Acute Liver Research Articles

Mo1772 Kinase Inhibition Normalizes Mutation-Mediated Keratin Filament Disruption and Predisposition to Apoptosis and Liver Injury

Objective: Mutations in members of the cytoskeletal intermediate filament proteins (IFs), keratin polypeptides 8 and 18 (K8/K18), predispose their carriers to acute and chronic liver disease progression. Keratin associated diseases are among .70 human diseases that are caused or predisposed to by IF mutations, with the defining feature in many of these diseases being disruption of the normal IF organization within the cytoplasm. In contrast to microfilaments and microtubules, no drugs are available for IFs that can modulate their organization and potentially provide a sorely lacking therapy. Methods: We used an imagingbased drug screening platform consisting of lung carcinoma A549 cells transduced with lentiviruses containing GFP-tagged K18 R90C and wild-type K8. The cells were screened against several compound libraries, and a select compound was further tested in cell culture and in livers of mice that express mutant K18 in order to define its mechanism of action. Results: Screening of a 320-compound kinase inhibitor library showed that the inhibitors of protein kinase C (PKC) and tyrosine kinases, particularly MIDOSTAURIN, led to significant correction of the defective organization of K18 R90C puncta towards normal filaments after 24 h with a maximal effect at 48 h. The findings were confirmed using multiple MIDOSTAURIN concentrations and in another cell line, Hacat, that expresses mutant K14. While MIDOSTAURIN diminished the expression of the mutant keratin 14 in Hacat cells, the effect in the A549 cells was independent of K18 expression but causes changes in K18 phosphorylation that are known to regulate filament formation. MIDOSTAURIN significantly prevented interferon and FAS ligand-induced apoptosis in A549 cells. The protective effect occurs after 48 h but not after 1 h treatment with MIDOSTAURIN, suggesting that the drug effect is more likely to be related to the correction of filament organization. In mice, administration of MIDOSTAURIN is well tolerated and its administration prior to exposure to FAS results in dramatic protection from liver injury. Conclusion: The kinase inhibitor MIDOSTAURIN alleviates cytoskeletal structural defects caused by K18 R90C mutation and also protects from apoptosis in cultured and in the livers of mice exposed to FAS ligand. MIDOSTAURIN may be a potential therapeutic for IF diseases and, notably, it is presently undergoing clinical trials in patients with acute myeloid leukemia and gastrointestinal stromal tumors.

Mo1775 Epigenetic Inactivation of Claudin 3 in Hepatocellular Carcinoma and Its Functional Consequences

Objective: Mutations in members of the cytoskeletal intermediate filament proteins (IFs), keratin polypeptides 8 and 18 (K8/K18), predispose their carriers to acute and chronic liver disease progression. Keratin associated diseases are among .70 human diseases that are caused or predisposed to by IF mutations, with the defining feature in many of these diseases being disruption of the normal IF organization within the cytoplasm. In contrast to microfilaments and microtubules, no drugs are available for IFs that can modulate their organization and potentially provide a sorely lacking therapy. Methods: We used an imagingbased drug screening platform consisting of lung carcinoma A549 cells transduced with lentiviruses containing GFP-tagged K18 R90C and wild-type K8. The cells were screened against several compound libraries, and a select compound was further tested in cell culture and in livers of mice that express mutant K18 in order to define its mechanism of action. Results: Screening of a 320-compound kinase inhibitor library showed that the inhibitors of protein kinase C (PKC) and tyrosine kinases, particularly MIDOSTAURIN, led to significant correction of the defective organization of K18 R90C puncta towards normal filaments after 24 h with a maximal effect at 48 h. The findings were confirmed using multiple MIDOSTAURIN concentrations and in another cell line, Hacat, that expresses mutant K14. While MIDOSTAURIN diminished the expression of the mutant keratin 14 in Hacat cells, the effect in the A549 cells was independent of K18 expression but causes changes in K18 phosphorylation that are known to regulate filament formation. MIDOSTAURIN significantly prevented interferon and FAS ligand-induced apoptosis in A549 cells. The protective effect occurs after 48 h but not after 1 h treatment with MIDOSTAURIN, suggesting that the drug effect is more likely to be related to the correction of filament organization. In mice, administration of MIDOSTAURIN is well tolerated and its administration prior to exposure to FAS results in dramatic protection from liver injury. Conclusion: The kinase inhibitor MIDOSTAURIN alleviates cytoskeletal structural defects caused by K18 R90C mutation and also protects from apoptosis in cultured and in the livers of mice exposed to FAS ligand. MIDOSTAURIN may be a potential therapeutic for IF diseases and, notably, it is presently undergoing clinical trials in patients with acute myeloid leukemia and gastrointestinal stromal tumors.

Mo1774 BCL6B Inhibits Hepatocellular Carcinoma Metastases In Vitro and in Mice

Objective: Mutations in members of the cytoskeletal intermediate filament proteins (IFs), keratin polypeptides 8 and 18 (K8/K18), predispose their carriers to acute and chronic liver disease progression. Keratin associated diseases are among .70 human diseases that are caused or predisposed to by IF mutations, with the defining feature in many of these diseases being disruption of the normal IF organization within the cytoplasm. In contrast to microfilaments and microtubules, no drugs are available for IFs that can modulate their organization and potentially provide a sorely lacking therapy. Methods: We used an imagingbased drug screening platform consisting of lung carcinoma A549 cells transduced with lentiviruses containing GFP-tagged K18 R90C and wild-type K8. The cells were screened against several compound libraries, and a select compound was further tested in cell culture and in livers of mice that express mutant K18 in order to define its mechanism of action. Results: Screening of a 320-compound kinase inhibitor library showed that the inhibitors of protein kinase C (PKC) and tyrosine kinases, particularly MIDOSTAURIN, led to significant correction of the defective organization of K18 R90C puncta towards normal filaments after 24 h with a maximal effect at 48 h. The findings were confirmed using multiple MIDOSTAURIN concentrations and in another cell line, Hacat, that expresses mutant K14. While MIDOSTAURIN diminished the expression of the mutant keratin 14 in Hacat cells, the effect in the A549 cells was independent of K18 expression but causes changes in K18 phosphorylation that are known to regulate filament formation. MIDOSTAURIN significantly prevented interferon and FAS ligand-induced apoptosis in A549 cells. The protective effect occurs after 48 h but not after 1 h treatment with MIDOSTAURIN, suggesting that the drug effect is more likely to be related to the correction of filament organization. In mice, administration of MIDOSTAURIN is well tolerated and its administration prior to exposure to FAS results in dramatic protection from liver injury. Conclusion: The kinase inhibitor MIDOSTAURIN alleviates cytoskeletal structural defects caused by K18 R90C mutation and also protects from apoptosis in cultured and in the livers of mice exposed to FAS ligand. MIDOSTAURIN may be a potential therapeutic for IF diseases and, notably, it is presently undergoing clinical trials in patients with acute myeloid leukemia and gastrointestinal stromal tumors.

Su1283 Risk of Ischemic Hepatitis Among Hospitalized Patients With Hemodynamic Instability: A Nationwide Analysis

we identified seven exhaled breath compounds that were significantly elevated in patients with liver disease compared to healthy control. Those compounds include: 2-propanol, acetaldehyde, ammonia, ethanol, acetone, pentane and trimethylamine (TMA). The mean concentration of TMA, in particular, in the exhaled breath was remarkably higher in patients with AH compared to patients with other forms of liver disease and to healthy volunteers (p,0.001). With the use of ROC curve, a cutoff value of breath TMA of 31 ppb provides 94% specificity and 98% sensitivity for the diagnosis of AH [AUC 95% CI: 0.990 (0.9771)]. Among patients with AH, the concentration of TMA in the exhaled breath correlates with the MELD score [rho (95% CI): 0.53 (0.04, 1.0); p=0.035]. Conclusion: TMA breath analysis can identify patients with AH from other forms of acute or chronic liver disease and may provide a novel diagnostic test for this disease.

Liver Biopsy in the Management of Autoimmune Hepatitis Acute Severe Onset

I read with interest the excellent review by Czaja on acute severe autoimmune hepatitis (AIH) [1], which greatly increased my awareness of the clinical and therapeutic characteristics of this extremely serious disease. The precise and extensive description of this medical problem provided by Czaja can be of upmost importance for the management of this specific subgroup of patients by clinicians. AIH is a chronic hepatitis of unknown etiology, and its clinical manifestations are highly variable, sometimes following a fluctuating course [2, 3]. An acute, severe, abrupt presentation is also possible and is secondary to variable causes [1]. The diagnosis of AIH is based on a clinical, biochemical, serological and histological patterns, and sometimes has to be confirmed by the response of the disease to immunosuppressive treatment [1–4]. Compared to patients presenting with a more chronic course, those presenting with acute onset more frequently have interface and centrilobular hepatitis, lobular disarray and central necrosis, but perhaps less fibrosis and cirrhosis [1, 2, 5]. Liver biopsy features in the acute presentation of AIH may be difficult to distinguish from those seen in acute hepatitis due to the effects of the virus or drugs [1, 5]. It is not yet clear whether the clinical presentation of AIH in the setting of drugs used for other diseases unmask and/or induce a typical AIH or simply result in a drug-induced hepatitis with features that mimic AIH [6]. Therefore, the histological assessment of acute and chronic liver is based on an accurate description of the pattern of injury within the tissue specimen. Interpretation of the injury pattern requires review of the characteristic clinical and laboratory features, and the morphological findings only achieve relevance if they satisfactorily explain the clinical syndrome [7]. Thus, the importance of the results of the histological analysis of liver samples with respect to the diagnosis and clinical management of severe acute AIH may be unclear and, occasionally, biopsy results may actually delay therapeutic decisions; in other words, biopsy results may be unnecessary for the rapid initiation of treatment given that early steroid therapy may save lives [8–11]. It has recently been reported that patients with typical laboratory features of AIH rarely need liver biopsy for diagnosis. Most patients presenting with features of AIH based on laboratory tests are likely to have a compatible liver histology, and few patients have atypical histology. Consequently, tissue findings may have little impact on patient management. Thus, biopsy samples might not need to be collected from patients who meet other typical criteria for AIH [9]. When the scoring system based upon titers of autoantibodies, immunoglobulin G (IgG) levels and the exclusion of viral hepatitis is used as a diagnostic tool, a score of six points can be used as a simplified criterium of a probable diagnosis of AIH [5]. At admission, the recommended analyses include general biochemistry, conventional autoantibodies, quantification of immunoglobulins and IgG serum levels. It is important to exclude other virus infections, such as hepatitis A and E viruses, hepatitis C virus (serum HCV-RNA R. Moreno-Otero (&) Liver Unit, and Instituto de Investigacion Sanitaria (IIS-IP), Digestive Diseases Service (Planta 3), Hospital Universitario La Princesa, Universidad Autonoma de Madrid, Diego de Leon, 62, 28028 Madrid, Spain e-mail: rmoreno.hlpr@salud.madrid.org; rmorenoo@salud.madrid.org

50 Jahre Hepatologie - Vom therapeutischen Nihilismus zur gezielten Therapie

Over the past 50 years significant progress has been made in the whole field of hepatology. Part of this is translation of basic research (biochemistry, immunology, virology, molecular biology and others) into clinical hepatology. This enabled us to understand more about the pathogenesis of liver diseases and led to the discovery of the five major hepatotropic viruses, the identification of hepatocellular autoantigens, and to the development of specific therapies for chronic hepatitis B, C and D. In addition, the molecular basis of most genetic liver diseases has been identified. Significant progress was made in the development of medical therapies for various liver diseases with different underlying etiologies. Surgery significantly contributed to the progress in the management of liver diseases; examples are laparoscopic cholecystectomy and the development of liver transplantation. A multimodal therapeutic algorithm has been established for the therapy of hepatocelluar carcinoma (HCC); with Sorafenib "targeted therapy" has entered the area of HCC. The progress made over the last 50 years not only led to an aetiological differentiation of acute and chronic liver diseases but also to specific therapies based on the identification and understanding of the underlying etiology.

Blockade of CCR5 to protect the liver graft in HIV/HCV co-infected patients

Background: Graft-versus-host disease (GVHD) is a major barrier to successful allogeneic hematopoietic stem-cell transplantation (HSCT). The chemokine receptor CCR5 appears to play a role in alloreactivity. We tested whether CCR5 blockade would be safe and limit GVHD in humans. Methods: We tested the in vitro effect of the CCR5 antagonist maraviroc on lymphocyte function and chemotaxis. We then enrolled 38 high-risk patients in a single-group phase 1 and 2 study of reduced-intensity allogeneic HSCT that combined maraviroc with standard GVHD prophylaxis. Results: Maraviroc inhibited CCR5 internalization and lymphocyte chemotaxis in vitro without impairing T-cell function or formation of hematopoietic-cell colonies. In 35 patients who could be evaluated, the cumulative incidence rate (±SE) of grade II to IV acute GVHD was low at 14.7 ± 6.2% on day 100 and 23.6 ± 7.4% on day 180. Acute liver and gut GVHD were not observed before day 100 and remained uncommon before day 180, resulting in a low cumulative incidence of grade III or IV GVHD on day 180 (5.9 ± 4.1%). The 1-year rate of death that was not preceded by disease relapse was 11.7 ± 5.6% without excessive rates of relapse or infection. Serum from patients receiving maraviroc prevented CCR5 internalization by CCL5 and blocked T-cell chemotaxis in vitro, providing evidence of antichemotactic activity. Journal of Hepatology 20

Irreversible Liver Failure

Liver transplantation is the only established, causally directed treatment for irreversible chronic or acute liver failure. This review is based on papers retrieved by a selective search in the PubMed database, the index of randomized controlled trials of the European Society of Organ Transplantation, and the Cochrane database, along with an analysis of data from the authors' own center. 1199 liver transplantations were performed in Germany in 2011. The most common indications were alcoholic cirrhosis (28%), cirrhosis of other causes (24%), and intrahepatic tumors (20%). Among recipients, the sex ratio was nearly 1:1 and the median age was just under 50. Across Europe, the 1-, 5-, and 10-year survival rates after liver transplantation were 82%, 71% and 61%. In our own center, the Charité in Berlin, the corresponding rates were 90.4%, 79.6% and 70.3%, based on an experience of 100 to 120 cases per year. The current rate of functioning transplants five years after liver transplantation is 52.6% in Germany and 66.2% internationally. Standard immunosuppression consists of a calcineurin inhibitor, tacrolimus or cyclosporine A, and steroids. Early complications include primary functional failure of the transplant, hemorrhage, thrombosis, acute rejection, and biliary complications. Over the long term, complications that can impair the outcome include chronic rejection, biliary strictures, cardiovascular and metabolic adverse effects, nephrotoxicity, neurotoxicity, and opportunistic infections and malignancies. Liver transplantation is a successful and well-established form of treatment that is nonetheless endangered by a shortage of donor organs and other structural and organizational difficulties.

Ernährung in der Intensivmedizin

Therapy of intensive care patients is often complicated by co-morbidities or complex systemic disorders such as sepsis. The necessity to generate an individualized nutritional regime has gained in importance in recent years as this essential part of supportive care has a direct impact on the prognosis of the patient. In the present article a special focus is put on particular questions of nutritional aspects of intensive care patients. The current guidelines and study data on disorders relevant in intensive care medicine, such as acute or chronic renal and liver failure, acute respiratory distress syndrome and sepsis are presented and discussed. Another focus is the establishment of an adequate nutritional regime for patients after operations or suffering from multiple trauma.

Profile of liver biopsy in tertiary care hospital–1 year data

S 21 ANNUAL CONFERENCE—2013 M isella n eo u s patients. Intrahepatic cholestasis of pregnancy (IHCP) was diagnosed by raised serum bile acid (>10mmol/L). Preeclampsia and Acute fatty liver of pregnancy were diagnosed by standard criteria. Results: Seventy eight patients were studies (median age 27 year range 20-42years). Eighty three percent (65/78) were in the third trimester of pregnancy. Spectrum of liver diseases was as follows IHCP was 58.9% (46/78), viral hepatitis 21.7% (17/78), preeclampsia 10.2% (8/78), hyperemesis 2.5%(2/78), AFLP 2.5%(2/78), Chronic Liver disease 2.5%(2/78). Among 17 patients with viral hepatitis, 11 had incidental HBsAg positivity, 5 with hepatitis E and 2 with hepatitis A presented as acute hepatitis. Maternal mortality was seen in 4 cases (2 AFLP, 2 preeclampsia) in the third trimester. Preterm delivery was observed in 8 patients, out of which 6 had IHCP (17.3%). Low birth weight was seen in 22 patients of whom 18 had IHCP (39.1%). There were 6 fetal deaths (2-AFLP, 2-preclampsia, 1-acute liver failure (hepatitis E related) and 1-CLD). The overall maternal and fetal death was 5.22 % and 7.69% respectively and was seen in AFLP, preeclampsia and ALF patients. Conclusions: IHCP is the commonest cause of deranged LFT during pregnancy. Preterm labor is common in IHCP. AFLP and preeclampsia were associated with poor maternal and fetal outcome. Corresponding author: Mohan B Goyal. E-mail: mohandr77@gmail.com PROFILE OF LIVER BIOPSY IN TERTIARY CARE HOSPITAL–1 YEAR DATA Pravir Amrutrao Gambhire, Ashok Rohidas Mohite, Samit Sampat Jain, Dharmesh Kirit Shah, Amol Ajit Khot, Piyush Somani, Pravin Rathi, Anjali Amrapurkar Departments of Gastroenterology and Pathology, BYL Nair Hospital

Protective effects of Lipocalin-2 (LCN2) in acute and chronic liver injury indicate a novel function in liver homeostasis

Protective effects of Lipocalin-2 (LCN2) in acute and chronic liver injury indicate a novel function in liver homeostasis

Expression of the novel stem/progenitor cell marker Neighbor of Punc E 11 after acute and chronic liver injury in the adult mouse liver

Expression of the novel stem/progenitor cell marker Neighbor of Punc E 11 after acute and chronic liver injury in the adult mouse liver

Intestinal Alkaline Phosphatase Prevents Acute Alcohol-Induced Liver Injury

Intestinal Alkaline Phosphatase Prevents Acute Alcohol-Induced Liver Injury

Plasmapheresis in Acute Fatty Liver of Pregnancy: An Effective Treatment

Acute fatty liver of pregnancy (AFLP) is an idiopathic disorder with an unknown cause occurring in late pregnancy. The treatment in these patients is often immediate termination of pregnancy, and plasmapheresis provides an effective treatment option. In this paper, we introduce three pregnant women treated with plasmapheresis. The first case was a 22-year-old primigravida woman treated with 22 sessions of plasmapheresis due to AFLP, hepatic and renal failure, coagulopathy, and ventilator-dependent respiratory failure. The second case was a 23-year-old woman in her second pregnancy treated with 4 plasmapheresis sessions due to AFLP, hepatic and renal failure, coagulopathy, and hypoglycemia. The third patient was a 23-year-old primigravida woman treated with 3 plasmapheresis sessions due to AFLP, renal failure, and coagulopathy. Plasmapheresis can be a life-saving treatment in patients with AFLP and is strongly recommended for patients with severity of their disease accompanied by other organ disorders. In addition, shortening the time interval between the termination of pregnancy and initializing plasmapheresis improves the outcome and reduces the duration of hospital stay and sessions of plasmapheresis.

Acute Oral Toxicity and Kinetic Behaviors of Inorganic Layered Nanoparticles

Layered double hydroxide (LDH) nanoparticles, also known as anionic clays, have attracted a great deal of interest for their potential as delivery carriers. Recent studies showed that LDH nanoparticles can efficiently deliver drugs or bioactive molecules into cells, which are highly related to their endocytic pathway. However, the efficient cell permeation capacity of LDH may also raise concern about their toxicity potential. In this study, the acute oral toxicity of LDH nanoparticles was assessed, and their kinetic behaviors, such as plasma concentration‐time curve, tissue distribution, and excretion, were also evaluated in mice. No significant effects of oral LDH nanoparticles on behaviors, body weight gain, survival rate, and organosomatic index were observed up to the dose of 2000 mg/kg for 14 days. Serum biochemical parameters did not significantly increase, indicating that LDH nanoparticles did not cause acute liver or kidney injury. Plasma concentration of LDH nanoparticles rapidly decreased within 30 min depending on exposure doses, but they did not accumulate in any specific organ. Their excretion via urine and feces was observed within 24 h. These findings suggest that LDH nanoparticles do not exhibit acute oral toxicity and favorable kinetic behaviors in mice and, therefore, will be promising candidates for biological and pharmaceutical applications.

A Case of Epistaxis with Toxic Shock Syndrome

A 50-year-old man was referred to our hospital with epistaxis from the right nostril. His left nostril had been packed with several pieces of gauze for a long time. Acute liver and kidney failure made control of the epistaxis difficult. Toxic shock syndrome (TSS) is associated with complications such as multiple organ failure or shock due to exotoxins, such as toxic shock syndrome toxin-1 (TSST-1), produced by Staphylococcus aureus. Definitive diagnosis of TSS is difficult, because the diagnostic criteria of the Centers for Disease Control and Prevention (CDC) are stringent. Although our present patient did not fulfill the diagnostic criteria, TSS was strongly suspected. TSS after sinus surgery has been frequently reported. Few studies have been published about TSS caused by gauze packing for epistaxis. It is necessary to be borne in mind the possibility of TSS not only after surgery, but also after gauze packing for epistaxis.

Effect of Entamoeba histolytica calreticulin in the immune response of patients with acute amoebic liver abscess

Effect of Entamoeba histolytica calreticulin in the immune response of patients with acute amoebic liver abscess

Clinical profile of cases of alcoholic liver disease

Background: Alcohol is most common substance abused. Alcoholic liver disease is a major health care problem in India. Alcohol consumption is directly associated with liver disease mortality and accounts for increased social and economic costs. Alcoholic liver disease may take the forms of acute involvement (alcoholic hepatitis) or chronic liver disease (steatosis, steatohepatitis, fibrosis and cirrhosis. The severity and prognosis of alcohol- induced liver disease depends on the amount, pattern and duration of alcohol consumption, as well as on the presence of liver inflammation, diet, nutritional status and genetic predisposition of an individual. While steatosis is complete benign disease, liver cirrhosis is associated with marked morbidity, mortality and life expectancy shortening. Aims & Objective: To study alcoholic liver disease, clinical presentation, to access the severity of alcoholic liver disease and its complications and their treatment response and overall outcome among patients using laboratory and radiological parameters. Material and Methods: A total of 50 patients were studied and their clinical profile, laboratory parameters and radiological investigations were taken. Results: Among 50 patients 58 % belonged to age group 40-49 years. All were male. 78 % of patients from urban areas, with 60 % belonging to middle class.60 % of patients have chief complaint of abdominal distension and melena each. Jaundice (60%) and ascites (60%) were commonest finding. All patients had raised SGPT, SGOT, S.AlPO4 and S. bilirubin suggesting liver damage. Prolonged PT and reduced S. albumin suggested reduced protein synthesis because of liver disease. Alcoholic hepatitis was in24% cases, while 40% had fatty liver and 36 % had alcoholic cirrhosis. Overall mortality rate was 20 %. , most common cause is encephalopathy (40%), coagulopathy leading to DIC (40%) and hepatorenal syndrome (20%). Conclusion: Alcoholic liver disease was seen among the productive age group with high morbidity and mortality. Mortality and morbidity associated with this disease is matter of serious economic loss to the nation and grief for the society

Efficacy of lamivudine combined with adefovir dipivoxil in management of chronic hepatitis B with acute or sub-acute liver failure

Efficacy of lamivudine combined with adefovir dipivoxil in management of chronic hepatitis B with acute or sub-acute liver failure

Potential Applications of Tanshinones in Gastrointestinal and Hepatic Diseases

The biological activities of tanshinones, the lipophilic components isolated from Salvia miltiorrhiza (Danshen) have been extensively investigated in the past decades. Numerous studies have investigated the versatile abilities of tanshinones in a variety of diseases including acute ischemic stroke, cardiac arrhythmias, atherosclerosis, hypertension, obesity, metabolic syndrome, liver fibrosis, chronic hepatitis and different kinds of cancers. In this regard, the underlying mechanisms of action and potential clinical applications of tanshinones especially in drug adjuvant therapy have been evaluated. The present review briefly summarizes recent studies on the pharmacological activities and possible mechanisms of action for tanshinones as therapeutic agents in different gastrointestinal and hepatic diseases with emphasis on tanshinone IIA, the most abundant tanshinone found in Danshen.