High Fat Research Articles

Overweight Leads to an Increase in Vitamin E Absorption and Status in Mice.

This study investigates whether vitamin E (VE) deficiency in subjects with obesity could, at least partly, be due to a defect in VE intestinal absorption. Mice follow either a high-fat (HF) or a control (CTL) diet for 12 weeks. The study evaluates their VE status, the expression of genes encoding proteins involved in lipid and fat-soluble vitamin intestinal absorption, and VE absorption using a γ-tocopherol-rich emulsion. HF mice have a weight (+23.0%) and an adiposity index (AI, +157.0) superior to CTL mice (p<0.05). α-Tocopherol concentrations are higher in both plasma (+45.0%) and liver (+116.9%) of HF mice compared to CTL mice (p<0.05). α-Tocopherol concentration in the adipose tissue of HF mice is higher than that of CTL mice after correction by the AI (+72.4%, p<0.05). No difference is found in the expression of genes coding for proteins involved in intestinal lipid metabolism in fasting mice. After force-feeding, γ-tocopherol plasma concentration is higher in HF mice compared to CTL mice (+181.5% at 1.5h after force-feeding, p<0.05). HF mice display higher status and more efficient absorption of VE than CTL mice. VE absorption is thus likely not impaired in the early stages of obesity.

Computational Identification of Milk Trait Regulation Through Transcription Factor Cooperation in Murciano-Granadina Goats

The Murciano-Granadina goat (MUG) is a renowned dairy breed, known for its adaptability and resilience, as well as for its exceptional milk traits characterized by high protein and fat content, along with low somatic cell counts. These traits are governed by complex biological processes, crucial in shaping phenotypic diversity. Thus, it is imperative to explore the factors regulating milk production and lactation for this breed. In this study, we investigated the genetic architecture of seven milk traits in MUGs, employing a two-step computational analysis to examine genotype–phenotype associations. Initially, a random forest algorithm identified the relative importance of each single-nucleotide polymorphism (SNP) in determining the traits of interest. The second step applied an information theory-based approach to exploring the complex genetic architecture of quantitative milk traits, focusing on epistatic interactions that may have been overlooked in the first step. These approaches allowed us to identify an almost distinct set of candidate genes for each trait. In contrast, by analyzing the promoter regions of these genes, we revealed common regulatory networks among the milk traits under study. These findings are crucial for understanding the molecular mechanisms underlying gene regulation, and they highlight the pivotal role of transcription factors (TFs) and their preferential interactions in the development of these traits. Notably, TFs such as DBP, HAND1E47, HOXA4, PPARA, and THAP1 were consistently identified for all traits, highlighting their important roles in immunity within the mammary gland and milk production during lactation.

Impact of L-carnitine supplementation on gastric emptying and bowel function in pediatric ketogenic diet therapy: a clinical trial.

Ketogenic diet (KD) is an excess fat, enough protein, and minimal carbohydrate diet. The high fat content in KD lowers the oesophageal sphincter tone, slows gastric emptying, and decreases intestinal transit time. The primary aim of the current clinical trial was to study the effect of L-carnitine supplementation on gastric emptying in children with drug resistant epilepsy (DRE) on KD. Assessment of the protective effect of L-carnitine on bowel function and habits in those patients was a secondary aim. The current study recruited 30 patients aged 12 months to 18 years newly diagnosed with DRE assigned to start KD who were following up at the Pediatric Clinical Nutrition and Neurology Outpatient Clinics or were admitted due to DRE at the Pediatric Neurology Inpatient Department, Children's Hospital, Ain Shams University (Egypt). Participants were assigned randomly into 2 arms; arm I: received KD with L-carnitine supplementation, arm II: received KD only. Patients were assessed at baseline and after 3 months of starting KD, the assessments of children included: 24-hour dietary recall, Chalfont Seizures Severity Scale, gastrointestinal symptoms score and Bristol stool chart, frequency of defecation per week, anthropometric measurements assessment, fasting serum lipid profile and measurement of the antral length by ultrasound. There was significant increase in antral length in the patients who received KD with L-carnitine supplementation compared to the non-supplemented group. The antral length showed a significant negative correlation with GI symptoms score in all cases and the L-carnitine supplemented group. It also showed a significant positive correlation with Bristol stool score in all patients and a significant positive correlation with stool frequency in the L-carnitine supplemented group only. L-carnitine supplementation to children with DRE on KD has a significant role in improving gastric motility and it increases the frequency of defecation. Further studies are recommended to explore additional benefits, meanwhile it is prudent to advise L-carnitine supplementation for such patients.

Ethanol extract of cassia seed alleviates metabolic dysfunction-associated steatotic liver disease by acting on multiple lipid metabolism-related pathways

Abstract Background and objective In northern China’s cold regions, the prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) exceeds 50%, significantly higher than the national and global rates. MASLD is an important risk factor for cardiovascular and cerebrovascular diseases, including coronary heart disease, stroke, and tumors, with no specific therapeutic drugs currently available. The ethanol extract of cassia seed (CSEE) has shown promise in lowering blood lipids and improving hepatic steatosis, but its mechanism in treating MASLD remains underexplored. This study aims to investigate the therapeutic effects and mechanisms of CSEE. Methods MASLD models were established in male Wistar rats and golden hamsters using a high fat diet (HFD). CSEE (10, 50, 250 mg/kg) was administered via gavage for six weeks. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), as well as liver TC and TG, were measured using biochemical kits. Histopathological changes in the liver were evaluated using Oil Red O staining, Hematoxylin-eosin (H&amp;E) staining, and transmission electron microscopy (TEM). HepG2 cell viability was assessed using the cell counting kit-8 (CCK8) and Calcein-AM/PI staining. Network pharmacology was used to analyze drug-disease targets, and western blotting was used to confirm these predictions. Results CSEE treatment significantly reduced serum levels of TC, TG, LDL-C, ALT, and AST, and improved liver weight, liver index, and hepatic lipid deposition in rats and golden hamsters. In addition, CSEE alleviated free fatty acid (FFA)-induced lipid deposition in HepG2 cells. Molecular biology experiments demonstrated that CSEE increased the protein levels of p-AMPK, p-ACC, PPARα, CPT1A, PI3K P110 and p-AKT, while decreasing the protein levels of SREBP1, FASN, C/EBPα, and PPARγ, thus improving hepatic lipid metabolism and reducing lipid deposition. The beneficial effects of CSEE were reversed by small molecule inhibitors of the signaling pathways in vitro. Conclusion CSEE improves liver lipid metabolism and reduces lipid droplet deposition in Wistar rats and golden hamsters with MASLD by activating hepatic AMPK, PPARα, and PI3K/AKT signaling pathways.

Optimization of the Combined Use of Z-Sep Plus and EMR-Lipid in QuEChERS Procedure for the Analysis of Eight Pesticides in Real Milk Samples

AbstractOne of the most applied procedures for the determination of trace analytes in complex matrices is QuEChERS (an acronym for Quick, Easy, Cheap, Effective, Rugged, and Safe). QuEChERS procedures include an extraction step followed by a dispersive solid-phase extraction (dSPE) for analytes cleaning-up from the matrix components. A challenging task in QuEChERS procedures is extracting and determining pesticides from samples of high fat such as milk samples. This challenge induced the innovation of new adsorbents for the clean-up step such as Z-Sep Plus® and EMR-Lipid® to enable removal of fatty matrix components without affecting the recovery of hydrophobic analytes. This work aims to apply experimental design to optimize the combined application of both QuEChERS clean-up adsorbents; Z-Sep Plus® and EMR-Lipid® in addition to other QuEChERS parameters in the determination of eight pesticides: hexachlorocyclohexane, dichlorodiphenyldichloroethane, dichlorodiphenyltrichloroethane, primiphos ethyl, diazinon, malathion, endrin, and dimethoate in milk matrix. This was augmented by optimization of GC–MS/MS and UPLC-MS/MS to detect and determine analytes in extracts. The experimental design of QuEChERS procedure enabled the optimization of Z-Sep Plus®- and EMR-Lipid®-added adsorbent amounts with other method parameters to enable the maximum recovery of analytes. Furthermore, the optimized methods enabled low detection limits of the studied pesticides within a short analysis time (28 min for GC and 12 min for LC methods, respectively). The procedure was validated according to European SANTE/11312/2021 Guideline. Quantitation limit ranged from 1.7 to 3.2 ng/mL for GC–MS/MS method and from 1.7 to 3 ng/mL for UPLC-MS/MS method. Greenness assessment of the methods followed four approaches indicating an excellent value of greenness for the proposed methods. Furthermore, 45 real milk samples collected from the Egyptian market were tested with the developed procedure for the presence of pesticides.

Effect of very low-carbohydrate high-fat diet and high-intensity interval training on mental health-related indicators in individuals with excessive weight or obesity.

Very low carbohydrate high fat (VLCHF) diet and high-intensity interval training (HIIT) are widely utilized for weight reduction and cardiorespiratory fitness improvement, respectively. To assess the acceptability of these approaches, it is essential to examine mental health-related indicators. This secondary analysis of a randomized controlled trial investigated the isolated and synergistic effects of VLCHF and HIIT on mental health-related indicators in individuals with excessive weight or obesity. Sixty-eight participants (age = 42 ± 10.2; 20-60 years; BMI = 29.8 ± 3.7) were analysed across four groups: HIIT (n = 15, 4 males, 11 females), VLCHF (n = 19, 4 males, 15 females), VLCHF + HIIT (n = 19, 4 males, 15 females), and control (n = 15, 4 males, 11 females). The 12-week intervention, involved VLCHF diet or HIIT sessions, depending on group affiliation and completing online questionnaires via Qualtrics software before and after the intervention. The questionnaires included the 12-item Short Form Survey (SF-12) for mental (MHS) and physical health scores (PHS), the Satisfaction with Life Scale (SWLS), and the Perceived Stress Scale (PSS). Using the Kruskal-Wallis test, we found no significant differences in mental health-related indicators between groups after 12 weeks, except for SWLS (p = 0.031; ES = 0.133; medium), which improved significantly in the VLCHF + HIIT group compared to the HIIT group. Our findings indicate that HIIT and VLCHF, alone or combined, do not significantly affect mental health-related indicators.

Complete inhibition of liver acetyl-CoA carboxylase activity is required to exacerbate liver tumorigenesis in mice treated with diethylnitrosamine.

The metabolic pathway of de novo lipogenesis (DNL) is upregulated in fatty liver disease and liver cancer. Inhibitors of DNL are in development for the treatment of these disorders; however, our previous study showed that blocking DNL unexpectedly exacerbated liver tumorigenesis when liver acetyl-CoA carboxylase (ACC) 1 and 2 enzymes were deleted in mice treated with diethylnitrosamine (DEN) and fed high fat diet. Herein, we used 3 new approaches including ACC1 vs. ACC2 isotype-selective inhibition, delaying ACC inhibition until after carcinogen treatment, and feeding mice normal chow diet to better understand the impact of ACC inhibition on liver tumorigenesis. Six genotypes of female C57BL/6J mice with floxed ACC1 and/or ACC2 alleles were injected with DEN at 2 weeks of age followed by liver-specific knockout of ACC genes at 9 weeks. Mice were fed a normal chow diet and evaluated at 52 weeks for liver tumours. Compared to the wildtype control group, no genotype decreased tumour multiplicity or burden; however, mice completely lacking liver ACC1 and ACC2 had > 5-fold increases in liver tumour multiplicity and burden. ACC inhibition exacerbated DEN-induced liver tumorigenesis only when both ACC isotypes were completely inhibited. The pro-tumour phenotype of ACC inhibition was strongly reproducible irrespective of chow or high fat feeding, and irrespective of ACC inhibition prior to or after DEN treatment. Retaining partial ACC activity at either isotype prevented tumour exacerbation in mice at risk for developing liver tumours.

Contextual effects of color on food choices: Red ambient color induces indulgence

AbstractThis research examines how red ambient color in restaurants/cafeterias influences food choices. Prior research shows that red directly related to a food product (such as on nutrition labels or plates/cups) leads to avoidance of unhealthy foods. Yet, many successful fast‐food restaurants (e.g., McDonald's and Dairy Queen) use red in their ambiance, suggesting that in the context of food/eating, ambient red may have a different meaning than product‐related red. Indeed, the current research shows that consumers associate ambient red (e.g., wall color) with unhealthy restaurants. The presence of ambient red (vs. blue, gray, or white) leads to greater preference for unhealthy (i.e., high calorie, high fat, and indulgent) food options because consumers draw on ambient factors to make inferences about products sold in a retail establishment and then choose contextually appropriate products. The effect is moderated by the extent to which a consumer associates red with unhealthy restaurants. This research highlights how, in the context of food/eating, the placement of red influences its meaning and ultimately whether consumers approach or avoid unhealthy foods.

Abstract 4136786: Prediabetes increases platelet activation and thrombotic susceptibility via mitochondrial oxidants

Recent studies suggest that prediabetes is not only a prelude to diabetes, but also is an independent risk factor for cardiovascular thrombotic events in young and aged people. However, the mechanisms that may promote platelet activation and thrombosis during prediabetes have not been studied. We hypothesized that in prediabetes, dysregulation of electron transfer and accumulation of mitochondrial oxidants promotes platelet activation and thrombosis, and that aging exacerbates the phenotype. We recruited young (18-50 year) and middle-aged (51-64 year) Veterans with prediabetes and age- and BMI- matched healthy Veterans. Compared to healthy cohort, platelets from prediabetes exhibited enhanced agonist induced platelet activation, mitochondrial oxidants, membrane potential and thrombus formation ex vivo in both young and middle-aged cohorts to a similar extent. Preincubation of platelets with either a mitochondria targetable superoxide dismutase (SOD) mimetic or MitoQ (transfers electrons from Complex I/II to complex III to lower superoxide generation within mitochondria), rescued the prothrombotic phenotype. We confirmed these platelet phenotypes in C57BL6/J mice fed high fat (HF) diet for 2 weeks (model of early diabetes). Importantly, HF-fed mice showed increased susceptibility to carotid artery and pulmonary thrombosis in vivo . Pharmacological treatment of HF-fed mice with SOD-mimetic or MitoQ, or genetic overexpression of catalase within mitochondria, rescued the prothrombotic state. This is the first report showing platelet hyperactivity and enhanced thrombotic susceptibility in prediabetes. Further, our findings suggest that enhanced mitochondrial oxidants due to the premature electron escape contribute to the phenotype. Our data imply a therapeutic potential of SOD-mimetic and MitoQ in lowering thrombotic burden in prediabetes.

Abstract 4136294: Carbohydrate intakes and cardiovascular disease risk factors in South Asians: Results from 56,024 participants in Bangladesh, India, Pakistan, and Sri Lanka

Background: Evidence on effects of carbohydrate intake with cardiovascular disease risk factors is inconsistent and has limited generalizability to diverse populations such as South Asians, whose carbohydrate intakes are typically high. Aims: We examined the association of the quantity and types of carbohydrate intake with two risk factors for major cardiovascular events– hypertension (HT) and hypercholesterolemia (HC). Methods: We analyzed cross-sectional data from participants aged ≥18 years in the South Asia Biobank in Bangladesh, India, Pakistan, and Sri Lanka. Using dietary data assessed with interviewer-led 24h diet recalls, we calculated intakes of total and subtypes of carbohydrates and several indices of carbohydrate quality. Primary outcomes were HT and HC, defined by medication use, doctor diagnosis, blood pressure and blood lipid levels. With mixed effects Poisson regression, HT and HC was regressed on quantiles of carbohydrate intakes by country to estimate prevalence ratios (PR) and 95% confidence intervals (CI), followed by meta-analysis pooling country-specific estimates. We examined interactions by diabetes status and adiposity. Results: Of 56,024 adults, 36.6% had HT, 16.1% had HC. In pooled multivariable analyses comparing extreme quantiles, total carbohydrate intake was not associated with HT or HC prevalence. Fructose, free sugar and free sugar-to-fiber ratio were associated with higher prevalence of HT (PRs Q5vsQ1 : 1.03-1.15). Higher intakes of cereal fiber (PR Q4vsQ1 : 0.96, 95% CI: 0.95-0.98) and legume fiber (PR Q4vsQ1 : 0.98, 95% CI: 0.97-0.98) were associated with lower prevalence of HT and higher cereal fiber intake with lower HC prevalence (PR Q5vsQ1 : 0.80, 95% CI: 0.75-0.85). Diabetes and adiposity were observed as effect-modifiers (p&lt;0.001). There were null associations in adults with diabetes but in non-diabetic adults, free sugar intake was related positively with HT (PR Q5vsQ1 : 1.07, 95% CI: 1.05-1.09) while fiber intake was inversely related (PR Q5vsQ1 : 0.98, 0.95-0.97). Starch, sugar and free sugar intake were associated positively with HT prevalence in adults with high body fat (≥20% for men; ≥30% for women) and visceral fat (≥10%) levels (PRs Q5vsQ1 : 1.03-1.20); associations were null for low body and visceral fat levels. Associations varied by country (between-country heterogeneity I 2 &gt;60%). Conclusion: Associations of dietary carbohydrates with HT and HC depended on the quality and type of intake in South Asians.

Abstract 4135301: Cardiac specific gene therapy to treat diabetic heart failure with preserved ejection fraction

Background: We have previously demonstrated that diabetes mellitus (DM) causes cardiac inflammation leading to heart failure with preserved ejection fraction (HFpEF). Arachidonate 5-lipoxygenase (5-LOX), encoded by the ALOX5 gene, is an enzyme that is highly expressed in leukocytes. 5-LOX synthesizes specialized pro-resolving mediators (SPMs) including Resolvin D1 (RvD1), which reduce inflammation. Hypothesis: We hypothesize that cardiac-specific ALOX5 upregulation could prevent diabetes-associated HFpEF without inducing systemic immunosuppression. Methods: DM was induced by feeding mice with a high fat diet (HFD, 60% fat by kcal) starting from 6 weeks of age for 22-24 weeks. At 24 weeks of age, DM mice were treated with RvD1, by intraperitoneal injection at 100 ng/mouse for 2 weeks. To specifically upregulate 5-LOX in the heart, another cohort of DM mice were injected with either AAV9-plain or AAV9- ALOX5 viral vectors under the control of the α-myosin heavy chain (MHC) at 22 weeks old. At 28 weeks, we performed echocardiographic measurement of diastolic function and heart extraction. Results: HFD-induced DM mice showed lower cardiac 5-LOX expression (0.72 ± 0.05 a.u. vs. 1.09 ± 0.12, p &lt; 0.05) and RvD1 levels (1 ± 0.08 vs. 1.9 ± 0.14 fold change over DM mice, p&lt;0.0001) than the mice with normal diet. RvD1 treatment significantly improved DM-associated diastolic function (19.7 ± 1.4 a.u. vs. 22.9 ± 0.4, p&lt;0.05). With cardiac specific upregulation of 5-LOX, cardiac 5-LOX expression (0.89 ± 0.06 vs. 0.70 ± 0.04 a.u., p&lt;0.05) and RvD1 levels were increased (2.1 ± 0.3 a.u. vs. 1.2 ± 0.2, normalized to AAV9-plain, p&lt;0.05). AAV9- ALOX5 also reduced NLRP3 inflammasome activity (0.80 ± 0.02 a.u. vs. 0.90 ± 0.02, p&lt;0.01) and improved DM-associated diastolic dysfunction (17.0 ± 1.2 a.u. vs. 20.5 ± 1.0, p&lt;0.05) without affecting glucose metabolism and obesity in DM mice. Moreover, no alterations of cardiac macrophage infiltration, inflammatory IL1β signaling, or macrophage phenotype were observed in response to cardiac-specific ALOX5 upregulation. Conclusion: In conclusion, both direct RvD1 treatment and cardiac-specific gene therapy to enhance resolution of inflammation in cardiomyocytes reversed DM-associated HFpEF. Gene therapy avoided off-target immunosuppression. Therefore, AAV-directed cardiac ALOX5 upregulation represents a novel cardiac-specific gene therapy for HFpEF that avoids systemic immunosuppression.

Abstract 4132968: CCR5 modulation enhances M-MDSCs functionality in early atherosclerosis

Background: The role of immune suppressive microenvironment, promulgated by the presence of Myeloid Derived Suppressor Cells (MDSC) have been the major focus of research delineating the underlying mechanisms of cardiovascular diseases (CVD). CCR5, a chemokine receptor, has been associated with both immunosuppressive and inflammatory phenotypes, however, the possible role of CCR5 pertaining to MDSCs in the development of atherosclerosis has not been elucidated yet. Therefore, we investigated the therapeutic potential of CCR5 modulation in early atherosclerosis progression. Methods: CCR5 receptor-ligand expression was studied with flow cytometry, real time PCR and immunocytochemistry in 65 individuals (25 Healthy controls, 20 young hypercholesterolemic individuals, and 20 stable [Coronary Artery Disease] CAD patients). To induce atherosclerosis, mice were fed high fat diet (HFD) for 6 weeks following partial ligation of left carotid artery in C57BL6 mice. MDSCs phenotype and functionality were assessed using CCR5 inhibitor in vitro (10 -8 M) and in vivo (i.p 3ng/day for 15 days) . M-MDSCs phenotype switch, atherosclerotic lesion development and plaque phenotype was studied in vivo . Results: We observed CCR5 elevation on Monocytic-MDSCs in the early phase of atherosclerosis. M-MDSCs displayed an intermediate phenotype with expression of both pro and anti-inflammatory mediators in early atherosclerosis, a similar trend was observed in inflammatory cytokines stimulated M-MDSCs ( in vitro ). Notably, DAPTA treatment shifted phenotype of M-MDSCs to anti-inflammatory profile with elevated expression of IL-10 and reduced expression of IFNγ, IL4, and IL22. Further, CCR5 modulation expanded IL10+M-MDSC in vivo . Additionally, DAPTA reduced the migratory potential, reduced cholesterol uptake and improved the functionality (suppression of T cell) of M-MDSCs. Blocking CCR5 with DAPTA led to the formation of stable plaque in HFD-fed mice. Conclusion: Increased expression of CCR5 coupled with inflammatory cytokine secretion by regulatory cells lead to dysfunctional M-MDSCs in CVD risk factor population. Possibly this dysfunctionality contributes to the development and progression of CVD including atherosclerosis. Our findings show targeting of CCR5 with DAPTA reduces atherogenesis, increases plaque stabilization and corrects phenotypic alterations in immune suppressive M-MDSCs. The functionality and phenotype of M-MDSCs in atherosclerosis, may be ameliorated by targeting CCR5.

Abstract 4138344: High fat diet induced the premature senescence of white adipose tissue through downregulating SIRT1 expression and promoting its S-nitroslyation modification

Background and Objectives: The dysfunction of subcutaneous white adipose tissue is one of the early manifestations of obesity, which was associated with accelerated cellular process in aging induced by high fat-diet. The premature senescence of adipose-derived mesenchymal stem cells (AMSCs) is the key factor contributing to the aging of subcutaneous white adipose tissue. However, the underlying mechanism was not clear. Here, we explored the role and mechanism of obesity on the aging of white adipose tissue and AMSCs, and evaluated the anti-aging effect of resveratrol on this process. Methods: Subcutaneous white adipose tissue in non-elderly obese individuals and mice fed with high fat-diet was obtained to detect the TG metabolism and premature senescence. Then the postprandial triglyceride-rich lipoproteins (TRL) after high-fat diet was extracted, and the role and mechanism of TRL on the senescence of AMSCs were explored. Finally, the anti-aging effect of resveratrol on the white adipose tissue and AMSCs was evaluated. Results: Postprandial TG metabolism was abnormal in non-elderly obese humans and mice, and high-fat diet remarkably aggravated white adipose tissue senescence in mice (Fig. 1). TRL induced the premature senescence of AMSCs, with the downregulation of of silent mating-type information regulation 2 homolog 1 (SIRT1) via protein kinase B/forkhead box protein O1 pathway. Besides, the S-nitrosylation of SIRT1 was increased which was regulated by ten-eleven translocation 1/S-nitrosoglutathione reductase pathway. Resveratrol, a SIRT1 agonist, inhibited the TRL-induced AMSCs senescence, resulting in the amelioration of white adipose tissue senescence in obese mice (Fig. 2). Conclusions: These findings established a new mechanism for high-fat diet-induced white adipose tissue senescence and provided a potential therapeutic strategy for high-fat diet induced white adipose tissue senescence and metabolic disorders.

Abstract Sa1108: The Aberrant Autophagy-Apoptosis regulation Mediates the Severe Neurological Dysfunction after Cardiac Arrest in Diabetic Mice

Background: Patients with diabetes have lower survival and worse neurological outcome after cardiopulmonary resuscitation (CPR) compared with non-diabetes. Previous studies have mentioned the aberrant autophagy in diabetic mice. However, whether this abnormal autophagy exacerbates neuronal apoptosis and neurological outcome in diabetic mice remains unclear. Methods: Patients were divided into non-diabetic cardiac arrest (CA) patients and diabetic CA patients, and the survival and neurological outcomes at discharge and day 28 after ROSC were compared between these two groups. Diabetic mice were fed with high fat diet (HFD), and CA model was established by inducing ventricular fibrillation electrically. The neurological prognosis was assessed by open field test (OPT), novel object recognition (NOR) and Y-maze. Neuronal apoptosis and autophagy were evaluated by immunofluorescence and western blotting, including TUNEL, cleaved caspase 3, LC3B, p62 and beclin1. Results: Compared with non-diabetic patients, the survival at discharge and at day 28 after ROSC are significantly lower in IHCA patients with diabetes. Meanwhile, the neurological deficits at discharge and at day 28 after ROSC are much severer in diabetic-CA patients. In addition, the lower survival and worse neurological function have been discovered, and the cognition impairment is more significant after ROSC in HFD mice. Neuronal apoptosis is severer in HFD mice. In ND mice, the activated autophagy has been revealed after CA-CPR-ROSC, however, autophagy can not be activated after ROSC in HFD mice since the baseline autophagy is at a higher level when compared with ND mice. Neuronal apoptosis is exacerbated after autophagy inhibition by 3-MA in ND mice, while after autophagy activation by rapamycin in HFD mice, apoptosis is attenuated after ROSC. Conclusion: Diabetes is the risk factor for neurological outcome in IHCA patients. The aberrant autophagy regulation in cortex neurons contributes to the severe apoptosis and neurological deficits after ROSC in diabetic mice.

Abstract 4147602: The Paradox Role of Sirtuin 6 In Coronary Microvascular Function under Metabolic Stress

Coronary microvascular dysfunction (CMD), which is associated with diabetic cardiomyopathy, Takotsubo cardiomyopathy, andheart failure with preserved ejection fraction (HFpEF), is understudied. CMD is characterized by impaired endothelial-dependent vasodilation, but detailed mechanisms have yet to be elucidated. Nuclear Sirtuin 6 (SIRT6) plays essential roles in gene transcriptional, stress tolerance, DNA repair, inflammation, and aging. SIRT6 is strongly associated with cardiovascular pathologies, but how SIRT6 regulates endothelial metabolisms and homeostasis under metabolic stress and the underlying mechanism remains poorly understood. It might be because global Sirt6 knockout mice are perinatally lethal caused by hypoglycemia, suggesting the essential role of SIRT6 in glucose metabolism. In our preliminary studies, we generated inducible global Sirt6 knockout mice by crossing with Sirt6 f/f mice with CAG-cre (Sirt6 f/f, CAG ), and mice were viable with normal glucose levels. However, they showed impaired endothelial-dependent dilation (EDD) and impaired coronary flow reserve (CFR), an index clinically used to diagnose CMD. It suggests that deletion of Sirt6 might cause EC dysfunction because Sirt6 is reported to protect EC from premature senescence and oxidative stress by sustaining high eNOS levels. Surprisingly, when we studied non-inducible Sirt6 endothelial-specific knockout (Sirt6 f/f, tie-2 cre ) and inducible Sirt6 endothelial-specific knockout (Sirt6 f/f, Cdh5-cre/ERT2 ) and wild-type (WT) mice, Sirt6 f/f, Tie-2 and Sirt6 f/f, Cadh5 mice do not phenocopy the inducible global SIRT6 knockout mice, they had normal EDD and CFR. When the mice were fed a high fat and high sugar (HFHS) diet, the Sirt6 f/f, Tie-2 and Sirt6 f/f, Cadh5 had impaired EDD, suggesting Sirt 6 functioned differently in the mice fed with chow diet or HFHS diet. We hypothesize Sirt 6 deficiency causes coronary endothelial dysfunction and contributes to CMD; activating Sirt6 will ameliorate CMD. EDD was assessed using myography (DMT). Myocardial blood flow (MBF) was measured by Doppler. Our preliminary data show that the mediator of coronary vasodilation switched from NO to H 2 O 2 in the Sirt6 knockout mice with impaired EDD. Interestingly, when the mice fed on HFHS were treated with Sirt 6 activator MDL-800, the coronary microvascular function was improved, and the blood glucose level was decreased. The underlying mechanism and the pathways involved will be elucidated.

A multivariate model for the prediction of pregnancy following laparoscopic artificial insemination of sheep.

The causes of variation in the success of laparoscopic artificial insemination (AI) in sheep are not well understood. As such, this study incorporated the contributions of multiple male and female factors relevant to the success of AI into a comprehensive prediction model for pregnancy success. Data from Merino ewes (N = 30 254) including age, uterine tone (1; pale/flaccid-5; turgid/pink), intra-abdominal fat (1; little to no fat present-5; high fat), time of insemination and sire used, were recorded during AI. A subset of semen per sire (N = 388) was thawed and assessed for volume, subjective motility, sperm concentration, and morphology. Sperm motility (CASA), viability and acrosome integrity (FITC-PNA/PI), membrane fluidity (M540/Yo-Pro), mitochondrial superoxide production (Mitosox Red/Sytox Green), lipid peroxidation (Bodipy C11), level of intracellular reactive oxygen species (H2DCFDA) and DNA fragmentation (Acridine Orange) were also assessed 0, 3 and 6h post-thaw. Logistic binomial regression revealed sperm concentration (P < 0.001), CASA parameters at 0h (PCA3; P = 0.03), viable acrosome intact sperm at 6h (P = 0.02), abnormal morphology (P < 0.001), uterine tone (P < 0.001) and intra-abdominal fat (P = 0.03) of ewes influenced likelihood of pregnancy. Results generated will help standardise the pre-screening and selection of semen and ewes prior to artificial breeding programs, reducing variation in the success of sheep AI.

Weight loss and metabolic effects of an ALDH1A1-specific inhibitor, FSI-TN42, in a diet induced mouse model of obesity.

Retinoic acid (RA) participates in weight regulation and energy metabolism. Mice lacking ALDH1A1, one of the major enzymes responsible for RA biosynthesis, are resistant to diet-induced obesity. Previously, we identified FSI-TN42 (N42) as an ALDH1A1-specific inhibitor and reported its pharmacokinetics and pharmacodynamics as well as its efficacy in weight suppression. In the first study, C57BL/6 J male mice were fed a high fat diet for 8 weeks to induce obesity. Mice were then divided into three groups and fed (1) moderate fat diet (MFD), (2) MFD + WIN 18,446 (1 g/kg diet), or (3) MFD + N42 (1 g/kg diet) for 8 weeks. A control group of mice were fed a low-fat diet for the entire period. Mice were weighed weekly and fasting glucose was determined every 4 weeks. Tissues were examined for potential toxicity using histopathology and complete blood counts. In the second study, we examined influences of N42 on energy balance and/or appetite by determining food intake, activity and energy expenditure in mice with obesity treated with MFD or MFD + N42. Lastly, we tested fertility with a mating study. N42 significantly accelerated weight loss compared to MFD alone in mice with obesity by reducing fat mass without decreasing lean mass. N42 did not alter food intake or activity levels. While mice treated with N42 lost significantly more weight, they maintained a similar level of energy expenditure compared to mice fed MFD only. Mice fed N42 preferentially used fat postprandially, especially under thermoneutral or mild cold challenge. N42 did not affect male fertility. N42 promotes weight loss when used with MFD in mice with diet-induced obesity without causing significant organ toxicity or male infertility. Future studies will determine if N42 can be used to promote further weight loss if combined with current weight loss drugs.

CTNI-64. TRIAL IN PROGRESS: DIET2TREAT -- A RANDOMIZED MULTICENTER PHASE 2 TRIAL OF A KETOGENIC DIET VS STANDARD DIETARY GUIDANCE IN COMBINATION WITH STANDARD-OF-CARE TREATMENT FOR PATIENTS WITH NEWLY DIAGNOSED GLIOBLASTOMA

Abstract BACKGROUND Many glioblastoma (GBM) patients inquire about the potential therapeutic benefits of a high fat / low carbohydrate ketogenic diet (KD). Preclinical models and early-phase trials have shown encouraging results, but definitive evidence of clinical efficacy is lacking. METHODS We are conducting a multicenter randomized controlled phase 2 clinical trial (NCT05708352) of KD vs standard dietary guidance (SD) for patients with newly diagnosed GBM. All patients receive standard-of-care chemoradiation. Inclusion criteria: newly diagnosed GBM, age ≥ 18 years, Karnofsky Performance Status ≥ 70. Exclusion criteria: inability to wean steroids below 8 mg dexamethasone/day, Body Mass Index &amp;lt; 21 kg/m2, food preferences incompatible with KD. Patients are randomized 1:1 to implement either KD or SD for a study period of 18 weeks, starting alongside chemoradiation. The primary outcome is overall survival for patients randomized to the KD arm compared to those randomized to the SD arm. Secondary outcomes include progression-free survival, health-related quality-of-life (FACT-BR, FACIT-F), cognitive performance (HVLT-R, Trail-Making Test A/B), and physical activity (modified Godin leisure questionnaire, continuous activity monitoring (Fitbit)). Patients randomized to the KD arm will be provided a fingerstick monitor to check blood ketone and glucose levels twice daily. Glucose and ketone levels are also periodically monitored for patients on the SD arm. Blood, tumor, and stool samples are being collected for high throughput profiling to assess the effects of KD on metabolic markers and immune response. Target accrual is 170 patients over 5 years. Enrollment began in July 2023 and is ongoing. As of May 2024, the trial is open for enrollment at Cedars-Sinai and UCSF, and will be opening soon at Duke, St. John’s Cancer Institute, and the Medical College of Wisconsin. 13 patients have been randomized to date. Funding provided by NIH R01CA276919 and the Foundation for Metabolic Cancer Therapies.

Association between dietary diversity and lipid indices among Indian Kho-Kho players

Purpose Dietary diversity (DD) is a proxy indicator of nutrient adequacy and has been propagated as a health beneficial component of dietary behavior. This study aims to understand DD and its association with lipid indices among Indian Kho-Kho players. Design/methodology/approach In all, 50 Indian Kho-Kho players aged between 16 years and 31 years belonging to different regions of India were recruited. Dietary diversity score (DDS; food groups/day) and food variety score (FVS; number of food items/day) were considered to assess the DD. The United Nations Food and Agricultural Organization food frequency questionnaire was used as a tool. Serum total cholesterol (TC), triglycerides (TG), LDL, VLDL and HDL were measured. Findings The DDS and FVS showed that athletes were consuming approximately 7 of 9 food groups (Mean ± SD: 6.5 ± 1.3) and 27 varieties of foods (Mean ± SD:27.0 ± 8.6) from 9 food groups per day. This study could not find significant differences for DDS and FVS among athletes belonging to different regions. FVSs of organ meat, eggs and spices and condiments were significantly positively associated with TG (p &lt; 0.036, p &lt; 0.020 and p &lt; 0.001) and VLDL (p &lt; 0.054, p &lt; 0.021 and p &lt; 0.001). LDL was inversely significantly associated with FVS of cereals, white root and tubers (p &lt; 0.032). Research limitations/implications DDS and FVS were not associated with lipid indices among Kho-Kho players. However, individual FVS of organ meat, eggs and spices and condiments may increase TG and VLDL. Optimum consumption of cereals and root tubers may help to maintain LDL levels among Kho-Kho players. Originality/value It was found that daily consumption of organ meat and eggs may increase TG and VLDL, and optimum consumption of cereals and white roots and tubers may help to maintain LDL. High energy and fat foods may act as mediators along with spices and condiments in increasing TG and VLDL.

Histone demethylase JMJD1C advances macrophage foam cell formation and atherosclerosis progression by promoting the transcription of PCSK9.

Macrophage is considered as a critical driving factor in the progression of atherosclerosis (AS), and epigenetic heterogeneity contributes important mechanisms in this process. Here, we identified that a histone demethylase jumonji domain-containing protein 1C (JMJD1C) is a promising biomarker for atherosclerotic cerebral infarction through clinical analysis. Then, AOPE-/- mice fed with a high fat diet and RAW264.7 cells induced by oxidized low-density lipoprotein (ox-LDL) were used as AS models to verify the function of JMJD1C in AS development in vivo and in vitro. JMJD1C knockdown significantly reduced plaque area, inflammation and endothelial damage in AS model mice, and also alleviated foam cell formation, inflammatory cytokines production and cell apoptosis in ox-LDL-treated RAW264.7 cells. Mechanistically, JMJD1C promoted the transcription of proprotein convertase subtilisin/kexin type 9 (PCSK9) through mediating H3 Lysine 9 demethylation. The effects of JMJD1C knockdown on ox-LDL-induced macrophages were blocked by PCSK9 overexpression. Altogether, our study proves that JMJD1C advances macrophage foam cell formation, inflammation and apoptosis to accelerate AS progression through H3 demethylation of PCSK9. The findings underscore the important role of JMJD1C-mediated histone modification in macrophage regulation and AS progression, which brings a new insight into the pathobiology of AS.