Dietary conjugated linoleic acid affects blood parameters, liver morphology and expression of selected hepatic genes in laying hens.

The objective of this research were to investigate the effect of a conjugated linoleic acid (CLA)-enriched diet on Isa Brown laying hen health status and to provide a comprehensive analysis of changes in blood parameters, liver morphology and selected hepatic gene expression. Hens were allocated to the control and experimental group (diet enriched with 0.75% CLA) for a total period of 4 m. At the end of the experiment half of the hens from each group were slaughtered for analyses. The remaining hens were transferred to an organic farm for the next 5 m and fed on the diet without CLA supplementation. The CLA-enriched diet resulted in significant changes in blood and serum parameters; specifically, haematocrit (HCT), mean corpuscular volume (MCV) and white blood cells (WBC) count were decreased compared to the control. The total cholesterol (TC) was not significantly affected while the triacylglycerol's (TG) concentration was elevated. The activity of alanine aminotransferase (ALT) was significantly increased in the CLA-supplemented group, while aspartate aminotransferase (AST) showed an increasing tendency. Liver biopsies showed pathological changes classified as non-alcoholic fatty liver disease (NAFLD). Additionally, the expression of hepatic genes involved in fatty acids synthesis (ME1, ACLY, ACC, FASN, SCD1), oxidation (CPT1α, PPARA), detoxification processes (Cytochrome P450, CYP, Flavin-containing monooxygenase, FMO3), oxidative stress (NOX4, XbP1) and inflammation (IL6, TNFα) were elevated. Cessation of CLA supplementation for 5 m of organic farming resulted in normalisation of blood and hepatic parameters to the levels observed in control hens. The results of this study indicate that dietary CLA triggers an integrated stress response in laying hens and activates mechanisms involved in liver detoxification.

Disturbances in some gene expression in T regulatory cells separated from children with metabolic syndrome.

The metabolic syndrome (MS) is defined as a cluster of risk factors, including abdominal obesity, dyslipidaemia, glucose intolerance and hypertension, which increase the risk for coronary heart disease. The immunological aspects of obesity and MS, including the role of T regulatory cells, have been intensively investigated. The aim of this study was to determine whether there is any disturbance in T regulatory cells number and/or function in children with MS. The percentages of T regulatory cells in the peripheral blood of children fulfilling the International Diabetes Federation criteria of the disease (n = 47) were assessed. Treg cells were also separated for further analysis of multiple genes important in their function with the use of real-time RT-PCR. We did not observe any difference in Treg percentages between study and control group but there was lower expression of some molecules including transforming growth factor-beta and interleukin-12 family members in Treg cells separated from children with MS compared to the healthy subjects. Our study is the first to report significant disturbances in some gene expression in T regulatory cells separated from children with MS. The results should be useful for further research in this field, including immunotherapeutic interventions.

Liver cell dysplasia in liver cirrhosis and hepatocellular carcinoma.

The aim of the study was to assess the incidence of both types of liver cell dysplasia and concomitance with cirrhosis, hepatocellular carcinoma (HCC) and positive reaction for HBsAg in the autopsy material and an attempt to determine a relationship between these two types of liver cell dysplasia and hepatocellular carcinoma. Autopsy material included 102 cases of hepatocellular carcinoma, 101 cases of hepatocirrhosis without accompanying cancer and 106 control cases. Histological specimens stained with HE were analyzed for the presence of large liver cell dysplasia (LLCD) according to Anthony et al., small liver cell dysplasia (SLCD) according to Watanabe et al., the presence of macroregenerative nodules (< 8 mm) and antigen HBs (stained with orcein according to Shikata). The detected LLCD were also assessed semiquantitatively taking into account the number of dysplastic areas in a given case. Statistical significance of the results was tested with the chi square test. LLCD was most frequently detected in HCC with concomitant cirrhosis (55.3%), then in cirrhosis without HCC (40.6%), and in HCC without cirrhosis only in 12.5%. LLCD was found significantly more frequently (p < 0.05) in cirrhosis with HCC than in cirrhosis without HCC. Antigen HBs was found in 25.6% of cirrhoses and/or HCC. No significant differences in the presence of HBsAg were seen between the analyzed groups. The incidence of LLCD and HBsAg in controls was significantly lower than in other groups. A mean age at death in case of cirrhosis with HCC subdivided into that with or without LLCD was not significantly different, whereas in case with cirrhosis with LLCD age at death was 10.8 years higher (the difference statistically significant). Analysis of material with respect to gender revealed a high proportion of men in case of HCC with concomitant cirrhosis but without LLCD (13:1). A strong relationship was seen between the presence of positive reaction for HBsAg and LLCD (p < 0.001). Also the intensity of LLCD positively correlated with the presence of HBsAg. Furthermore, a positive correlation was seen between the presence of LLCD and macronodular cirrhosis (posthepatitic). The present findings suggest a closer relation between HBV infection and LLCD than between cirrhosis or HCC and LLCD. Also morphological patterns of LLCD foci do not confirm the hypothesis of some investigators about the precancerous character of these lesions. In the whole current material only seven cases of SLCD were detected. They were all present in cirrhotic livers with concomitant HCC. Both the morphological pattern of these lesions and their sometimes discerned close spatial relation with HCC foci indicate that SLCD is an alternative way of HCC development.

[Humoral response in hepatitis C infections: types of anti-HCV and reactivity evaluated using BLOT-index]. / Odpowiedz humoralna w zakazeniach wirusem C zapalenia watroby; rodzaje anty-HCV i typy reaktywnosci oceniane wskaznikiem BLOT-indeks.

In this paper an attempt has been made at quantitative evaluation of reactivity of various types anti-HCV using proposed BLOT-index. We based our research on 20 patients diagnosed with hepatitis-C (clinical, biochemical, serological and enzymatical criteria--repeatedly positive results of second generation screening tests EIA ABBOTT HCV and UBI HCV Monoelisa Organon Teknika) who were anti-HCV determined at the beginning the acute phase and after 3 and 6 months. Multiantigen tests were used veryfying LIATEK-HCV 2 and LIATEK-HCV 3 Organon Teknika. Value of BLOT-index was algebraic sum of "pluses" for particular anti-HCV. Three models of dynamics of BLOT-index were observed: increase, plateau and decrease. Statistically significant differences between values of BLOT-index were shown at the beginning of the acute phase of hepatitis-C and after 6 months. There were no differences observed between convalescents and patients who developed chronic hepatitis-C (test t-Student's and f-Fisher's). Reactivity of anti-HCV, evaluated using BLOT-index, shown increasing trend, which is a dominant pattern, however quantitative and qualitative changes do not occur more frequently than every 6 months. The LIATEK-HCV 3 gives fewer undetermined reactions and detects anti-HCV earlier than a former generation of this test.

[The influence of ursodeoxycholic acid on some biochemical, immunologic and histopathologic parameters in patients with primary biliary cirrhosis]. / Wplyw kwasu ursodezoksycholowego na niektóre parametry biochemiczne, immunologiczne oraz obraz histopatologiczny u chorych z pierwotna zólciowa marskoscia watroby.

UNLABELLED: Primary biliary cirrhosis (PBC) is a chronic, inflammatory disease, which affects the small and medium size bile ducts and is characterized by chronic cholestasis. Ursodeoxycholic acid (UDCA), that had earlier been used in treatment of cholesterol cholelithiasis, was introduced to pharmacotherapy of PBC 10 years ago. Our study was carried out in 30 patients, who were suffering from PBC and were treated at Department of Gastroenterology, Collegium Medicum, Jagiellonian University in Cracow. Beside the routine biochemical and enzymatic tests all patients had the level of immunoglobulin checked and selected ones acute phase protein. The percutaneous liver biopsy was carried out in each case; the morphological changes were classified according to Ludwig scale. All above tests were carried out before implementing UDCA as well as after one year from the beginning of the treatment. The group of patients is still closely monitored. CONCLUSIONS: 1. By using the UDCA in treatment of PBC biochemical parameters of cholestasis improved, also in some cases the inflammatory reaction decreased; 2. UDCA used in monotherapy had no influence on the process of fibrosis.

Syncope in children and adolescents living in north-eastern Poland - scope of causes.

PURPOSE: To investigate the diagnostic yield of different tests and asses the scope of causes in children referring to the hospital with a syncope in north-eastern Poland. METHODS: A review of 386 consecutive patients (age 2-18 years) living in north-eastern Poland presenting to the cardiology department with a new onset syncope (which appeared to be neurally mediated by history) was undertaken. The patients underwent physical examination, laboratory tests, electrocardiography, 24-holter monitoring, head-up tilt-test, exercise test, echocardiography and electroencephalography. All the tests were performed in most of the patients, without ending the diagnostics after finding the first probable cause of loss of consciousness. RESULTS: 229 potential causes of syncope were found in 191 patients (49.4%), with 2 possible causes in 32 patients and 3 potential causes in 3 patients. The top 3 tests with the highest diagnostic yield were: head-up tilt test (41.4%), 24-holter monitoring (14.5%) and echocardiography (8.4%). Electroencephalography was useful in 3 patients (1.5%) and exercise test did not help in any patient. CONCLUSION: In some children more than one potential cause of syncope was diagnosed. That might confirm multiple factors' causality of syncope. Head-up tilt-test has the highest diagnostic yield in children with syncope. If the diagnostic protocol is not very strict, one might find some crucial conditions in about 8.3% of patients.

Effects of peripherally and centrally applied ghrelin in the pathogenesis of ischemia-reperfusion induced injury of the small intestine.

Ghrelin is an important hormone involved in the control of the human appetite center. Recently, protective properties of this hormone have been recognized in various models of impairment of the gastric mucosa, including stress, ischemia and reperfusion (I/R). Ghrelin is predominantly secreted by the gastric mucosa of stomach, but there are other sources of ghrelin, for example in the hypothalamus and various parts of the central nervous system (CNS) that should be taken into consideration. This hormone exerts biological effects via the activation of growth hormone secretagogue receptor (GHSR), the presence of which was confirmed in different parts of the gastrointestinal (GI) tract and midbrain structures. Although substantial evidence of the divergent biological effects of ghrelin and the mechanism of its action has been emphasized, the precise mechanisms of ghrelin which affords GI protection is still unclear. Particularly, there is a sparse amount of evidence concerning its action on the GI system. The major aim of the present study was to evaluate the importance of peripherally and centrally administered ghrelin at different times of the ischemia and reperfusion (I/R period in the modulation of resistance of the intestinal mucosa to the injury induced by ischemia and subsequent reperfusion. Secondly, we wanted to evaluate the possible mechanism of the action of ghrelin with a particular focus on its influence on the intestinal blood flow. Male Wistar rats were divided into 4 series (A-D) of the experimental groups (n=7). In series A the importance of peripherally administered ghrelin at different time of I/R period was studied. In series B the importance of centrally administered ghrelin at different time of I/R period was evaluated. In series C and D, the mechanisms of peripherally and centrally administered hormone were examined, respectively. Two models of the I/R period were selected: short lasting (30/60 min) and long lasting (60/120 min). The following drugs were used: ghrelin (50 µg/kg i.p. or 1 nmol in 10 µl i.c.v.), 6 hydroxy dopamine (50 mg/kg i.p.), nadolol (0.5 mg/kg i.p.), calcitonin gene related peptide fragment (CGRP(8-37), 100 µg /kg i.p.), capsaicin (5-10 mg/100 ml solution s.c.). The mesenteric blood flow (MBF-ml/min), the intestinal microcirculatory blood flow (LDBF-PU), the arterio-venous oxygen difference (AVO(2)-ml/O(2)/100 ml blood), and the intestinal oxygen uptake (VO(2)) in ml O(2)/min were measured. Mucosal impairment was assessed planimetrically with the use of a digital photo analyzer (LA) and histologically with the use of the six-point Park/Chiu scale. Peripheral administration of ghrelin evoked marked increase of MBF and LDBF by 42% and 48%, respectively, with significant reduction of LA by 38%. When ghrelin was administered at the beginning of the reperfusion period during the short I/R period or prior to the long lasting I/R period, the vascular reactions and protective effects were reduced, but not completely abolished. The central administration of ghrelin before the short I/R period significantly increased the MBF and LDBF by about 32% and 35%, respectively, as well as LA reduction by about 20% in comparison to the control group. However, when ghrelin was administered prior to the long I/R period or after the onset of completed ischemia, neither vascular nor protective effects were noticed. Sensory denervation and the blockade of the CGRP1 receptors totally blocked the protective and hyperemic effects of the peripherally administered ghrelin. Selective blockade of the adrenergic system or blunting of the vagal nerves (vagotomy) significantly but not totally eliminated the effects of centrally applied ghrelin, which were abolished when both adrenergic and parasympathetic pathways were ablated. These results indicate that ghrelin applied centrally or peripherally markedly increases resistance of the intestinal tissue during the I/R period induced mucosal and hyperemic impairment evoked by I/R. Ghrelin is an important mediator of the increase in the intestinal microcirculation and elevation of the intestinal metabolism, which seems to be, at least in part, responsible for the observed protection of the intestine subjected to I/R. Impairment of this microvasculature response due to I/R seems to be responsible for a markedly observed weaker effect of ghrelin when this hormone was administered after the ischemic period. The lack of a protective effect observed after central administration of this peptide against a long lasting I/R period is probably due to damage of neural pathways caused by I/R. Finally, the peripheral activity of ghrelin in the intestine is mediated by the sensory neurons with a prominent role of CGRP released from their endings. However, this peripheral action of ghrelin depends upon the proper functioning of both the sympathetic and parasympathetic system.

Met-enkephalin in the liver as a marker of hepatocellular damage in chronic viral hepatitis type B and C.

PURPOSE: The aim of the study was to assess the liver Met-enkephalin concentration in chronic viral hepatitis type B and C as well as in liver cirrhosis in order to estimate the role of opioid system in pathogenesis of liver disease. MATERIAL AND METHODS: The concentration of Met-enkephalin was examined in liver tissue of 103 consecutive patients with chronic hepatitis type B and C. Control group consisted of uninfected patients. Met-enkephalin concentration was analyzed in relation to the degree of hepatic necroinflammatory activity and the extent of fibrosis estimated by histopathological examination of liver bioptates and compared to such parameters as age, sex and concomitant diseases. RESULTS: Significant differences in Met-enkephalin concentration were found between cases with advanced fibrosis (stage 3 and 4 acc. to Batts and Ludwig classification) and cases with fibrosis classified as stage 2 (p < 0.05). Met-enkephalin concentration was higher in HCV infected patients in comparison to HBV infected patients (p < 0.05) and uninfected controls (0.05 < p < 0.1). There wasn't found any correlation between Met-enkephalin level and necroinflammatory activity in the liver, age, sex and concomitant diseases. CONCLUSIONS: Met-enkephalin concentration measurement in the liver tissue seams to be a useful method for differentiation of stage 2 from stages 3 and 4 of severe liver fibrosis. There is increased concentration of Met-enkephalin in liver tissue in HCV infected patients in comparison to HBV infected or uninfected individuals. The degree of necroinflammatory activity in the liver as well as sex and age of patients with chronic hepatitis do not correlate with changes in opioid system.

Colorectal polyps in autopsy material. Part I. Adenomatous polyps.

In prospective studies specimens of the large bowel were obtained from 733 autopsy cases. After fixing they were examined under illuminated magnifying lens and all polypous lesions were excised for histological analysis and classified according to Correa's criteria (8). Adenomatous polyps were found in 280 cases (38.2%) and their prevalence increased with age in both sexes. Adenomatous polyps, also multiple were found most frequently in the transverse colon, and then ascending colon, sigmoid, descending colon and rectum. The mean number of adenomatous polyps per positive specimen was 2.33 for men and 2.60 for women. In 6.8% their diameter was > or = 10 mm. The prevalence of polyps with severe degree of dysplasia (III degree) increased from proximal to distal segments of the colon in both sexes and their prevalence is significantly higher in the ascending colon in women. In the whole series there were 6 adenocarcinomas of 5-18 mm in size, all in women over 80 years of age. The results of the present study were compared with the data concerning other populations of the world. In the light of a repeatedly confirmed a generally accepted relationship between adenoma and adenocarcinoma of the large intestine the results of our study seem to underestimate the prevalence of colon adenocarcinoma in the population of Cracow and Cracow district.