, at least in part, in the liver. However, little is known about their levels in various hepatic diseases. METHODS: This study was carried out on 147 patients including chronic hepatitis B (n = 35), hepatitis C (n = 52) and liver cirrhosis (n = 60) of class A (n = 38), class B (n = 15) and class C (n = 7) (Child-Pugh classification) and age- and sex-matched healthy controls (n = 65). The diagnosis of chronic hepatitis was confirmed by liver biopsy in all patients, whereas the diagnosis of liver cirrhosis was based on clinical and laboratory findings. Liver biopsy was done in 38 out of 60 patients. Blood samples were collected under basal conditions and separated plasma samples were kept frozen at -70 degrees C until radioimmunoassay of progastrin and its products, including bioactive amidated gastrins. RESULTS: Median (range) plasma concentrations of total progastrin product and amidated gastrin in control subjects were 147.5 (73-345) pM and 33 (15-65), respectively. These concentrations in hepatitis B and C were not significantly different from those in controls. In cirrhosis (classes A, B and C), the concentrations of the progastrin and of gastrin were significantly (P < 0.05) higher than in controls reaching, respectively, 253.5 (135-683 pM) and 47.5 (17-385) pM. Both progastrin and gastrin levels were significantly higher in H. pylori-positive than in negative cirrhotic patients. Antibodies against H. pylori were present in about 50% of controls, 68% of hepatitis B, 57% of hepatitis C and in 83% in cirrhosis patients. The difference in H. pylori prevalence between cirrhosis and controls was statistically significant. CONCLUSIONS: Plasma levels of progastrin and gastrin are significantly increased in cirrhotic patients and this could be attributed to reduced metabolism of these peptides in liver cirrhosis and to their increased release due to H. pylori infection rate in this disease.

Reduction of CD45RA isoform expression and decrease in CD4 and CD8 receptor density in lymphocytes of patients with primary biliary cirrhosis.

BACKGROUND: The immunological background of primary biliary cirrhosis (PBC) remains largely obscure. METHODS: Using double colour flow cytometry, we estimated the distribution of functionally different lymphocyte subpopulations in the peripheral blood of 25 PBC patients and 18 controls. We examined: 1) the expression of CD3, CD4, CD8, CD19 and CD56 surface receptors, 2) the distribution of lymphocyte subsets bearing 'naive' (CD45RA+) and 'memory' (CD45RO+) phenotypes in both CD4+ and CD8+ cell populations, 3) the expression of an early activation marker (CD69), 4) the distribution of C1.7 mAb binding cytotoxic effectors in CD3+, CD8+ and CD56+ cells. The surface marker expression was evaluated in terms of percentage of positive cells and receptor density. RESULTS: We found: 1) a decrease in the percentage of total CD3+ and CD4+ cells, an unchanged proportion of CD8+ cells but elevated proportion of CD19+ cells and NK lymphocytes; 2) a reduction in the percentage of 'naive' CD4+ but normal proportion of 'naive' CD8+ as well as CD4+ and CD8+ 'memory' cell subsets; 3) a decrease in the density of CD4 and CD8 receptors in the subsets of 'naive' and 'memory' T cells, 4) an increase in the percentage of CD69 receptor bearing T cells but unchanged proportion of C1.7 mAb. CONCLUSIONS: It is concluded that the reduction in number of 'suppressor-inducer-like 'naive' CD4+ T-cell subsets in association with the decrease in fluorescence intensity for CD4 and CD8 may significantly contribute to the mechanisms that could account for a development of PBC.

Reduction of CD45RA Isoform Expression and Decrease in CD4 and CD8 Receptor Density in Lymphocytes of Patients with Primary Biliary Cirrhosis.

BACKGROUND: The immunological background of primary biliary cirrhosis (PBC) remains largely obscure. METHODS: Using double colour flow cytometry, we estimated the distribution of functionally different lymphocyte subpopulations in the peripheral blood of 25 PBC patients and 18 controls. We examined: 1) the expression of CD3, CD4, CD8, CD 19 and CD56 surface receptors, 2) the distribution of lymphocyte subsets bearing 'naive' (CD45RA+) and 'memory' (CD45RO+) phenotypes in both CD4+ and CD8+ cell populations, 3) the expression of an early activation marker (CD69), 4) the distribution of C1.7 mAb binding cytotoxic effectors in CD3+, CD8+ and CD56+ cells. The surface marker expression was evaluated in terms of percentage of positive cells and receptor density. RESULTS: We found: 1) a decrease in the percentage of total CD3+ and CD4+ cells, an unchanged proportion of CD8+ cells but elevated proportion of CD 19+ cells and NK lymphocytes; 2) a reduction in the percentage of 'naive' CD4+ but normal proportion of 'naive' CD8+ as well as CD4+ and CD8+ 'memory' cell subsets; 3) a decrease in the density of CD4 and CD8 receptors in the subsets of 'naive' and 'memory' T cells, 4) an increase in the percentage of CD69 receptor bearing T cells but unchanged proportion of C1.7 mAb. CONCLUSIONS: It is concluded that the reduction in number of 'suppressor-inducer-like 'naive' CD4+ T-cell subsets in association with the decrease in fluorescence intensity for CD4 and CD8 may significantly contribute to the mechanisms that could account for a development of PBC.

Positive and negative strands of HCV-RNA in sera and peripheral blood mononuclear cells of chronically hemodialyzed patients.

BACKGROUND: Hepatitis C virus (HCV) RNA can be detected in sera and peripheral blood mononuclear cells (PBMC) of patients undergoing chronic hemodialysis. However, the natural course of HCV infection in this group of patients is not fully known. Although the exact mechanism of HCV replication is not completely explained, there is evidence that HCV replicate through synthesis of complementary negative (-) RNA strand, whereas positive (+) RNA strand serves as a template. Thus, the detection of negative HCV-RNA strand can be regarded as a marker of ongoing viral replication. MATERIAL AND METHODS: We investigated the prevalence of (+) and (-) strands of HCV-RNAs in sera and PBMC of 45 chronically hemodialyzed patients using PCR methods. We also determined HCV genotypes and their subtypes by Inno-LIPA method. RESULTS: Eight (17.8%) of analyzed patients were anti-HCV positive. In this group, we detected HCV-RNA (+) strands in sera and PBMC in 2 and 4 cases respectively, whereas HCV-RNA (-) ones were found in PBMC of 4 patients. Among the remaining 37 anti-HCV negative patients we found HCV-RNA positive strands in sera and PBMC in 2 and 3 cases respectively, whereas HCV-RNA negative strand was present in PBMC in one of them. CONCLUSIONS: Our results indicate that HCV actively replicate in PBMC in chronically hemodialyzed patients. In number of patients HCV-RNAs can be detected only in PBMC without concomitant presence of viremia or anti-HCV in sera. We did not find any correlation between genotypes of HCV and presence of HCV-RNAs strands in PBMC of the patients.

Short-term changes of serum IL-2 and IL-6 induced by interferon alpha-2b in patients with chronic hepatitis C.

BACKGROUND: The standard therapy of chronic hepatitis C with interferon alpha (IFN alpha) and ribavirin has established but limited efficacy. The prognostic factors of treatment are still under investigation. IL-2 and IL-6 are key cytokines involved in activation of B and T lymphocytes and thus in humoral and cellular responses; they are also deeply involved in generation and maintenance of inflammatory processes. The aim of the study was to evaluate the short-term influence of INF alpha-2b on serum IL-2 and IL-6 levels in sustained responders (SR) and non-responders (NR). MATERIAL AND METHODS: Altogether 12 patients (7 males and 5 females) chronically infected with HCV (anti-HCV positive, HCV-RNA positive by PCR) were enrolled to the study. Patients were treated with IFN 3 MU tiw for 6 months and then they were followed for another 6 months. Five patients responded to the treatment (sustained responders-SR)-Group I, seven patients did not respond (non-responders-NR)-Group II. Serum concentrations of IL-2 and IL-6 were assessed by ELISA before ['0'] and at 1st ['1'], 2nd ['2'], 3rd ['3'], 6th ['4'] and 12th ['5'] hour after the first IFN injection. CONCLUSIONS: Interferon alpha-2b induced short-term increase of serum IL-2 concentrations in SR but not in NR. Serum IL-6 level increased both in SR and NR but this effect was more pronounced and persisted longer in sustained responders.

Interferon versus interferon and UDCA combined therapy in chronic hepatitis C.

BACKGROUND: Interferon alpha (IFN) has been shown to have established efficacy in the treatment of chronic hepatitis C but its effectiveness is unsatisfactory. Combined therapies with IFN and other antiviral or immunomodulatory drugs are under evaluation. A combination of interferon alpha and ursodeoxycholic acid UDCA has been reported to give better results than interferon alone. The aim of the study was to assess the efficacy of IFN monotherapy versus IFN and UDCA therapy in patients with chronic hepatitis C. MATERIAL AND METHODS: We studied 38 patients (25 males and 13 females) chronically infected with HCV (anti-HCV positive, HCV-RNA positive by PCR). Seventeen of them were treated with IFN 3 MU tiw for 6 months--Group I. The remaining 21 patients were treated with IFN, at the same dosage, plus UDCA (10 mg/kg/day) also for 6 months--Group II. Patients were followed for 6 months. 6 months after the end of therapy, laboratory biochemical parameters, HCV viremia and proportion as well as time to relapse were assessed. CONCLUSIONS: In contrast to previous reports we did not find any differences neither in proportion of HCV reactivation nor in the time of its appearance among patients treated because of chronic hepatitis B with IFN alone or with IFN plus UDCA combined therapy. We also did not find any difference in initial and late response to the treatments in both groups.

Assessment of HBV DNA levels in human plasma and peripheral blood mononuclear cells by TaqMan Q-PCR assay.

Hepatitis B virus DNA (HBV DNA) levels were quantitatively determined by a TaqMan quantitative polymerase chain reaction (TaqMan Q-PCR) in the plasma and in the peripheral blood mononuclear cells (PBMCs) of 120 patients with biopsy proven chronic hepatitis. The number of HBV DNA copies ranged from 10(1) to 10(10)/ml of the blood. There were no significant differences between plasma and PBMCs levels of HBV DNA in neither of HBV infected alone nor HCV co-infected patients. The number of HBV DNA copies was higher in the both compartments in blood of patients infected with HBV only, in comparison to those co-infected with HBV and HCV. In patients chronic hepatitis caused by both viruses (co-infection), decrease of HBV DNA copies in the PBMCs preceded its disappearance from the plasma. Our results suggest that assessment of HBV DNA in PBMCs does not produce important advantage in diagnosis of chronic hepatitis, and it seems to have the prognostic significance of positive outcome among the patients with chronic hepatitis induced by HBV and HCV co-infection. Our results also indirectly indicate that HCV may reciprocally inhibit replication of HBV.

Increased AST and GGT activity as marker of RT-PCR inhibition in RNA extracts from peripheral blood.

The presence of HCV RNA in PMBC, with simultaneous absence of the virus in the plasma (7.8%), suggests that blood is better material for HCV RNA detection than plasma or serum in the diagnostic procedures of patients with chronic hepatitis C as well as in monitoring the antiviral therapy. We studied 111 patients with chronic hepatitis C (anti-HCV+) and elevated level of at least one biochemical marker: AST, ALAT, GGT, AP and bilirubin. Inhibition of amplification was 2% in plasma and 34% in whole blood samples. We applied modification of extraction to reduce the inhibitory effect on PCR, by introducing additional purification of the RNA extract in the Chomczynski method. After our modification of extraction was applied, inhibition was reduced to 1%. In attempt to identify such inhibitory markers that would label the samples, in which additional RNA extract purification should be applied, we analysed the activity of AST and ALAT enzymes, the key markers for parenchymal liver damage; GGT and AP, the markers for cholestatic hepatitis as well as bilirubin. We observed that the increased GGT and AST activities were correlated with the inhibition of RT-PCR. This correlation was statistically significant; for AST (Mann-Whitney test p = 0.09654 and Kolmogorow-Smirnow test p = 0.01543) and for GGT (Mann-Whitney test p = 0.02419 and Kolmogorow-Smirnow test p = 0.01921).

Dynamics of HCV replication in patients with chronic hepatitis C during interferon and ribavirin combined therapy.

Objective of the present study was to determine the effect of combined therapy with interferon alfa-2b (3 MU; thrice weekly s.c.) and ribavirin (1200 mg daily p.o.) on the number of copies of positive and negative strands of the hepatitis C virus RNA (HCV RNA) in patients with biopsy-proven chronic hepatitis C. Number of copies of both strands was determined by a TaqMan reverse transcription quantitative polymerase chain reaction (TaqMan RT Q-PCR) in the whole blood before treatment, and 4, 12 or 24 weeks after introduction of the treatment. Before the treatment positive strand of HCV RNA was more frequently detectable and its level was higher compared to that of the negative strand. In several patients, we observed therapy-induced transient appearance or increase in the number of copies of the negative HCV RNA strand. As a result of 24-week treatment, the negative strand of HCV RNA was eliminated from the blood more effectively than the positive strand. Our results suggest that the assessment of dynamics of changes of the positive and negative strands levels of HCV RNA in interferon-naive patients with chronic hepatitis C may be useful in monitoring short-term effectiveness of combined antiviral therapy. They also indicate that TaqMan RT Q-PCR appears to be a valuable tool in monitoring the therapy in these patients.

Do fibrinolytic proteins of human bile derive exclusively from gall bladder?

BACKGROUND: In this study we addressed the issue of whether fibrinolytic proteins are presented in gall bladder bile only or in choledochus bile as well. MATERIAL AND METHODS: Gall bladder bile was obtained from 20 patients (Group I) undergoing laparoscopic cholecystectomy. Bile from common bile duct was aspirated after insertion Kehr drainage from 9 patients (Group II). The concentrations of t-PA, u-PA, PAI-1 and PAI-2 were measured by ELISA. RESULTS: We have shown that in cholecystectomized patients fibrinolytic proteins can be detected in bile both from gall bladder and from choledochus. Mean concentrations of t-PA, u-PA, PAI-1 were lower in Group II (5.69 ng/ml vs 15.7; 0.46 ng/ml vs 0.7; 16.82 ng/ml vs 26.16 ng/ml) or nearly equal for PAI-2 (343.53 ng/ml vs 341.02). All differences were insignificant (p > 0.05). CONCLUSIONS: Based on these results we concluded that the entire biliary tree produces the fibrinolytic proteins thus this production is not restricted to the gall bladder as it was earlier reported [1].

Lymphocyte subsets in peripheral blood and bronchoalveolar lavage in patients with primary biliary cirrhosis.

BACKGROUND: The objective of this study was to examine the hypothesis that total blood and bronchoalveolar lavage (BAL) lymphocytes and their subsets in patients with primary biliary cirrhosis (PBC) with coexistent alveolitis are different from those in patients with PBC alone. Total blood and BAL lymphocytes and their subsets were evaluated in both types of patients and compared with a control group of normal healthy subjects. MATERIAL AND METHODS: 17 patients with well-documented PBC were divided into two groups: Group A--patients with coexisting alveolitis and Group B--patients with no alveolitis. Diagnosis was based on bronchoalveolar lavage. The control group consisted of healthy subjects. The lymphocyte subsets, including CD3, CD4, CD8, CD20 and CD56 were identified in blood and BAL cytospin smears using specific monoclonal antibodies. RESULTS: The percentage of CD3 and CD8 lymphocytes in BAL was significantly lower in a Group A, whereas the percentage of NK was significantly increased in group B as compared with the reference group. The percentage of blood CD4 lymphocytes in both subgroups of the PBC patients was significantly decreased as compared with the controls. The percentage of NK in both groups was significantly increased as compared with the controls. CONCLUSIONS: The relationship between the blood and BAL lymphocyte subsets in both studies groups were similar. Therefore, we suggest that PBC is a systemic disease in which lymphocytic infiltrations are not limited to the liver but may also involve lungs and probably other organs.

[Effectiveness of antiviral treatment of patients with chronic hepatitis C (a Polish multicenter study)]. / Efektywnosc leczenia przeciwwirusowego przewleklego zapalenia watroby typu C (wieloosrodkowe badania pòlskie).

Interferon alpha (INF) is routine treatment in patients with chronic hepatitis C. Many controlled investigations were evaluated to establish the optimal schedule of treatment with sustained virological and biochemical response. Recently, multicentre meta-analyses suggest that combination therapy (INF + Ribavirin) was more effective than treatment with interferon alone. The aim of this study was to compare the efficacy of four schedules of antiviral treatment in 445 patients with chronic hepatitis C. Combination therapy (INF + Ribavirin) given for 6 mo. and monotherapy (INF) for 18 mo. were more effective than interferon alone given for 6 mo. Treatment with INF alone for 6 mo. was demonstrated to be insufficient.

Cardiac sympathovagal balance during endoscopic retrograde cholangiopancreatography.

BACKGROUND AND STUDY AIMS: Cardiopulmonary complications have been reported during upper gastrointestinal endoscopy and endoscopic retrograde cholangiopancreatography (ERCP). The aim of this study was to evaluate the sympathovagal response to the stretching of the common bile duct caused by contrast medium injection during ERCP. PATIENTS AND METHODS: The 16 patients included in the study were assigned to two groups according to the size of common bile duct. Group A consisted of patients with normal-sized bile ducts, while Group B patients had dilated common bile ducts. The heart rate variability (HRV) technique was used to assess the sympathovagal balance. The HRV data were gathered using the Holter technique, and frequency domain analysis revealed two main spectral components of HRV: low frequency (LF) and high frequency (HF). RESULTS: In patients with normal-sized bile ducts, contrast injection initiated a rapid increase in the power spectra (ms2) of both the HF and LF components. In patients with dilated common bile ducts, this phenomenon was not observed. CONCLUSIONS: During ERCP, contrast injection into a dilated common bile duct does not stimulate the autonomic nervous system in the same way as it does when the common bile duct is of normal size.

Accuracy of t-PA, u-PA, PAI-1 and PAI-2 estimation in human bile by ELISA kits.

UNLABELLED: The aim of this study was to establish the accuracy of determination of fibrinolytic proteins in human bile using commercial kits. MATERIAL AND METHOD: Gall bladder bile was obtained from patient undergoing laparoscopic cholecystectomy. The concentrations of tissue plasminogen activator (tPA), urokinase (uPA) and plasminogen activator inhibitor-1 (PAI-1) and inhibitor-2 (PAI-2) in bile were measured by enzyme linked immunoassay. Afterwards we conducted so called 'recovery test'. RESULTS: Recovery percents of t-PA, u-PA, PAI-1, PAI 2 were very high i.e. 91.94, 94.07, 93.95, 91.2. We concluded that human fibrinolytic proteins in bile could be detected very accurately using commercial kits.

Isolation and evaluation of susceptibility to sulphasalazine of Desulfovibrio desulfuricans strains from the human digestive tract.

Various genera of sulphate reducing bacteria (SRB) have been found in the human digestive tract. It is suggested that some of SRB species may be responsible for the development of the clinical symptoms of ulcerative colitis and other disease of large intestine. Sulphasalazine (salicyl-azo-sulphapyridine, SAS) is commonly used to treat patients with ulcerative colitis and Crohn disease. Above 30 samples of faeces or biopsy specimens from 25 patients (age 45 +/- 14 years; M/F, 13/12) suffering from gastrointestinal disorders were used for isolation of Desulfovibrio desulfuricans species. The morphological, physiological and biochemical characteristics of isolated strains and also their susceptibility to SAS was determined. D. desulfuricans isolates were obtained from 5 amongst all patients assayed. Some abnormal, cigar-shaped cells were detected as accompanying the cells represented by rods, curved rods and vibrios. After strains purification, two types of colonies were present on the solid Postgate's medium B (containing lactate as a carbon source and sulphate for energy conservation): the black colonies growing in bulk of agar medium and the transparent, surface-growing mucous colonies. These two types of D. desulfuricans colonies may be a result of different iron availability for bacterial cells. High metabolic activity of strain was not always accompanied by the presence of H2S gas lock in the test tube, although the H2S odor was perceptible. All tested strains multiplied inconsiderably slowly in the presence of SAS at concentrations 10, 20, 40 and 60 mg/cm3. The growing concentrations of SAS did not cause a proportional decrease of the bacterial cells number. Taking into account the positive results of using SAS to treat patients with some colonic diseases and the indicated resistance to SAS of intestinal D. desulfuricans strains, it appears probable, that this SRB species isn't responsible for the development of mentioned diseases.