Histological Examination Of Liver Research Articles

Mechanical ventilation and volutrauma: study in vivo of a healthy pig model.

Mechanical ventilation is essential in intensive care units. However, it may itself induce lung injury. Current studies are based on rodents, using exceptionally large tidal volumes for very short periods, often after a "priming" pulmonary insult. Our study deepens a clinically relevant large animal model, closely resembling human physiology and the ventilator setting used in clinic settings. Our aim was to evaluate the pathophysiological mechanisms involved in alveolo/capillary barrier damage due to mechanical stress in healthy subjects. We randomly divided 18 pigs (sedated with medetomidine/tiletamine-zolazepam and anesthetised with thiopental sodium) into three groups (n=6): two were mechanically ventilated (tidal volume of 8 or 20 ml/kg), the third breathed spontaneously for 4 hours, then animals were sacrificed (thiopental overdose). We analyzed every 30' hemogasanalysis and the main circulatory and respiratory parameters. Matrix gelatinase expression was evaluated on bronchoalveolar lavage fluid after surgery and before euthanasia. On autoptic samples we performed zymographic analysis of lung, kidney and liver tissues and histological examination of lung. Results evidenced that high Vt evoked profound alterations of lung mechanics and structure, although low Vt strategy was not devoid of side effects, too. Unexpectedly, also animals that were spontaneously breathing showed a worsening of the respiratory functions.

Toxicity studies of alkaloids of seeds of Datura stramonium and synthesis alkaloids in male rats

The effects of acute, subacute and chronic administration of alkaloids atropine and scopolamine, the main constituents of the active principle of Datura stramonium, with toxic properties, were studied in male Albino-Wistar rats. After acute i.p administration of dose 100 mg/kg (1/4 DL 50 ) of total alkaloids to the seeds of D. stramonium , there were no remarkable changes in general appearance and no deaths occurred in any experimental group. 24 h after total alkaloids of seeds, a significant reduction in tissues (liver, spleen and brain) was observed. The red blood cells (RBC), Hematocrit (HCT), Hemoglobin (HGB) and white blood cells (WBC) were significantly higher in the treated groups than the control group. There were no statistical differences in Glutamic-oxaloacetic transaminase (GOT), Glutamic-Pyruvic Transaminase (GPT) and alkaline phosphatase (ALP) observed between groups. Histological examination of liver showed no histopathological changes. Subacute study for four weeks showed no resulting mortality or signs of toxicity. The relative weight of kidneys showed a significant decrease, however, these doses of synthetic alkaloids (5.2 mg/kg of atropine and 2.6 mg/kg of scopolamine) produced significant increase of lungs in comparison with the control group. RBC, HBG, HCT and PLT values of control group were significantly higher than those of the treated group. The enzyme activities of GOT, GPT and ALP were significantly increased. The microscopic examination of liver showed normal conservative lobular architecture and necrotic areas. In chronic study, the synthetic alkaloids administered i.p at daily doses 4.2 mg/kg of atropine and 1.6 mg/kg of scopolamine, did not produce death, However the diarrhoea and hypoactivity were observed. The relative weight of liver was significantly less than that of the control group. The haematological analysis revealed a significant decrease in RBC, HCT, HBG and WBC and we observed manifold centrolobular necrotic areas, and blood congestion and dilated central veins in treated groups.

The changing role of liver biopsy in diagnosis and management of haemochromatosis

Liver biopsy with histological examination of liver tissue was for many years the cornerstone of the diagnosis of haemochromatosis, allowing assessment of the degree of iron overload and examination of liver histology for the acute and chronic effects of iron overload. In the past two decades the role of liver biopsy in haemochromatosis has changed dramatically. Liver biopsy is rarely requested for two main reasons: (1) genetic testing for human haemochromatosis (HFE) mutations has proved to be very reliable in the diagnosis of haemochromatosis in Caucasian populations, and (2) the majority of patients with haemochromatosis are now diagnosed at an early stage well before permanent tissue damage occurs, so the need to assess tissue and organ damage has diminished. Liver biopsy continues to have a very important role in a small number of haemochromatosis patients for whom it has both diagnostic and prognostic implications. Liver biopsy is essential for the accurate assessment of patients with non-HFE haemochromatosis and in patients who have dual pathology. It is also useful where there appears to be a discrepancy between HFE genotypes and iron studies, particularly in HFE heterozygotes. Finally, liver biopsy is currently the 'gold standard' for the diagnosis of fibrosis and cirrhosis, although this is changing as non-invasive methods for assessing fibrosis become more reliable and available. Therefore, it is important that pathologists maintain their knowledge and skills in the use of liver biopsy in haemochromatosis and other iron storage disorders.

Effect of Metformin and Glimepiride on Liver and Kidney Functions in Alloxan-Induced Diabetic Rats

Metformin and glimepiride have been marked as potential hypoglycemic agents for the treatment of type 2 diabetes. The purpose of this study was to investigate the effect of daily oral administration of metformin (500 mg/kg) or glimepiride (0.5 mg/kg) alone or in combination for 28 days on some physiological parameters and histological alterations in liver and kidney of the diabetic rats. Diabetes was induced by a single intraperitoneal dose of alloxan (150 mg/kg) Data showed that metformin and glimepiride alone or in combination induced a significant decrease in serum glucose levels in the diabetic rats. Treating the diabetic rats with glimepiride increased the AST activity significantly. Meanwhile, administration of glimepiride or metformin had no significant effect on the activity of ALT. On the other hand, the combination of both drugs exhibited a significant reduction in the activities of AST and ALT as compared to that in the diabetic control rats. Glimepiride treatment revealed that total protein and globulin levels were increased, while metformin or the combined drug resulted in a significant decrease in albumin level which almost reached the normal value and an increase in the globulin than that of the diabetic rats. Creatinine level improved significantly in all the treated groups in comparison to that of the diabetic group and reached the normal values. On the other hand, no sign of improvement in the levels of urea was observed in the rats treated with metformin or the combined drug, while treatment with glimepiride decreased urea levels significantly. Histological examination of liver in the diabetic rats showed extensive necrosis of hepatocytes, cytoplasmic vacuolation, distended sinusoids with massive congestion. In addition, the kidney glomeruli increased in size and the renal tubules degenerated. The treatment with metformin or glimepiride ameliorated the hepatic injury, at the same time; the combined drug caused severe destruction in the liver cells. The treated rats also exhibited shrinking in the glomerular capillaries with widening of Bowman’s space, mesangial matrix expansion, necrosis and vacuolation of renal tubules.In conclusion, the combination of biochemical and histological biomarkers provides useful and sensitive tools in the investigation of chronic effects induced by diabetes and anti-diabetic drugs.

A critical life-supporting role for cystathionine γ-lyase in the absence of dietary cysteine supply

This study examined the important relationship between cystathionine γ-lyase (CSE) functionality and cysteine supply for normal growth and life span. Mice with a targeted deletion of the CSE gene (CSE-KO) were fed a cysteine-limited diet and their growth and survival patterns as well as levels of cysteine, homocysteine, glutathione, and hydrogen sulfide (H2S) were measured. CSE-KO mice fed a cysteine-limited diet exhibited growth retardation; decreased levels of cysteine, glutathione, and H2S; and increased plasma homocysteine level. However, histological examinations of liver did not reveal any abnormality and plasma levels of aspartate aminotransferase, alanine aminotransferase, and albumin were normal in these animals. No CSE-KO mice survived after 12weeks of feeding with the cysteine-limited diet. Supplementation of H2S to the CSE-KO mice failed to reverse the aforementioned abnormalities. On the other hand, supplementation of cysteine in the drinking water of the CSE-KO mice significantly increased plasma cysteine and glutathione levels. This eventually led to an increase in body weight and rescued the animals from death. In conclusion, CSE is critical for cysteine biosynthesis through the transsulfuration pathway and the combination of CSE deficiency and lack of dietary cysteine supply would threaten life sustainability.

Differential expression of MAPK-pathway genes during liver regeneration of mouse

To investigate the different expression profiles of MAPK pathway genes and their corresponding functions during liver regeneration, we used a CCl4 induced mouse liver regeneration model in this study. Mouse was injected with CCl4 in the abdominal cavity to cause damage in the liver and followed by liver histology examination and measurement of serum ALT levels in blood sample collected at 0, 0.5, 1.5, 4.5, and 7 d after CCl4 injection. Differentially expressed genes in the MAPK pathway during liver regeneration were analyzed using mouse cDNA microarray method (Affymetrix). The results obtained were further subjected to hierarchical clustering study and were validated with real-time PCR. Microarray hybridization identified 31 out of the 93 MAPK pathway component genes, which have significantly altered their expression levels during liver regeneration. Among them, both up- and down-regulated genes were classified into various groups according to clustering studies and functional analysis. At the initial stage of liver regeneration, the number of up-regulated genes was greater than the down-regulated genes, while at the late stage the situation was reversed. Our results suggest that MAPK pathway might play different regulatory roles in responding to different stages of liver regeneration.

Hepatoprotective and Antioxidant Effects of Silybum marianum Plant in Rats

The plant phenolic compounds such as flavonoids and isoflavonoids have an important role in the treatment of many diseases and some of them induce a potent hepatoprotective effect. In the present study, the hepatoprotective and antioxidant effects of ethanolic extract of Silybum marianum (Milk thistle) plant were evaluated by measuring liver function; tissue antioxidant enzymes and histological examination of liver. Antioxidant enzymes as SOD, CAT and GSH were measured in liver homogenate. In vitro determination of the extract activity was compared to DPPH and measured spectrophotometrically. Oral administration of Silybum marianum extract significantly (P < 0.05) decreased liver enzyme activity when given in repeated doses. The small and large doses increased the activity of antioxidant enzyme. The obtained results proved the protective effect of ethanolic extract on liver cells. The protective effect of this extract may be attributed to presence of flavonoids compounds and their antioxidant effects. It could be concluded that ethanol extract of Silybium marianum have a significant hepatoprotective and antioxidant activity and may be useful for patients who suffer from liver diseases.

Mechanical ventilation and volutrauma: study in vivo of a healthy pig model

Mechanical ventilation is essential in intensive care units. However, it may itself induce lung injury. Current studies are based on rodents, using exceptionally large tidal volumes for very short periods, often after a "priming" pulmonary insult. Our study deepens a clinically relevant large animal model, closely resembling human physiology and the ventilator setting used in clinic settings. Our aim was to evaluate the pathophysiological mechanisms involved in alveolo/capillary barrier damage due to mechanical stress in healthy subjects. We randomly divided 18 pigs (sedated with medetomidine/tiletamine-zolazepam and anesthetised with thiopental sodium) into three groups (n=6): two were mechanically ventilated (tidal volume of 8 or 20 ml/kg), the third breathed spontaneously for 4 hours, then animals were sacrificed (thiopental overdose). We analyzed every 30' hemogasanalysis and the main circulatory and respiratory parameters. Matrix gelatinase expression was evaluated on bronchoalveolar lavage fluid after surgery and before euthanasia. On autoptic samples we performed zymographic analysis of lung, kidney and liver tissues and histological examination of lung. Results evidenced that high Vt evoked profound alterations of lung mechanics and structure, although low Vt strategy was not devoid of side effects, too. Unexpectedly, also animals that were spontaneously breathing showed a worsening of the respiratory functions.

The Protective Function of Neutrophil Elastase Inhibitor in Liver Ischemia/Reperfusion Injury

BACKGROUND.: A neutrophil elastase (NE) inhibitor, Sivelestat, has been approved for the treatment of acute lung injury associated with systemic inflammation in humans. Some reports have also shown its protective effects in liver inflammatory states. We have recently documented the importance of NE in the pathophysiology of liver ischemia/reperfusion injury, a local Ag-independent inflammation response. This study was designed to explore putative cytoprotective functions of clinically available Sivelestat in liver ischemia/reperfusion injury. METHODS.: Partial warm ischemia was produced in the left and middle hepatic lobes of C57BL/6 mice for 90 min, followed by 6 or 24 hr of reperfusion. The mice were given Sivelestat (100 mg/kg, subcutaneous) at 10 min before ischemia, 10 min before reperfusion, and at 1 and 3 hr of reperfusion thereafter. RESULTS.: Sivelestat treatment significantly reduced serum alanine aminotransferase levels and NE activity, when compared with controls. Histological liver examination has revealed that unlike in controls, Sivelestat ameliorated the hepatocellular damage and decreased local neutrophil activity and infiltration. The expression of proinflammatory cytokines (tumor necrosis factor-alpha and interleukin-6), chemokines (CXCL-1, CXCL-2, and CXCL-10), and toll-like receptor 4 was significantly reduced in the treatment group, along with diminished apoptosis through caspase-3 pathway. Moreover, in vitro studies confirmed downregulation of proinflammatory cytokine and chemokine programs in mouse macrophage cell cultures, along with depression of innate toll-like receptor 4 signaling. CONCLUSION.: Sivelestat-mediated NE inhibition may represent an effective therapeutic option in liver transplantation and other inflammation disease states.

Abstract 1886: Selenium prevents specific iron-mediated hepatic cellular changes associated with antioxidant defense and proliferation

Abstract Hepatocellular carcinoma (HCC), one of the most common causes of cancer death, is on the rise in the United States. HCC is a slow process occurring in the setting of liver cell injury that can be induced by oxidative stress leading to DNA damage and resulting in mutations that accumulate with time. Although many risk factors for HCC are known, the molecular pathogenesis of HCC is not well understood. In many chronic liver diseases that culminate in HCC, including hereditary hemochromatosis and chronic hepatitis C, excess iron deposition is observed. However, little is known about the effects of iron overload in the liver on cellular pathways. We generated a mouse model of chronic hepatic iron overload of liver by supplementing the diet with the iron donor 3,5,5-trimethyl-hexanoyl ferrocene (TMHF). Histological examination of livers from mice fed a TMHF diet for as long as 9 weeks showed no signs of inflammation or necrosis. Hepatocytes underwent expected physiological responses to iron in that ferritin levels were increased and hemosiderin accumulated with time. TMHF treated mice developed hepatomegaly and had elevated hepatic cyclin D1 and PCNA expression. TMHF treatment also resulted in a time-dependent decrease in hepatic levels of the antioxidant protein glutathione peroxidase 1 (GPX-1) but had no effect on Cu/Zn superoxide dismutase (SOD) or Mn-SOD. TMHF treatment also led to activation of β-catenin, induction of matrilysin (MMP7), and increased levels of cyp2b10. In this study, we tested the ability of selenium to specifically restore molecular and cellular changes induced by chronic hepatic iron overload. Mice were fed a diet supplemented with or without TMHF in the presence or absence of selenium for periods of time ranging from one to nine weeks. Selenium was provided as sodium selenite (15 ppm) or the synthetic organoselenium 1,4-phenylenebis (methylene) selenocyanate, p-XSC (15 ppm). Treatment of control mice, not fed TMHF, with selenium had no effect on the measured end points. As expected, supplementation with selenium in either form had no effect on TMHF induced ferritin expression or hemosiderin accumulation. However, dietary supplementation with selenium, simultaneously with TMHF treatment, markedly reduced TMHF mediated induction of cyclin D1 and PCNA expression and prevented iron mediated decrease in GPX-1 levels. Selenium had no effect on iron-induced hepatomegaly, activation of β-catenin, induction of MMP7, or increased cyp2b10. We conclude that (1) selenium restores the levels of the antioxidant defense enzyme GPX-1 and reduces iron induced stimulation of proliferation in a mouse model of hepatic iron overload; (2) the effects of selenium on these pathways occur by a mechanism other than simply reduction in hepatic iron levels; and (3) the effects of selenium are specific to some and not all iron-mediated changes in hepatic cellular signaling. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1886.

A canine portal hypertension model induced by intra‐portal administration of Sephadex microsphere

Big animal models of portal hypertension are important for the research into this disease. The aim of this study was to establish a canine portal hypertension model by intra-portal administration of microspheres. Sixteen mongrel dogs were assigned to control group and experimental group randomly. The catheterization of portal vein was performed by laparotomy and the outer end of the catheter was fixed subcutaneously in the abdominal wall. The dogs of the experimental group were given intra-portal injections of microspheres at a five-day interval, six times in total. Portal hemodynamics, blood cell counting, liver and renal function test, portography, gastroscopy, liver, spleen and lung histological examination were taken to evaluate the model. 1, 2, 3 and 4 months after initial injection of microspheres, portal venous pressure rose from baseline 8.7 +/- 0.7 mmHg to 24.3 +/- 1.6, 20.6 +/- 2.1, 19.0 +/- 1.8 and 17.7 +/- 2.0 mmHg, respectively (P < 0.01). The diameter of portal vein increased from 7.6 +/- 0.3 to 8.6 +/- 0.3 mm, calculated portal resistance increased from 0.46 +/- 0.06 to 1.06 +/- 0.20 (mmHg/mL/min/kg body weight); velocity of portal blood flow decreased from 35.1 +/- 1.7 to 26.1 +/- 2.4 cm/s (P < 0.01, respectively). The animals of experimental group developed marked splenomegaly and profuse portosystemic collateral circulations with normal liver and renal function. Repeated intra-portal administration of microspheres can induce stable and reproducible chronic portal hypertension in dogs with normal liver and renal functions. This model can meet multiple demands of both basic and clinical research of portal hypertension.

Sero-biochemical studies in sheep fed with Bt cotton plants

An experimental study was conducted to evaluate the toxicological effects, if any, due to feeding of Bt (Bacillus thuringiensis) cotton plants to sheep. A total of 32 sheep of one year of age belonging to Deccani breed were randomly divided into four groups, consisting of eight sheep in each group. Group 1 was maintained on basal diet (concentrate feed at the rate of 300 g + green fodder at the rate of 3 kg/sheep/day), group 2 on non-Bt cotton plant at the rate of 1.5 kg + green fodder at the rate of 1.5 kg + concentrate feed at the rate of 300 g/sheep/day, group 3 on Bt cotton plants (50%) at the rate of 1.5 kg + green fodder at the rate of 1.5 + concentrate feed at the rate of 300 g/sheep/day, and group 4 on Bt cotton plants ad libitum + concentrate feed at the rate of 300 g/sheep/day. All the groups of sheep were maintained for three months and various hemato-biochemical parameters were studied at monthly intervals. The activity of aspartate transaminase, gamma glutamyltransferase, and creatine kinase in sera samples, and the concentration of blood urea nitrogen and creatinine did not differ significantly among different groups at different time intervals. The histological examination of liver and kidney did not reveal any significant changes in Bt and non-Bt cotton-fed groups. In conclusion, the results of the present investigation enunciated that feeding of genetically modified (Bt) cotton plants to sheep was without detrimental effects in the biological system of sheep.

Viewpoints on the Way to a Consensus Session

Insulin resistance and the insulin resistance syndrome refer to a constellation of anthropometric and metabolic features that may be summarized as overweight/obesity, glucose intolerance, dyslipidemia, and hypertension. These anthropometric and metabolic abnormalities are associated with type 2 diabetes, cardiovascular disease, polycystic ovary syndrome, and nonalcoholic fatty liver disease. The liver plays a central role in the regulation of whole-body glucose, fatty acid, and amino acid metabolism. It is the main source of endogenous glucose production, it is a major site of fatty acid disposal (esterification and oxidation) and of amino acid metabolism, and it is the primary site of insulin degradation. ### Tissue-specific insulin receptor knockout mice Studies using tissue-specific insulin receptor knockout have demonstrated that mice lacking the muscle insulin receptor alone (MIRKO) are characterized by reduced insulin-stimulated muscle glucose uptake, but total-body glucose homeostasis remained normal (1). Adipose tissue insulin receptor knockout mice (FIRKO) had impaired adipose tissue glucose uptake but were protected against obesity, glucose intolerance, and dyslipidemia and manifested a prolonged lifespan (1). In contrast, knockout of the insulin receptor in the liver resulted in both fasting and postprandial hyperglycemia and the subsequent development of peripheral (muscle) insulin resistance. Collectively, these observations support the view that hepatic insulin resistance is the primary event and leads to the subsequent development of peripheral (muscle and adipose) tissue insulin resistance. ### The model of nonalcoholic fatty liver disease Nonalcoholic fatty liver disease (NAFLD) is part of the broad spectrum of nonalcoholic fatty liver diseases, which also includes steatosis. The distinction between fatty liver and steatohepatitis only can be made by examination of liver histology, which allows the assessment of inflammatory infiltrate, cellular degeneration, and necrosis and fibrosis, and cannot be made on the basis of clinical or laboratory parameters. Despite the weak correlation between liver function tests and the severity of liver disease, epidemiological studies have shown that transaminases, and, in particular, elevated …

A controlled safety study of diindolylmethane in the immature rat model

Diindolylmethane (DIM), a natural product from cruciferous vegetables, has been shown to be a dietary component that has inhibitory effects on some tumors (e.g., laryngeal papilloma). However, current evidence to support its safety is based on adult humans or mature animals. There is little to show on its safety in children. This study is designed to assess safety in the young rat model. Prospective controlled animal study. Forty rats were separated into four treatment groups of 10 rats each, based on the amount of study drug they received in their daily food: 1) immature rats fed a low dose of DIM, (0.6 mg/kg/day); 2) immature rats fed a high dose of DIM (6.0 mg/kg/day); 3) immature rats fed no DIM (control); and 4) adult rats fed a high dose of DIM (6.0 mg/kg/day). At the conclusion of the study we collected blood to compare serum chemistries and vitamin D levels, and harvested organs to observe for any gross or histological changes among the groups. Statistical methods involved one-way analysis of variance and pairwise comparisons with Tukey multiple comparison adjustment. Although our numbers do not allow for statistical significance, there was no appreciable difference in rat weights among the immature groups, nor was there appreciable difference in serum chemistries, or gross or histological examination of liver, kidney, and bone. Diindolylmethane seems to have no adverse affects on the rat even when given in doses 3x what we propose to be therapeutic. This adds evidence to the safety of this drug in the pediatric population as a treatment option for recurrent respiratory papilloma.

Assessment of Liver Fibrosis by Transient Elastography in Persons with Hepatitis C Virus Infection or HIV–Hepatitis C Virus Coinfection

Transient elastography is a novel, noninvasive method for staging liver fibrosis. We compared elastography with histologic methods among hepatitis C virus (HCV)-infected and human immunodeficiency virus (HIV)-HCV-coinfected participants in an urban, predominantly black study population. Participants recruited from the AIDS Linked to the Intravenous Experience and the Johns Hopkins HIV Clinical Cohort studies underwent elastography to determine liver stiffness measurements. Liver biopsy specimens were staged F0-F4 in accordance with the Metavir score. Diagnostic accuracy and determination of liver stiffness cutoff values, compared with histologic methods, were determined by receiver operating characteristic analysis. Logistic regression methods identified parameters associated with discordant classification status. Of 192 participants, 139 (72%) were coinfected with HIV and HCV, 121 (63%) had insignificant fibrosis, and 48 (25%) had cirrhosis. Overall, the area-under-the-curve receiver operating characteristic was 0.87 for detection of both significant fibrosis (95% confidence interval, 0.82-0.92) and cirrhosis (95% confidence interval, 0.81-0.93). With use of cutoff values of 9.3 kPa for fibrosis and 12.3 kPa for cirrhosis, 79%-83% of participants were correctly classified by liver stiffness measurement (compared with histologic methods); accuracy appeared to be higher among HIV-uninfected participants than among HIV-infected participants. Most discordance occurred when liver stiffness measurements indicated liver disease and histologic examination did not (in 16% of participants); the patients with these discordant results were more likely to have attributes that increased the odds of significant fibrosis, such as elevated serum fibrosis markers or HIV-related immunosuppression, compared with persons in whom low fibrosis was predicted by both examination of a biopsy specimen and elastography. For most HCV-infected persons, fibrosis stage predicted by elastography is similar to that predicted by examination of a biopsy specimen. Elastography-based measurement of liver stiffness holds promise to expand liver disease screening and monitoring, particularly among injection drug users.

Multicentric occurrence and spread of hepatocellular carcinoma in whole explanted end‐stage liver

Hepatocellular carcinoma (HCC) arising from the end stages of liver cirrhosis is a fair indication for liver transplantation (LT). To pathologically investigate the multicentric occurrence of relatively early staged HCC in cirrhosis, we studied whole explanted livers. Fourteen explanted livers from patients undergoing living donor LT (LDLT) were examined. The stage of the HCCs was judged to be within the Milan criteria (M-C; a single HCC less than 5 cm or three HCCs less than 3 cm). Histological examination was performed using serially sectioned specimens 5-7 mm in width. Characterization of preoperatively detectable and undetectable lesions was also performed. In nine patients (64.3%), a total of 34 nodules were found after whole liver histological examination (WLHE). In five patients (31%), the results exceeded the M-C. The characteristics of undetectable HCCs included a minute (median size 6 mm), well-differentiated appearance (80%), with indistinct margins (85.3%) and without vascular invasion (94%). There was no recurrence in any patients at the time of follow up (median follow-up period, 30.1 months). A multicentric occurrence of HCCs was demonstrated in cirrhotic livers with HCCs within the M-C. Undetectable HCCs in cirrhotic livers may have no impact on recurrence after LT.

Major Histocompatibility Complex Class I-related Chains A and B (MIC A/B): A Role in the Pathogenesis of Non-alcoholic Steatohepatitis

Hepatocellular apoptosis is a key mechanism of liver injury in the pathogenesis of non-alcoholic steatohepatitis (NASH). Recently, soluble forms of major histocompatibility complex (MHC) class I-related chains A and B (MIC A/B) were reported to be increased in the sera of patients with chronic liver disease and hepatocellular malignancy. However, the role of MIC A and B in the pathogenesis of NASH has not been explored. We thus aimed to investigate the impact of these stress-induced ligands on liver injury, apoptosis and hepatic fibrosis in patients with NASH. Methodology: Blood and liver tissue were obtained from 40 patients with NASH undergoing gastric banding for obesity. The control group consisted of 10 healthy volunteers. Real-time PCR was used to measure mRNA transcripts of MIC A/B and the death receptors CD95/Fas, TRAIL-DR4 and DR5. Intrahepatic liver cell apoptosis was quantified by TUNEL assay, and on the systemic level, apoptosis was determined by the caspase-cleaved biomarker M30. Liver injury was assessed by histopathology using the NAFLD activity score (NAS) and serum ALT/AST determinations. Hepatic fibrosis was assessed by Sirius-Red staining, and mRNA transcript for α-SMA. Results: Our preliminary data from patients with NASH following gastric banding revealed significant increases (p&lt;0.05) (i) in hepatic MIC A/B mRNA by &gt;2.8; (ii) in CD95/Fas, DR4 and DR5 mRNAs; (iii) in TUNEL+ hepatocytes by &gt;4.1; and (iv) in systemic M30 levels. Consistent with this data, histological examination of livers from NASH patients revealed relevantly increased NAS scores. Serum ALT and AST values were significantly higher in NASH patients. Hepatic fibrosis was &gt;2.6 increased in patients with NASH. Conclusion: In patients with NASH, hepatocyte apoptosis and hepatic fibrosis are increased. Our findings suggest a role for MIC A/B-mediated liver injury. Thus, therapeutic intervention aimed at reducing MIC A/B may have a beneficial effect on the progression of NASH.

Potential Ameliorative Role Of Silymarin Against Methyl Parathion-Induced Oxidative Stress And Hepato-Renal Toxicity In Albino Rats

Methyl parathion (MP) (C8H10NO5PS) is an organophosphate insecticide that has been used in agriculturefor several years. The people who are at the greatest risk of being exposed to MP are farm workers,chemical sprayers, and people who work in factories handling MP. The main route of human exposureis inhalation, but dermal contact and inadvertent ingestion can also be substantial. The present workwas planned to study the toxic effects of MP on some antioxidative markers as well as the liver and kidney.The influence of silymarin (Sily) on MP induced toxicity has not been studied, So, the possible protectiveeffect of (Sily) had been also evaluated. The study was conducted on one hundred male albino ratsdivided into five main equal groups and each of them were subdivided equally into two subgroups.Group-I (A&B), control group: received no treatment, group-II (A&B): vehicle group (1 ml/kg/day ofcorn oil, P.O.), group-III (A&B): Sily group (100 mg/kg/day, P.O.), group-IV (A&B): MP group (0.28mg/kg/day, P.O.), group-V (A&B): Sily+MP. At the end of 4th and 8th weeks, blood samples, renal andhepatic tissues were collected for biochemical and histological examinations. Biomarkers selected for oxidativestress were antioxidant defense system superoxide dismutase (SOD) and reduced glutathione(GSH) as well as malondialdehyde (MDA). In addition, liver profile (AST, ALT, and ALP) and kidneyprofile (Blood urea and creatinine) were assessed. Also, histological examination of liver and kidney wasdone. The results showed that administration of MP resulted in oxidative stress as evidenced by significantdecrease in SOD activity and GSH levels accompanied with significant increase in MDA levels. Atthe same time, there was hepato-nephrotoxicity as manifested by significant increase in liver and kidneyfunction tests when compared with control rats. Comparing with MP-treated rats, Sily supplementation tothe rats, 30 minutes before MP administration, resulted in a significant increase in SOD activity and GSHlevels. However, MDA levels, liver and kidney enzymes showed significant decrease. The histopathologicalresults confirmed the biochemical results. In conclusion, the obtained biochemical and histologicalresults of this study revealed hepatic and renal toxic effects induced by MP in a time-dependent manner.Sily supplementation resulted in a remarkable protective effect against MP-induced oxidative stress andhepato-renal toxicity.

Chronic administration of ursodeoxycholic acid decreases portal pressure in rats with biliary cirrhosis

Liver cirrhosis is characterized by increased IHR (intrahepatic resistance) and lipid peroxidation, and decreased antioxidative defence. The present study investigates the effects of administration for 1 month of the antioxidant UDCA (ursodeoxycholic acid) in BDL (bile-duct-ligated) cirrhotic rats. Splanchnic haemodynamics, IHR, hepatic levels of TBARS (thiobarbituric acid-reacting substances), GSH (glutathione), SOD (superoxide dismutase) activity, nitrite, PIIINP (N-terminal propeptide of type III procollagen) and collagen deposition, histological examination of liver, mRNA expression of PIIIP-alpha1 (type III procollagen) and TGF-beta1 (transforming growth factor-beta1), protein expression of TXS (thromboxane synthase) and iNOS (inducible NO synthase), and TXA2 (thromboxane A2) production in liver perfusates were measured. The results showed that portal pressure and IHR, hepatic levels of PIIINP, hepatic collagen deposition, mRNA expression of PIIIP-alpha1 and TGF-beta1, protein expression of iNOS and TXS, and production of TXA2 in liver perfusates were significantly decreased in UDCA-treated BDL rats. The increased levels of hepatic GSH and SOD activity and decreased levels of TBARS and nitrite were also observed in UDCA-treated BDL rats. In UDCA-treated BDL rats, the reduction in portal pressure resulted from a decrease in IHR, which mostly acted through the suppression of hepatic TXA2 production and lipid peroxidation, and an increase in antioxidative defence, leading to the prevention of hepatic fibrosis.

β-Glycosphingolipids improve glucose intolerance and hepatic steatosis of the Cohen diabetic rat

A link between altered levels of various gangliosides and the development of insulin resistance was described in transgenic mice. Naturally occurring glycosphingolipids were shown to exert immunomodulatory effects in a natural killer T (NKT) cell-dependent manner. This study examined whether glycosphingolipid-induced modulation of the immune system may reduce pancreatic and liver steatosis and stimulate insulin secretion in the Cohen diabetes-sensitive (CDS) rat, a lean model of non-insulin-resistant, nutritionally induced diabetes. Four groups of CDS rats fed a diabetogenic diet were treated with daily intraperitoneal injections of glycosphingolipids beta-glucosylceramide, beta-lactosylceramide, a combination of both (IGL), or vehicle (PBS) for up to 45 days. Immune modulation was assessed by fluorescence-activated cell sorting analysis of intrahepatic and intrasplenic lymphocytes. Steatosis was assessed by MRI imaging and histological examination of liver and pancreas, Blood glucose and plasma insulin concentrations were assessed during an oral glucose tolerance test. Administration of glycosphingolipids, particularly IGL, increased intrahepatic trapping of CD8 T and NKT lymphocytes. Pancreatic and liver histology were markedly improved and steatosis was reduced in all treated groups compared with vehicle-treated rats. Insulin secretion was restored after glycosphingolipid treatment, resulting in improved glucose tolerance. The immunomodulatory effect of beta-glycosphingolipids improved the beta-cell function of the hyperglycemic CDS rat. Thus our results suggest a role for the immune system in the pathogenesis of diabetes in this model.