Fatty Liver Stage Research Articles

Disrupted Post-Transcriptional Regulation of Gene Expression as a Hallmark of Fatty Liver Progression

It is known that both transcriptional and post-transcriptional mechanisms control messenger RNA (mRNA) levels. Compared to transcriptional regulations, our understanding of how post-transcriptional regulations adapt during fatty liver progression at the whole-transcriptome level is unclear. While traditional RNA-seq analysis uses only reads mapped to exons to determine gene expression, recent studies support the idea that intron-mapped reads can be reliably used to estimate gene transcription. In this study, we analyzed differential gene expression at both the exon and intron levels using two liver RNA-seq datasets from mice that were fed a high-fat diet for seven weeks (mild fatty liver) or thirty weeks (severe fatty liver). We found that the correlation between gene transcription and mature mRNA levels was much lower in mice with mild fatty liver as compared with mice with severe fatty liver. This result indicates broad post-transcriptional regulations for early fatty liver and such regulations are compromised for severe fatty liver. Specifically, gene ontology analysis revealed that genes involved in synapse organization and cell adhesion were transcriptionally upregulated, while their mature mRNAs were unaffected in mild fatty liver. Further characterization of post-transcriptionally suppressed genes in early fatty liver revealed that their mRNAs harbor a significantly longer 3′ UTR, one of the major features that may subject RNA transcripts to nonsense-mediated RNA decay (NMD). We further show that the expression of representative genes that were post-transcriptionally suppressed were upregulated in mice with a hepatocyte-specific defect of NMD. Finally, we provide data supporting a time-dependent decrease in NMD activity in the liver of a diet-induced metabolic-dysfunction-associated fatty liver disease mouse model. In summary, our study supports the conclusion that NMD is essential in preventing unwanted/harmful gene expression at the early stage of fatty liver and such a mechanism is lost due to decreased NMD activity in mice with severe fatty liver.

Unlocking the Therapeutic Potential of Ellagic Acid for Non-Alcoholic Fatty Liver Disease and Non-Alcoholic Steatohepatitis.

Obesity is in epidemic proportions in many parts of the world, contributing to increasing rates of non-alcoholic fatty liver disease (NAFLD). NAFLD represents a range of conditions from the initial stage of fatty liver to non-alcoholic steatohepatitis (NASH), which can progress to severe fibrosis, through to hepatocellular carcinoma. There currently exists no treatment for the long-term management of NAFLD/NASH, however, dietary interventions have been investigated for the treatment of NASH, including several polyphenolic compounds. Ellagic acid is one such polyphenolic compound. Nutraceutical food abundant in ellagic acid undergoes initial hydrolysis to free ellagic acid within the stomach and small intestine. The proposed mechanism of action of ellagic acid extends beyond its initial therapeutic potential, as it is further broken down by the gut microbiome into urolithin. Both ellagic acid and urolithin have been found to alleviate oxidative stress, inflammation, and fibrosis, which are associated with NAFLD/NASH. While progress has been made in understanding the pharmacological and biological activity of ellagic acid and its involvement in NAFLD/NASH, it has yet to be fully elucidated. Thus, the aim of this review is to summarise the currently available literature elucidating the therapeutic potential of ellagic acid and its microbial-derived metabolite urolithin in NAFLD/NASH.

Molecular insights on intracellular Wnt/β-catenin signaling in alcoholic liver disease.

Alcoholic liver disease (ALD) is one of the most common health problems worldwide, especially in developing countries caused by chronic consumption of alcohol on a daily basis. The ALD spectrum is initiated with the early stages of alcoholic fatty liver (steatosis), progressing to alcoholic steatohepatitis, followed by the later stages of fibrosis and in some cases, cirrhosis and hepatocellular carcinoma (HCC). The Wnt/β-catenin signaling required for healthy liver development, function, and regeneration is found to be aberrated in ALD, attributed to its progression. This review is to elucidate the association of Wnt/β-catenin signaling with various stages of ALD progression. Alcohol causes downregulation of Wnt/β-catenin signaling components and thereby suppressing the pathway. Reports have been published that aberrated Wnt/β-catenin signaling, especially the absence of β-catenin, results in decreased alcohol metabolism, causing steatosis followed by steatohepatitis via lipid accumulation, lipid peroxidation, liver injury, increased oxidative stress and apoptosis of hepatocytes, contributing to the advancement of ALD. Contrastingly, the progression of later stages of ALD like fibrosis and HCC depends on the increased activation of Wnt/β-catenin signaling and its components. Existing studies reveal the varied expression of Wnt/β-catenin signaling in ALD. However, the dual role of the Wnt/β-catenin pathway in earlier and later stages of ALD is not clear. Therefore, studies on the Wnt/β-catenin pathway and its components in various manifestations of ALD might provide insight in targeting the Wnt/β-catenin pathway in ALD treatment.

Comparison of deep learning algorithms for quantitative diagnosis of fatty liver using multidimensional-moments heatmaps

Non-alcoholic fatty liver disease (NAFLD) carries a high risk of progressing to non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma. Therefore, the quantitative diagnosis of NAFLD, i.e., determining hepatic fat deposition, is desired during ultrasound inspection. It is known that the deposition of fat droplets within hepatocytes increases the attenuation in ultrasound images. Because the texture of the attenuated ultrasound image becomes a homogeneous speckle, the probability density function of the echo envelopes can be approximated by a Rayleigh distribution. The diagnosis methods of NAFLD using echo-envelope statistics of ultrasound images have been reported. In this study, we focus on the distribution and variance of echo-envelope statistics and propose a new diagnosis method to analysis the characteristics of echo-envelope statistics using a deep learning algorithm (DLA). In the proposed method, first- and third-order moments as echo-envelope statistics are calculated at each pixel in the ultrasound image. Then, the heatmap is formed from the distributions of both moments in each region of interest within the liver. The heatmaps are classified into the stages of fatty liver using DLA such as a convolutional neural network or a vision transformer. In this presentation, a comparative study on the accuracies by various DLA are reported.

The Role of ElastPQ in Assessing Liver Stiffness for Non-Alcoholic Fatty Liver Disease in Patients Treated with Atypical Antipsychotic Drugs.

To evaluate the role of elastography point quantification (ElastPQ) for the quantitative assessment of stiffness in the fatty liver disease in mental disorder patients and to provide a noninvasive detection method for non-alcoholic fatty liver (NAFLD) caused by atypical antipsychotics drugs (AAPDs). A total number of 168 mental disorder patients treated with AAPDs and 58 healthy volunteers were enrolled in this study. All the subjects underwent ultrasound and ElastPQ tests. The basic data of the patients were analyzed. BMI, liver function, and the value of ElastPQ were considerably higher in the patient group than that in the healthy volunteers. The values of liver stiffness obtained by ElastPQ were increased gradually from 3.48(3.14-3.81) kPa in the normal liver to 8.15(6.44-9.88) in the severe fatty liver. The receiver operating characteristic (ROC) for the diagnosis of fatty liver with ElastPQ were 0.85, 0.79, 0.80, and 0.87 for the diagnosis of normal, mild, moderate, and severe steatosis, respectively, with a sensitive/specificity of 79%/76.4%, 85.7%/78.3%, 86.2%/73%, and 81.3%/82.1%, correspondingly. Moreover, ElastPQ in the olanzapine group was higher than those in the risperidone and aripiprazole groups (5.11(3.83-5.61) kPa vs 4.35(3.63-4.98) kPa, P < 0.05; 5.11(3.83-5.61) kPa vs 4.79(4.18-5.24) kPa, P < 0.05). After one-year treatment, the value of ElastPQ was 4.43(3.85-5.22) kPa, but it was 5.81(5.09-7.33) kPa in patients treated for more than three years. This value increased with treatment prolongation (P < 0.05). ElastPQ is a real-time, quantitative method for assessing the stiffness of NAFLD. The liver stiffness value could be varied in the different stages of fatty liver. Olanzapine has a considerable influence on liver stiffness. The long-term use of AAPDs can increase the stiffness value of fatty liver.

Etiopathogenesis of Yakruth Vriddi w.s.r ALD

Yakruth plays an important role in the Chayapachay and Ranjana of Rasa Dhatu. Yakruth and Pleeha are formed by the Raktadhatu. Yakruth is situated at the Dakshina bhaga and Pleeha at the Vaama bahga. Yakruth and Pleeha are the Moola Sthana of the Raktavaha Srothas. Madya is considered as alcohol it’s having qualities of Amla Usha laghu Tikshna, Sukshma, Vyavayi Ruksha Vikasi and Vishada. Madya vitiates Pitta as well as Rakta and that leads to Yakruth Vriddi. Rakta is one of the main Dushyas in Yakruth Vriddi. Diet and lifestyle are major factors that influence susceptibility to liver disorders. Alcohol use is quite common in India. According to recent data published by NHPI 74% of men and 48% of women are alcoholics. Alcohol disorders cover the spectrum of disorders beginning from the fatty liver, jaundice, hepatomegaly, ascites, and cirrhosis advanced and irreversible forms of liver injury related to the consumption of alcohol. There are three histopathological stages of alcoholic liver disease alcoholic fatty liver, alcoholic hepatitis, and, alcoholic cirrhosis. These alcoholic liver disorders’ pathogenesis can be understood in Ayurveda with the help of Kamala, Yakruth Vriddhi, Shchutha, and Ashchutha Yakruthodara. Hence an attempt is made to understand the etiopathogenesis of ALD w.s.r Yakruth Vriddi.

Hepatocytes demarcated by EphB2 contribute to the progression of nonalcoholic steatohepatitis.

Current therapeutic strategies for treating nonalcoholic steatohepatitis (NASH) have failed to alleviate liver fibrosis, which is a devastating feature leading to hepatic dysfunction. Here, we integrated single-nucleus transcriptomics and epigenomics to characterize all major liver cell types during NASH development in mice and humans. The bifurcation of hepatocyte trajectory with NASH progression was conserved between mice and humans. At the nonalcoholic fatty liver (NAFL) stage, hepatocytes exhibited metabolic adaptation, whereas at the NASH stage, a subset of hepatocytes was enriched for the signatures of cell adhesion and migration, which were mainly demarcated by receptor tyrosine kinase ephrin type B receptor 2 (EphB2). EphB2, acting as a downstream effector of Notch signaling in hepatocytes, was sufficient to induce cell-autonomous inflammation. Knockdown of Ephb2 in hepatocytes ameliorated inflammation and fibrosis in a mouse model of NASH. Thus, EphB2-expressing hepatocytes contribute to NASH progression and may serve as a potential therapeutic target.

Effect of Pioglitazone on Nonalcoholic Fatty Liver Disease in Morbid Obese Patients; a Randomized Controlled Trial.

Nonalcoholic fatty liver disease (NAFLD) is a common obesity-related disease. In this study, we aimed to investigate the effects of pioglitazone on NAFLD in morbid obese patients. This is a randomized controlled trial study that was performed in 2020-2021 on 44 patients who had grade 3 NAFLD. At the beginning of the study, we collected the following data: age, gender, body mass index (BMI), fasting blood glucose (FBS), lipid profile, aspartate aminotransferase, alanine aminotransferase (ALT), and the total size and volume of the liver and the left lobe of the liver. Patients in the control group were given a special diet. For patients in the treatment group, pioglitazone 15 mg tablets were administered twice daily for 4 months. At the beginning of the study, all patients in both groups had grade 3 of NAFLD. After the treatments, 50% of the pioglitazone group had grade 1 NAFLD, and 50% of other patients had grade 2 that showed significant improvements in patients (P < 0.001). We also found significant improvements in the following items in the intervention group: liver size (P < 0.001), size of the left liver lobe (P < 0.001), FBS (P = 0.036), ALT (P = 0.011), and BMI (P < 0.001). No significant improvements were found in the control group (P > 0.05). The use of pioglitazone for 4 months resulted in improvements in fatty liver stage, liver size, BMI, FBS, and lipid profile. These data show the effectiveness of pioglitazone in NAFLD.

IMPACT OF LIVER DISEASES ON PREGNANCY AND PUERPERIUM STAGE: A SYSTEMATIC REVIEW

Purpose: Changes in the liver function tests (LFTs) are a serious complication during pregnancy and require proper analysis in order to avoid the risk factors in the diagnosis. Pregnancy-specific diseases are the most common reason for abnormal liver function tests while pregnant, especially in the third trimester.&#x0D; Materials and Methods: In this review article, pregnancy-related liver disease articles indexed in various databases were used. The collection of articles was evaluated by using keywords including liver diseases, pregnancy, eclampsia, fatty liver, liver enzymes and puerperium stage.&#x0D; Results: Liver cirrhosis, autoimmune infections, primary biliary redness, Wilson’s unwellness are few examples of pre-existing liver diseases which are exceptionally rare because pregnant women are often young and healthy. Intrahepatic cholestasis of pregnancy, the HELLP syndrome (haemolysis, increased liver enzymes, low platelets), eclampsia and acute fatty liver are all liver conditions that are specific to pregnancy. These abnormalities may cause foetal distress, serious liver damage, and even hepatic failure; as a result, rapid diagnosis and treatment are required. Serum transaminases, alkaline phosphatase, bilirubin etc. are frequently used diagnostic procedures.&#x0D; Conclusion: This review mainly focuses on the pregnancy-associated liver disorders with several biochemical tests, and their pathophysiology to interpret the abnormal condition of the liver during pregnancy. It also highlights the current scenario of Maternal Mortality Ratio in India. The effects of alcohol on unsuccessful pregnancies in females are also discussed. In this, so far bleak situation, changes in lifestyle, early and timely collaborative care by the obstetric and medical teams can produce the best results.

ESTIMATION OF LIPID PROFILE AND GLYCEMIC STATUS AND LIVER ENZYMES, AMONG PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE

Aim: The aim of the study is mainly to evaluate the association of obesity, liver enzymes, lipid profile and glycemic status with Non -alcoholic Fatty Liver Disease (NAFLD). Methods: This cross-sectional observational study Results: This study shows that characteristics of studied population (n = 400) as control, The study population was categorized into three age groups viz. 18-35 yrs, 36-60 yrs and &gt;60 yrs. Out of the 400 subjects studied more than 50% belonged to the age group 36-60 years, 27.5% to the age group18-35 years and the remainder to the group above 60 years. The genders wise distribution of study group is presented 53% of the subjects were females and 47% subjects were males. 59.5% of the subjects were obese according to the WHOs classification of BMI for Asians. 18.5% were overweight and the remaining 22% were normal. 25% of the subjects were diabetics and 20 % were hypertensives. The association between the prevalence of NAFLD and the age categories mentioned previously, gender, BMI &amp; comorbidities was tested using Chi-Square test. There was a significant association with age, BMI, DM and HTN with a P-value of 0.003, 0.0.000, 0.000 and 0.000 respectively. No significant association was found with gender. The results of Chi-square tests performed to find out the association between NAFLD &amp; serum levels of liver enzymes. There was no significant association between NAFLD and liver enzymes. The results of the Chi-square tests performed to find out the association between NAFLD &amp; serum lipid levels. A significant association was found between the prevalence of NAFLD and total cholesterol, triglycerides, LDL-C and VLDL-C. No significant association was observed between NAFLD and HDL-C. The association between NAFLD and the glycemic status was tested. There was a significant association between plasma fasting glucose level and NAFLD. The percentage of subjects diagnosed with NAFLD in each of the various stages according to their Fibroscan results. Among the 75 NAFLD patients, 80 % of patients were in fatty liver stage, 17,3% had had progressed to NASH and 2.7% progressed to the fibrosis stage. None of the patients were found to have liver cirrhosis. There was a significant association between the HBA1c and the progression of NAFLD with a P-value of 0.001. A significant association was found to exist between the various stages of NAFLD and the plasma fasting glucose level. The P-value was 0.025. The results are summarized in different tables. Conclusion: The Prevalence of NAFLD in the study population was 18.8%.The prevalence of NAFLD was higher among males. Age, BMI and co-morbidities like diabetes and hypertension were significantly associated with NAFLD. A significant association between NAFLD and total cholesterol, triglycerides, LDL-C, VLDL-C &amp; fasting glucose was observed. There was a significant association between disease progression and glucose intolerance.

Ferroptosis resistance cooperates with cellular senescence in the overt stage of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis.

Cellular senescence and ferroptosis are the two main, fine-tuned processes in tissue damage restraint; however, they can be overactivated in pathologies such as nonalcoholic fatty liver disease/nonalcoholic steatohepatitis (NAFLD/NASH), becoming dangerous stimuli. Senescence is characterized by a decline in cell division and an abnormal release of reactive oxygen species (ROS), and ferroptosis is represented by iron deposition associated with an excessive accumulation of ROS. ROS and cellular stress pathways are also drivers of NAFLD/NASH development. The etiology of NAFLD/NASH lies in poor diets enriched in fat and sugar. This food regimen leads to liver steatosis, resulting in progressive degeneration of the organ, with a late onset of irreversible fibrosis and cirrhosis. Few studies have investigated the possible connection between senescence and ferroptosis in NAFLD/NASH progression, despite the two events sharing some molecular players. We hypothesized a possible link between senescence and ferroptosis in a NAFLD background. To thoroughly investigate this in the context of “Western-style” diet (WSD) abuse, we used an amylin-modified liver NASH mouse model. The main NASH hallmarks have been confirmed in this model, as well as an increase in apoptosis, and Ki-67 and p53 expression in the liver. Senescent beta-galactosidase-positive cells were elevated, as well as the expression of the related secretory molecules Il-6 and MMP-1. Features of DNA damage and iron-overload were found in the livers of NASH mice. Gpx4 (glutathione peroxidase 4) expression, counteracting ferroptotic cell death, was increased. Notably, an increased number of senescent cells showing overexpression of gpx4 was also found. Our data seem to suggest that senescent cells acquire a gpx4-mediated mechanism of ferroptosis resistance and thus remain in the liver, fostering the deterioration of liver fitness.

Combined Transcriptomic and Lipidomic Analysis Reveals Dysregulated Genes Expression and Lipid Metabolism Profiles in the Early Stage of Fatty Liver Disease in Rats.

Non-alcoholic fatty liver disease develops from simple steatosis to non-alcoholic steatohepatitis (NASH), which then potentially develops into liver cirrhosis. It is a serious threat to human health. Therefore, investigating the formation and development mechanism of non-alcoholic fatty liver disease (NAFLD) is of great significance. Herein, an early model of NAFLD was successfully established by feeding rats with a high-fat and choline-deficient diet. Liver tissue samples were obtained from rats in the fatty liver model group (NAFL) and normal diet control group (CON). Afterward, transcriptome and lipidomic analysis was performed. Transcriptome results revealed that 178 differentially expressed genes were detected in NAFL and CON groups. Out of which, 105 genes were up-regulated, 73 genes were downregulated, and 8 pathways were significantly enriched. A total of 982 metabolites were detected in lipidomic analysis. Out of which 474 metabolites were significantly different, 273 were up-regulated, 201 were downregulated, and 7 pathways were significantly enriched. Based on the joint analysis, 3 common enrichment pathways were found, including cholesterol metabolism and fat digestion and absorption metabolic pathways. Overall, in the early stage of NAFLD, a small number of genetic changes caused a strong response to lipid components. The strongest reflection was glycerides and glycerophospholipids. A significant increase in fatty acid uptake accompanied by cholesterol metabolism is the most prominent metabolic feature of the liver in the early stage of NAFLD. In the early stage of fatty liver, the liver had shown the characteristics of NASH.

Effect of Chronic Western Diets on Non-Alcoholic Fatty Liver of Male Mice Modifying the PPAR-γ Pathway via miR-27b-5p Regulation.

Western diets contribute to metabolic diseases. However, the effects of various diets and epigenetic mechanisms are mostly unknown. Here, six week-old C57BL/6J male and female mice were fed with a low-fat diet (LFD), high-fat diet (HFD), and high-fat high-fructose diet (HFD-HF) for 20 weeks. We determined that HFD-HF or HFD mice experienced significant metabolic dysregulation compared to the LFD. HFD-HF and HFD-fed male mice showed significantly increased body weight, liver size, and fasting glucose levels with downregulated PPARγ, SCD1, and FAS protein expression. In contrast, female mice were less affected by HFD and HFD-HF. As miR-27b contains a seed sequence in PPARγ, it was discovered that these changes are accompanied by male-specific upregulation of miR-27b-5p, which is even more pronounced in the HFD-HF group (p < 0.01 vs. LFD) compared to the HFD group (p < 0.05 vs. LFD). Other miR-27 subtypes were increased but not significantly. HFD-HF showed insignificant changes in fibrosis markers when compared to LFD. Interestingly, fat ballooning in hepatocytes was increased in HFD-fed mice compared to HFD-HF fed mice, however, the HFD-HF liver showed an increase in the number of small cells. Here, we concluded that chronic Western diet-composition administered for 20 weeks may surpass the non-alcoholic fatty liver (NAFL) stage but may be at an intermediate stage between fatty liver and fibrosis via miR-27b-5p-induced PPARγ downregulation.

Co-Culture of Human Mesenchymal Stromal Cells and Primary Mouse Hepatocytes.

Human mesenchymal stromal cells (MSC) are adult stem cells, which feature hepatotropism by supporting liver regeneration through amelioration of hepatic inflammation and lipid accumulation in a mouse model of non-alcoholic steatohepatitis (NASH), a more advanced stage of fatty liver. It remains open, how MSC impact on hepatocytic lipid metabolism. To study MSC actions on fatty liver mechanistically, we established an in vitro model of co-culture comprising MSC and isolated mouse hepatocytes at a ratio of 1:1. Lipid storage in hepatocytes was induced by the treatment with medium deficiency of methionine and choline (MCD). The protocol can be adapted for the use of other lipid storage-inducing agents such as palmitic acid and linoleic acid. This co-culture model allows to study, e.g., whether MSC act indirectly via MSC-born paracrine mechanisms or through direct physical interactions between cells beside others. The protocol allows us to detect the formation of extensions (filopodia) from MSC to contact the fatty hepatocytes or other MSC within 24h of co-culture. These structures may represent tunneling nanotubes (TNT), allowing for long-range intercellular communication.

Effects of Probiotics on Clinical, Biochemical and Radiological parameters in Obese Children with NASH /NAFLD in Combination with Dietary and Lifestyle Modification: A Randomised Control Trial

Background: Childhood obesity is a global problem with metabolic abnormalities including NAFLD/ NASH. The gut liver axis is thought to play a major role in pathogenesis and altering it could affect the pathophysiological process. Probiotics are known to alter the gut microbiota and therefore could be a therapeutic option in the management of childhood obesity related complications. A double-blind, randomised, placebo control trial was conducted to evaluate the effects of probiotics on metabolic derangements in obese children with NAFLD/ NASH. Methods and Results: Obese children from the nutrition clinic conducted by University Paediatric Unit at Lady Ridgeway Hospital, Colombo with NAFLD/ NASH were recruited. Anthropometry, body fat, metabolic derangements and ultrasound scan (USS) of liver were evaluated at the beginning and at the end of 6 months. Transient elastography (Fibroscan®) was performed in a subsample of patients. Eighty-four patients were recruited and randomised into probiotics group (43) and placebo group (41). Mean age of the probiotic group was 11.3 years (+/-1.9 SD) and placebo group was 12.1 years (+/-1.5 SD). Baseline parameters including USS stage of the liver, body fat percentage, fasting blood sugars, lipid profile, liver functions and CRP showed no statistical difference. In the Probiotic group, statistically significant reduction of BMI was noted from baseline. However, the reduction was not significant when compared with the Placebo group. There was significant reduction in triglycerides, AST, ALT, AST/ ALT and ALP in placebo group over treatment period. Even though the USS stage of fatty liver showed improvement from stage II - III to stage I in a small number in the probiotic treated group, transient elastography performed in a subsample did not demonstrate significant improvement in either group. Conclusion: Our results indicate that there does not seem to be an advantage of probiotics over lifestyle modifications in improving obesity associated metabolic derangement in children

Gene Expression and Histochemical Analyses in the Fatty Livers of Rats Fed a Histidine-Excess Diet.

Triglyceride (TG) and cholesterol accumulation are known to occur in the liver of rats fed a histidine-excess (5%) diet, but there are few studies reporting histochemical and molecular biological analyses of the rat liver. The aim of this study was to elucidate the molecular basis of this lipid-accumulation mechanism. Lipid accumulations, tissue section images, and gene expression levels were compared in the livers of rats fed a control or histidine-excess diet for 5 wk (n=8/group). Serum levels of TGs, free fatty acids, total cholesterol, high-density lipoprotein cholesterol, glucose, albumin, and the enzyme activities of aspartate aminotransferase and alanine aminotransferase were also analyzed. In the livers of rats fed a histidine-excess diet, histochemical analyses showed what appeared to be a preliminary stage of nonalcoholic fatty liver, characterized by lipid accumulation around the central vein area and minor fibrosis. However, there were no changes in serum TG or free fatty acid levels. Quantitative PCR analyses showed the up-regulation of FAT/CD36, which is related to the uptake of fatty acids into cells, and the downregulation of two apolipoprotein genes, ApoC3 and ApoE. The mRNA levels of PPARγ, LXRα, and AMPKα in the liver were also reduced by excess histidine intake. The results of this study suggest that steatosis caused by excess histidine intake may be the result of an imbalance between lipid transport from the liver and the uptake of free fatty acids into hepatocytes.

2310-PUB: Chronic Western Diets Initiate Fibrosis Progression in Nonalcoholic Fatty Liver of Male Mice Modifying the PPARγ Pathway via Mir-27b-5p Regulation

Increased fructose intake in Western diets contributes to metabolic diseases. However, the epigenetic mechanisms are mostly unknown. Here, six week-old C57BL/6J male and female mice were fed with a low-fat diet (LFD), high-fat diet (HFD), and high-fat high-fructose diet (HFD-HF) for 20 weeks. We determined that HFD-HF or HFD mice experienced significant metabolic dysregulation compared to the LFD. Metabolic changes in male mice were significantly dysregulated compared to female mice. HFD-HF and HDF male-specific responses included physiological/metabolic changes such as significant increased body weight, increased liver size, increased fasting glucose levels and downregulated PPARγ (P &amp;lt; 0.001), SCD1 (P &amp;lt; 0.05 to P &amp;lt; 0.01), and FAS (P &amp;lt; 0.05) protein expression. In contrast, female mice were less affected by HFD and HFD-HF. Interestingly, none of the groups exhibited less insulin sensitivity after 20 weeks of feeding when compared to LFD. As miR-27b contains a seed sequence in PPARγ, it was discovered that these changes are accompanied by male-specific upregulation of miR-27b-5p, which is even more pronounced in the HFD-HF group (P &amp;lt; 0.01 vs. LFD) compared to the HFD group (P &amp;lt; 0.05 vs. LFD). Other miR-27 subtypes were increased but not significantly. Fat ballooning in hepatocytes were increased in HFD fed mice compared to HFD-HF fed mice, however, the HFD-HF liver showed an increase in the number of small cells. Here, we concluded that our chronic western diet-composition administered for 20 weeks surpassed the nonalcoholic fatty liver (NAFL) stage and initiated liver fibrosis via miR-27b-5p-induced PPARγ downregulation. Disclosure J. Zhang: None. C. Powell: None. M.K. Kay: None. R. Sonkar: None. S. Meruvu: None. M. Choudhury: None. Funding Morris L. Lichtenstein, Jr. Medical Research Foundation

Quantitative Assessment of Fatty Liver using Ultrasound withNormalized Local Variance Technique.

To assess the diagnostic performance of the normalized local variance (NLV) ultrasound technique in the detection of the fatty liver using histopathology as a reference standard. We prospectively enrolled 194 consecutive patients with clinical suspicion of diffuse liver disease or history of liver transplantation. Conventional grayscale ultrasound and NLV examinations were performed and immediately followed by liver biopsies. The degrees of fatty liver, necroinflammatory activity, and fibrosis stage were evaluated by histopathological assessment. The diagnostic performance of the NLV values in detecting each grade of fatty liver was determined using receiver operating characteristics analyses, and multivariate linear regression analyses were performed to identify variables significantly associated with the NLV values. The number of patients in each degree of fatty liver and hepatic fibrosis was 118/37/26/13 and 81/68/24/6/14 for none/mild/moderate/severe steatosis and F0 / F1/F2 / F3/F4 fibrosis on histopathological examinations, respectively. The area under the receiver operating characteristics curve and optimal cut-off NLV value for detecting fatty liver of varying degrees were 0.911 and 1.095 for ≥ S1, 0.974 and 1.055 for ≥ S2, and 0.954 and 1.025 for ≥ S3, respectively. Multivariate analyses revealed that not fibrosis or inflammation but rather the degree of steatosis was associated with the NLV value. The NLV value demonstrated excellent diagnostic performance for detecting varying degrees of fatty liver, and the degree of steatosis on histopathological examinations was the only significant factor affecting the NLV value.

Three-dimensional Visualization of Ultrasound Backscatter Statistics by Window-modulated Compounding Nakagami Imaging.

In this study, the window-modulated compounding (WMC) technique was integrated into three-dimensional (3D) ultrasound Nakagami imaging for improving the spatial visualization of backscatter statistics. A 3D WMC Nakagami image was produced by summing and averaging a number of 3D Nakagami images (number of frames denoted as N) formed using sliding cubes with varying side lengths ranging from 1 to N times the transducer pulse. To evaluate the performance of the proposed 3D WMC Nakagami imaging method, agar phantoms with scatterer concentrations ranging from 2 to 64 scatterers/mm3 were made, and six stages of fatty liver (zero, one, two, four, six, and eight weeks) were induced in rats by methionine-choline-deficient diets (three rats for each stage, total n = 18). A mechanical scanning system with a 5-MHz focused single-element transducer was used for ultrasound radiofrequency data acquisition. The experimental results showed that 3D WMC Nakagami imaging was able to characterize different scatterer concentrations. Backscatter statistics were visualized with various numbers of frames; N = 5 reduced the estimation error of 3D WMC Nakagami imaging in visualizing the backscatter statistics. Compared with conventional 3D Nakagami imaging, 3D WMC Nakagami imaging improved the image smoothness without significant image resolution degradation, and it can thus be used for describing different stages of fatty liver in rats.

Corneal endothelial alterations in alcohol dependence syndrome

Background/aimsTo evaluate the corneal endothelial changes in patients with alcohol dependence syndrome (ADS).MethodsA total of 322 corneas of 322 subjects were studied, and two groups were formed. The first, the...