Increased fecal ethanol and enriched ethanol-producing gut bacteria Limosilactobacillus fermentum, Enterocloster bolteae, Mediterraneibacter gnavus and Streptococcus mutans in nonalcoholic steatohepatitis.

Background: Non-alcoholic steatohepatitis (NASH) has become a major public health issue as one of the leading causes of liver disease and transplantation worldwide. The instrumental role of the gut microbiota is emerging but still under investigation. Endogenous ethanol (EtOH) production by gut bacteria and yeasts is an emerging putative mechanism. Microbial metagenomics and culture studies targeting enterobacteria or yeasts have been reported, but no culturomics studies have been conducted so far. Aim: To assess fecal EtOH and other biochemical parameters, characterize NASH-associated dysbiosis and identify EtOH-producing gut microbes associated with the disease, fecal samples from 41 NASH patients and 24 controls were analyzed. High-performance liquid chromatography (HPLC) was used for EtOH, glucose, total proteins, triglyceride and total cholesterol. Viable bacteria were assessed with microbial culturomics. Microbial genetic material was assessed using 16S metagenomics targeting the hypervariable V3V4 region. Results: Fecal EtOH and glucose was elevated in the stools of NASH patients (p < 0.05) but not triglyceride, total cholesterol or proteins. In culturomics, EtOH-producing Enterocloster bolteae and Limosilactobacillus fermentum were enriched in NASH. V3V4 16S rRNA amplicon sequencing confirmed the enrichment in EtOH-producing bacteria including L. fermentum, Mediterraneibacter gnavus and Streptococcus mutans, species previously associated with NASH and other dysbiosis-associated diseases. Strikingly, E. bolteae was identified only by culturomics. The well-known Lacticaseibacillus casei was identified in controls but never isolated in patients with NASH (p < 0.05). Conclusion: Elevated fecal EtOH and glucose is a feature of NASH. Several different EtOH-producing gut bacteria may play an instrumental role in the disease. Culturomics and metagenomics, two complementary methods, will be critical to identify EtOH-producing bacteria for future diagnostic markers and therapeutic targets for NASH. Suppression of EtOH-producing gut microbes and L. casei administration are options to be tested in NASH treatment.

Development of bile acid activated receptors hybrid molecules for the treatment of inflammatory and metabolic disorders.

The farnesoid-x-receptor (FXR) and the G protein bile acid activated receptor (GPBAR)1 are two bile acid activated receptors highly expressed in entero-hepatic, immune, adipose and cardiovascular tissues. FXR and GPBAR1 are clinically validated targets in the treatment of metabolic disorders and FXR agonists are currently trialled in patients with non-alcoholic steato-hepatitis (NASH). Results of these trials, however, have raised concerns over safety and efficacy of selective FXR ligands suggesting that the development of novel agent designed to impact on multiple targets might have utility in the treatment of complex, multigenic, disorders. Harnessing on FXR and GPBAR1 agonists, several novel hybrid molecules have been developed, including dual FXR and GPBAR1 agonists and antagonists, while exploiting the flexibility of FXR agonists toward other nuclear receptors, dual FXR and peroxisome proliferators-activated receptors (PPARs) and liver-X-receptors (LXRs) and Pregnane-X-receptor (PXR) agonists have been reported. In addition, modifications of FXR agonists has led to the discovery of dual FXR agonists and fatty acid binding protein (FABP)1 and Leukotriene B4 hydrolase (LTB4H) inhibitors. The GPBAR1 binding site has also proven flexible to accommodate hybrid molecules functioning as GPBAR1 agonist and cysteinyl leukotriene receptor (CYSLTR)1 antagonists, as well as dual GPBAR1 agonists and retinoid-related orphan receptor (ROR)γt antagonists, dual GPBAR1 agonist and LXR antagonists and dual GPBAR1 agonists endowed with inhibitory activity on dipeptidyl peptidase 4 (DPP4). In this review we have revised the current landscape of FXR and GPBAR1 based hybrid agents focusing on their utility in the treatment of metabolic associated liver disorders.

Psoas Muscle Index: A Simple and Reliable Method of Sarcopenia Assessment on Computed Tomography Scan in Chronic Liver Disease and its Impact on Mortality.

Objectives: Psoas muscle parameters have been proposed as a simple and quick method for sarcopenia assessment. The aim of this study was to assess sarcopenia in cirrhotics by psoas muscle on computed tomography and its impact on mortality. Methods: One hundred and fifty patients (75 cirrhotics, 75 subjects) were assessed for psoas muscle on CT scan. Psoas muscle index (PMI) was calculated as 'total psoas muscle area/(height of subject)2'. Cut off values for sarcopenia diagnosis were derived from local subjects (n = 75) who did not have cirrhosis/other causes of sarcopenia. Results: Sarcopenia assessed by PMI was seen in 36% (n = 27) of the cirrhotics. Sarcopenia was significantly higher in patients having Child-Pugh C. Ascites, hepatic encephalopathy (HE) and gastro-intestinal bleed were seen in 48%, 18.7% and 24%, respectively. Sarcopenia was significantly associated with ascites and HE (P < 0.05). Out of the 75 cases, 53 cases completed the follow-up period of 1 year. Among the 20 cases who had sarcopenia, 35% (n = 7) succumbed to liver-related illness during 1 year follow-up, and out of the 33 cases without sarcopenia, only 6% (n = 2) died. The association of sarcopenia and 1 year mortality was statistically significant (P = 0.01). Conclusions: The PMI, a simple method for sarcopenia assessment detected sarcopenia in 36% of cirrhotics. Patients with sarcopenia had a significantly higher 1 year mortality rate and appropriate prognostication of such patients is needed.

Non-invasive diagnosis of severe alcoholic hepatitis: Usefulness of cross-sectional imaging.

PURPOSE: To describe the computed tomography (CT) and magnetic resonance imaging (MRI) features of severe acute alcoholic hepatitis (SAAH) and estimate the capabilities of CT and MRI in differentiating SAAH from alcoholic cirrhosis and non-alcoholic steato-hepatitis (NASH) cirrhosis. MATERIALS AND METHODS: Fifty patients with pathologically proven SAAH (SAAH group) who underwent CT or MRI examinations up to 30 days before or 15 days after liver biopsy between January 2008 and June 2018 were retrospectively included. There were 31 men and 29 women with a mean age of 52±9 (SD) years (range: 33-67 years). Imaging features of the SAAH group were compared to those obtained in two control groups including 62 patients with alcoholic cirrhosis without acute alcoholic hepatitis (control group 1) and 19 patients with NASH cirrhosis (control group 2) by two independent radiologists blinded to the final diagnosis. Univariate analyses were performed to compare imaging characteristics between the three groups, followed by diagnostic performance analysis for the diagnosis of SAAH of the main CT features. RESULTS: Heterogeneous steatosis was significantly more frequent in SAAH group than in the control groups (41/50; 82% vs. 7/62; 10% and 1/19; 5% in control groups 1 and 2, respectively for reader 1 and 34/50; 68% vs. 8/62; 13% and 1/19; 5% in control groups 1 and 2, respectively for reader 2; both P=0.01). Transient perfusion disorders were more frequent in SAAH group than in the control groups (35/50; 70% vs. 12/62; 21% and 5/19; 26% in control groups 1 and 2, respectively for reader 1 and 39/50; 78% vs. 14/62; 23% and 13/19; 6% in control groups 1 and 2, respectively for reader 2; both P=0.01). The combination of these two findings yielded 100% specificity (45/45; 95% CI: 92-100) for readers 1 and 2 for the diagnosis of SAAH vs. alcoholic cirrhosis and NASH cirrhosis. CONCLUSION: The imaging features of SAAH are specific and mainly associate transient heterogeneous steatosis and liver perfusion disorders. CT/MRI may be useful to differentiate SAAH from alcoholic cirrhosis and NASH cirrhosis.

Multiple target tissue effects of GLP-1 analogues on non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

Accumulating experimental and clinical evidences over the last decade indicate that GLP-1 analogues have a series of central nervous system and peripheral target tissues actions which are able to significantly influence the liver metabolism. GLP-1 analogues pleiotropic effects proved to be efficacious in T2DM subjects not only reducing liver steatosis and ameliorating NAFLD and NASH, but also in lowering plasma glucose and liver inflammation, improving cardiac function and protecting from kidney dysfunction. While the experimental and clinical data are robust, the precise mechanisms of action potentially involved in these protective multi-target effects need further investigation. Here we present a systematic review of the most recent literature data on the multi-target effects of GLP-1 analogues on the liver, on adipose and muscular tissue and on the nervous system, all capable of influencing significant aspects of the fatty liver disease physiopathology. From this analysis, we can conclude that the multi-target beneficial action of the GLP-1 analogues could explain the positive effects observed in animal and human models on progression of NAFLD to NASH.

Gut-Liver Axis Derangement in Non-Alcoholic Fatty Liver Disease.

Non-alcoholic fatty liver disease (NAFLD) is the most frequent type of chronic liver disease in the pediatric age group, paralleling an obesity pandemic. A "multiple-hit" hypothesis has been invoked to explain its pathogenesis. The "first hit" is liver lipid accumulation in obese children with insulin resistance. In the absence of significant lifestyle modifications leading to weight loss and increased physical activity, other factors may act as "second hits" implicated in liver damage progression leading to more severe forms of inflammation and hepatic fibrosis. In this regard, the gut-liver axis (GLA) seems to play a central role. Principal players are the gut microbiota, its bacterial products, and the intestinal barrier. A derangement of GLA (namely, dysbiosis and altered intestinal permeability) may promote bacteria/bacterial product translocation into portal circulation, activation of inflammation via toll-like receptors signaling in hepatocytes, and progression from simple steatosis to non-alcoholic steato-hepatitis (NASH). Among other factors a relevant role has been attributed to the farnesoid X receptor, a nuclear transcriptional factor activated from bile acids chemically modified by gut microbiota (GM) enzymes. The individuation and elucidation of GLA derangement in NAFLD pathomechanisms is of interest at all ages and especially in pediatrics to identify new therapeutic approaches in patients recalcitrant to lifestyle changes. Specific targeting of gut microbiota via pre-/probiotic supplementation, feces transplantation, and farnesoid X receptor modulation appear promising.

Routine Liver Biopsy During Bariatric Surgery: an Analysis of Evidence Base.

Non-alcoholic fatty liver disease and non-alcoholic steato-hepatitis are common in patients undergoing bariatric surgery. Non-alcoholic steato-hepatitis can progress to cirrhosis of the liver and hepatocellular carcinoma. Non-invasive methods of diagnosing non-alcoholic steato-hepatitis are not as accurate as liver biopsy, and bariatric surgery presents a unique opportunity to carry out a simultaneous liver biopsy. Routine liver biopsy can help early and accurate diagnosis of obesity-associated liver conditions. This has led some surgeons to argue for routine liver biopsy at the time of bariatric surgery. However, most bariatric surgeons remain unconvinced and liver biopsy is currently not routine practice with bariatric surgery. This review examines published scientific literature to ascertain the usefulness of routine liver biopsy at the time of bariatric surgery.

Reprint of: Nutrition in the Management of Cirrhosis and its Neurological Complications.

Malnutrition is a common feature of chronic liver diseases that is often associated with a poor prognosis including worsening of clinical outcome, neuropsychiatric complications as well as outcome following liver transplantation. Nutritional assessment in patients with cirrhosis is challenging owing to confounding factors related to liver failure. The objectives of nutritional intervention in cirrhotic patients are the support of liver regeneration, the prevention or correction of specific nutritional deficiencies and the prevention and/or treatment of the complications of liver disease per se and of liver transplantation. Nutritional recommendations target the optimal supply of adequate substrates related to requirements linked to energy, protein, carbohydrates, lipids, vitamins and minerals. Some issues relating to malnutrition in chronic liver disease remain to be addressed including the development of an appropriate well-validated nutritional assessment tool, the identification of mechanistic targets or therapy for sarcopenia, the development of nutritional recommendations for obese cirrhotic patients and liver-transplant recipients and the elucidation of the roles of vitamin A hepatotoxicity, as well as the impact of deficiencies in riboflavin and zinc on clinical outcomes. Early identification and treatment of malnutrition in chronic liver disease has the potential to lead to better disease outcome as well as prevention of the complications of chronic liver disease and improved transplant outcomes.

Autophagy modulation as a potential therapeutic target for liver diseases.

Autophagy is a critical intracellular pathway which maintains cellular function by lysosomal degradation of damaged proteins and organelles besides elimination of invading pathogens. Its primary function is to prevent cell death. Autophagy has diverse physiological functions namely; starvation adaptation, prevention of tumorigenesis, energy homeostasis, intracellular quality control and degradation of abnormal intracellular protein aggregates. Understanding the molecular mechanisms of autophagy has given key insights into the pathogenesis of various diseases like Non Alcoholic Steato-Hepatitis, Hepatitis B and C infections, Alpha-1 antitrypsin deficiency and hepatocellular carcinoma. Pharmacological modulation of autophagy may have a therapeutic potential in management of these liver diseases.

Nutrition in the management of cirrhosis and its neurological complications.

Malnutrition is a common feature of chronic liver diseases that is often associated with a poor prognosis including worsening of clinical outcome, neuropsychiatric complications as well as outcome following liver transplantation. Nutritional assessment in patients with cirrhosis is challenging owing to confounding factors related to liver failure. The objectives of nutritional intervention in cirrhotic patients are the support of liver regeneration, the prevention or correction of specific nutritional deficiencies and the prevention and/or treatment of the complications of liver disease per se and of liver transplantation. Nutritional recommendations target the optimal supply of adequate substrates related to requirements linked to energy, protein, carbohydrates, lipids, vitamins and minerals. Some issues relating to malnutrition in chronic liver disease remain to be addressed including the development of an appropriate well-validated nutritional assessment tool, the identification of mechanistic targets or therapy for sarcopenia, the development of nutritional recommendations for obese cirrhotic patients and liver-transplant recipients and the elucidation of the roles of vitamin A hepatotoxicity, as well as the impact of deficiencies in riboflavin and zinc on clinical outcomes. Early identification and treatment of malnutrition in chronic liver disease has the potential to lead to better disease outcome as well as prevention of the complications of chronic liver disease and improved transplant outcomes.

Relación entre el diagnóstico morfológico de EHNA (esteatohepatitis no alcohólica) y pruebas de función hepática en un grupo de pacientes con obesidad extrema / Relationship between morphological diagnosis of NASH (Non-Alcoholic SteatoHepatitis) and liver function test in a group of patients with morbid obesity. Cases study

The non-alcoholic steato-hepatitis (NASH) is a common disorder in obese, type 2 diabetics, female and patients with dislipidaemia. Hepatic biochemical test are abnormal. Despite the lack of its own morphological characteristics, NASH can be differentiated from other pathologies, the gold standard for diagnosis is liver biopsy. Material and methods: We designed a retrolective, comparative, observational and cross-sectional study. Thirty-five obese subjects (11 men and 24 women) who underwent to bariatric surgery and liver biopsy were included. Data were taken from clinical files, such as anthropometric and biochemical test. Those who had clinical history of related alcohol ingestión or liver damage related to drugs were excluded. A experimented pathologist classified the biopsies according to Brunt classification. Liver slides were classified according to 1) presence of NASH; 2) Inflammation and 3) Fibrosis. Differences between groups were analysed by ANOVA and Spearman correlation. Results: We found differences between women (w) and men (m) for height (m: 1.71 ± 0.9 vs. w:1.60 ± 0.09m); weight (m: 172.5 ± 39.1 vs. w: 126.9 ± 24.1kg) and BMI (m:58.2 ± 9.8 vs. w:49.8 ± 9) , but not for NASH frequency. Nevertheless subjects with NASH (n = 29, 82.8%) were older than those without NASH (38.3 ± 9.6 vs. 29.5 ± 5.2) and had higher aminotrasferases serum levels (AST: 33.1 ± 19.2 vs. 23.7 ± 6.3 UL/L; ALT: 36.5 ± 19.8 vs. 20.3 ± 7.6UL/L). NASH patíents and those with higher grade of histological inflammation had increment of transaminases and albumin levels. Fibrosis showed correlation only with AST (p = 0.020) and ALT (p = 0.002). Conclusion.The NASH frequency in patients who underwent to bariatric surgery for weight reduction is very high (82.8%) and exists correlation among liver test and histological findings but not with clinical because the clinical diagnosis is complicated.

La esteatohepatitis no alcohólica (EHNA) es una alteración hepática frecuente en obesos, diabéticos tipo 2, mujeres y personas con dislipidemia. Clínicamente se acompaña de alteraciones en las pruebas de función hepática (PFH), y aunque carece de características morfológicas distintivas, puede ser razonablemente diferenciada de otras entidades, el método diagnóstico por excelencia es la biopsia hepática. Material y métodos. Se diseñó un estudio retrolectivo, comparativo, observacional y transversal en el que se incluyeron 35 pacientes obesos (11 hombres y 24 mujeres) sometidos a tratamiento quirárgico para reducción de peso, con biopsia hepática en el periodo transoperatorio. Se obtuvieron, del expediente clínico, datos antropométricos y de laboratorio. Se excluyeron los pacientes con antecedentes de ingestión de alcohol y medicamentos asociados a la presencia de EHNA. Se recabaron muestras de las biopsias hepáticas que fueron analizadas por un patólogo experimentado, empleando la clasificación de Brunt para estratificación de EHNA. Los datos se clasificaron de acuerdo con: 1) Presencia de EHNA, 2) Grado de inflamación, 3) Presencia de fibrosis. Las diferencias entre los grupos fueron analizadas con Krusskal Wallis y correlación de Spearman. Resultados. Se encontró diferencias entre hombres (H) y mujeres (M) en estatura: (H:1.71 ± 0.9. vs. M:1.60 ±0.09 m); peso (H: 172.5 ± 39.1 vs. M:126.9 ± 24.1 kg) e índice de masa corporal (H:58.2 ± 9.8 vs. M: 49.8 ± 9); no hubo diferencias en la frecuencia de EHNA por género. Los sujetos con diagnóstico morfológico de EHNA (n = 29, 82.8%) mostraron una edad promedio mayor que el grupo sin EHNA (38.3 ± 9.6 vs. 29.5 ± 5.2, respectivamente) del mismo modo, la concentración de transaminasas fue mayor para el grupo con EHNA (AST: 33.1 ± 19.2 vs. 23.7 ± 6.3 UL/ L; ALT: 36.5 ± 19.8 vs. 20.3 ± 7.6 UL/L). Los pacientes con EHNA y con mayor grado de inflamación histológica mostraron mayor elevación de transaminasas y albámina. La presencia de fibrosis correlacionó con la elevación de aspartato aminotransferasa (AST p = 0.020) y alanino aminotransferasa (ALT p = 0.002). Conclusión. Este estudio demuestra que la frecuencia de EHNA en pacientes obesos sometidos a cirugía para reducción de peso en la clínica de obesidad del Instituto es alta (82.8%) y que existe una buena correlación entre las pruebas de función hepática y las alteraciones morfológicas; sin embargo, las anteriores no correlacionan con las manifestaciones clínicas por lo que el diagnóstico clínico temprano es difícil.

Effect of total flavonoids of Chinese hawthorn leaf on expression of NF-?B and its inhibitor in rat liver with non-alcoholic steato-hepatitis / 中华中医药杂志

Objective:To study the effect of NF-?B and its inhibitor in rats liver during the development of non-alcoholic steato-hepatitis and observe the effect of total flavonoids of Chinese hawthorn leaf on expressions of NF-?B and its inhibitor,and explore the prevention and treatment mechanism of non-alcoholic steato-hepatitis. Methods:Non-alcoholic steato-hepatitis SD rats model were established by administering a high-fat diet for 12 weeks and were treated with total flavonoids of Chinese hawthorn leaf at dosages of 250mg/kg body weight /d,125 mg/kg body weight /d and with Essentiale at a dosage of 195.4mg /kg body weight /d. Pathological changes of liver tissues were observed with HE,the serum MDA,SOD and TNF-? were measured,the protein and mRNA expressions of NF-?B and I?B in rats liver were detected with RT-PCR and immunohistochemical method. Results:There were severe steatosis inflammatory cell infiltration and necrotic foci in non-alcoholic steato-hepatitis rat. Compared with the control group,contents of the serum MDA and TNF-? in non-alcoholic steato-hepatitis group obviously increased,the activity of SOD decreased,the mRNA and protein expressions of NF-?B P65 and I?B? in liver obviously strengthened. There was a positive relativity of the protein expression of NF-?B p65,I?B? with the contents of serum TNF-?,MDA,but a negative relativity with the activity of serum SOD. Conclusion:Total flavonoids of Chinese hawthorn leaf can obviously lower lipid peroxidation in rats,lower the harm of cytokine to hepatocytes,and regulate the expression of protein and mRNA of NF-?B and I?B. It maybe the important mechanism of total flavonoids of Chinese hawthorn leaf for preventing and treating the development of non-alcoholic steato-hepatitis.