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Background and Aims: Several chronic multifactorial diseases originate from energy unbalance between food intake and body energy expenditure, including non-alcoholic fatty liver disease (NAFLD), diabetes, and cardiovascular disorders. Vascular endothelium plays a central role in body homeostasis, and NAFLD is often associated with endothelial dysfunction (ED), the first step in atherosclerosis. Both sugars and fatty acids (FAs) are fuel sources for energy production, but their excess leads to liver steatosis which may trigger ED through a network of mechanisms which need to be clarified. Here, we investigated the crosstalk pathways between in vitro cultured steatotic hepatocytes (FaO) and endothelial cells (HECV) being mediated by soluble factors.Methods and Results: We employed the conditioned medium approach to test how different extent and features of hepatic steatosis distinctively affect endothelium leading to ED. The steatogenic media collected from steatotic hepatocytes were characterized by high triglyceride content and led to lipid accumulation and fat-dependent dysfunction in HECV cells. We found a parallelism between (i) extent of hepatocyte steatosis and level of lipid accumulation in HECV cells; (ii) type of hepatocyte steatosis (with macro- or microvesicular LDs) and extent of oxidative stress, lipid peroxidation, nitric oxide release and expression of ED markers in HECV cells.Conclusions: The present findings seem to suggest that, in addition to triglycerides, other soluble mediators should be released by steatotic hepatocytes and may influence lipid accumulation and function of HECV cells. Further studies need to depict the exact profile of soluble factors involved in steatotic hepatocyte-endothelium crosstalk. [ABSTRACT FROM AUTHOR]

[Display omitted] • Thymbra spicata extracts as dietary supplements for Drosophila melanogaster flies differentiated by age and gender. • The T. spicata aqueous extract (TW) extends the lifespan of young females. • The pro-longevity potential is associated with a reduced body-weight gain and improved antioxidant defense. • Thymbra spicata potential as nutraceutics for anti-ageing strategies. Aging represents a decline in function over time. Polyphenols are dietary antioxidants that could exert beneficial effects on aging. This study investigated the effects of dietary supplementation with two extracts from Thymbra spicata L. aerial parts on the lifespan, body-weight control, and antioxidant responses of Drosophila melanogaster cohorts distinguished for age and sex. The aqueous extract extended the lifespan of young female flies, and this pro-longevity potential was associated with a reduction in the body-weight gain. Furthermore, the aqueous extract promoted the antioxidant defense when young females were exposed to Paraquat by reducing oxidative stress and mortality. This study provides the first insight into the beneficial effects of T. spicata supplementation in an animal model, suggesting that the healthy effect could mainly depend on strengthening the antioxidant defenses which reflect on the lifespan. Further investigations will better clarify the mechanisms and potential of T. spicata as nutraceutics in anti-aging strategies. [ABSTRACT FROM AUTHOR]

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease often associated with overnutrition. Number and morphometry of lipid droplets (LDs) define micro vs macrovesicular steatosis, influence the morphology and function of hepatocytes and possibly their stiffness. The link between grade and features of steatosis and biomechanical properties of single hepatocytes requires deeper investigations. In vitro NAFLD models with distinct steatosis conditions were set by exposing FaO hepatoma cells to single or combined fructose (Fru), fatty acids (FA), and tumor necrosis factor (TNF)α. Single Cell Force Spectroscopy and Quantitative Phase Microscopy quantified the single cell stiffness and a series of morphometric parameters; the mRNA expression of genes involved in lipid metabolism was quantified by real-time PCR. In our models, LD size and number increased with Fru and FA as single agents, and more with combined Fru/FA (macrovesicular steatosis), while FA/TNFα combination increased LD number with a reduction in their size (microvesicular steatosis). We found that the changes in LD size and number influenced cell stiffness and morphometry as follows: (i) single cell elasticity increased in macrovesicular steatosis (maximally with combined Fru/FA); (ii) FA-induced steatosis resulted in cells thinner and larger, whereas combined FA/TNFα shrunk the hepatocytes. Taken together the data on hepatocyte biomechanics show that, in addition to extent of lipid accumulation, cell stiffness is mainly influenced by LD size, while cell morphometry directly relates to LD number. Our findings suggest that a novel mechanobiology perspective might provide future contributions in NAFLD research. Image 1 • Single cell biomechanics of in vitro models of NAFLD is assessed using a biophysical approach. • Single cell stiffness is influenced by lipid droplet size in steatotic hepatocytes. • Single cell morphometry directly relates to lipid droplet number in steatotic hepatocytes. • Single cell elasticity might be exploited as a label-free biomarker of NAFLD severity. • Mechanobiology perspective might contribute to the knowledge of NAFLD evolution. [ABSTRACT FROM AUTHOR]

Highlights • Phytocompounds influence molecular, cellular and systemic pathways. • Phytocompounds also modulate Aquaporins, a family of membrane channel proteins. • Gender differences govern the interaction between phytochemicals and AQPs. • Phytocompounds acting on AQP expression and function will make clinical benefits. Abstract A series of plant-derived bioactive compounds belonging to the class of polyphenols, terpenes and capsaicinoids, interact with important pathophysiological pathways at a molecular, cellular and systemic level. Mechanisms of action include altering cell growth and differentiation, apoptosis, autophagy, inflammation, redox balance and metabolic and energy homeostasis. These effects might also involve the expression and function of Aquaporins (AQPs), a family of membrane channel proteins, involved in several body functions. The ultimate translational beneficial effect of such phytocompounds on AQPs in health and disease is a matter of intensive research. Results might provide novel therapeutic approaches to a number of human diseases. Here, we give an updated overview of this fast growing and promising field, discussing a number of phytocompounds and their action on AQPs and related potential clinical achievements. [ABSTRACT FROM AUTHOR]

Patients with irritable bowel syndrome (IBS) often associate their symptoms to certain foods. In congenital sucrase-isomaltase deficiency (CSID), recessive mutations in the SI gene (coding for the disaccharidase digesting sucrose and 60% of dietary starch)1 cause clinical features of IBS through colonic accumulation of undigested carbohydrates, triggering bowel symptoms.2 Hence, in a previous study,3 we hypothesized that CSID variants reducing SI enzymatic activity may contribute to development of IBS symptoms. We detected association with increased risk of IBS for 4 rare loss-of-function variants typically found in (homozygous) CSID patients, because carriers (heterozygous) of these rare variants were more common in patients than in controls.1,4 Through a 2-step computational and experimental strategy, the present study aimed to determine whether other (dys-)functional SI variants are associated with risk of IBS in addition to known CSID mutations. We first aimed to identify all SI rare pathogenic variants (SI-RPVs) on the basis of integrated Mendelian Clinically Applicable Pathogenicity (M-CAP) and Combined Annotation Dependent Depletion (CADD) predictive (clinically relevant) scores; next, we inspected genotype data currently available for 2207 IBS patients from a large ongoing project to compare SI-RPV case frequencies with ethnically matched population frequencies from the Exome Aggregation Consortium (ExAC). [ABSTRACT FROM AUTHOR]

Summary Background & aims Malnutrition is emerging as a multidimensional concern of ageing with a high prevalence among nursing home residents. This study investigated the extent of malnutrition among old subjects in family practice and its relationship with major complications. Methods Over 75 years old subjects ( n = 274) filled the Mini Nutritional Assessment questionnaire. Appearance of major events in the following 6 months were registered. Results MNA scored were 11.5 ± 3.1, with 175 (64%) subjects showing no malnutrition, 69 (25%) resulted at risk, and 30 (11%) malnourished. Within at risk group, 1.4% was resident, 7% bed rested, 8% had a history of major bone fracture, 33% was demented and 24.6% hospitalized at least once in the last year. Among malnourished patients, 10% was resident, ten bed rested with 70% showing multiple bedsores, 20% have had bone fractures, 60% were demented and 13% hospital admitted in the previous year. In over 90% of them, malnutrition had neither diagnosed nor considered before. During follow-up, a significantly higher number of major events including death occurred in the malnourished group. By multivariate logistic regression, n = 56 (20.4%) patients resulted at risk of major complications. The sensitivity of the questionnaire in identifying these patients was 84% with the cut-off value of 7 associated with the highest prediction (positive predictive value, 0.92; negative predictive value, 0.71) yielding a specificity of 92%. Conclusions The prevalence of malnutrition is high among older subjects in the setting of family practice. The Mini Nutritional Assessment allows to identify malnourished subjects better than BMI and effectively predicts the risk of major events. [ABSTRACT FROM AUTHOR]

Background & aims: Lieber-DeCarli diet has been used to induce obesity and non-alcoholic steatohe-patitis (NASH). As scarce anatomical and clinical-related information on this diet model exists and being exercise an advised strategy to counteract metabolic diseases, we aimed to analyze the preventive (voluntary physical activity - VPA) and therapeutic (endurance training - ET) effect of exercise on clinical/anatomical features of rats fed with Lieber-DeCarli diet. Methods: In the beginning of the protocol, Sprague-Dawley rats were divided into standard-diet sedentary (SS, n = 20), standard-diet VPA (SVPA, n = 10), high-fat diet sedentary (HS, n = 20) and high-fat diet VPA (HVPA, n = 10) groups. After 9-weeks, half (n = 10) of SS and HS groups were engaged in an ET program (8 wks/5 d/wk/60 min/day). At this time, a blood sample was collected for biochemical analysis. At the end of protocol (17-weeks) anatomic measures were assessed. Heart, liver, femur and visceral fat were weighted and blood was collected again. Liver section was used for histopathological examination. Results: At 17-weeks, high-fat diet increased visceral adiposity (HS vs. SS), which was counteracted by both exercises. However, ET was the only intervention able to diminished obesity-related measures and the histological features of NASH. Moreover, blood analysis at 9 weeks showed that high-fat diet increased ALT, AST, cholesterol and HDL while VLDL and TG levels were decreased (HS vs. SS). Notably, although these parameters were counteracted after 9-weeks of VPA, they were transitory and not observed after 17-weeks. Conclusions: ET used as a therapeutic tool mitigated the clinical/anatomical-related features induced by Liber-DeCarli diet, thus possibly contributing to control obesity and metabolic disorders. [ABSTRACT FROM AUTHOR]

Background & Aims: SCN5A encodes the α-subunit of the voltage-gated sodium channel NaV1.5. Many patients with cardiac arrhythmias caused by mutations in SCN5A also have symptoms of irritable bowel syndrome (IBS). We investigated whether patients with IBS have SCN5A variants that affect the function of NaV1.5. Methods: We performed genotype analysis of SCN5A in 584 persons with IBS and 1380 without IBS (controls). Mutant forms of SCN5A were expressed in human embryonic kidney-293 cells, and functions were assessed by voltage clamp analysis. A genome-wide association study was analyzed for an association signal for the SCN5A gene, and replicated in 1745 patients in 4 independent cohorts of IBS patients and controls. Results: Missense mutations were found in SCN5A in 13 of 584 patients (2.2%, probands). Diarrhea-predominant IBS was the most prevalent form of IBS in the overall study population (25%). However, a greater percentage of individuals with SCN5A mutations had constipation-predominant IBS (31%) than diarrhea-predominant IBS (10%; P < .05). Electrophysiologic analysis showed that 10 of 13 detected mutations disrupted NaV1.5 function (9 loss-of-function and 1 gain-of-function function). The p. A997T-NaV1.5 had the greatest effect in reducing NaV1.5 function. Incubation of cells that expressed this variant with mexiletine restored their sodium current and administration of mexiletine to 1 carrier of this mutation (who had constipation-predominant IBS) normalized their bowel habits. In the genome-wide association study and 4 replicated studies, the SCN5A locus was strongly associated with IBS. Conclusions: About 2% of patients with IBS carry mutations in SCN5A. Most of these are loss-of-function mutations that disrupt NaV1.5 channel function. These findings provide a new pathogenic mechanism for IBS and possible treatment options. [Copyright &y& Elsevier]

• FMF is the most common autoinflammatory disease with colchicine confirmed as the most effective first-line therapy. • The presence of a prevalent lympho-myeloid infiltrate in synovitis, might confer colchicine resistance to FMF arthritis. • Detailed information about histopathological pattern of synovitis in FMF patients may help to better understand the pathogenic mechanisms leading to arthritis, thus allowing a suitable treatment choice. [ABSTRACT FROM AUTHOR]

[Display omitted] • Plant-based healthy foods generate health-promoting effects, especially on chronic and cardio-metabolic diseases. • Za'atar mixture is commonly employed in the Mediterranean diet and is rich of bioactive compounds. • Za'atar components modulate gut microbiota, oxidative stress, chronic inflammation, and obesity. • The combined action of Za'atar components might generate beneficial effects for human health. Interest in plant-based food has grown in recent years due to their primary prevention potential. Za'atar, an ancient and popular Lebanese herbal mixture, might disclose relevant clinical interest, due to the well-known intrinsic properties of its individual components. Za'atar mixture contain Origanum syriacum (Lebanese thyme), Thymbra spicata (Wild thyme), Rhus coriaria (Sumac), and Sesamum indicum (Sesame). Here we explored the history, composition, general employment, and bio-active aspects of Za'atar through available in vitro , animal, and clinical trials evidence to depict its possible role as an innovative nutraceutical tool. The combined action of Za'atar constituents is able to generate comprehensive beneficial effects on several common pathogenic pathways underlying chronic cardio-metabolic diseases and cancer. However, main available evidence derives from animal and in vitro studies. Thus, further human studies are needed to fully characterize Za'atar as a preventive and curative tool. [ABSTRACT FROM AUTHOR]

Abstract: An essential function of the liver is the formation and secretion of bile, a complex aqueous solution of organic and inorganic compounds essential as route for the elimination of body cholesterol as unesterified cholesterol or as bile acids. In bile, a considerable amount of otherwise insoluble cholesterol is solubilized by carriers including two other classes of lipids, namely phospholipid and bile acids. Formation of bile and generation of bile flow are driven by the active secretion of bile acids, lipids and electrolytes into the canalicular and bile duct lumens followed by the parallel movement of water. Thus, water has to cross rapidly into and out of the cell interior driven by osmotic forces. Bile as a fluid, results from complicated interplay of hepatocyte and cholangiocyte uptake and secretion, concentration, by involving a number of transporters of lipids, anions, cations, and water. The discovery of the aquaporin water channels, has clarified the mechanisms by which water, the major component of bile (more than 95%), moves across the hepatobiliary epithelia. This review is focusing on novel acquisitions in liver membrane lipidic and water transport and functional participation of aquaporin water channels in multiple aspects of hepatobiliary fluid balance. Involvement of aquaporins in a series of clinically relevant hepatobiliary disorders are also discussed. [Copyright &y& Elsevier]

Abstract: Background: Despite chronic liver diseases represent an important cause of illness in Italy, data from family practice are poor. Aim: To assess the management of chronic liver diseases by general practitioners in a large area of Southern Italy. Methods: This was a 5-year retrospective analysis from 104 physicians in charge of a population of 143,159 adult subjects. Results: Amongst 6550 patients with chronic liver disease (4.7%, 3400 M, median age 57 years), 1330 (20.3%) had HCV infection, 226 (3.4%) HBV infection, and 293 (4.5%) liver cirrhosis (25 alcohol-related). The prevalence of alcohol consumption, recorded by 90% of physicians, was 20.4%. Hypertransaminasemia and liver steatosis had a prevalence of 6.7% and 2.4%, respectively. Although transaminases were checked 3 times over 5 years in 80% of cases, few patients were investigated for viral infection, and less than 50% underwent ultrasonography and consultation, leaving undefined a consistent number of cases. Alcohol consumption, body mass index and ultrasonography were poorly checked even in hypertransaminasemic patients. Conclusions: This study shows that data recording by general practitioners in chronic liver disease patients lacks homogeneity and can miss important information. One unmet need is therefore the integration between theoretical knowledge and practice to share similar behaviours and improve the management of these patients. [Copyright &y& Elsevier]

Non-alcoholic fatty liver disease (NAFLD), the most common liver disorder in the Western world, is a clinico-histopathological entity in which excessive triglyceride accumulation in the liver occurs. Non-alcoholic steatohepatitis (NASH) represents the necroinflammatory form, which can lead to advanced liver fibrosis, cirrhosis, and hepatocellular carcinoma. The pathogenesis of NAFLD/NASH is complex but increased visceral adiposity plus insulin resistance with increased free fatty acids release play an initial key role for the onset and perpetuation of liver steatosis. Further events in the liver include oxidative stress and lipid peroxidation, decreased antioxidant defences, early mitochondrial dysfunction, iron accumulation, unbalance of adipose-derived adipokines with a chronic proinflammatory status, and gut-derived microbial adducts. New gene polymorphisms increasing the risk of fatty liver, namely APOC3 and PNPLA3, have been lately identified allowing further insights into the pathogenesis of this condition. In our review pathophysiological, genetic, and essential diagnostic and therapeutic aspects of NAFLD are examined with future trends in this field highlighted. [ABSTRACT FROM AUTHOR]

Abstract: Background: Standardization of the 13C-octanoic acid breath test is still lacking. Aim: To evaluate the accuracy of the 13C-octanoic acid breath test using a new standardized ready-to-eat, gluten-, glucose-, and lactose-free muffin. Methods: Healthy subjects were recruited and sorted by sex and age. Patients with diabetic gastroparesis and untreated celiac disease with known gastric motility disorders were also tested with the new labelled muffin. Expired breath 13CO2 was analysed and t 1/2 was calculated. Results: Overall, 131 healthy subjects were enrolled. The reference range of t 1/2 was 88±29min with the value of 146min as the upper limit of normal range. No significant difference in t 1/2 was found among subjects sorted by sex or age. The within-subject variability of t 1/2 was 17%. Mean (±standard deviation) t 1/2 values were 179±50min in patients with diabetic gastroparesis (n =8) and 151±20min in those with untreated celiac disease (n =11) (p ≤0.001 vs controls). Conclusions: A new standardized test meal simplifies the execution of the 13C-octanoic acid breath test, is not influenced by sex or age, has low intra-individual variability, is palatable, does not cause side effects, and is able to evaluate gastric emptying in both patients and healthy controls. Moreover, it can be used in subjects with lactose intolerance, diabetes, and celiac disease. [Copyright &y& Elsevier]

Dietary choline deprivation (CD) is associated with behavioral changes, but mechanisms underlying these detrimental effects are not well characterized. For instance, no literature data are available concerning the CD effects on brain mitochondrial function related to impairment in cognition. Therefore, we investigated brain mitochondrial function and redox status in male Wistar rats fed a CD diet for 28 d. Moreover, the CD behavioral phenotype was characterized. Compared with rats fed a control diet (CTRL), CD rats showed lower NAD-dependent mitochondrial state Ill and state IV respiration, 40% lower complex I activity, and significantly higher reactive oxygen species production. Total glutathione was oxidatively consumed more in CD than in CTRL rats and the rate of protein oxidation was 40% higher in CD than in CTRL rats, reflecting an oxidative stress condition. The mitochondrial concentrations of cardiolipin, a phospholipid required for optimal activity of complex I, was 20% lower in CD rats than in CTRL rats. Compared with CTRL rats, the behavioral phenotype of CD rats was characterized by impairment in motor coordination and motor learning assessed with the rotarod/accelerod test. Furthermore, compared with CTRL rats, CD rats were less capable of learning the active avoidance task and the number of attempts they made to avoid foot shock was fewer. The results suggest that CD induced dysfunction in brain mitochondria may be responsible for impairment in cognition and underline that, similar to the liver, the brain also needs an adequate choline supply for its normal functioning. [ABSTRACT FROM AUTHOR]

Abstract: Background: Alteration in intestinal permeability may be an important factor in the pathogenesis of both the progression of some chronic liver diseases and the onset of some complications in patients with liver cirrhosis. Aims: To investigate the relationships between intestinal permeability, portal hypertension, alcohol use, plasma levels of pro-inflammatory cytokines, and nitric oxide, expressed as s-nitrosothiols, and nitrite levels in patients with various types and degrees of chronic liver diseases. Methods: 134 healthy volunteers and 83 patients with chronic liver damage entered the study. Intestinal permeability was assessed with the lactulose/mannitol test. Plasma levels of tumour necrosis factor-alpha, interleukin-6, and nitrite and total s-nitrosothiols were determined. Results: Intestinal permeability was altered in patients with advanced liver disease and impaired in 15–35% of patients without cirrhosis. Independent factors for intestinal permeability alteration were age, portal hypertension, alcohol use, and diabetes. Plasma levels of inflammatory cytokines and nitrosothiols were significantly higher in patients with altered intestinal permeability. Conclusions: An intestinal permeability evaluation in patients with chronic liver diseases might clarify the significance of intestinal permeability in the pathophysiology of both the progression of liver damage, and the occurrence of complications that accompany liver cirrhosis. [Copyright &y& Elsevier]

Abstract: We investigated the effect of gallbladder hypomotility on cholesterol crystallization and growth during the early stage of gallstone formation in CCK knockout mice. Contrary to wild-type mice, fasting gallbladder volumes were enlarged and the response of gallbladder emptying to a high-fat meal was impaired in knockout mice on chow or the lithogenic diet. In the lithogenic state, large amounts of mucin gel and liquid crystals as well as arc-like and tubular crystals formed first, followed by rapid formation of classic parallelogram-shaped cholesterol monohydrate crystals in knockout mice. Furthermore, three patterns of crystal growth habits were observed: proportional enlargement, spiral dislocation growth, and twin crystal growth, all enlarging solid cholesterol crystals. At day 15 on the lithogenic diet, 75% of knockout mice formed gallstones. However, wild-type mice formed very little mucin gel, liquid, and solid crystals, and gallstones were not observed. We conclude that lack of CCK induces gallbladder hypomotility that prolongs the residence time of excess cholesterol in the gallbladder, leading to rapid crystallization and precipitation of solid cholesterol crystals. Moreover, during the early stage of gallstone formation, there are two pathways of liquid and polymorph anhydrous crystals evolving to monohydrate crystals and three modes for cholesterol crystal growth. [Copyright &y& Elsevier]

[Display omitted] • Suma c is a spicy ingredient commonly employed in the Mediterranean diet and folk medicine. • Sumac contains (poly)phenolic compounds, organic acids, essential oils, and vitamins. • NAFLD is often associated with metabolic disorders and increased risk of atherosclerosis. • Evidence suggests that sumac has beneficial effects on metabolically related diseases. Sumac (Rhus coriaria L.) is a commonly used spice in the Mediterranean region and considered as healthy food ingredients. The beneficial value of sumac is well documented in folk medicine. Accumulating data explored the phytochemical, nutritional and therapeutic proprieties suggesting sumac as a potential functional food. Here, we discuss the general and scientific aspects of sumac. Sumac is rich in different polyphenolic compounds such as flavonoids, tannins, and phenolic acids. The potential therapeutic effects of sumac have been studied in various cellular and animal models, as well as in human. These reports suggest that Sumac has potential effect against oxidative stress, inflammation, obesity, hyperglycemia, hypercholesterolemia, and hyperlipidemia, which represent key pathogenic mechanisms contributing to cardio-metabolic, liver, and cancer diseases. Clinical studies using sumac or its major compounds, suggest that this herbal product may represent a useful therapeutic tool in the management of metabolic-related conditions such as liver-atherosclerosis complications. [ABSTRACT FROM AUTHOR]

Abstract: Mechanisms by which hydrophobic bile salts cause tissue changes below their critical micellar concentration (CMC, 1–2mM) and above (4–8mM) remain poorly understood. In this study, rat colonic mucosa was exposed to different concentrations of taurodeoxycholate (TDC), t-butyl-hydroperoxide (t-BH) or glutathione ester with or without pre-incubation with 2mM TDC. Exposure to 2mM TDC was associated with 10% higher tissue levels of total glutathione (GSH, basal values: 33.7±3.3nmol/mgprot). With TDC 8mM, GSH decreased to 16.4±2.3nmol/mgprot (P <0.05), oxidized glutathione (GSSG) increased by 60% (P <0.05), glutathione peroxidase (GSH-Px) and reductase activities were threefold increased, protein carbonyls fourfold increased, protein sulfhydrils decreased by 78%, lactate dehydrogenase (LDH) and GSSG release in the incubation medium were sixfold higher. In 2mM TDC pre-treated tissues, the subsequent incubation with 8mM TDC induced a lower loss of tissue GSH, and a lower release of LDH and GSSG. Pre-incubation with 2mM TDC partly protected against t-BH toxicity, while glutathione ester protected against 8mM TDC toxicity. In conclusion, TDC exposure causes opposite effects depending on CMC: induction of antioxidant protective systems including glutathione system (pre-conditioning effect) was observed with TDC below CMC, oxidative damages pointing to decreased mucosal detoxification potential with above CMC. [Copyright &y& Elsevier]

Background & Aims: Cholesterol cholelithiasis is one of the most prevalent and most costly digestive diseases in developed countries and its incidence has increased markedly in Asian countries owing to the adoption of Western-type dietary habits. Because animal experiments showed that high efficiency of intestinal cholesterol absorption contributes to gallstone formation, we explored whether the potent cholesterol absorption inhibitor ezetimibe could prevent gallstones and promote gallstone dissolution in mice and reduce biliary cholesterol content in human beings. Methods: Male gallstone-susceptible C57L mice were fed a lithogenic diet and concomitantly administered with ezetimibe at 0, 0.8, 4, or 8 mg/kg/day for 8 or 12 weeks. Gallbladder biles and gallstones were examined by microscopy. Gallbladder emptying in response to cholecystokinin octapeptide was measured gravimetrically. Biliary lipid outputs were analyzed by physical–chemical methods. Cholesterol absorption efficiency was determined by fecal dual-isotope ratio and mass balance methods. Lipid changes in gallbladder biles of gallstone patients vs overweight subjects without gallstones were examined before (day 0) and at 30 days after ezetimibe treatment (20 mg/day). Results: Ezetimibe prevented gallstones by effectively reducing intestinal cholesterol absorption and biliary cholesterol secretion, and protected gallbladder motility function by desaturating bile in mice. Treatment with ezetimibe promoted the dissolution of gallstones by forming an abundance of unsaturated micelles. Furthermore, ezetimibe significantly reduced biliary cholesterol saturation and retarded cholesterol crystallization in biles of patients with gallstones. Conclusions: Ezetimibe is a novel approach to reduce biliary cholesterol content and a promising strategy for preventing or treating cholesterol gallstones by inhibiting intestinal cholesterol absorption. [Copyright &y& Elsevier]

Abstract: Mitochondrial dysfunction and oxidative stress play a central role in the pathophysiology of nonalcoholic fatty liver disease (NAFLD). This study aimed to elucidate the mechanism(s) responsible for mitochondrial dysfunction in nonalcoholic fatty liver. Fatty liver was induced in rats with a choline-deficient (CD) diet for 30 days. We examined the effect of CD diet on various parameters related to mitochondrial function such as complex I activity, oxygen consumption, reactive oxygen species (ROS) generation and cardiolipin content and oxidation. The activity of complex I was reduced by 35% in mitochondria isolated from CD livers compared with the controls. These changes in complex I activity were associated with parallel changes in state 3 respiration. Hydrogen peroxide (H2O2) generation was significantly increased in mitochondria isolated from CD livers. The mitochondrial content of cardiolipin, a phospholipid required for optimal activity of complex I, decreased by 38% as function of CD diet, while there was a significantly increase in the level of peroxidized cardiolipin. The lower complex I activity in mitochondria from CD livers could be completely restored to the level of control livers by exogenously added cardiolipin. This effect of cardiolipin could not be replaced by other phospholipids nor by peroxidized cardiolipin. It is concluded that CD diet causes mitochondrial complex I dysfunction which can be attributed to ROS-induced cardiolipin oxidation. These findings provide new insights into the alterations underlying mitochondrial dysfunction in NAFLD. [Copyright &y& Elsevier]