IL-33 expression is altered in hepatitis C virus-related pathologies

Hepatitis C virus (HCV) currently affects approximately 71 million people worldwide and causes extensive liver damage that can transition into hepatocellular carcinoma (HCC), a deadly malignant liver cancer. Unfortunately, patients tend to seek diagnostic testing late, when more severe symptoms exist, indicating advanced disease with severe fibrosis. There is a gap in knowledge about the immunologic process of HCV-induced fibrosis and cirrhosis development. Interleukin-33 (IL-33) is a cytokine implicated in various inflammatory and fibrotic diseases. We hypothesize that IL-33 plays a role in the pathogenesis of HCV, particularly in fibrosis and cirrhosis development leading to malignant transformation and thus may serve as an inflammatory biomarker. We studied livers from normal subjects - no liver-related diagnoses (n=6), subjects with HCV-cirrhosis (n=7), and subjects with HCV/HCC (n=7). We performed immunohistochemistry on paraffin-embedded liver tissue sections using a human IL-33 antibody and DAB staining system. To quantify our results, we performed image analyses quantifying IL-33 chromogen stain using an algorithm to calculate the signal strength of an image to determine the amount of antibody-specific chromogen per pixel, expressed in energy units per pixel (eu/px). We found that IL-33 is normally expressed in human hepatocytes; it is expressed at higher levels in HCV-cirrhosis patients and at lower levels in HCV-HCC patients. We plan to expand this study by including more patients in the future

Proteomics study reveals a gender-based ribosomal inflammatory biomarker in Hepatitis C virus induced cirrhosis

Hepatitis C Virus (HCV) infection is a major cause of Hepatocellular Carcinoma (HCC). Further, HCC is a leading cause of liver-related death world-wide linked to liver cirrhosis and chronic liver disease. Clinical evidence suggests that pre-menopausal women, with elevated levels of circulating estrogen, clear HCV infection faster than males, and show low incidence of HCC. Our studies have shown gender-based differential estrogen receptor expression in the normal liver, which could contribute to protection in pre-menopausal women against chronic liver diseases including HCC. There is a gap in the current knowledge of biomarkers that could be used for early detection of HCV-related cirrhosis and HCC development. Further, biomarkers that account for gender differences in HCV-related pathogenesis have not yet been identified.The purpose of this study was to identify early cancer biomarkers in HCV-related cirrhosis by analyzing liver proteins-specifically focusing on ribosomal family of proteins (RPs). Ribosomal proteins play an important role in cellular apoptosis, cell proliferation, and cancer development. Ribosomes convert mRNA into proteins using various RPs and have been shown to change in different tissues and tumors. Ribosomal gene FAU (Finkel-Biskis-Reilly murine sarcoma virus) has recently been identified as an apoptosis regulatory gene involved in ovarian, breast, and prostate cancer. To identify gender-based protein differences in male and female liver cirrhosis patients, we investigated RPs via proteomic study. Normal and HCV diseased liver tissue extracts were processed and sent to NIH IDeA National Resource for Quantitative Proteomics core lab for DIA proteomic analysis. We had four experimental groups with 10 samples in each group. The four groups included were normal control male, normal control female, HCV cirrhosis male, and HCV cirrhosis female. A total of 4,443 proteins were identified of which 132 were ribosomal proteins. Within the ribosomal protein cohort, 78 exhibited differential expression in liver cirrhosis compared to normals, with 13 showing gender-based significance.FAU gene encodes for the 40S ribosomal protein S30 in the cytoplasmic ribosome and illustrated gender-based differences in cirrhosis group. FAU was significantly upregulated (logFC 1.04050553) in HCV females compared to HCV male liver tissues (p-value .0012) and was significantly downregulated in HCV male liver tissues compared to normal control males. No differences were observed in remaining experimental groups (HCV F vs. normal control F and normal control M vs. normal control F). This study identified the ribosomal protein FAU and its potential to serve as a novel gender-based biomarker in liver cirrhosis and cancer development.This finding could be essential in understanding why males may be more susceptible to developing HCC. To our knowledge, this is the first report showing FAU ribosomal protein in HCV-related cirrhosis with significant gender-based differences. FAU could serve as a valuable prognostic biomarker to monitor HCC disease progression and response to treatment

Proteomics study reveals a hydroxysteroid dehydrogenase (HSD) as a sex-based biomarker in Hepatitis C virus induced cirrhosis

Hepatitis C virus (HCV) infection-related inflammation, liver fibrosis and cirrhosis often lead to development of hepatocellular carcinoma (HCC). In the United States, 4.6 million people are infected with HCV. Studies show that chronic HCV infections are more prevalent in males and progress more rapidly to cirrhosis and cancer development as compared to females. In contrast, pre-menopausal females and women on hormone replacement therapy have been associated with less-severe disease through all stages of HCV infection. We have previously identified sex-based differences in the expression of estrogen receptors (ERs)in normal livers and dysregulated mRNA and protein expression of ER subtypes in both HCV-related cirrhosis and HCC suggesting a possible role in its pathogenesis. Sex based differential biomarkers could serve as an early prognostic tool for HCV cirrhosis and HCC development. The enzyme families including hydroxysteroid dehydrogenases (HSDs) contribute largely to the synthesis and degradation of steroid hormones such as testosterone and estrogen sex-hormones, as well as cholesterol and fatty acid metabolism. Chronic inflammation due to HCV infection in the liver may alter HSD enzyme regulation and affect hormone metabolism. We hypothesized that chronic HCV infection leads to dysregulated HSDs in male HCV cirrhosis patients leading to development of HCC. Our current study utilized proteomic analysis to determine sex-based differences in HSD protein expression in male and female HCV cirrhosis. We studied a total 40 liver tissues that included both sexes from HCV-related cirrhosis and normal controls. The liver tissues extracts were prepared and sent to NIH IDeA National Resource of Quantitative Proteomics core lab for DIA proteomic analysis. We were able to profile 4,443 genes belonging to different protein families that were differentially expressed in HCV cirrhosis and controls. Within the HSD protein cohort 7 exhibited differential expression in the liver cirrhosis groups compared to healthy controlss. Of these 7 proteins, only HSD17B13 demonstrated a significant sex-based differential expression between male and female HCV cirrhosis groups. More specifically, HCV cirrhosis females demonstrated a positive logFC value of 2.241 (p < 0.01) when compared to HCV cirrhosis males, suggesting HSD17B13 may serve as sex-based pre-cancerous biomarker. Further, HSD17B13 showed downregulation in HCV cirrhosis males compared to normal control males, but not significantly. No differences were observed in the remaining experimental groups (HCV F vs. normal control F and normal control M vs. normal control F). HSD17B13 protein may serve as a sex-based biomarker in liver cirrhosis and cancer development. To the best of our knowledge this is the first report showing sex-based differences in HSD proteins in premalignant HCV-related cirrhosis. Further, detailed studies may lead us to new sex-based tailored clinical therapies in halting cirrhosis and HCC progression in males

A comparative study of functional outcome of external fixation and volar plating in unstable distal radius fractures

Background: Distal radius fractures are amongst the most common injuries seen in the emergency department. It is imperative to restore the anatomy as much as possible in order to restore wrist function. Identifying a fracture as ‘unstable’ is critical in predicting the final outcome of treatment. The present study compares two primary modalities of treatment: external fixation and volar plating in terms of the functional outcome post-surgery.  Aim of the study was to compare the functional outcome of fixation of unstable distal radius fractures by external fixation to that of volar plating by the Disabilities of the Arm, Shoulder and Hand (DASH) scoring system and analysis of recovery of grip strength and range of motion. Settings and design: Hospital-based; randomized control trial.Methods: 80 patients presenting with unstable distal radius fractures were randomized into two groups of 40 each. One group received external fixation and the second received open reduction with volar plate fixation as the primary intervention. DASH scores were obtained and compared at specified time intervals following surgery.Results: The results of our study show a better improvement in the mean DASH scores at 3, 6 and 12 months in the volar plating group as compared to the external fixation group.Conclusions: Volar plating, with its definitive advantage of direct fracture visualization, has an overall better functional outcome with greater postoperative wrist motion and a lower incidence of complications.

Estrogen receptor expression in chronic hepatitis C and hepatocellular carcinoma pathogenesis

AIM: To investigate gender-specific liver estrogen receptor (ER) expression in normal subjects and patients with hepatitis C virus (HCV)-related cirrhosis and hepatocellular carcinoma (HCC).METHODS: Liver tissues from normal donors and patients diagnosed with HCV-related cirrhosis and HCV-related HCC were obtained from the NIH Liver Tissue and Cell Distribution System. The expression of ER subtypes, ERa and ERB, were evaluated by Western blotting and real-time RT-PCR. The subcellular distribution of ERa and ERB was further determined in nuclear and cytoplasmic tissue lysates along with the expression of inflammatory [activated NF-KB and IKB-kinase (IKK)] and oncogenic (cyclin D1) markers by Western blotting and immunohistochemistry. The expression of ERa and ERB was correlated with the expression of activated NF-KB, activated IKK and cyclin D1 by Spearman's correlation.RESULTS: Both ER subtypes were expressed in normal livers but male livers showed significantly higher expression of ERa than females (P < 0.05). We observed significantly higher mRNA expression of ERa in HCV-related HCC liver tissues as compared to normals (P < 0.05) and ERB in livers of HCV-related cirrhosis and HCV-related HCC subjects (P < 0.05). At the protein level, there was a significantly higher expression of nuclear ERa in livers of HCV-related HCC patients and nuclear ERB in HCV-related cirrhosis patients as compared to normals (P < 0.05). Furthermore, we observed a significantly higher expression of phosphorylated NF-KB and cyclin D1 in diseased livers (P < 0.05). There was a positive correlation between the expression of nuclear ER subtypes and nuclear cyclin D1 and a negative correlation between cytoplasmic ER subtypes and cytoplasmic phosphorylated IKK in HCV-related HCC livers. These findings suggest that dysregulated expression of ER subtypes following chronic HCV-infection may contribute to the progression of HCV-related cirrhosis to HCV-related HCC.CONCLUSION: Gender differences were observed in ERa expression in normal livers. Alterations in ER subtype expression observed in diseased livers may influence gender-related disparity in HCV-related pathogenesis.Peer reviewedBiochemistry and MicrobiologyHealth Care AdministrationStatistic

17β-HSD13 has sex-based differential expression in Hepatitis C virus-induced cirrhosis and hepatocellular carcinoma

Background: Sex-based differences are observed in chronic hepatitis C virus (HCV) infections leading to cirrhosis and hepatocellular carcinoma (HCC). We previously showed that liver estrogen receptor (ER-) mediated sex-based differences exist in cirrhosis and HCC. Liver ER-binding may lead to protective effects in pre-menopausal women. This study aimed to determine sex-based differential role of 17βHSD13 in development of cirrhosis and HCC. We hypothesized that chronic HCV infection leads to dysregulated 17β-HSD13 in male cirrhosis and progression to HCC.Methods: 65 (normal, cirrhosis, HCC) liver tissues were obtained from NIH Liver Tissue Bank. DIA proteomics mapped 4445 proteins, including 17β-HSD13. Clinical correlation with bilirubin, AST, ALP, and creatinine was determined (spearman’s). Immunohistochemistry validated 17β-HSD13 protein expression in tissues.Results: 17β-HSD13 had significantly lower expression in male cirrhosis group than females (P<0.05). In contrast, 17β-HSD13 expression in normal males was significantly greater than normal females (P<0.05). In HCC group, the expression in males was down-regulated compared to HCC females (P<0.05). Bilirubin values showed negative correlation with 17β-HSD13 expression (P<0.05) between cirrhosis and HCC (males alone and combined sex data).Conclusions: Low 17β-HSD13 levels may predict worse disease in males with cirrhosis or HCC serving as disease biomarker. This novel report shows sex-based differences in 17β-HSD13 during HCV-induced cirrhosis development

‘It Takes Two Hands to Clap’: How Gaddi Shepherds in the Indian Himalayas Negotiate Access to Grazing

This article examines the effects of state intervention on the workings of informal institutions that coordinate the communal use and management of natural resources. Specifically it focuses on the case of the nomadic Gaddi shepherds and official attempts to regulate their access to grazing pastures in the Indian Himalayas. It is often predicted that the increased presence of the modern state critically undermines locally appropriate and community-based resource management arrangements. Drawing on the work of Pauline Peters and Francis Cleaver, I identify key instances of socially embedded ‘common’ management institutions and explain the evolution of these arrangements through dynamic interactions between individuals, communities and the agents of the state. Through describing the ‘living space’ of Gaddi shepherds across the annual cycle of nomadic migration with their flocks I explore the ways in which they have been able to creatively reinterpret external interventions, and suggest how contemporary arrangements for accessing pasture at different moments of the annual cycle involve complex combinations of the formal and the informal, the ‘traditional’ and the ‘modern’

Piperidinols that show anti-tubercular activity as inhibitors of arylamine N-acetyltransferase: an essential enzyme for mycobacterial survival inside macrophages

Latent M. tuberculosis infection presents one of the major obstacles in the global eradication of tuberculosis (TB). Cholesterol plays a critical role in the persistence of M. tuberculosis within the macrophage during latent infection. Catabolism of cholesterol contributes to the pool of propionyl-CoA, a precursor that is incorporated into cell-wall lipids. Arylamine N-acetyltransferase (NAT) is encoded within a gene cluster that is involved in the cholesterol sterol-ring degradation and is essential for intracellular survival. The ability of the NAT from M. tuberculosis (TBNAT) to utilise propionyl-CoA links it to the cholesterol-catabolism pathway. Deleting the nat gene or inhibiting the NAT enzyme prevents intracellular survival and results in depletion of cell-wall lipids. TBNAT has been investigated as a potential target for TB therapies. From a previous high-throughput screen, 3-benzoyl-4-phenyl-1-methylpiperidinol was identified as a selective inhibitor of prokaryotic NAT that exhibited antimycobacterial activity. The compound resulted in time-dependent irreversible inhibition of the NAT activity when tested against NAT from M. marinum (MMNAT). To further evaluate the antimycobacterial activity and the NAT inhibition of this compound, four piperidinol analogues were tested. All five compounds exert potent antimycobacterial activity against M. tuberculosis with MIC values of 2.3-16.9 µM. Treatment of the MMNAT enzyme with this set of inhibitors resulted in an irreversible time-dependent inhibition of NAT activity. Here we investigate the mechanism of NAT inhibition by studying protein-ligand interactions using mass spectrometry in combination with enzyme analysis and structure determination. We propose a covalent mechanism of NAT inhibition that involves the formation of a reactive intermediate and selective cysteine residue modification. These piperidinols present a unique class of antimycobacterial compounds that have a novel mode of action different from known anti-tubercular drugs