Targeting Heat Shock Protein 90 Alters Epigenetic Genes in Alcoholic Liver Disease

Chronic alcohol induces acetylation and methylation of chromatin-associated histones that influence transcriptional activity of genes. Molecular chaperone heat shock protein 90 (hsp90) is being recognized as mediator of chromatin remodeling and is upregulated in alcoholic liver disease. We hypothesize that hsp90 plays a pivotal role in altered expression of chromatin modifying enzymes during alcoholic liver injury. To test our hypotheses, C57Bl/6 mice were fed Lieber-deCarli diet with 5% v/v ethanol for 10 days followed by a binge containing 20%v/v alcohol. A single injection of hsp90 inhibitor, 17-DMAG [17-Dimethylamino-ethylamino-17-demethoxygeldanamycin] was administered (30-50 mg/kg BW) i.p. before the binge. Epigenetic PCR array analyzing chromatin modifying enzyme expression was employed to determine effect of chronic alcohol and 17-DMAG treatment. Elevated ALT, triglycerides and steatosis confirmed alcoholic liver injury. Results show significant up-regulation of 5 genes including, ATF2 (p=0.0003), a transcription factor with histone acetyltransferase activity, PRMT6 (p=0.0001) and SETD7 (p=0.002), protein methyltransferases, RPS6KA3 (p=4.4 e-6) kinase and HDAC3 (p=0.0001) whereas HDAC9 (p=0.002) was decreased in alcoholic whole livers. Further, ATF2 was exclusively up-regulated in Kupffer cells (KCs) while PRMT6 increased in alcoholic hepatocytes. HDAC3, SETD7 and RPS6KA3 were increased in both, alcoholic KCs and hepatocytes. HDAC9 was exclusively down regulated in alcohol exposed hepatocytes but not in KCs. Inhibition of hsp90 by 17-DMAG after chronic alcohol exposure alleviated liver injury as noted by significantly lowered serum ALT, TBARS and liver triglycerides. Interestingly, 17-DMAG treatment prevented upregulation of ATF2 (p=3.13 e-5), PRMT6 (p=0.001) and HDAC3 (p=0.001) and inhibited down regulation of HDAC9 (p=2.6 e-8) without any effect on expression of SETD7 and RPS6KA3 genes in liver. In summary, our data indicate that chronic alcohol exposure regulates chromatin modifying epigenetic genes in a cell-specific manner likely via molecular chaperone, hsp90. Inhibition of hsp90 leads to modulation of chromatin modifying enzymes with resolution of liver injury. (Supported by the NIH/NIAAA # AA179086

Cellular Signaling Pathways in Alcoholic Liver Disease

Introduction: The pathogenesis of acute and chronic alcohol consumption is complex with diverse consequences in different tissues. Alcohol abuse is associated with a continuum of liver abnormalities ranging from steatosis or fat deposition, steatohepatitis or fat plus inflammation to cirrhosis and hepatocellular carcinoma. The progression of alcohol-induced liver damage involves both parenchymal and non-parenchymal cells of the liver. The signaling pathways affected by direct or indirect alcohol exposure range from oxidative stress mechanims, metabolism related effects, inflammation, and apoptosis. Understanding the interactions of inter- and intra-cellular signaling pathways in the liver during alcohol exposure will aid in identification of new integrative approaches as it relates to alcoholic liver disease and provide potential new directions to develop therapeutic target intervention. The goal of this chapter is to review signaling pathways related to oxidative stress and inflammatory responses modulated by alcohol in parenchymal and nonparenchymal cells of the liver important to ALD

Age at menarche and the menstrual pattern of secondary school adolescents in central India

Background: Population studies on characteristics of menstrual cycles are scarce. Knowledge on this variability is necessary for patient education and to guide clinical evaluation.Methods: A cross sectional study was conducted and 622 school girls were selected randomly. A pretested questionnaire was used to gather data.Results: Mean age of participants was 16.9 ± 1 years. About 92.2% had attained menarche. Probit analysis of the status-quo data yielded median age at menarche of 14.8 (13.9-15.3) years and by recall method to be 15.8 ± 1 years. The mean age at menarche was, significantly, 0.3 years younger for urban females compared with rural ones. Cycle length between 21-35 days was observed in 70.3% of the girls. Mean duration of flow was 4 ± 1.3 days. Menstrual cycles were irregular in 42.8% of the subjects. Overall prevalence of dysmenorrhoea was 72%. and of PMS was 75.4%.Conclusions: Age of menarche was found to be significantly delayed. Considerable number of students complained of dysmenorrhoea and premenstrual symptoms

Design and Implementation Of Vehicle Tracking System Using GPS

Surveillance system using phone line for security and tracking. Based on the above statement, it is targeted that this project will serve as good indication of how important it is to curb car theft in the country. Surveillance is specified to car alarm system and the means of sending the data to the owner of the vehicle using SMS when the alarm is triggered. Due to the inefficient conventional car security system, the possibility of the car can be stolen is high. The main reason is that the alarm is limited to the audible distance. Somehow if there is another way of transmitting the alarm to the car owner ,tracking the vehicle ,knowing the exactly that the car is been stolen at the same time that is not limited to the audible and line of sight, the system can be upgraded. SMS is a good choice of the communication to replace the conventional alarm, because it can be done and does not require much cost. Although most of people know  GPS can provide more security for the car but the main reason people does not apply it because the cost. Advance car security system is too expensive. Cost for the gadget is too high. Beside that, people also must pay for the service monthly. Tracking systems were first developed for the shipping industry because they wanted to determine where each vehicle was at any given time. Passive systems were developed in the beginning to fulfill these requirements. For the applications which require real time location information of the vehicle, these systems can't be employed because they save the location information in the internal storage and location information can only be accessed when vehicle is available. To achieve automatic Vehicle Location system that can transmit the location information in real time. Active systems are developed. Real time vehicular tracking system incorporates a hardware device installed in the vehicle (In-Vehicle Unit) and a remote Tracking server. The information is transmitted to Tracking server using GSM/GPRS modem on GSM network by using SMS or using direct TCP/IP connection with Tracking server through GPRS. Tracking server also has GSM/GPRS modem that receives vehicle location information via GSM network and stores this information in database. This information is available to authorized users of the system via website over the internet. Keywords: GPS,GPRS,Sensor

FICO score - ROLE IN MODERN FINANCIAL SYSTEM

FICO assessment business is a corner portion in the budgetary administrations field. In the post-changes period, with expanded action in the Indian Financial division both existing and new organizations are selecting money from the capital business sector. The opposition among firms for a cut of the reserve funds cake has expanded. FICO score business in India is a sweet spot as it is on the cusp of vigorous development potential, driven by three triggers: Strong capex cycle in Indian economy, lower infiltration of corporate security market and administrative push because of execution of Basel II standards. FICO score helps in the improvement of money related markets. FICO score is a financial specialist administration and a rating organization is required to keep up the most astounding conceivable level of expository capability and trustworthiness. The diagnostic structure of rating manages assessment of both the business and monetary dangers connected with that element. Other than subjective perspectives like administration abilities additionally assume an extensive part in deciding a rating. FICO assessments set up a connection between danger and return. They in this manner give a measuring stick against which to quantify the danger characteristic in any instrument. Explanatory structure of rating manages assessment of both the business and money related dangers connected with that substance. The Reserve Bank of India liaises with SEBI, on the issue of rating offices' adherence to IOSCO Code of Conduct Fundamentals.Given the droop confronted by economies all around and the ascent in the quantity of defaultees, the time has come for the channel had a solid FICO score framework set up to guarantee smooth operation for the whole chain. The most noteworthy change in late identifies with accentuation on their responsibility and more imperative, the alert in controllers' utilization of evaluations

Alcoholic hepatitis accelerates early hepatobiliary cancer by increasing stemness and miR-122-mediated HIF-1alpha activation

Alcohol-related hepatocellular carcinoma (HCC) develops with advanced alcoholic liver disease and liver fibrosis. Using adult mice, we evaluate the effect of alcoholic steatohepatitis on early hepatobiliary carcinoma after initiation by diethyl-nitrosamine (DEN). Here we show that alcohol-fed DEN-injected mice have higher ALT and liver-to-body weight ratio compared to pair-fed DEN-injected mice. Alcohol feeding results in steatohepatitis indicated by increased pro-inflammatory cytokines and fibrotic genes. MRI and liver histology of alcohol+DEN mice shows hepatobiliary cysts, early hepatic neoplasia and increase in serum alpha-fetoprotein. Proliferation makers (BrdU, cyclin D1, p53) and cancer stem cell markers (CD133 and nanog) are significantly up-regulated in livers of alcohol-fed DEN-injected mice compared to controls. In livers with tumors, loss of miR-122 expression with a significant up-regulation of miR-122 target HIF-1alpha is seen. We conclude that alcoholic steatohepatitis accelerates hepatobiliary tumors with characteristic molecular features of HCC by up-regulating inflammation, cell proliferation, stemness, and miR-122 loss

Thanatochemistry: Study of synovial fluid potassium

Death interval estimation is one of the major attributes obtained from thanatochemistry. Body fluids like vitreous humour have been consistently used to estimate death interval from regression equations, but these may be useless when dealing with eye trauma, ocular disorders or in mutilated remains. Until recently, there was no consistent assessment of the reliability of measurements of the synovial fluid for death interval estimation. The purpose of this paper is to test previously developed regression formulae for estimating death interval based on synovial fluid potassium and to assess its reliability in estimating death interval. Synovial fluid potassium was measured on a sample of 308 individuals. Death interval was regressed on synovial fluid potassium and a regression formula was obtained. The regression model provided a 95% confidence interval of ±3.0 h and a correlation coefficient of 0.739. Compared to other studies, regression formulae based on the synovial fluid potassium in the present study provided considerably less standard errors.KEYWORDS Thanatochemistry; Cadaveric; Synovial fluid; Potassium; Postmortem interva

Association between various antioxidants in normals and chronic obstructive pulmonary disease, their alteration and impact of smoking and disease on levels of antioxidants

Background: There is total alteration of various antioxidants in response to the oxidative stress, which is one of the major patho-physiologic hallmarks in chronic obstructive pulmonary disease (COPD) development. This study aims to establish the correlation between different antioxidants in normals and COPD, study the alteration in the correlation due to COPD and smoking as well as the impact of COPD and smoking on antioxidants levels.Methods: Study comprises of 96 normals as group I and 96 COPD patients as group II. The antioxidants albumin (Alb), bilirubin (Bil), uric acid (UA) ceruloplasmin (Cp), glutathione peroxidase (GSHPx), catalase (CAT) and superoxide dismutase 3 (SOD3) were estimated.Results: Significant lower serum Alb, UA, SOD3 and increased serum Cp and GSHPx were found in Group II. Significant correlation was found between Alb and UA (r=0.24); Bil and UA (r=0.26); Alb and CAT (r=0.211) and SOD3 and CAT (r=0.318) in normals. However, these correlations were altered in COPD where Alb correlates with Bil (r=0.235); UA with CAT (r=0.203) and SOD3 with GSHPx (r=-0.27). The correlation between SOD3 and CAT remained unaltered. Similar correlation of UA with Alb and Bil was observed in nonsmoker normals and between SOD3 and CAT in smoker normals. In COPD, no correlation was seen in nonsmokers, while in smokers Alb correlates with Bil (r=0.316) and SOD3 with CAT (r=0.317).Conclusions: These alterations may have clinical ramifications in further understanding the pathogenesis of COPD and developing therapeutic approaches

FXR and TGR5 Agonists Ameliorate Liver Injury, Steatosis, and Inflammation After Binge or Prolonged Alcohol Feeding in Mice

Bile acids (BAs) activate various dedicated receptors, including the farnesoid X receptor (FXR) and the Takeda G protein-coupled receptor 5 (TGR5). The FXR agonist obeticholic acid (OCA) is licensed for the treatment of primary biliary cholangitis and has shown promising results in NASH patients, whereas TGR5 agonists target inflammation and metabolism. We hypothesized that FXR and/or TGR5 agonists may be therapeutic in early alcoholic liver disease (ALD) in mice, in which hepatic inflammation plays a major role. OCA, INT-777, and INT-767 are BA derivatives with selective agonist properties for FXR, TGR5, or both, respectively. These compounds were tested in two mouse models (3-day binge model and prolonged Lieber DeCarli diet for 12 days) of early ALD. Serum alanine aminotransferase and liver histology were used to assess liver injury, Oil Red O staining of liver sections to assess steatosis, and real-time polymerase chain reaction to assess changes in gene expression. In the ethanol binge model, treatment with OCA and INT-777 decreased hepatic macrovesicular steatosis and protected from ethanol-induced liver injury. After prolonged ethanol administration, mice treated with OCA, INT-767, or INT-777 showed decreased hepatic steatosis, associated with reduced liver fatty acid synthase protein expression, and protection from liver injury. Treatment with BA receptor agonists in both models of ethanol administration modulated lipogenic gene expression, and decreased liver interleukin-1beta mRNA expression associated with increased ubiquitination of NLRP3 inflammasome through cyclic adenosine monophosphate-induced activation of protein kinase A. Conclusion: OCA, INT-767, or INT-777 administration is effective in reducing acute and chronic ethanol-induced steatosis and inflammation in mice, with varying degrees of efficacy depending on the duration of ethanol administration, indicating that both FXR and TGR5 activation can protect from liver injury in ALD models

Progression of non-alcoholic steatosis to steatohepatitis and fibrosis parallels cumulative accumulation of danger signals that promote inflammation and liver tumors in a high fat-cholesterol-sugar diet model in mice

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is becoming a pandemic. While multiple \u27hits\u27 have been reported to contribute to NAFLD progression to non-alcoholic steatohepatitis (NASH), fibrosis and liver cancer, understanding the natural history of the specific molecular signals leading to hepatocyte damage, inflammation and fibrosis, is hampered by the lack of suitable animal models that reproduce disease progression in humans. The purpose of this study was first, to develop a mouse model that closely mimics progressive NAFLD covering the spectrum of immune, metabolic and histopathologic abnormalities present in human disease; and second, to characterize the temporal relationship between sterile/exogenous danger signals, inflammation, inflammasome activation and NAFLD progression. METHODS: Male C57Bl/6 mice were fed a high fat diet with high cholesterol and a high sugar supplement (HF-HC-HSD) for 8, 27, and 49 weeks and the extent of steatosis, liver inflammation, fibrosis and tumor development were evaluated at each time point. RESULTS: The HF-HC-HSD resulted in liver steatosis at 8 weeks, progressing to steatohepatitis and early fibrosis at 27 weeks, and steatohepatitis, fibrosis, and tumor development at 49 weeks compared to chow diet. Steatohepatitis was characterized by increased levels of MCP-1, TNFalpha, IL-1beta and increased liver NASH histological score. We found increased serum levels of sterile danger signals, uric acid and HMGB1, as early as 8 weeks, while endotoxin and ATP levels increased only after 49 weeks. Increased levels of these sterile and microbial danger signals paralleled upregulation and activation of the multiprotein complex inflammasome. At 27, 49 weeks of HF-HC-HSD, activation of M1 macrophages and loss of M2 macrophages as well as liver fibrosis were present. Finally, similar to human NASH, liver tumors occurred in 41% of mice in the absence of cirrhosis and livers expressed increased p53 and detectable AFP. CONCLUSIONS: HF-HC-HSD over 49 weeks induces the full spectrum of liver pathophysiologic changes that characterizes the progression of NAFLD in humans. NAFLD progression to NASH, fibrosis and liver tumor follows progressive accumulation of sterile and microbial danger signals, inflammasome activation, altered M1/M2 cell ratios that likely contribute to NASH progression and hepatic tumor formation