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PDF file - 292K, Table S1 shows FACS analysis results of MET, CD133, and CD15 in the freshly isolated patient GBM cells, and Table S2 describes the list of patient-derived GBM cells and their derivatives used in this study. Patient information of 14 matched pre- and postradiated GBM specimens is shown in Table S3

PDF file - 6.4MB, Figure S1: High MET mRNA expression in GBM specimens portends poor patient survival. Figure S2: Dual immunofluorescence of MET and GSC enrichment markers (CD133 and CD15) on the frozen sections of GBM specimens. Figure S3: HGF/MET signaling in GBMs. Figure S4: METhigh cells generated xenograft tumors consisting of both METhigh and METlow/- cells. Figure S5: Immunohistochemical analysis of HGF protein in orthotopic xenograft tumors derived of from GBM 578 (A and C) and 464 (B and D). Figure S6: Effects of MET blockade on cell survival and clonogenicity of GSCs. Figure S7: In vitro migration and invasion assays using GSCs with MET targeting. Figure S8: Immunohistochemical analysis using human nuclei-specific antibody on the sections of GBM xenografts. Figure S9: Limiting dilution sphere forming assay to determine the clonogenic potentials of 822 GSCs after treatments of radiation and SU11274

Flavin adenine dinucleotide (FAD) interacts with flavoproteins to mediate oxidation-reduction reactions required for cellular energy demands. Not surprisingly, mutations that alter FAD binding to flavoproteins cause rare inborn errors of metabolism (IEMs) that disrupt liver function and render fasting intolerance, hepatic steatosis, and lipodystrophy. In our study, depleting FAD pools in mice with a vitamin B2 deficient diet (B2D) caused phenotypes associated with organic acidemias and other IEMs, including reduced body weight, hypoglycemia, and fatty liver disease. Integrated discovery approaches revealed B2D tempered fasting activation of target genes for the nuclear receptor PPARα, including those required for gluconeogenesis. Treatment with the PPARα agonist fenofibrate activated the integrated stress response and refilled amino acid substrates to rescue fasting glucose availability and overcome B2D phenotypes. Overall, these findings reveal PPARα governs metabolic responses to FAD availability and nominate its pharmacologic activation as strategies for organic acidemias.

In-memory computing has been widely used to process data quickly in memory, but it is no longer able to cope with the current data explosion. This has occurred because modern computers have structural constraints in terms of both memory capacity and bandwidth, fundamentally limiting the amount of data that can be processed in memory. In order to overcome these limitations, the memory-centric computing concept was proposed, and as an attempt to make this concept feasible, nonvolatile memory, next-generation interconnects, and memory-centric operating system technologies are being actively studied. Although, however, there have been many studies of these concepts, the essential problem of both efficiently and safely using the extremely large memory space has not been completely solved. This paper proposes what is termed Global-Heap which solves the above essential problem, thus supporting the core technology of memory-centric computing. In this technology, a heap-level abstraction which can be switched very lightly between processes is provided in an application-friendly manner as a global resource. Global-Heap also provides an effective means by which safely to use this vast address space based on Intel memory protection keys. We implemented Global-Heap on Linux and developed three useful use cases applicable to data-centric applications. Our primary evaluation shows that Global-Heap enables reorganization-free hashing. In addition, Global-Heap can improve the performance of an in-memory key-value store, Redis, by at least 7.56 times, and it can reduce the running time of graph applications executed in a pipeline approach by at least 48%.

Our early-life environment has a profound influence on developing organs that impacts metabolic function and determines disease susceptibility across the life-course. Using a rat model for exposure to an endocrine disrupting chemical (EDC), we show that early-life chemical exposure causes metabolic dysfunction in adulthood and reprograms histone marks in the developing liver to accelerate acquisition of an adult epigenomic signature. This epigenomic reprogramming persists long after the initial exposure, but many reprogrammed genes remain transcriptionally silent with their impact on metabolism not revealed until a later life exposure to a Western-style diet. Diet-dependent metabolic disruption was largely driven by reprogramming of the Early Growth Response 1 (EGR1) transcriptome and production of metabolites in pathways linked to cholesterol, lipid and one-carbon metabolism. These findings demonstrate the importance of epigenome:environment interactions, which early in life accelerate epigenomic aging, and later in adulthood unlock metabolically restricted epigenetic reprogramming to drive metabolic dysfunction., Early life exposure to environmental stressors, including endocrine disrupting chemicals (EDCs), can impact health later in life. Here, the authors show that neonatal EDC exposure in rats causes epigenetic reprogramming in the liver, which is transcriptionally silent until animals are placed on a Western-style diet.

Cirrhotic cardiomyopathy is a clinical syndrome in patients with liver cirrhosis characterized by blunted cardiac contractile responses to stress and/or heart rate-corrected QT (QTc) interval prolongation. Caveolin-3 (Cav-3) plays a critical role in cardiac protection and is an emerging therapeutic target for heart disease. We investigated the protective role of cardiac-specific overexpression (OE) of Cav-3 in cirrhotic cardiomyopathy. Biliary fibrosis was induced in male Cav-3 OE mice and transgene negative (TGneg) littermates by feeding a diet containing 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC; 0.1%) for 3 wk. Liver pathology and blood chemistries were assessed, and stress echocardiography, telemetry, and isolated heart perfusion studies to assess adrenergic responsiveness were performed. Cav-3 OE mice showed a similar degree of hyperdynamic contractility, pulmonary hypertension, and QTc interval prolongation as TGneg mice after 3 wk of DDC diet. Blunted systolic responses were shown in both DDC-fed Cav-3 OE and TGneg hearts after in vivo isoproterenol challenge. However, QTc interval prolongation after in vivo isoproterenol challenge was significantly less in DDC-fed Cav-3 OE hearts compared with DDC-fed TGneg hearts. In ex vivo perfused hearts, where circulatory factors are absent, isoproterenol challenge showed hearts from DDC-fed Cav-3 OE mice had better cardiac contractility and relaxation compared with DDC-fed TGneg hearts. Although Cav-3 OE in the heart did not prevent cardiac alterations in DDC-induced biliary fibrosis, cardiac expression of Cav-3 reduced QTc interval prolongation after adrenergic stimulation in cirrhosis. NEW & NOTEWORTHY Prevalence of cirrhotic cardiomyopathy is up to 50% in cirrhotic patients, and liver transplantation is the only treatment. However, cirrhotic cardiomyopathy is associated with perioperative morbidity and mortality after liver transplantation; therefore, management of cirrhotic cardiomyopathy is crucial for successful liver transplantation. This study shows cardiac myocyte specific overexpression of caveolin-3 (Cav-3) provides better cardiac contractile responses and less corrected QT prolongation during adrenergic stress in a cirrhotic cardiomyopathy model, suggesting beneficial effects of Cav-3 expression in cirrhotic cardiomyopathy.

Inactivating mutations in the copper transporter Atp7b result in Wilson’s disease. The Atp7b −/− mouse develops hallmarks of Wilson’s disease. The activity of several nuclear receptors decreased in Atp7b −/− mice, and nuclear receptors are critical for maintaining metabolic homeostasis. Therefore, we anticipated that Atp7b −/− mice would exhibit altered progression of diet-induced obesity, fatty liver, and insulin resistance. Following 10 wk on a chow or Western-type diet (40% kcal fat), parameters of glucose and lipid homeostasis were measured. Hepatic metabolites were measured by liquid chromatography–mass spectrometry and correlated with transcriptomic data. Atp7b −/− mice fed a chow diet presented with blunted body-weight gain over time, had lower fat mass, and were more glucose tolerant than wild type (WT) littermate controls. On the Western diet, Atp7b −/− mice exhibited reduced body weight, adiposity, and hepatic steatosis compared with WT controls. Atp7b −/− mice fed either diet were more insulin sensitive than WT controls; however, fasted Atp7b −/− mice exhibited hypoglycemia after administration of insulin due to an impaired glucose counterregulatory response, as evidenced by reduced hepatic glucose production. Coupling gene expression with metabolomic analyses, we observed striking changes in hepatic metabolic profiles in Atp7b −/− mice, including increases in glycolytic intermediates and components of the tricarboxylic acid cycle. In addition, the active phosphorylated form of AMP kinase was significantly increased in Atp7b −/− mice relative to WT controls. Alterations in hepatic metabolic profiles and nuclear receptor signaling were associated with improved glucose tolerance and insulin sensitivity as well as with impaired fasting glucose production in Atp7b −/− mice.

Background and aims Hepatic endoplasmic reticulum (ER) stress, whether triggered by intrinsic or extrinsic factors, can be resolved by the unfolded protein response (UPR). Sustained UPR activation leads to cell death and inflammatory response and contributes to liver disease progression. Hepatic tissue macrophages are key players in orchestrating liver inflammation, and ER stress can enhance macrophage activation. However, it is not well defined how the interplay between ER stress and inflammation is regulated during hepatic stress response. Approach and results Here we demonstrate that vitamin D receptor (VDR) activation mitigates hepatic ER stress response, whereas VDR knockout mice undergo persistent UPR activation and apoptosis in response to chemical ER stress inducer. Moreover, VDR deficiency promotes hepatic macrophage infiltration and increases gene expression and systematic levels of proinflammatory cytokines, including interleukin (IL)-1β, IL-6, and tumor necrosis factor α. VDR expression is induced in hepatic macrophages by ER stress, and VDR plays a dual regulatory role in macrophages by protecting against ER stress and promoting anti-inflammatory polarization. Co-culture with VDR-activated bone marrow-derived macrophages suppresses UPR target genes in primary hepatocytes treated with ER stress inducers. Thus, the immunomodulatory functions of VDR in macrophages are critical in hepatic ER stress resolution in mice. Conclusions VDR signaling in macrophages regulates a shift between proinflammatory and anti-inflammatory activation during ER stress-induced inflammation to promote hepatic ER stress resolution.

An extensive literature base combined with advances in sequencing technologies demonstrate microRNA levels correlate with various metabolic diseases. Mechanistic studies also establish microRNAs regulate central metabolic pathways and thus play vital roles in maintaining organismal energy balance and metabolic homeostasis. This review highlights research progress on the roles and regulation of microRNAs in the peripheral tissues that confer insulin sensitivity. We discuss sequencing technologies used to comprehensively define the target spectrum of microRNAs in metabolic disease that complement studies reporting physiologic roles for microRNAs in the regulation of glucose and lipid metabolism in animal models. We also discuss the emerging roles of exosomal microRNAs as endocrine signals to regulate lipid and carbohydrate metabolism.

Low-grade, sustained inflammation in white adipose tissue (WAT) characterizes obesity and coincides with type 2 diabetes mellitus (T2DM). However, pharmacological targeting of inflammation lacks durable therapeutic effects in insulin-resistant conditions. Through a computational screen, we discovered that the FDA-approved rheumatoid arthritis drug auranofin improved insulin sensitivity and normalized obesity-associated abnormalities, including hepatic steatosis and hyperinsulinemia in mouse models of T2DM. We also discovered that auranofin accumulation in WAT depleted inflammatory responses to a high-fat diet without altering body composition in obese wild-type mice. Surprisingly, elevated leptin levels and blunted beta-adrenergic receptor activity achieved by leptin receptor deletion abolished the antidiabetic effects of auranofin. These experiments also revealed that the metabolic benefits of leptin reduction were superior to immune impacts of auranofin in WAT. Our studies uncover important metabolic properties of anti-inflammatory treatments and contribute to the notion that leptin reduction in the periphery can be accomplished to treat obesity and T2DM.

SUMMARYLow-grade, sustained inflammation in white adipose tissue (WAT) characterizes obesity and frequently coincides with insulin resistance and type 2 diabetes (T2D). However, pharmacological targeting of WAT inflammation lacks durable therapeutic effects. Through a computational screen, we identified the FDA-approved rheumatoid arthritis drug auranofin is a putative small molecule for obesity treatment. We discovered that allometrically scaled safe auranofin doses homed to WAT and improved insulin sensitivity in obese wild-type mice. Auranofin treatment also normalized other obesity-associated abnormalities, including hepatic steatosis and hyperinsulinemia. Surprisingly, the anti-diabetic effects of auranofin required leptin lowering and beta-adrenergic receptors in WAT. These metabolic benefits of leptin reduction were superior to any immune impacts of auranofin in WAT. Our studies reveal important metabolic properties of anti-inflammatory treatments and contribute to the notion that leptin reduction in the periphery can be accomplished to treat obesity and T2D.

With the growing importance of in-memory data processing, cloud service providers have launched large memory virtual machine services to accommodate memory intensive workloads. Such large memory services using low volume scaled-up machines are far less cost-efficient than scaled-out services consisting of high volume commodity servers. By exploiting memory usage imbalance across cloud nodes, disaggregated memory can scale up the memory capacity for a virtual machine in a cost-effective way. Disaggregated memory allows available memory in remote nodes to be used for the virtual machine requiring more memory than its locally available memory. It supports high performance with the faster direct memory while satisfying the memory capacity demand with the slower remote memory. This paper proposes a new hypervisor-integrated disaggregated memory system for cloud computing. The hypervisor-integrated design has several new contributions in its disaggregated memory design and implementation. First, with the tight hypervisor integration, it investigates a new page management mechanism and policy tuned for disaggregated memory in virtualized systems. Second, it restructures the memory management procedures and relieves the scalability concern for supporting large virtual machines. Third, exploiting page access records available to the hypervisor, it supports application-aware elastic block sizes for fetching remote memory pages with different granularities. Depending on the degrees of spatial locality for different regions of memory in a virtual machine, the optimal block size for each memory region is dynamically selected. The experimental results with the implementation integrated to the KVM hypervisor, show that the disaggregated memory can provide on average 6 percent performance degradation compared to the ideal local-memory only machine, even though the direct memory capacity is only 50 percent of the total memory footprint.

Background and Objectives: This study investigated the various impulse effects of whole-body electromyostimulation (WB-EMS) on psychophysiological responses and adaptations. Materials and Methods: The participants included fifty-four men between 20 and 27 years of age who practiced isometric exercises for 20 min, three days a week, for 12 weeks while wearing WB-EMS suits, which enabled the simultaneous activation of eight muscle groups with three types of impulse intensities. Participants were allocated to one of four groups: control group (CON), low-impulse-intensity group (LIG), mid-impulse-intensity group (MIG), and high-impulse-intensity group (HIG). Psychophysiological conditions were measured at week 0, week 4, week 8, and week 12. Results: Compared with the CON, (1) three psychological conditions in LIG, MIG, and HIG showed positive tendencies every four weeks, and the analysis of covariance (ANCOVA) test revealed that body image (p = 0.004), body shape (p = 0.007), and self-esteem (p = 0.001) were significantly different among the groups. (2) Body weight, fat mass, body mass index, and percent fat in the CON showed decreasing tendencies, whereas those in LIG, MIG, and HIG showed a noticeable decrease, which revealed that there were significant differences among the groups. Specifically, a higher impulse intensity resulted in a greater increase in muscle mass. (3) Although there was no interaction effect in the abdominal visceral fat area, there were significant interactions in the abdominal subcutaneous fat (ASF) and total fat (ATF) areas. Both the ASF and ATF in the CON showed decreasing tendencies, whereas those in other groups showed a noticeable decrease. The ANCOVA revealed that the ASF (p = 0.002) and ATF (p = 0.001) were significantly different among the groups. In particular, the higher the impulse intensity, the greater the decrease in abdominal fat. Conclusions: This study confirmed that high-impulse-intensity EMS can improve psychophysiological conditions. In other words, healthy young adults felt that the extent to which their body image, body shape, and self-esteem improved depended on how intense their EMS impulse intensities were. The results also showed that higher levels of impulse intensity led to improved physical conditions.

Bile acids have recently emerged as key metabolic hormones with beneficial impacts in multiple metabolic diseases. We previously discovered that hepatic bile acid overload distally modulates glucose and fatty acid metabolism in adipose tissues to exert anti-obesity effects. However, the detailed mechanisms that explain the salutary effects of serum bile acid elevation remain unclear. Here, proteomic profiling identified a new hepatokine, Orosomucoid (ORM) that governs liver-adipose tissue crosstalk. Hepatic ORMs were highly induced by both genetic and dietary bile acid overload. To address the direct metabolic effects of ORM, purified ORM proteins were administered during adipogenic differentiation of 3T3-L1 cells and mouse stromal vascular fibroblasts. ORM suppressed adipocyte differentiation and strongly inhibited gene expression of adipogenic transcription factors such as C/EBPβ, KLF5, C/EBPα, and PPARγ. Taken together, our data clearly suggest that bile acid-induced ORM secretion from the liver blocks adipocyte differentiation, potentially linked to anti-obesity effect of bile acids.

To gain insights into mechanisms underlying coagulopathy in malnutrition, we fed mice a low‐protein, low‐fat diet and assessed hepatic gene expression and plasma coagulation indices. Malnourished males were affected more profoundly than females, and exhibited coagulopathy, decreased expression of fibrinogen‐alpha and factor 11, decreased FXR activation, and decreased FXR binding to regulatory elements on these two coagulation factor genes. These findings are associated with profoundly depleted PPAR‐alpha protein, peroxisome loss, and decreased concentrations of FXR agonist bile acids in malnourished males but not females., Liver dysfunction, including coagulopathy, is a prominent feature of protein‐energy malnutrition. To identify mechanisms underlying malnutrition‐associated coagulopathy, we administered a low‐protein low‐fat diet to lactating dams and examined hepatic transcription and plasma coagulation parameters in young adult weanlings. Malnutrition impacted body composition to a greater extent in male versus female mice. Transcriptional profiles suggested opposing effects of nutrient‐sensing nuclear receptors, namely induction of peroxisome proliferator‐activated receptor α (PPARα) targets and repression of farnesoid‐X‐receptor (FXR) targets. Coagulopathy with decreased synthesis of fibrinogen‐α (FGA) and factor 11 (F11) was observed in malnourished male animals but not female animals. In primary mouse hepatocytes, FXR agonist increased and PPARα agonist decreased Fga and F11 messenger RNA expression. Nuclear receptor DNA response elements were identified in the Fga and F11 gene regulatory regions, and opposing effects of FXR and PPARα were confirmed with luciferase assays. Unexpectedly, hepatic PPARα protein was markedly depleted in malnourished male liver and was not enriched on Fga or F11 response elements. Rather, there was loss of FXR binding at these response elements. Reduced PPARα protein was associated with loss of hepatocyte peroxisomes, which are necessary for bile acid biosynthesis, and with decreased concentrations of bile acids that function as FXR ligands, most notably the FXR agonist chenodeoxycholic acid. Conclusion: Malnutrition impairs growth and liver synthetic function more severely in male mice than in female mice. Malnourished male mice are coagulopathic and exhibit decreased hepatocyte peroxisomes, FXR agonist bile acids, FXR binding on Fga and F11 gene regulatory elements, and coagulation factor synthesis. These effects are absent in female mice, which have low baseline levels of PPARα, suggesting that nutrient‐sensing nuclear receptors regulate coagulation factor synthesis in response to host nutritional status in a sex‐specific manner.

Obesity fosters low-grade inflammation in white adipose tissue (WAT) that may contribute to the insulin resistance that characterizes type 2 diabetes mellitus (T2DM). However, the causal relationship of these events remains unclear. The established dominance of signal transducer and activator of transcription 1 (STAT1) function in the immune response suggests an obligate link between inflammation and the co-morbidities of obesity. To this end, we sought to determine how STAT1 activity in white adipocytes affects insulin sensitivity. STAT1 expression in WAT inversely correlated with fasting plasma glucose in both obese mice and humans. Metabolomic and gene expression profiling established STAT1 deletion in adipocytes (STAT1a-KO­­­) enhanced mitochondrial function and accelerated TCA cycle flux coupled with reduced fat cell size in subcutaneous WAT depots. STAT1a-KO reduced WAT inflammation, but insulin resistance persisted in obese mice. Rather, elimination of type I cytokine interferon gamma (IFNg) activity enhanced insulin sensitivity in diet-induced obesity. Our findings reveal a permissive mechanism that bridges WAT inflammation to whole-body insulin sensitivity.

ObjectiveWhite adipose tissue (WAT) expansion regulates energy balance and overall metabolic homeostasis. WAT absence or loss occurring through lipodystrophy and lipoatrophy contributes to the development of dyslipidemia, hepatic steatosis, and insulin resistance. We previously demonstrated the sole small ubiquitin-like modifier (SUMO) E2-conjuguating enzyme Ubc9 represses human adipocyte differentiation. Germline and other tissue-specific deletions of Ubc9 frequently cause lethality in mice. As a result, the role of Ubc9 during WAT development remains unknown.MethodsTo determine how Ubc9 impacts body composition and energy balance, we generated adipocyte-specific Ubc9 knockout mice (Ubc9a-KO). CRISPR/Cas9 gene editing inserted loxP sites flanking exons 3 and 4 at the Ubc9 locus. Subsequent genetic crosses to AdipoQ-Cre transgenic mice allowed deletion of Ubc9 in white and brown adipocytes. We measured multiple metabolic endpoints that describe energy balance and carbohydrate metabolism in Ubc9a-KO and littermate controls during postnatal growth.ResultsTo our surprise, Ubc9a-KO mice developed hyperinsulinemia and hepatic steatosis. Global energy balance defects emerged from dysfunctional WAT marked by pronounced local inflammation, loss of serum adipokines, hepatomegaly, and near absence of major adipose tissue depots. We observed progressive lipoatrophy that commences in the early adolescent period.ConclusionsOur results demonstrate that Ubc9 expression in mature adipocytes is essential for maintaining WAT expansion. Deletion of Ubc9 in fat cells compromised and diminished adipocyte function that provoked WAT inflammation and ectopic lipid accumulation in the liver. Our findings reveal an indispensable role for Ubc9 during white adipocyte expansion and endocrine control of energy balance.

White adipose tissue (WAT) is an endocrine organ that dynamically expands and contracts to meet the metabolic demands of the organism. However, prolonged excessive caloric intake overwhelms WAT depots, resulting in peripheral fat deposition, systemic glucose dysregulation, tissue fibrosis, insulin resistance, and type 2 diabetes mellitus. The chronic triggers that degrade WAT function remain undefined, hindering the development of effective strategies to treat obesity and prevent its co-morbidities. We have identified a microRNA, miR-30a, that potently stimulates subcutaneous adipose tissue metabolic flexibility and enhances systemic insulin sensitivity. In follow-up studies, we created a genetically engineered mouse model to conclusively show miR-30a expression in white adipocytes is antidiabetic. Obese mice modified to express miR-30a at elevated levels in adipose tissues maintain insulin sensitivity coupled with reduced hepatic steatosis. Proteomic profiling indicated that miR-30a targets Plasminogen Activator Inhibitor-1 (PAI-1) to limit pro-fibrotic programs that would otherwise restrict subcutaneous WAT expansion and decrease insulin sensitivity. While visceral WAT depots remain a site of significant fibrosis in mice, subcutaneous WAT fibrosis is epidemiologically linked to metabolic disease risk in adult humans. In obese humans, miR-30a expression negatively correlates with PAI-1 levels in subcutaneous WAT, supporting an anti-fibrotic role for miR-30a that ultimately defends insulin sensitivity. Collectively, our observations linking miR-30a expression to insulin sensitivity suggest a new method that restores WAT homeostasis. Disclosure N. Chernis: None. P. Saha: None. J.B. Felix: None. P.M. Masschelin: None. L. Lian: None. A. Cox: None. K. Kim: None. H. Wu: None. S.M. Hartig: None. Funding American Diabetes Association (1-18-IBS-105 to S.M.H.); National Institutes of Health (R01DK114356)

Obesity fosters low-grade inflammation in white adipose tissue (WAT) that may contribute to the insulin resistance that characterizes type 2 diabetes mellitus (T2DM). However, the causal relationship of these events remains unclear. The established dominance of signal transducer and activator of transcription 1 (STAT1) function in the immune response suggests an obligate link between inflammation and the co-morbidities of obesity. To this end, we sought to determine how STAT1 activity in white adipocytes affects insulin sensitivity. STAT1 expression in WAT inversely correlated with fasting plasma glucose in both obese mice and humans. Metabolomic and gene expression profiling established STAT1 deletion in adipocytes (STAT1a-KO) enhanced mitochondrial function and accelerated TCA cycle flux coupled with reduced fat cell size in subcutaneous WAT depots. STAT1a-KO reduced WAT inflammation, but insulin resistance persisted in obese mice. Rather, elimination of type I cytokine interferon gamma (IFNγ) activity enhanced insulin sensitivity in diet-induced obesity. Our findings reveal a permissive mechanism that bridges WAT inflammation to whole-body insulin sensitivity.

Inactivating mutations in the copper transporter Atp7b result in Wilson’s disease. The Atp7b(−/−) mouse develops hallmarks of Wilson’s disease. The activity of several nuclear receptors decreased in Atp7b(−/−) mice, and nuclear receptors are critical for maintaining metabolic homeostasis. Therefore, we anticipated that Atp7b(−/−) mice would exhibit altered progression of diet-induced obesity, fatty liver, and insulin resistance. Following 10 wk on a chow or Western-type diet (40% kcal fat), parameters of glucose and lipid homeostasis were measured. Hepatic metabolites were measured by liquid chromatography–mass spectrometry and correlated with transcriptomic data. Atp7b(−/−) mice fed a chow diet presented with blunted body-weight gain over time, had lower fat mass, and were more glucose tolerant than wild type (WT) littermate controls. On the Western diet, Atp7b(−/−) mice exhibited reduced body weight, adiposity, and hepatic steatosis compared with WT controls. Atp7b(−/−) mice fed either diet were more insulin sensitive than WT controls; however, fasted Atp7b(−/−) mice exhibited hypoglycemia after administration of insulin due to an impaired glucose counterregulatory response, as evidenced by reduced hepatic glucose production. Coupling gene expression with metabolomic analyses, we observed striking changes in hepatic metabolic profiles in Atp7b(−/−) mice, including increases in glycolytic intermediates and components of the tricarboxylic acid cycle. In addition, the active phosphorylated form of AMP kinase was significantly increased in Atp7b(−/−) mice relative to WT controls. Alterations in hepatic metabolic profiles and nuclear receptor signaling were associated with improved glucose tolerance and insulin sensitivity as well as with impaired fasting glucose production in Atp7b(−/−) mice.

Bile acid (BA) homeostasis is tightly regulated by multiple transcription factors, including farnesoid X receptor (FXR) and small heterodimer partner (SHP). We previously reported that loss of the FXR/SHP axis causes severe intrahepatic cholestasis, similar to human progressive familial intrahepatic cholestasis type 5 (PFIC5). In this study, we found that constitutive androstane receptor (CAR) is endogenously activated in Fxr:Shp double knockout (DKO) mice. To test the hypothesis that CAR activation protects DKO mice from further liver damage, we generated Fxr;Shp;Car triple knockout (TKO) mice. In TKO mice, residual adenosine triphosphate (ATP) binding cassette, subfamily B member 11 (ABCB11; alias bile salt export pump [BSEP]) function and fecal BA excretion are completely impaired, resulting in severe hepatic and biliary damage due to excess BA overload. In addition, we discovered that pharmacologic CAR activation has different effects on intrahepatic cholestasis of different etiologies. In DKO mice, CAR agonist 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP; here on TC) treatment attenuated cholestatic liver injury, as expected. However, in the PFIC2 model Bsep knockout (BKO) mice, TC treatment exhibited opposite effects that reflect increased BA accumulation and liver injury. These contrasting results may be linked to differential regulation of systemic cholesterol homeostasis in DKO and BKO livers. TC treatment selectively up-regulated hepatic cholesterol levels in BKO mice, supporting de novo BA synthesis. Conclusion: CAR activation in DKO mice is generally protective against cholestatic liver injury in these mice, which model PFIC5, but not in the PFIC2 model BKO mice. Our results emphasize the importance of the genetic and physiologic background when implementing targeted therapies to treat intrahepatic cholestasis.

Progressive familial intrahepatic cholestasis (PFIC) is a genetically heterogeneous disorder of bile flow disruption due to abnormal canalicular transport or impaired bile acid (BA) metabolism, causing excess BA accumulation and liver failure. We previously reported an intrahepatic cholestasis mouse model based on loss of function of both farnesoid X receptor (FXR; NR1H4) and a small heterodimer partner (SHP; NR0B2) [double knockout (DKO)], which has strong similarities to human PFIC5. We compared the pathogenesis of DKO livers with that of another intrahepatic cholestasis model,Bsep−/−, which represents human PFIC2. Both models exhibit severe hepatomegaly and hepatic BA accumulation, but DKO showed greater circulating BA and liver injury, andBsep−/− had milder phenotypes. Molecular profiling of BAs uncovered specific enrichment of cholic acid (CA)–derived BAs in DKO livers but chenodeoxycholate-derived BAs inBsep−/− livers. Transcriptomic and proteomic analysis revealed specific activation of CA synthesis and alternative basolateral BA transport in DKO but increased chenodeoxycholic acid synthesis and canalicular transport inBsep−/−. The constitutive androstane receptor (CAR)/pregnane X receptor (PXR)–CYP2B/CYP2C axis is activated in DKO livers but not in other cholestasis models. Loss of this axis inFxr:Shp:Car:Pxr quadruple knockouts blockedCyp2b/Cyp2c gene induction, impaired bilirubin conjugation/elimination, and increased liver injury. Differential CYP2B expression in DKO andBsep−/− was recapitulated in human PFIC5 and PFIC2 livers. In conclusion, loss of FXR/SHP results in distinct molecular pathogenesis and CAR/PXR activation, which promotesCyp2b/Cyp2c gene transcription and bilirubin clearance. CAR/PXR activation was not observed inBsep−/− mice or PFIC2 patients. These findings provide a deeper understanding of the heterogeneity of intrahepatic cholestasis., Activation of the CAR/PXR-CYP2B/CYP2C axis is specific for molecular pathogenesis of the PFIC5 mouse model (Fxr−/−;Shp−/−), whereas it is impaired in the PFIC2 mouse model (Bsep−/−).

The present study addresses the aqueous phase removal of hexavalent chromium [Cr(VI)] using a composite adsorbent. An adsorbent was prepared by immobilizing hydrous zirconium oxide (HZO) into alginate beads (HZO@AB). The material was characterized via scanning electron microscopy (SEM), which showed an increase in surface roughness after HZO immobilization. The impregnation of alginate beads with HZO and successful adsorption of Cr(VI) onto HZO@AB were analyzed by X-ray photoelectron spectroscopy (XPS). The adsorption of Cr(VI) was well described by a pseudo-second-order kinetic model (R2 > 0.99). The Langmuir model provided the best correlation with adsorption of Cr(VI) on HZO@AB (R2 > 0.99), supporting the nature of adsorption as a monolayer. The adsorption of Cr(VI) [as CrO42− and HCrO4−)] was effective between pH 3 and 6 and decreased at higher pH. The maximum equilibrium sorption capacity of HZO@AB for Cr(VI) was 9.8 mg g-1 at pH 5. The thermodynamic parameters (ΔG°, ΔH°, ΔS°) were evaluated, and the process of Cr(VI) adsorption on HZO@AB was identified as endothermic, thermodynamically feasible, and spontaneous (‒ΔG°). The adsorbent was satisfactorily reused for three consecutive cycles. The adsorption capacity of regenerated composite adsorbent for Cr(VI) was 6.92 mg g-1 in the third cycle, and the adsorbent retained approximately 73% of its original Cr(VI) adsorption capacity. The results of this study indicate that the developed adsorbent (HZO@AB) could be a potential candidate for treating wastewater containing Cr(VI).

This study investigated the influence of physical education class (PEC) as an intervention method for aggression, sociality, stress, and physical fitness levels in children from multicultural families. The hypothesis was that participating in PEC would result in reduced aggression and stress and improved sociality and physical fitness in multicultural children. A three-item questionnaire, a body composition test, and physical fitness tests were given three times. Eighty-four subjects were divided into four groups: multicultural children who participated in PEC (multi-PEG, n=12), multicultural children who did not participate in PEC (multi-NPEG, n=13), single-cultural children who participated in PEC (sing-PEG, n=11), and single-cultural children who did not participate in PEC (sing-NPEG, n=12), respectively. Parametric and nonparametric statistical methods were conducted on the collected data with a significance level set a priori at P

Most protocols for testing and rehabilitation for recovery and improvement of muscular endurance have been set at 180°/sec, 240°/sec, and 300°/sec. These protocols can cause confusion to clinical providers or other researchers. This study was aimed at investigating the optimal isokinetic angular speed for measuring or developing muscular endurance after assessing the relationship between cardiopulmonary responses and isokinetic moments. This study was conducted with 31 male and female college students. Graded exercise test and body composition were measured as well as the isokinetic moments of the knee muscles at three angular speeds: 180°/sec, 240°/sec, and 300°/sec. The specific isokinetic moments of knee muscles that were measured included: peak torque (PT) and total work (TW) on extensor (e) and flexor (f) of knee joints, which were denoted as ePT180, fPT180, eTW180, fTW180, ePT240, fPT240, eTW240, fTW240, ePT300, fPT300, eTW300, and fTW300 according to the three angular speeds. Spearman correlation test was used to examine the relationship between the sum means of cardiopulmonary responses and the variables of isokinetic moments. This study confirmed that the optimal angular speed for testing or training for muscular endurance was 180°/sec, which showed a stronger relationship between cardiopulmonary responses and isokinetic moments. Therefore, this angular speed is recommended for testing and training for muscular endurance of the knee joints.

Background and aims Liver receptor homolog-1 (LRH-1; NR5A2) is a nuclear receptor that regulates metabolic homeostasis in the liver. Previous studies identified phosphatidylcholines as potential endogenous agonist ligands for LRH-1. In the liver, distinct subsets of phosphatidylcholine species are generated by two different pathways: choline addition to phosphatidic acid through the Kennedy pathway and trimethylation of phosphatidylethanolamine through phosphatidylethanolamine N-methyl transferase (PEMT). Approach and results Here, we report that a PEMT-LRH-1 pathway specifically couples methyl metabolism and mitochondrial activities in hepatocytes. We show that the loss of Lrh-1 reduces mitochondrial number, basal respiration, beta-oxidation, and adenosine triphosphate production in hepatocytes and decreases expression of mitochondrial biogenesis and beta-oxidation genes. In contrast, activation of LRH-1 by its phosphatidylcholine agonists exerts opposite effects. While disruption of the Kennedy pathway does not affect the LRH-1-mediated regulation of mitochondrial activities, genetic or pharmaceutical inhibition of the PEMT pathway recapitulates the effects of Lrh-1 knockdown on mitochondria. Furthermore, we show that S-adenosyl methionine, a cofactor required for PEMT, is sufficient to induce Lrh-1 transactivation and consequently mitochondrial biogenesis. Conclusions A PEMT-LRH-1 axis regulates mitochondrial biogenesis and beta-oxidation in hepatocytes.

Objective White adipose tissue (WAT) expansion regulates energy balance and overall metabolic homeostasis. The absence or loss of WAT occurring through lipodystrophy and lipoatrophy contributes to the development of hepatic steatosis and insulin resistance. We previously demonstrated that sole small ubiquitin-like modifier (SUMO) E2-conjugating enzyme Ube2i represses human adipocyte differentiation. The role of Ube2i during WAT development remains unknown. Methods To determine how Ube2i impacts body composition and energy balance, we generated adipocyte-specific Ube2i knockout mice (Ube2ia-KO). CRISPR/Cas9 gene editing inserted loxP sites flanking exons 3 and 4 at the Ube2i locus. Subsequent genetic crosses to Adipoq-Cre transgenic mice allowed deletion of Ube2i in white and brown adipocytes. We measured multiple metabolic endpoints that describe energy balance and carbohydrate metabolism in Ube2ia-KO and littermate controls during postnatal growth. Results Surprisingly, Ube2ia-KO mice developed hyperinsulinemia and hepatic steatosis. Global energy balance defects emerged from dysfunctional WAT marked by pronounced local inflammation, loss of serum adipokines, hepatomegaly, and near absence of major adipose tissue depots. We observed progressive lipoatrophy that commences in the early adolescent period. Conclusions Our results demonstrate that Ube2i expression in mature adipocytes allows WAT expansion during postnatal growth. Deletion of Ube2i in fat cells compromises and diminishes adipocyte function that induces WAT inflammation and ectopic lipid accumulation in the liver. Our findings reveal an indispensable role for Ube2i during white adipocyte expansion and endocrine control of energy balance., Highlights • A new mouse model reveals that Ube2i loss in fat cells impacts body composition. • Ube2i fat-specific knockout (Ube2ia-KO) causes fatty liver and hyperinsulinemia. • Ube2ia-KO mice develop metabolic inflexibility and cold intolerance. • Inflammation and caspase activation of cell death occur in Ube2ia-KO adipocytes.