Alcoholic Hepatitis Patients Research Articles

Loss of long-chain acyl-CoA synthetase 1 promotes hepatocyte death in alcohol-induced steatohepatitis

BackgroundAlcohol consumption has been shown to disrupt hepatic lipid homeostasis. Long-chain acyl-CoA synthetase 1 (ACSL1) critically regulates hepatic fatty acid metabolism and lipid homeostasis by channeling fatty acids to lipid metabolic pathways. However, it remains unclear how ACSL1 contributes to the development of alcohol-associated liver disease (ALD). MethodsWe performed chronic alcohol feeding animal studies with hepatocyte-specific ACSL1 knockout (ACSL1Δhep) mice, hepatocyte-specific STAT5 knockout (STAT5Δhep) mice, and ACSL1Δhep based-STAT5B overexpression (Stat5b-OE) mice. Cell studies were conducted to define the causal role of ACSL1 deficiency in the pathogenesis of alcohol-induced liver injury. The clinical relevance of the STAT5-ACSL1 pathway was examined using liver tissues from patients with alcoholic hepatitis (AH) and normal subjects (Normal). ResultsWe found that chronic alcohol consumption reduced hepatic ACSL1 expression in AH patients and ALD mice. Hepatocyte-specific ACSL1 deletion exacerbated alcohol-induced liver injury by increasing free fatty acids (FFA) accumulation and cell death. Cell studies revealed that FFA elicited the translocation of BAX and p-MLKL to the lysosomal membrane, resulting in lysosomal membrane permeabilization (LMP) and thereby initiating lysosomal-mediated cell death pathway. Furthermore, we identified that the signal transducer and activator of transcription 5 (STAT5) is a novel transcriptional regulator of ACSL1. Deletion of STAT5 exacerbated alcohol-induced liver injury in association with downregulation of ACSL1, and reactivation of ACSL1 by STAT5 overexpression effectively ameliorated alcohol-induced liver injury. In addition, ACSL1 expression was positively correlated with STAT5 and negatively correlated with cell death was also validated in the liver of AH patients. ConclusionsACSL1 deficiency due to STAT5 inactivation critically mediates alcohol-induced lipotoxicity and cell death in the development of ALD. These findings provide insights into alcohol-induced liver injury.

Emergence of highly profibrotic and proinflammatory Lrat+Fbln2+ HSC subpopulation in alcoholic hepatitis.

Relative roles of HSCs and portal fibroblasts in alcoholic hepatitis (AH) are unknown. We aimed to identify subpopulations of collagen type 1 alpha 1 (Col1a1)-expressing cells in a mouse AH model by single-cell RNA sequencing (scRNA-seq) and filtering the cells with the HSC (lecithin retinol acyltransferase [Lrat]) and portal fibroblast (Thy-1 cell surface antigen [Thy1] and fibulin 2 [Fbln2]) markers and vitamin A (VitA) storage. Col1a1-green fluorescent protein (GFP) mice underwent AH, CCl 4 , and bile duct ligation (BDL) procedures to have comparable F1-F2 liver fibrosis. Col1a1-expressing cells were sorted via FACS by VitA autofluorescence and GFP for single-cell RNA sequencing. In AH, approximately 80% of Lrat+Thy1-Fbln2- activated HSCs were VitA-depleted (vs. ~13% in BDL and CCl 4 ). Supervised clustering identified a subset co-expressing Lrat and Fbln2 (Lrat+Fbln2+), which expanded 44-fold, 17-fold, and 1.3-fold in AH, BDL, and CCl 4 . Lrat+Fbln2+ cells had 3-15-times inductions of profibrotic, myofibroblastic, and immunoregulatory genes versus Lrat+Fbln2- cells, but 2-4-times repressed HSC-selective genes. AH activated HSCs had up-regulated inflammatory (chemokine [C-X-C motif] ligand 2 [Cxcl2], chemokine [C-C motif] ligand 2), antimicrobial (Il-33, Zc3h12a), and antigen presentation (H2-Q6, H2-T23) genes versus BDL and CCl 4 . Computational deconvolution of AH versus normal human bulk-liver RNA-sequencing data supported an expansion of LRAT+FBLN2+ cells in AH; AH patient liver immunohistochemistry showed FBLN2 staining along fibrotic septa enriched with LRAT+ cells; and in situ hybridization confirmed co-expression of FBLN2 with CXCL2 and/or human leukocyte antigen E in patient AH. Finally, HSC tracing in Lrat-Cre;Rosa26mTmG mice detected GFP+FBLN2+ cells in AH. A highly profibrotic, inflammatory, and immunoregulatory Lrat+Fbln2+ subpopulation emerges from HSCs in AH and may contribute to the inflammatory and immunoreactive nature of AH.

S1409 Improving Provider Adherence to Nutritional and Metabolic Recommendations in Patients With Acute Alcoholic Hepatitis: A Quality Improvement Initiative

Introduction: Alcoholic hepatitis (AH) is an acute, inflammatory liver disease associated with high morbidity and mortality, both short and long term. Current medical therapy for severe alcoholic hepatitis relies on corticosteroids, which have modest efficacy. Alcohol abstinence is of critical importance but recidivism is high. Efficacious medical treatments for alcoholic hepatitis are lacking. Preliminary and Preclinical studies include use of Antibiotics, IL-1 inhibitors, GCSF and antioxidants. Malnutrition and sarcopenia are common among hospitalized AH patients with negative impact on outcome.[1] Current guidelines generally recommend daily protein intake of 1.2–1.5 g/kg/day and caloric intake of 30- 40 kcal/kg/day in alcoholic hepatitis patients. [2][3] Our objective is to implement methods to facilitate adherence to nutritional recommendations in hospitalized patients with AH. Methods: We conducted a 12-month retrospective study followed by a 3-month prospective interventional study at Presbyterian Medical Center. Chart review was performed to evaluate adherence with nutritional recommendations in patients with AH. A 3-month intervention was performed to improve compliance. This included the creation of a customized best practice advisory alert (BPA) in the Electronic medical record (EMR). The triggers included elevated ALT/AST, bilirubin >3 and documented alcohol use within 8 weeks. Alert encouraged physicians to request a nutrition consult if the diagnosis of AH was appropriate. Results: 105 patients with 82 in the pre- and 23 post-intervention periods were reviewed. No significant difference in patient characteristics and demographics was noted between the 2 groups. The intervention implemented revealed a 79% increase in adherence to nutritional recommendation (P < 0.023). Adherence was quantified as modified diet orders in EMR to include high protein diet ( minimum of 60 grams/daily ) and/or addition of supplemental high protein milk shakes to meals and/or electronic consult to nutritionist. Conclusion: AH has high morbidity and mortality. Current pharmacological options remain limited. Patients with AH are commonly malnourished and sarcopenic. System based auto-generated EMR alerts help increase compliance with the nutritional recommendations as well as enforce evidence-based practices in this select patient population.

Beneficial Shifts in Gut Microbiota by Lacticaseibacillus rhamnosus R0011 and Lactobacillus helveticus R0052 in Alcoholic Hepatitis.

Gut microbiota performs indispensable functions in the pathophysiology of alcoholic hepatitis (AH). We investigated the effects of Lacticaseibacillus rhamnosus R0011 and Lactobacillus helveticus for gut microbial restoration toward eubiosis in patients with AH. A multicenter, double-blind, and randomized trial was conducted. Probiotics (n = 44) and placebo (n = 45) groups received, during 7 days, L. rhamnosus R0011/L. helveticus R0052 at 120 mg/day and placebo. All patients were hospitalized to ensure abstinence. Liver function, lipopolysaccharide level, and stool analysis were evaluated in patients before and after 7 days of treatment. At baseline, the dominant bacteria were Gram-negative in both groups which decreased after the probiotics treatment and exhibited a significant reduction in lipopolysaccharide level (p < 0.001). The probiotics ameliorated the Child–Pugh scores (p < 0.001). Furthermore, the probiotics group showed a decline in the levels of alanine aminotransferase and gamma-glutamyltranspeptidase (p < 0.05). The probiotics changed the gut microbial composition at various taxonomical levels. The proportion of Bacteroidetes (147%) was increased after 7 days of probiotics supplementation while Proteobacteria (30%) and Fusobacteria (0%) were decreased. Administration of L. rhamnosus R0011 and L. helveticus R0052 conceivably associated with restoration of gut microbiome in AH patients and improved AH by modulating the gut–liver axis.

Distinct histopathological phenotypes of severe alcoholic hepatitis suggest different mechanisms driving liver injury and failure.

Intrahepatic neutrophil infiltration has been implicated in severe alcoholic hepatitis (SAH) pathogenesis; however, the mechanism underlying neutrophil-induced injury in SAH remains obscure. This translational study aims to describe the patterns of intrahepatic neutrophil infiltration and its involvement in SAH pathogenesis. Immunohistochemistry analyses of explanted livers identified two SAH phenotypes despite a similar clinical presentation, one with high intrahepatic neutrophils (Neuhi), but low levels of CD8+ T cells, and vice versa. RNA-Seq analyses demonstrated that neutrophil cytosolic factor 1 (NCF1), a key factor in controlling neutrophilic ROS production, was upregulated and correlated with hepatic inflammation and disease progression. To study specifically the mechanisms related to Neuhi in AH patients and liver injury, we used the mouse model of chronic-plus-binge ethanol feeding and found that myeloid-specific deletion of the Ncf1 gene abolished ethanol-induced hepatic inflammation and steatosis. RNA-Seq analysis and the data from experimental models revealed that neutrophilic NCF1-dependent ROS promoted alcoholic hepatitis (AH) by inhibiting AMP-activated protein kinase (a key regulator of lipid metabolism) and microRNA-223 (a key antiinflammatory and antifibrotic microRNA). In conclusion, two distinct histopathological phenotypes based on liver immune phenotyping are observed in SAH patients, suggesting a separate mechanism driving liver injury and/or failure in these patients.

Trends of Readmissions for Gastrointestinal Bleeding After Alcoholic Hepatitis: Analysis of the Nationwide Readmission Database.

BackgroundAlcohol consumption is associated with numerous hepatic manifestations, including alcoholic fatty liver disease, alcoholic hepatitis (AH), and liver cirrhosis. AH is a common and serious complication of alcohol use. Gastrointestinal bleeding (GIB) remains one of the most common causes of death in these patients. In this article, we studied the trends of GIB after AH.MethodsThis was a retrospective interrupted trend study. We analyzed the 2010, 2012, 2014, 2016, and 2018 Nationwide Readmission Databases. The first AH hospitalization in the year was marked as index hospitalization. We identified subsequent hospitalizations with GIB within 30 days and marked them as readmissions. A multivariate regression analysis was used to calculate the risk-adjusted odds of trends for GIB readmissions, including esophageal varices bleeding (EVB), upper GIB, lower GIB, and all GIB.ResultsThe volume of index hospitalizations increased from 10,248 in 2010 to 16,479 in 2018. Similarly, all readmissions increased from 1,838 in 2010 to 3,908 in 2018. Of all readmissions, EVB increased from 3.9% in 2010 to 5.9% in 2018 (odds ratio (OR) trend 1.10; P < 0.001). Readmissions for upper GIB increased from 2.4% in 2010 to 7.8% in 2018 (OR trend 1.22; P < 0.001). On the other hand, lower GIB readmissions decreased from 7.2% in 2010 to 4.7% in 2018 (OR trend 0.95; P = 0.015). There was no statistically significant trend for all GIB readmissions (OR trend 1; P = 0.915).ConclusionFurther studies are needed to evaluate the patterns of lower GIB in patients with liver disease and the recent trends of corticosteroids use in AH patients.

A Novel Murine Model for Studying Alcohol‐associated Biliary Dysfunction

Alcoholic hepatitis (AH) is a severe, potentially life‐threatening form of alcohol‐associated liver disease (ALD) with limited therapeutic options. Existing evidence indicates that cholestasis and biliary dysfunction are common in AH patients and can worsen the prognosis. However, most ALD murine models used today can only recapitulate hepatic steatosis with limited features of AH, which significantly curbs research related to ALD pathogenesis. To develop a murine model that is feasible and less time‐consuming, we included a subtoxic dose of α‐naphthylisothiocyanate (ANIT), a well‐studied intrahepatic cholestasis inducer, in an alcohol feeding model, as a second hit. Mice were fed with an ethanol diet (EtOH) for 10 days or 4 weeks, followed by ANIT administration 48 hours before sample collection. Our results showed that ANIT did not cause overt liver injury in pair‐fed mice, but an observable increase in serum levels of AST in mice fed EtOH for 10 days. Remarkably, in the 4‐week ALD model, serum levels of ALT, AST, total bile acids (TBA), and hepatic levels of TBA were all significantly increased in mice given EtOH and ANIT, indicating a synergistic effect of EtOH and ANIT on liver injury that is dependent on EtOH feeding. In the liver, 4‐week EtOH‐fed mice displayed cholangitis, lytic cell death, and neutrophil infiltration around the periportal area following ANIT treatment as shown by H&amp;E and LY6G staining. In contrast, pair‐fed mice did not show abnormal liver histology following ANIT treatment. Expression of several inflammation‐related genes that are increased in livers from AH patients were analyzed. Consistent with histological observations, hepatic expression of genes like Cxcl1, Ccl2, Ccl20, and Lcn2 were significantly increased in 4‐week EtOH‐fed mice following ANIT treatment, suggesting that ANIT synergistically enhanced EtOH‐induced immune response in the liver. Taken together, our results suggest that EtOH‐dependent changes in the liver can cause higher susceptibility to biliary injury and that a subtoxic dose of ANIT pushes alcohol‐associated liver damage towards a more severe phenotype in mice. Overall, our results support a treatment scheme for alcohol‐associated cholestasis that is feasible, less time‐consuming, cost‐effective, and with modulable severity of phenotypes.

Impaired regulation of PMCA activity by defective CFTR expression promotes epithelial cell damage in alcoholic pancreatitis and hepatitis.

Alcoholic pancreatitis and hepatitis are frequent, potentially lethal diseases with limited treatment options. Our previous study reported that the expression of CFTR Cl- channel is impaired by ethanol in pancreatic ductal cells leading to more severe alcohol-induced pancreatitis. In addition to determining epithelial ion secretion, CFTR has multiple interactions with other proteins, which may influence intracellular Ca2+ signaling. Thus, we aimed to investigate the impact of ethanol-mediated CFTR damage on intracellular Ca2+ homeostasis in pancreatic ductal epithelial cells and cholangiocytes. Human and mouse pancreas and liver samples and organoids were used to study ion secretion, intracellular signaling, protein expression and interaction. The effect of PMCA4 inhibition was analyzed in a mouse model of alcohol-induced pancreatitis. The decreased CFTR expression impaired PMCA function and resulted in sustained intracellular Ca2+ elevation in ethanol-treated and mouse and human pancreatic organoids. Liver samples derived from alcoholic hepatitis patients and ethanol-treated mouse liver organoids showed decreased CFTR expression and function, and impaired PMCA4 activity. PMCA4 co-localizes and physically interacts with CFTR on the apical membrane of polarized epithelial cells, where CFTR-dependent calmodulin recruitment determines PMCA4 activity. The sustained intracellular Ca2+ elevation in the absence of CFTR inhibited mitochondrial function and was accompanied with increased apoptosis in pancreatic epithelial cells and PMCA4 inhibition increased the severity of alcohol-induced AP in mice. Our results suggest that improving Ca2+ extrusion in epithelial cells may be a potential novel therapeutic approach to protect the exocrine pancreatic function in alcoholic pancreatitis and prevent the development of cholestasis in alcoholic hepatitis.

Rising Trend in Waitlisting for Alcoholic Hepatitis With More Favorable Outcomes Than Other High Model for End-stage Liver Disease in the Current Era.

In the appropriate candidate, liver transplantation (LT) is a viable treatment for alcoholic hepatitis (AH). We compared the waitlisting trends and outcomes of AH patients in the context of others with high Model for End-stage Liver Disease (MELD) score. LT listings for AH between January 1, 2008, and June 12, 2020 were identified in the United Network for Organ Sharing database. Temporal trends in listings for AH were assessed. Covariate adjusted competing risks models evaluated waitlist mortality and LT rates between AH candidates and others with listing native MELD ≥30. Between 2008 and 2019, waitlist additions for AH increased 6.5-fold. Waiting time for AH candidates was short (median 10 d). Delisting for clinical improvement was infrequent in AH, albeit higher than MELD ≥30 patients (3.3% versus 0.8%; P < 0.001). Among 99 centers with ≥1 AH listing, AH patients accounted for 0.2%-18.2% of all alcohol-related listings and 0.6%-25.0% of those with native listing MELD ≥30. Overall listing volume was larger at these 99 centers than the 40 with no AH listings (P < 0.001). AH candidates in 2014-2020 experienced improved waitlist survival (adjusted subhazard ratio, 0.67; 95% confidence interval, 0.52-0.86; P = 0.002) and higher transplant rates (adjusted subhazard ratio, 1.14; 95% confidence interval, 1.04-1.25; P = 0.006) versus other MELD ≥30 candidates. There has been a rising trend in waitlisting patients with AH and high MELD score. Liver disease causes influence waitlist outcomes and those of AH candidates are more favorable. Further research and allocation adjustments may be needed to ensure equitable organ allocation, based on liver disease cause, for those on the LT waitlist.

Clinical Spectrum of Alcoholic Liver Disease in Subjects Attending Outpatient Department at a Tertiary Care Hospital

Background: Alcoholic liver disease is one of the primary medical complication of chronic ethanol abuse. It encloses a wide spectrum of diseases comprising of fatty liver, alcoholic hepatitis, alcoholic cirrhosis and hepatocellular carcinoma.&#x0D; Methodology: A prospective, observational study was done at AIG hospitals in the department of hepatology for a period of 6 months. A total of 200 patient’s diagnosed clinically and biochemically with various spectrum of ALD were recruited for the study.Non-invasive prognostic scores were calculated at the time of admission and correlated with severity of disease.&#x0D; Results: Among 200 study participants, 34.8% belongs to age group of 36-45 years. All were male patients with age group ranged from 25 to 73 years. We observed the high levels of alkaline phosphatese, aspartate aminotransferase/alanine aminotransferase ratio, mean corpuscular volume, total bilirubin and international normalized ratio in Severe Alcoholic hepatitis patients. Non-invasive prognostic markers like child-turcotte-pugh score,model for end stage liver disease-sodium and maddrey’s discriminant factor were assessed for all patients.Prevalence of different stages of ethanol mediated liver disease were-12% of alcoholic fatty liver,17.5% of alcoholic hepatitis,6% of acute on chronic liver failure,32.5% of chronic liver disease,24% of decompensated chronic liver disease,6% of past decompensated chronic liver disease with current living donor liver transplant and 2% cases of hepatocellular carcinoma.&#x0D; Conclusion: Hazardous ethanol abuse is more typical in adult males especially among productive age group. The subjects were cautioned on their personal basis regarding fallout of alcoholic liver disease, and guided for latency of alcohol not immediately but definitely.

Algorithms to Identify Alcoholic Hepatitis Hospitalizations in Patients with Cirrhosis.

Alcoholic hepatitis (AH) is a clinically diagnosed syndrome with high short-term mortality for which liver transplantation may be curative. A lack of validated algorithms to identify AH hospitalizations has hindered clinical epidemiology research. This was a retrospective cohort study of patients with cirrhosis using Veterans Health Administration (VHA) data from 2008 to 2015. We randomly sampled hospitalizations based upon abnormal liver tests and administrative codes for acute hepatitis or alcohol-associated liver disease (ALD). Hospitalizations were manually adjudicated for AH per society guidelines. A priori algorithms were evaluated to compute positive predicted value (PPV) and positive likelihood ratio (LR+), and were tested in an external University of Pennsylvania Health System (UPHS) cohort. Of 368 hospitalizations, 142 (38.6%) were adjudicated as AH. AH patients were younger (55 vs. 58years, p < 0.001), less likely to have prior cirrhosis decompensation (57% vs. 73.9%, p < 0.001), and had higher AST-to-ALT ratios (median 2.9 vs. 1.9mg/dL, p < 0.001) and higher bilirubin levels (median 2.9 vs. 1.9mg/dL, p < 0.001). Algorithms combining clinical laboratory criteria (AST > 85U/L but < 450 U/L, AST-to-ALT ratio > 2, total bilirubin > 5mg/dL) and administrative coding criteria yielded the highest PPV (96.4%, 95% CI 87.7-99.6) and the highest LR+ (43.0, 95% CI 10.6-173.5). Several algorithms demonstrated 100% PPV for definite AH in the UPHS external cohort. We have identified algorithms for AH hospitalizations with excellent PPV and LR+. These high-specificity algorithms may be used in VHA datasets to identify patients with high likelihood of AH, but should not be used to study AH incidence.

OXIDATIVE DAMAGE MARKERS IN ALCOHOLIC HEPATITIS

Alcohol hepatitis (AH) is a clinical syndrome in patients with excessive and chronic alcohol consumption. Ethanol metabolism produces free radicals: reactive oxygen species (ROS) and reactive nitrogen species (RNS). This contributes to cellular damage in AH. The malondialdehyde (MDA) and carbonylated proteins are markers of oxidative damage in lipids and proteins. Our objective is to evaluate the levels of serum MDA and carbonylated proteins in AH patients with. This transversal study included 147 individuals divided into two groups: the control group (n=100), which consists of individuals with alcohol consumption ≤ 10 g/day and AUDIT score ≤ 7, and the group with AH patients (n=47). We measured the serum levels of MDA (thiobarbituric acid method) and carbonylated proteins (DNPH reaction). The statistical analysis was performed by the Windows SPSS v22 software. Data were expressed as mean values ± SEM. Comparisons were carried out by analysis of variance (ANOVA). A p-value <0.05 was considered statistically significant. The control group (CT), with a mean of 30.47±0.52 years old, had a consumption of 2.32±0.21g of alcohol daily (gOH/day) and an AUDIT score of 2.24±0.10. The AH patients, with a mean of 41.68±1.3 years old, had a consumption of 354.25±39.54 gOH/day and an AUDIT score of 30±1.45. The AST, ALT, GGT, total and indirect bilirubin serum levels were higher in AH compared to CT (p<0.0001), with a ratio of AST/ALT ≥2. The albumin serum levels were lower (p<0.0001) in AH vs. CT. The carbonylated proteins serum levels were higher in patients with AH compared to CT (p<0.0001). No differences in MDA serum levels were found between both groups. We reported greater MDA serum levels (p=0.001) in alcoholic liver cirrhosis patients when compared to the control group; and an insignificant difference in carbonylated proteins serum level between both participant groups. The alcoholic hepatitis patients have an increase in carbonylated proteins oxidative damage while there is no lipidic damage. Our results suggest that carbonylated proteins may a damage oxidative marker in the AH Mexican population. This damage may increase the risk of malnutrition, susceptibility to infections and sepsis, deficient coagulation factors production, gastrointestinal bleeding, among other complications that increase mortality. It is necessary to counteract oxidative damage to improve and complement the actual treatment in alcoholic hepatitis. Competing interests. The authors declare they have no competing interests. This work was partially financed by CONACyT SALUD-2016-272579 and PAPIIT- UNAM TA200515

Influence of Pharmacokinetics in Responders and Non-Responders of Prednisolone Based Therapy in Alcoholic Hepatitis Patients

Influence of Pharmacokinetics in Responders and Non-Responders of Prednisolone Based Therapy in Alcoholic Hepatitis Patients

DETERMINATION OF LEUKOCYTE PROFILE IN CHRONIC ALCOHOL CONSUMPTION

Chronic alcohol consumption can induce Alcoholic Liver Disease (ALD) promoting biological alterations and liver damage; however, the immunological changes usually are underestimated. It has been reported, the increase of leucocytes in alcoholic hepatitis patients 1 , but the regulation of other cell lineages has not fully evaluated. To evaluate the leukocyte profile in patients with liver damage induces by chronic and acute alcohol consumption. A Cross-sectional study. Patients were classified as follow: (1) Controls with alcohol consumption <10g/day, AUDIT <8 (CT); (2) Chronic alcohol consumption AUDIT> 8, without clinical or biochemical data of liver damage (OH, alcoholism); (3) Cirrhotic patients due to alcohol (CiOH) and (4) Patients with alcoholic hepatitis (AH). Leukocytes, lymphocytes, monocytes, neutrophils, eosinophils, and basophils were determined by hematic biometry. U-Mann Whitney was used for statistical analysis, p <0.05 was considered significant. 570 patients were included. The mean in age was: 29.5±10.8 for CT; 31±12.6 for OH; 47.6±7.7 for CiOH and 41.2±9.2 years old (p<0.001). Alcohol consumption was higher in CiOH 240 (320, 120; p<0.001) and AH 320 (480, 160; p<0.05). Albumin decreases in CiOH 2.9 (3.5, 2.2; p<0.001) and AH 1.9 (2.3, 1.6; p<0.001). On the other hand, AST, ALT and GGT increase in CiOH and AH, 49.5 (75.3, 38; p<0.001), 155 (177, 121; p<0.001) for AST, 32.5 (47.3, 24; p<0.001), 49 (75, 35.3; p<0.001) for ALT and 91.5 (191.8, 48; p<0.001), 224 (525.5, 104; p<0.001) for GGT. There was no a significant difference in eosinophils and basophils. The statistical number of leukocyte profile is described in Table 1. Data is expressed as the median with interquartile values (Q3-Q1). a) Differences between Alcoholism and Controls; b) Cirrhosis and Controls; c) Alcoholic Hepatitis and Controls; d) Alcoholism and Cirrhosis; e) Alcoholism and Alcoholic Hepatitis; f) Cirrhosis and Alcoholic Hepatitis. €p<0.05; +p<0.01; *p<0.001. During alcoholism, lymphocytes decrease, whereas neutrophils increase; this could be related to a susceptibility to recurrent respiratory and gastrointestinal infections. Lymphocytes and neutrophils decrease in CiOH; the reduction in neutrophils could be explained because the stimuli in CiOH decrease. In patients with AH, monocytes and neutrophils increase, that in a consequence increases the inflammatory state, promoting liver fibrosis and mortality. Chronic alcohol consumption, liver cirrhosis and alcoholic hepatitis promote cellular alterations, this phenomenon is more evident in AH. Our findings can be used to design novel detection strategies for the treatments of chronic and acute alcohol consumption. Conflict of interest: The authors declare that there is no conflict of interest. This work was partially financed by CONACyT SALUD-2016-272579 y PAPIIT-UNAM TA200515

COMPARISON OF CXCL-8 and IFN-γ PRODUCTION IN ACUTE AND CHRONIC STAGES OF LIVER DISEASE

Hepatitis C virus and alcoholism are the main causes of Chronic Liver Disease. (1) The increase of CXCL-8 has been correlated with mortality in alcoholic hepatitis (AH), while in Chronic Hepatitis C (CHC) with the disease severity. (2) The IFN-γ has an anti-viral and anti-fibrotic function. There are few comparative studies in patients regarding the production of these mediators and their possible implication in liver damage. The objective is to evaluate the concentrations of CXCL-8 and IFN-γ in the serum of AH, alcoholic cirrhosis (CiOH) and CHC patients. A multicenter cross-sectional study was carried out. Four participant groups were included: AH, CiOH, CHC and control group (CT). For the quantification of CXCL-8 and IFN-γ a multiple suspension array assay (Milliplex®-MERCK©) was used. Statistical analysis was performed by the SPSS V.22 software using Mann-Whitney-U-test. A p-value<0.05 was considered statistically significant. Values were expressed as median (Q3, Q1). 110 individuals were included: AH (10), CiOH (25), CHC (25) and CT (50). In CXCL-8 quantification, significant differences were detected in AH vs. CT, CiOH vs. CT, CHC vs. CT, CiOH vs. AH and CHC vs. AH (p≤0.001). While in IFN-γ, the differences were detected in AH vs. CT, CiOH vs. CT, CHC vs. CT, CiOH vs. AH and CHC vs. AH (p≤0.001). Differences were detected in both molecules when comparing the chronic stages (CiOH and CHC) with the acute stage (HA), while no differences were found when comparing both chronic stages. The highest CXCL-8 concentration corresponds to AH, reflecting its importance in prognosis and mortality. (2) The increase of IFN-γ in CiOH and CHC may have a role in the regulation of fibrogenesis because of its anti-fibrotic function. The deregulation of mediators such as IL-8 and IFN-γ promotes systemic inflammation in both acute and chronic stages, being greater in AH. This may indicate an association with the susceptibility to persistent respiratory and gastrointestinal diseases. The authors declare that there is no conflict of interest. This work was partially financed by CONACyT SALUD-2016-272579 (GRG) and PAPIIT- UNAM TA200515 (GRG).

Co-relation between clinical parameters and histological parameters with outcome in severe alcoholic hepatitis patients

Background and Aim: Alcoholic hepatitis (AH) is characterized by recent onset of jaundice, with or without other signs of liver decompensation (ascites and/or encephalopathy), in patients with chronic alcohol abuse. Severe AH is defined by a modified Madey’s discriminant function of 32 or higher and has poor prognosis. Alcoholic hepatitis histologic score is a new approach which includes four histologic features (bridging fibrosis or cirrhosis, hepatocellular or canalicular/ductular bilirubinostasis, severe polymorphonuclear infiltration and megamitochonia) that allows patient classification into three groups with different risks of death within 90 days.In the present study,we aimed to correlate histologic and clinical parameters with outcomes in severe alcoholic hepatitis patients, which in turn will predict the prognosis of the patients.

Bile Acid-Mediated Activation of Brown Fat Protects From Alcohol-Induced Steatosis and Liver Injury in Mice.

Background & AimsAlcohol-associated liver disease (AALD) is one of the most common causes of liver injury and failure. Limited knowledge of the mechanisms underlying AALD impedes the development of efficacious therapies. Bile acid (BA) signaling was shown to participate in the progression of AALD. However, the mechanisms remain poorly understood.MethodsC57BL/6J wild-type (WT), Takeda G-protein–coupled bile acid receptor 5 (TGR5) knockout (KO) and brown adipose tissue (BAT)-specific TGR5 knockdown mice were subjected to ethanol feeding–induced AALD. Liver samples from alcoholic hepatitis patients were used to examine the BA circulation signaling. Human Embryonic Kidney Cells 293 were used for the TGR5 reporter assay. 23(S)-methyl-lithocholic acid was used as a molecular tool to confirm the regulatory functions of BAT in the AALD mouse model.ResultsEthanol feeding increased the expression of the thermogenesis genes downstream of TGR5 in BAT of WT, but not TGR5 KO, mice. TGR5 deficiency significantly blocked BAT activity and energy expenditure in mice after ethanol feeding. Alcohol increased serum BA levels in mice and human beings through altering BA transportation, and the altered BAs activated TGR5 signaling to regulate metabolism. Compared with ethanol-fed WT mice, ethanol-fed TGR5 KO mice showed less free fatty acid (FFA) β-oxidation in BAT, leading to higher levels of FFA in the circulation, increased liver uptake of FFAs, and exacerbated AALD. BAT-specific TGR5 knockdown mice showed similar results with TGR5 KO mice in AALD. Agonist treatment significantly activated TGR5 signaling in BAT, increased thermogenesis, reduced serum FFA level, and ameliorated hepatic steatosis and injury in AALD mice, while these effects were lost in TGR5 KO mice.ConclusionsBA signaling plays a protective role in AALD by enhancing BAT thermogenesis. Targeting TGR5 in BAT may be a promising approach for the treatment of AALD.

Efficacy of Phyllanthus niruri on improving liver functions in patients with alcoholic hepatitis: A double-blind randomized controlled trial.

INTRODUCTION:Phyllanthus niruri, a traditional herbal medicine, was found to be hepatoprotective as evidenced by several preclinical and clinical studies. However, to the best of our knowledge, there are no clinical trials available to date to evaluate its efficacy in alcoholic hepatitis.MATERIALS AND METHODS:The present study is a block randomized, double-blind, parallel-arm placebo-controlled trial that was designed to assess the efficacy of P. niruri on the liver and renal function parameters, total oxidant and antioxidant levels in alcoholic hepatitis patients in comparison to placebo over a 4-week period. Patients were screened by CAGE questionnaire, and those with a confirmed diagnosis of mild–moderate alcoholic hepatitis based on laboratory findings and Maddrey's discriminant function score were randomly allocated to treatment and placebo arms. Clinical assessments were done at baseline, 2 weeks, and 4 weeks. A total of 454 patients were screened and 100 eligible patients were recruited for the study, and 71 were analyzed using the modified intention-to-treat approach.RESULTS:Serum levels of liver and renal function parameters failed to demonstrate significant improvement with P. niruri. However, there was a statistically significant increase in the level of total antioxidants with P. niruri (P = 0.034) with an additional appetite stimulant activity (P = 0.03) in 4 weeks.CONCLUSION:A 4-week administration of P. niruri in mild–moderate alcoholic hepatitis patients showed an improvement in the total antioxidant levels with an appetite stimulant activity compared to a placebo.

Impact of the COVID-19 Pandemic on Hospitalizations for Alcoholic Hepatitis or Cirrhosis in Alberta, Canada

Background & AimsCoronavirus disease 2019 (COVID-19) pandemic lockdown and restrictions had significant disruption to patient care. We aimed to evaluate the impact of COVID-19 restrictions on hospitalizations of patients with alcoholic and nonalcoholic cirrhosis as well as alcoholic hepatitis (AH) in Alberta, Canada.MethodsWe used validated International Classification of Diseases (ICD-9 and ICD-10) coding algorithms to identify liver-related hospitalizations for nonalcoholic cirrhosis, alcoholic cirrhosis, and AH in the province of Alberta between March 2018 and September 2020. We used the provincial inpatient discharge and laboratory databases to identify our cohorts. We used elevated alanine aminotransferase or aspartate aminotransferase, elevated international normalized ratio, or bilirubin to identify AH patients. We compared COVID-19 restrictions (April–September 2020) with prior study periods. Joinpoint regression was used to evaluate the temporal trends among the 3 cohorts.ResultsWe identified 2916 hospitalizations for nonalcoholic cirrhosis, 2318 hospitalizations for alcoholic cirrhosis, and 1408 AH hospitalizations during our study time. The in-hospital mortality rate was stable in relation to the pandemic for alcoholic cirrhosis and AH. However, nonalcoholic cirrhosis patients had lower in-hospital mortality rate after March 2020 (8.5% vs 11.5%; P = .033). There was a significant increase in average monthly admissions in the AH cohort (22.1/10,000 admissions during the pandemic vs 11.6/10,000 admissions before March 2020; P < .001).ConclusionsBefore and during COVID-19 monthly admission rates were stable for nonalcoholic and alcoholic cirrhosis; however, there was a significant increase in AH admissions. Because alcohol sales surged during the pandemic, future impact on alcoholic liver disease could be detrimental.

Circulating high density lipoprotein distinguishes alcoholic hepatitis from heavy drinkers and predicts 90-day outcome: lipoproteins in alcoholic hepatitis

Alcohol-associated liver disease (ALD) and alcoholic hepatitis (AH) significantly impact the liver, an organ central to the lipid and lipoprotein metabolism. To define changes in the lipid and lipoprotein profiles in subjects with alcoholic hepatitis (AH) versus heavy drinkers with normal liver function and to determine the association of the AH-mediated lipoprotein phenotype with AH severity and outcomes. AH cases (n=196) and a heavy drinker control group (n=169) were identified in a multicenter, prospective cohort. The relationships between lipid panels and lipoprotein profiles among AH and heavy drinkers were interrogated using three common measurements: the conventional lipid panel, extended lipid panel by NMR, and NMR-based direct lipoprotein profiling. Predictive values for AH severity and mortality were determined using Harrell's C-Index. Lipid and lipoprotein profiles were significantly different in AH compared to heavy drinkers. Among them, high density lipoprotein (HDL) particle concentration exhibited the most significant reduction in AH compared to heavy drinkers (5.3 ± 3.4 vs 22.3 ± 5.4 μmol/L, p < 0.001). Within AH patients, HDL particle concentration was inversely associated with Maddrey's Discriminant Function (DF) (p < 0.001), and independently associated with mortality at both 90 and 365 days even after adjustment for DF (p=0.02, p=0.05 respectively). HDL particle concentration less than 3.5 μmol/L and total cholesterol ≤ 96 mg/dL identified AH patients with higher 90-day mortality. Lipid and lipoprotein profiles are profoundly altered in AH and can help in prognosticating disease severity and mortality.