High Sucrose Research Articles

Treatment with 1.25% cholesterol enriched diet produces severe fatty liver disease characterized by advanced fibrosis and inflammation and impaired autophagy in mice

Nonalcoholic fatty liver disease (NAFLD) is reaching pandemic proportions due to overnutrition. The understanding of advanced stages that recapitulate the human pathology is of great importance to get a better mechanistic insight. We hypothesized that feeding of WT (C57BL) mice with a diet containing a high content of fat (21%), sugar (41.5%) and 1.25% of cholesterol (called from now on high fat, sucrose and cholesterol diet, HFSCD) will reproduce the characteristics of disease severity. Analysis of 16 weeks HFSCD-fed mice demonstrated increased liver weight and plasmatic liver damage markers compared with control diet (CD)-fed mice. HFSCD-fed mice developed greater hepatic triglyceride, cholesterol and NEFA content, inflammation and NAFLD activity score (NAS) indicating an advanced disease. HFSCD-fed mice displayed augmented hepatic total CD3+ T and Th9 lymphocytes, as well as reduced Th2 lymphocytes and CD206 anti-inflammatory macrophages. Moreover, T cells and anti-inflammatory macrophages correlated positively and inversely, respectively, with intrahepatic cholesterol content. Consistently, circulating cytotoxic CD8+ T lymphocytes, Th1, and B cell levels were elevated in HFSCD-fed WT mice. Hepatic and adipose tissue expression analysis demonstrated changes in fibrotic and metabolic genes related with cholesterol, triglycerides, and fatty acid synthesis in HFSCD-fed WT. These mice also exhibited reduced antioxidant capacity and autophagy and elevated ERK signaling pathway activation and CHOP levels. Our results indicate that the feeding with a cholesterol-enriched diet in WT mice produces an advanced NAFLD stage with fibrosis, characterized by deficient autophagy and ER stress along with inflammasome activation partially via ERK pathway activation.

Intraspecific and interspecific resource partitioning between bumblebee workers and males related to nectar quantity and quality.

Bumblebees are important pollinators for many natural and agricultural systems in temperate regions. Interspecific and intraspecific variation in floral resource preferences have been proposed to influence bumblebee community structure. In particular, sexual dimorphism is a major source of intraspecific niche variation. Although interspecific resource partitioning is well studied, few studies have explored the intraspecific dynamics between workers and males. Here, we report a study on a total of 11528 workers and 2220 males of 14 bumblebee species recorded over 5years in the Hengduan Mountains of Southwest China. We first compared the potential for interspecific and intraspecific competition between workers and males using visitation records and resource partitioning indices (overlap index). We then evaluated the influence of nectar traits on flower preference, including nectar volume and the levels of hexose, sucrose and 10 essential amino acids (EAAs). We found that the niche overlap between intraspecific workers and males was higher than that between different species, and temporal overlap alone did not strongly determine diet overlap. Males of most species preferred flowers with high levels of EAAs and hexose, whereas workers of some species preferred flowers with high nectar volume and sucrose levels. This study suggests that there is floral resource partitioning among bumblebee species, and between workers and males, which may play a key role in alleviating interspecific and intraspecific competition. These findings also provide a useful guide for which kinds of plants might be most valuable for bumblebees, especially the understudied males, in this biodiversity hotspot.

Abstract Mo132: Pouring Salt in the Wound: Sex-specific Cardiometabolic Responses to Increased Dietary Fat, Sugar, and Salt.

Introduction: The general population is becoming more obese, and diabetes rates continue to rise, with overnutrition and consumption of processed foods likely contributing factors. Many people consume diets high in fat, sugar, and salt; however, this triad of factors is rarely examined in unison in studies. In addition, studies that examined diets high in fat and sugar in the context of heart failure with preserved ejection fraction and cardiometabolic outcomes have often performed short dietary interventions, with many studies performed in only male mice, and dietary salt content is often not considered or manipulated. Study Goal: This study aimed to assess the long-term impact of high-fat, high-sucrose diets with differing levels of salt (NaCl) on systemic and cardiovascular physiology in male and female mice. Methods and Results: 12-week-old C57BL/6J male and female mice ( n = 4/group) were fed one of the following diets for 25 weeks: normal chow (NC), low-fat diet (LFD; 10 kcal%), high-fat high sucrose (HFHS; 60 kcal%) with 0.33% NaCl, HFHS with 1% NaCl (HFHS1%), and HFHS with 8% NaCl (HFHS8%). Mice were subjected to basal tail blood metabolite assessment and weighed weekly. In week 24, mice were subjected to glucose tolerance testing, dual-energy X-ray absorptiometry followed by postmortem gravimetric, blood metabolite, histological, and biochemical analyses at 25 weeks. We found that irrespective of sex, body weight was significantly greater in mice receiving HFHS ( p <0.001) and HFHS1% ( p <0.001) versus the other diets. By 25 weeks, male mice were significantly larger than female mice with all diets except HFHS1% ( p <0.05), which correlated with increased percent body fat ( p <0.01), impaired glucose tolerance ( p <0.05) and increased ketone levels ( p <0.05) in female mice subjected to HFHS1% versus males on the same diet. There was also increased heart weight and atrial weights in female mice subjected to HFHS and HFHS1% versus the other females, which was not observed in male mice. Conclusions: Although previous short-term dietary studies have shown that males have altered cardiometabolic responses more than females, our findings together show that long-term dietary interventions with HFHS diets with salt may adversely impact female systemic and cardiovascular physiology.

The Synergistic Enhancement of Anti-Metabolic Diseases Function of Morus alba with the Combination of Cha (Camellia sinensis)

Background: Functional foods play an important role in the prevention and ameliora-tion of metabolic syndromes leading to type 2 diabetes. Plant resources that have anti-metabolic syndromes activity, such as Morus alba L. and Cha [Camellia sinensis L.], have been used in functional foods against diabetes. Since Morus and Cha have different mechanisms of action against metabolic syndromes, such as prevention of sugar uptake and lipidosis, respectively, and the combination of both resources will be a reliable approach for developing more efficient functional food against type 2 diabetes because certain synergism is expected in their functions. Methods: Male Wister Rats were fed the high fat-high sucrose [HFHS] diet for 12 weeks, with and without supplementation of Morus and Cha alone and their combination, and the effect of their supplementation on the markers of the metabolic syndrome such as obesity, lipidosis, and fatty liver formation, were examined. Results: Several metabolic syndrome markers, including body weight gain, lipid deposit, and fatty liver formation, were more significantly prevented by the diet supplemented with Morus and Cha combination compared to Morus or Cha given separately . Conclusion: Appropriate formulation of food resources with different functional mechanisms is a promising strategy for developing effective dietary treatment of type 2 diabetes that is a typical Mibyou.

Evaluation of the Efficacy of Targeted Microbiome Therapy in Non-Alcoholic Steatohepatitis Rat Model

Abstract Non-alcoholic steatohepatitis (NASH) is the clinically aggressive variant of non-alcoholic fatty liver disease. Hippo pathway dysregulation can contribute to NASH development and progression. The use of probiotics is effective in NASH management. Our aim is to investigate the efficacy of kefir Milk in NASH management via modulation of hepatic mRNA-miRNA based panel linked to NAFLD/NASH Hippo signaling and gut microbita regulated genes which was identified using bioinformatics tools. Firstly, we analyzed mRNAs (SOX11, SMAD4 and AMOTL2), and their epigenetic regulator (miR-6807) followed by validation of target effector proteins (TGFB1, IL6 and HepPar1). Molecular, biochemical, and histopathological, analyses were used to evaluate the effects of kefir on high sucrose high fat (HSHF) diet -induced NASH in rats. We found that administration of Kefir proved to prevent steatosis and development of the inflammatory component of NASH. Moreover, Kefir improved liver function and lipid panel. At the molecular level, kefir down-regulated the expression of miR 6807-5p with subsequent increase in the expression of SOX 11, AMOTL2 associated with downregulated SMAD4, resulting in reduction in the expression of the inflammatory and fibrotic markers, IL6 and TGF-β1 in the treated and prophylactic groups compared to the untreated rats. In conclusion, Kefir suppressed NASH progression and improved both fibrosis and hepatic inflammation. The produced effect was correlated with modulation of SOX11, SMAD4 and AMOTL2 mRNAs) – (miR-6807-5p) – (TGFB, IL6 and, HepPar1) expression.

Probiotic Growth Pattern and Physicochemical Evaluation of Water Kefir Fermentation

Probiotics are live-friendly microorganisms that can confer a health benefit on the host if it is consumed in sufficient amounts. Water kefir is a probiotic-rich fermented beverage that contains multi-species of live cultures. Brown sugar and palm sugar were used for water kefir fermentation due to their high sucrose and mineral contents. The objective of this study was to determine the probiotic growth pattern of water kefir and to evaluate the physicochemical parameters, including the pH changes, lactic acid content, reducing sugar content, and total soluble solids. The fermented water kefir was collected at every 6-hour interval, until the end of 72 hours of fermentation. The growth curve was determined by enumerated probiotics on De Man, Rogosa, and Sharpe (MRS) agar, Yeast Extract-Peptone-Dextrose (YPD) agar, and Gluconobacter (GM) agar plates, respectively. MRS, YPD, and GM agar plates were used to enumerate lactic acid bacteria, yeast, and acetic acid bacteria, respectively. The result showed increased probiotic growth as fermentation time increased with different phases observed from the growth curve. The stationary phase of probiotics was recorded at 30-42 h and was recommended as the optimal harvesting point. Besides, longer fermentation time produced lower pH values and lower total soluble solids while higher lactic acid and higher reducing sugars. At the end of fermentation, the concentration of lactic acid and reducing sugars were 2.16 ± 0.09 g/L and 13.66 ± 0.14 mg/mL, respectively. In conclusion, probiotics from water kefir fermentation are suggested to be best harvested between 30-42 hours and can be used for self-consume or downstream processing.

High sucrose diet-induced abnormal lipid metabolism in mice is related to the dysbiosis of gut microbiota

Background & AimsExcess sucrose intake induces metabolic syndrome. In human, abnormal lipids metabolism like obesity, hyperlipidemia and fatty liver are induced. However, excess sucrose causes different phenotypes in different species. Based on our previous study, excess sucrose induced fatty liver and hyperlipidemia in rats. The phenotypes and mechanism of abnormal lipid metabolism in mice is unclear. We investigated the different phenotypes in 5 strains of mice and the relationship between gut microbiome and abnormal lipid metabolism in C57BL/6N mice. MethodsWe examined the effect of a high sucrose diet in 5 different strains of mice. Besides, to find out the relationship between gut microbiome and metabolic disorder induced by excess sucrose, C57BL/6N mice were fed with a high sucrose diet with or without antibiotics cocktail. ResultsA high sucrose diet induced obesity and fatty liver in inbred mice, whereas did not induce hyperlipidemia in all strains of mice. Moreover, a high sucrose diet changed the composition of gut microbiota in C57BL/6N mice. Antibiotics treatment alleviated the abnormal lipid metabolism induced by high sucrose diet by changing the composition of gut short chain fatty acids. ConclusionsThese results indicates that the phenotypes of metabolic syndrome are influenced by genetic factors. Furthermore, the dysbiosis of gut microbiome caused by excess sucrose may contribute to the development of abnormal lipid metabolism via its metabolites.

Impact of Dietary Fiber on Soft Drinks with Varied Sweeteners

In the present work, the characteristics of the effect of dietary fiber in two types of soft drinks were compared, considering beverages with low and high sucrose content, using a 22 experiment design for the independent variables amount of dietary fiber and sucrose content. The dependent variable was the percentage of soluble solids, and experimental runs were carried out to establish the stability of the product during the process. The design mentioned above determined significant differences in physicochemical parameters, such as increased fiber percentage and sucrose amount in final brix degrees. Samples of the Beverage were taken during processing and evaluated over time to establish the behavior of soluble solids, density, and the amount of fiber during the pasteurization process, which was determined at 85ºC for 120 seconds. The behavior of the unpasteurized soft drink was also studied at refrigeration temperature for one day in samples with sucrose and partially substituted with sucralose, enriched with dietary fiber and without its addition, to evaluate and analyze if the thermal effect determines a significant difference in the quality of the finished product. The laboratory analyses were based on a quantitative, enzymatic-gravimetric analysis of dietary fiber and soluble solids. The best option was chosen considering the best variety of soft drinks with low sucrose content and enriched with dietary fiber, increasing the beverage's nutritional value. This selection was implemented in a soft drink factory, and the procedure was standardized at the industrial level. The development of this research contributed to technological improvements reflected in the product's final price, minimizing costs and highlighting that with a minimum investment, it is competitive. The results of the tests performed on the carbonated beverage to determine the response variables were statistically analyzed using STATGRAPHICS software. The triplicate results of each trial were initially evaluated for significant differences attributable to the treatments, and the validity of the data was verified by analysis of their distributions, considering a relative error of less than 10%. Finally, current computer programs automatically calculate the p-value, and for the interpretation of the various results, this calculated value is compared with a pre-established confidence level, which generally is 95%. Keywords: increased fiber; soluble solids; sucralose; sucrose.

Impact of Dietary Fiber on Soft Drinks with Varied Sweeteners

In the present work, the characteristics of the effect of dietary fiber in two types of soft drinks were compared, considering beverages with low and high sucrose content, using a 22 experiment design for the independent variables amount of dietary fiber and sucrose content. The dependent variable was the percentage of soluble solids, and experimental runs were carried out to establish the stability of the product during the process. The design mentioned above determined significant differences in physicochemical parameters, such as increased fiber percentage and sucrose amount in final brix degrees. Samples of the Beverage were taken during processing and evaluated over time to establish the behavior of soluble solids, density, and the amount of fiber during the pasteurization process, which was determined at 85ºC for 120 seconds. The behavior of the unpasteurized soft drink was also studied at refrigeration temperature for one day in samples with sucrose and partially substituted with sucralose, enriched with dietary fiber and without its addition, to evaluate and analyze if the thermal effect determines a significant difference in the quality of the finished product. The laboratory analyses were based on a quantitative, enzymatic-gravimetric analysis of dietary fiber and soluble solids. The best option was chosen considering the best variety of soft drinks with low sucrose content and enriched with dietary fiber, increasing the beverage's nutritional value. This selection was implemented in a soft drink factory, and the procedure was standardized at the industrial level. The development of this research contributed to technological improvements reflected in the product's final price, minimizing costs and highlighting that with a minimum investment, it is competitive. The results of the tests performed on the carbonated beverage to determine the response variables were statistically analyzed using STATGRAPHICS software. The triplicate results of each trial were initially evaluated for significant differences attributable to the treatments, and the validity of the data was verified by analysis of their distributions, considering a relative error of less than 10%. Finally, current computer programs automatically calculate the p-value, and for the interpretation of the various results, this calculated value is compared with a pre-established confidence level, which generally is 95%. Keywords: increased fiber; soluble solids; sucralose; sucrose.

Identification of male sterility-related genes in Saccharum officinarum and Saccharum spontaneum.

Candidate male sterility genes were identified in sugarcane, which interacts with kinase-related proteins, transcription factors, and plant hormone signaling pathways to regulate stamen and anther development. Saccharum officinarum is a cultivated sugarcane species that its predominant feature is high sucrose content in stems. Flowering is necessary for breeding new cultivars but will terminate plant growth and reduce sugar yield. The wild sugarcane species Saccharum spontaneum has robust and viable pollen, whereas most S. officinarum accessions are male sterile, which isa desirable trait of a maternal parent in sugarcane breeding. To study male sterility and related regulatory pathways in sugarcane, we carried out RNAseq using flowers in different developmental stages between male-sterile S. officinarum accession 'LA Purple' and fertile S. spontaneum accession 'SES208'. Gene expression profiles were used to detect how genes are differentially expressed between male sterile and fertile flowers and to identify candidate genes for male sterility. Weighted gene correlation networks analysis (WGCNA) was conducted to investigate the regulatory networks. Transcriptomic analyses showed that 988 genes and 2888 alleles were differentially expressed in S. officinarum compared to S. spontaneum. Ten differentially expressed genes and thirty alleles were identified as candidate genes and alleles for male sterility in sugarcane. The gene Sspon.03G0007630 and two alleles of the gene Sspon.08G0002270, Sspon.08G0002270-2B and Sspon.08G0014700-1A, were involved in the early stamen or carpel development stages, while the remaining genes were classified into the post-meiosis stage. Gibberellin, auxin, and jasmonic acid signaling pathways are involved in the stamen development in sugarcane. The results expanded our knowledge of male sterility-related genes in sugarcane and generated genomic resources to facilitate the selection of ideal maternal parents to improve breeding efficiency.

Oat protein modulates cholesterol metabolism and improves cardiac systolic function in high fat, high sucrose fed rats.

Oats are recognized to provide many health benefits that are mainly associated with its dietary fibre, β-glucan. However, the protein derived from oats is largely understudied with respect to its ability to maintain health and attenuate risk factors of chronic diseases. The goal of the current study was to investigate the metabolic effects of oat protein consumption in lieu of casein as the protein source in high fat, high sucrose (HF/HS) fed Wistar rats. Four-week-old rats were divided into three groups and were fed three different experimental diets: a control diet with casein as the protein source, an HF/HS diet with casein, or an HF/HS diet with oat protein for 16 weeks. Heart structure and function were determined by echocardiography. Blood pressure measurements, an oral glucose tolerance test, and markers of cholesterol metabolism, oxidative stress, inflammation, and liver and kidney damage were also performed. Our study results show that incorporation of oat protein in the diet was effective in preserving systolic heart function in HF/HS fed rats. Oat protein significantly reduced serum total and low-density lipoprotein cholesterol levels. Furthermore, oat protein normalized liver HMG-CoAR activity, which, to our knowledge, is the first time this has been reported in the literature. Therefore, our research suggests that oat protein can provide hypocholesterolemic and cardioprotective benefits in a diet-induced model of metabolic syndrome.

Isolation, identification, and tolerance analysis of yeast during the natural fermentation process of Sidamo coffee beans.

Yeast, which plays a pivotal role in the brewing, food, and medical industries, exhibits a close relationship with human beings. In this study, we isolated and purified 60 yeast strains from the natural fermentation broth of Sidamo coffee beans to screen for indigenous beneficial yeasts. Among them, 25 strains were obtained through morphological characterization on nutritional agar medium from Wallerstein Laboratory (WL), with molecular biology identifying Saccharomyces cerevisiae strain YBB-47 and the remaining 24 yeast strains identified as Pichia kudriavzevii. We investigated the fermentation performance, alcohol tolerance, SO2 tolerance, pH tolerance, sugar tolerance, temperature tolerance, ester production capacity, ethanol production capacity, H2S production capacity, and other brewing characteristics of YBB-33 and YBB-47. The results demonstrated that both strains could tolerate up to 3% alcohol by volume at a high sucrose mass concentration (400g/L) under elevated temperature conditions (40 ℃), while also exhibiting a remarkable ability to withstand an SO2 mass concentration of 300g/L at pH 3.2. Moreover, S. cerevisiae YBB-47 displayed a rapid gas production rate and strong ethanol productivity. whereas P. kudriavzevii YBB-33 exhibited excellent alcohol tolerance. Furthermore, this systematic classification and characterization of coffee bean yeast strains from the Sidamo region can potentially uncover additional yeasts that offer high-quality resources for industrial-scale coffee bean production.

Exercise protects the hypothalamus morphology from the deleterious effects of high sucrose diet consumption.

A growing body of evidence suggests that elevated sucrose intake may contribute to the development of neurological disorders. Recognizing that regular exercise has the potential to reduce the occurrence of neuromuscular disorders, the present research investigated the impact of exercise on the redox status of the hypothalamus in mitigating the adverse effects associated with high sucrose intake. Forty Wistar albino rats were subjected to a high sucrose diet, with some groups engaging in exercise for a duration of 3 months. The exercise regimen was found to sustain the redox balance in the hypothalamus. In summary, the consumption of a high sucrose diet resulted in the disturbance of the histological morphology of the hypothalamus, accompanied by an increased percentage of caspase-3 positive cells. Additionally, the high sucrose diet disrupted the oxidant/antioxidant ratio in favor of oxidants, leading to elevated levels of AOPPs and AGEP. Conversely, exercise was effective in restoring most of these values to levels approximating the control group, indicating a potential protective effect of regular exercise against the detrimental impacts of high sucrose dietary consumption on the hypothalamus. Graphical abstract.

Inositol and taurine ameliorate abnormal liver lipid metabolism induced by high sucrose intake

Previous attempts to limit fat intake failed to reduce metabolic syndrome incidence, spotlighting excessive sucrose consumption phenomenon. Recent studies challenge the traditional belief, actually fructose is predominantly metabolized in the small intestine, not just the liver. Evidence supports sucrose overconsumption leads to hepatic lipid accumulation via gut microbiome modulation, but the mechanisms and prevention strategies remain unclear. This study investigates the impact of osmolytes inositol and taurine, which are abundant in citrus fruits and seafood respectively, on lipid metabolism in rats consuming excessive sucrose. Five-week-old male Wistar rats fed with control diet, high sucrose diet, and high sucrose diet with inositol or taurine for 28 days. High sucrose diet-induced accumulation of hepatic triglyceride level was reduced only by inositol. Both inositol and taurine decreased hepatic SREBP1, ChREBP, lipogenesis enzymes mRNA levels increased by a high sucrose diet. Taurine also significantly increased the fatty acid uptake major player CD36 mRNA levels in liver. Expression of genes related to fatty acid degradation and transport remained unchanged. In cecal microbiome, high sucrose diet groups showed a decrease in alpha diversity and diverged in beta diversity from the starch diet group, while inositol and taurine did not significantly affect these aspects. Ten bacterial features including genera such as Bacteroides, Parabacteroides, Odoribacter, Streptococcus, etc., were filtered out related to the ameliorative effects of inositol and taurine on lipid metabolism. Overall, both inositol and taurine modulated hepatic lipogenesis and microbiome. However, due to taurine's enhancement of fatty acid uptake, only inositol improved steatosis in high sucrose diet rats.

Symplasmic phloem loading and subcellular transport in storage roots are key factors for carbon allocation in cassava.

Cassava (Manihot esculenta) is a deciduous woody perennial shrub that stores large amounts of carbon and water in its storage roots. Previous studies have shown that assimilating unloading into storage roots happens symplasmically once secondary anatomy is established. However, mechanisms controlling phloem loading and overall carbon partitioning to different cassava tissues remain unclear. Here, we used a combination of histological, transcriptional, and biochemical analyses on different cassava tissues and at different timepoints to better understand source-sink carbon allocation. We found that cassava likely utilizes a predominantly passive symplasmic phloem loading strategy, indicated by the lack of expression of genes coding for key players of sucrose transport, the existence of branched plasmodesmata in the companion cell/bundle sheath interface of minor leaf veins, and very high leaf sucrose concentrations. Furthermore, we showed that tissue-specific changes in anatomy and non-structural carbohydrate contents are associated with tissue-specific modification in gene expression for sucrose cleavage/synthesis, as well as subcellular compartmentalization of sugars. Overall, our data suggest that carbon allocation during storage root filling is mostly facilitated symplasmically and is likely mostly regulated by local tissue demand and subcellular compartmentalization.

Abstract 2061: Single-cell Analysis Reveals Insights Into Visceral Adiposity In Mice With Diabetes-associated Atherosclerosis

Introduction: Visceral white adipose tissue (vWAT), previously seen as passive and uniform, plays a dynamic role in metabolism and immune response. Dietary shifts affect its structure and function, impacting conditions like insulin resistance, dyslipidemia, and diabetes. vWAT is also linked to vascular diseases, but its role in atherosclerosis in mouse models is not fully understood. Methods: vWAT from male Ldlr -/- mice fed either a chow diet or high fat, high sucrose containing (HFSC) diet for 24 weeks were harvested. scRNA-seq, histology, and immunofluorescence were used to analyze the cellular and transcriptomic profiles in Chow and HFSC groups. Results: Distinct subpopulations of endothelial cells (ECs), immune cells, and adipose stem and progenitor cells (ASPCs) were identified, highlighting differences between healthy and diseased states. Notably, HFSC-fed mice exhibited an increase in immune cell composition, coupled with a decrease in ECs and progenitor cells. Cell subtype distribution in macrophages shifted notably, with more Lipid-associated (LAM), but fewer Perivascular-like macrophages (PVM). Novel markers Lyz2 and Trem2, linked to atherosclerosis in Ldlr -/- mice, were identified specific to LAM. Pathway analysis showed Pdgfa, S100a8, and Mmp13 upregulation in LAM, crucial for atherosclerosis development. The probability model of cell-cell communication was established and characterized as 3 distinct cell patterns that enriched in pathways (Chemokine, Wnt & BMP signaling) implicated in atherosclerosis progression. Conclusion: scRNA-seq atlas reveals interactions among adipose cell types in lean and obese state. Inflammation triggers cellular distribution changes, with overlapping markers and pathways in vWAT and atherosclerosis emerging in disease progression. This dataset serves as a valuable resource for exploring genes and cell types involved in vWAT function in diabetes-associated atherosclerosis.

Sucrose supplements modulate the Pseudomonas chlororaphis-Arabidopsis thaliana interaction via decreasing the production of phenazines and enhancing the root auxin response

Management of the plant microbiome may help support food needs for the human population. Bacteria influence plants through enhancing nutrient uptake, metabolism, photosynthesis, biomass production and/or reinforcing immunity. However, information into how these microbes behave under different growth conditions is missing. In this work, we tested how carbon supplements modulate the interaction of Pseudomonas chlororaphis with Arabidopsis thaliana. P. chlororaphis streaks strongly repressed primary root growth, lateral root formation and ultimately, biomass production. Noteworthy, increasing sucrose availability into the media from 0 to 2.4% restored plant growth and promoted lateral root formation in bacterized seedlings. This effect could not be observed by supplementing sucrose to leaves only, indicating that the interaction was strongly modulated by bacterial access to sugar. Total phenazine content decreased in the bacteria grown in high (2.4%) sucrose medium, and conversely, the expression of phzH and pslA genes were diminished by sugar supply. Pyocyanin antagonized the promoting effects of sucrose in lateral root formation and biomass production in inoculated seedlings, indicating that this virulence factor accounts for growth repression during the plant-bacterial interaction. Defence reporter transgenes PR-1::GUS and LOX2::GUS were induced in leaves, while the expression of the auxin-inducible, synthetic reporter gene DR5::GUS was enhanced in the roots of bacterized seedlings at low and high sucrose treatments, which suggests that growth/defence trade-offs in plants are critically modulated by P. chlororaphis. Collectively, our data suggest that bacterial carbon nutrition controls the outcome of the relation with plants.

Characterizing MetALD in a Nonhuman Primate Model using High Fat and Sucrose Diet and Chronic Binge Alcohol Administration

Mechanisms leading to metabolic and alcohol associated liver disease (MetALD) are not clearly delineated. This study examined the differential regulation of markers of hepatic mitochondrial function in nonhuman primates fed a high fat/high sucrose (HFSD) and administered chronic binge alcohol (CBA) or vehicle as compared to control-fed animals. We hypothesized that markers of hepatic dysregulation due to HFSD will be further altered by the combination of CBA/HFSD. Eight rhesus macaques were fed HFSD (27% sucrose and 42% fat, by kcal) with n=4 receiving vehicle and n=4 receiving 13-14g/kg/wk of ethanol through a gastric catheter. Rhesus macaques fed standard primate chow were used as reference controls (n=4-6). Liver RNA was isolated for determining expression of genes regulating mitochondrial biogenesis and function using RT-qPCR. Results were analyzed using one-way ANOVA and post-hoc pairwise multiple comparisons were performed using Tukey’s method, with alpha set to 0.05. Expression of inflammatory genes was significantly altered across the groups (p<0.05). HFSD increased tumor necrosis factor alpha (p=0.01), interleukin 1-beta (p=0.04), cluster of differentiation 38 (p=0.01), and transforming growth factor beta (p=0.01) expression compared to control. Both HFSD and CBA/HFSD increased interleukin-10 compared to control (p=0.02, p=0.03). Gene expression of mitochondrial function and oxidative stress regulators were significantly altered across the groups (p<0.05). CBA/HFSD decreased expression of peroxisome proliferator-activated receptor-gamma coactivator-1 beta (p=0.04) and nuclear respiratory factor 2 (p=0.04) compared to control. Both HFSD and CBA/HFSD decreased peroxisome proliferator-activated receptor alpha expression compared to control (p=0.01, 0.01). HFSD decreased superoxide dismutase 2 (SOD2) expression compared to control (p=0.01) and increased uncoupling protein 2 (p=0.01). SOD2 expression was increased only in CBA/HFSD compared to HFSD (p=0.01). These results reveal marked HFSD-mediated increases in markers of inflammation and CBA/HFSD-mediated changes in markers of mitochondrial function regulation and oxidative stress regulation. These findings suggest a greater impact of calorie-dense diets than CBA on hepatic inflammation and a combined impact of CBA and HFSD on mitochondrial regulation. Ongoing studies will examine oxidative phosphorylation complex protein activities and expression to elucidate and integrate HFSD or CBA mediated changes in hepatic bioenergetic capacity. Supported by F30AA030910, P60AA009803, and T32AA007577. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Development of an engineered Bacillus subtilis strain for antibiotic-free sucrose isomerase production.

Sucrose isomerase (SIase) catalyzes the hydrolysis and isomerization of sucrose into isomaltulose, a functional sugar extensively used in the food industry. However, the lack of safe and efficient heterologous expression systems for SIase has constrained its production and application. In this study, an engineered Bacillus subtilis strain for antibiotic-free SIase production was developed via a food-grade expression system. First, the B. subtilis strain TEA was modified through the CRISPR/Cas9 system, resulting in a mutant strain TEA4, which exhibited enhanced capabilities for recombinant protein expression. For efficient and safe production of SIase, different constitutive and inducible promoters were evaluated. The maltose-inducible promoter Poglv was found to have an extracellular SIase activity of 21.7 UmL-1 in engineered strain TEA4. Subsequent optimization of the culture medium further increased SIase activity to 26.4 UmL-1 during shake flask cultivation. Eventually, using the crude enzyme solution of the engineered strain in biotransformation reactions resulted in a high yield of isomaltulose under high concentrations sucrose, achieving a maximum yield of 83.1%. These findings demonstrated an engineered B. subtilis strain for antibiotic-free SIase production, paving the way for its scale-up industrial production and application.

SerpinA3N in Leptin Receptor Neurons Regulates Metabolic and Glucose Homeostasis

Obesity and associated conditions such as type 2 diabetes are among the most concerning issues for health in the U.S. Leptin receptor signaling in the brain plays a pivotal role in the regulation of food intake and energy expenditure. High fat diet induces inflammation within the hypothalamus, leading to leptin insensitivity which contribute to obesity. Previous studies have shown that a novel gene SerpinA3N, a serine protease inhibitor, which is highly expressed in the arcuate nucleus of the hypothalamus, where the leptin receptor is also highly expressed, is upregulated in diet-induced obesity and by leptin stimulation. However, the role of SerpinA3N in the leptin receptor expressing cells in the control of body weight and glucose homeostasis remain unknown. RNAseq analysis showed that SerpinA3N mRNA levels were significantly upregulated in the hypothalamus of mice fed high fat high sucrose diet (HFHSD) for 12 weeks. Upregulation of hypothalamic SerpinA3N in HFHSD fed mice was confirmed by Western blot. This increase in hypothalamic SerpinA3N appears to be caused by excess of nutrient as we found that exposure to fatty acid significantly increased SerpeinA3N promoter activity in mouse hypothalamic N39 cells. Using immunofluorescence staining, we confirmed that SerpinA3N is expressed the leptin receptor positive cells of the hypothalamus. Next, we investigated whether loss of SerpinA3N in leptin receptor expressing neurons affects metabolic and glucose homeostasis. For this, we crossed mice harboring floxed alleles of the SerpinA3N gene (SerpinA3Nfl/fl) with mice expressing Cre recombinase in leptin receptor cells (LepRCre). We found that female, but not male, conditional knockout (CKO, LepRCre/SerpinA3Nfl/fl) mice have significantly lower body weight when fed normal chow or HFHSD. This lower body weight was due to decreased fat mass, measured by NMR. However, no alteration in food intake was observed between female CKO mice and controls, indicating that increased energy expenditure may account for the lower body weight in female CKO mice. Interestingly, CKO mice displayed exaggerated weight loss and anorectic response following leptin treatment (1 mg/g, twice daily, IP), indicating that loss of SerpinA3N enhances leptin sensitivity. Female CKO mice fed normal chow diet also exhibited enhanced insulin sensitivity compared to control animals. On the other hand, male CKO mice displayed significantly improved glucose handling, but not change in insulin sensitivity, only when fed HFHSD. Finally, insulin-induced phosphorylation of AKT was elevated in skeletal muscle, but not in liver and white adipose tissue, of CKO mice relative to controls. These results demonstrate that SerpinA3N in leptin receptor-expressing neurons regulate body weight and glucose homeostasis in a sex-dependent manner. NIH and VA. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.