bluuebunny/crossref_metadata_2025_split huggingface dataset:
1405-P: Bacteroides thetaiotaomicron Fortifies the Gut Mucosa and Improves Host Metabolism in Dietary Obese Mice
Authors: LEE JENNIFER , WELLENSTEIN KERRY , KAHN BARBARA B. 
Published: May 2022

Disruption of the intestinal mucosa contributes to insulin resistance in obesity and Type 2 diabetes. Probiotic supplementation, which restores gut microbiota composition and fortifies intestinal barrier function, has therapeutic potential to treat obesity and metabolic disease. Treatment of mice on a chow diet or high fat diet (HFD) with Palmitic Acid Hydroxystearic Acids (PAHSAs) improves insulin sensitivity. These effects are mediated, at least in part, by changes in the gut microbiome. We used this paradigm to identify new microbes that may have beneficial metabolic effects. Metagenome sequencing of the cecum from mice treated with PAHSAs identified Bacteroides thetaiotaomicron (Bt) to be most strongly associated with insulin sensitivity. Here, we aimed to determine whether Bt supplementation in mice on a HFD diet has beneficial effects on energy balance and/or glucose homeostasis. Daily oral Bt supplementation in female mice fed a HFD resulted in less weight gain, lower adiposity, and improved glucose tolerance compared to vehicle-treated mice. Additionally, Bt supplementation preserved colonic mucus thickness, an indicator of gut health; increased the number of Goblet cells which are the cells that secrete mucin2, the predominant mucus protein that forms the intestinal mucosal barrier; and increased protein levels of mucin2 in the distal gut compared to vehicle-treated controls. These improvements in distal gut barrier architecture were associated with a reduction in circulating levels of pro-inflammatory lipopolysaccharide (LPS) and interleukin-6 (IL-6) . These beneficial effects of Bt were observed in female and not in male HFD-fed mice. These studies indicate that Bt has sex-specific effects to improve host metabolism in the setting of dietary obesity. Bt represents a next-generation probiotic and novel gut-based therapeutic strategy which could improve metabolic health in women.

Disclosure
J.Lee: None. K.Wellenstein: None. B.B.Kahn: Advisory Panel; Janssen Pharmaceuticals, Inc., Consultant; Mellicell, Other Relationship; Alnylam Pharmaceuticals, Inc.

Funding
NIH KDK114162; JPB Foundation
1417-P: Gut Microbiome Translocation to Visceral Adipose Tissue Promotes Recruitment of Adipose-Specific Neutrophils in Human Obesity
Authors: SHANTARAM DHARTI , BLASZCZAK ALECIA M. , HOYD REBECCA , SMITH ALAN , ANTWI LINDA , LIU JOEY Z. , WRIGHT VALERIE P. , BRADLEY DAVID , LIU YUNZHOU , WILLIAMS NYELIA F. , SPAKOWICZ DAN , HSUEH WILLA 
Published: May 2022

Neutrophils protect against invading bacteria, and their unique role in adipose tissue inflammation and insulin resistance (IR) in obesity is increasingly recognized. We found that visceral adipose tissue (VAT) of obese (n=83) compared to lean (n=13) individuals had higher neutrophil abundance, correlating with IR. The obese subjects had elevated circulating plasma zonulin, a marker of intestinal permeability, and lipopolysaccharide-binding protein (LPSBP) , a marker of bacterial presence, consistent with evidence suggesting that changes in the microbiome and gut permeability in obesity permits the entry of gut bacteria and bacterial products into the circulation. Therefore, we tested for bacterial presence in VAT under sterile conditions. 16s rRNA analyses revealed that the family Streptococcaceae and an unnamed genus in the family Ruminococaceae were enriched in obese VAT contrary to order Bacilliales and genus Marvinobryantia in lean VAT, majorly originating from the GI tract. To confirm whether higher bacterial load is causal for VAT neutrophil abundance in obesity, we gavaged high-fat diet fed mice with fecal material from an obese human, which increased neutrophil, but not macrophage recruitment to VAT. As VAT is highly vascularized, we used RNA-seq to compare the gene expression signature of VAT neutrophils to peripheral blood neutrophils to eliminate the possibility that VAT neutrophils were circulating cells. Evidently, VAT neutrophils had a differential activation state compared to the circulating neutrophils, with a distinct gene expression pattern representing upregulated inflammatory, chemotactic, extracellular matrix, and ROS generation pathways. Overall, we find that that increased gut bacterial translocation enhances the recruitment of VAT specific neutrophils in obesity offering novel targets for managing VAT inflammation and comorbidities of obesity.

Disclosure
D.Shantaram: None. N.F.Williams: None. D.Spakowicz: None. W.Hsueh: None. A.M.Blaszczak: None. R.Hoyd: None. A.Smith: None. L.Antwi: None. J.Z.Liu: None. V.P.Wright: None. D.Bradley: None. Y.Liu: None.
1423-P: Endoscopic Revision for Post-Bariatric Weight Regain Results in Less Weight Loss in People with Type 2 Diabetes
Authors: TEJANI SANAA , DUNN SAMUEL H. , TAVAKKOLI ANNA , MESSIAH SARAH E. , ALMANDOZ JAIME 
Published: May 2022

Background: Post-bariatric surgery weight regain is common and leads to recidivism of cardiometabolic diseases, including Type 2 Diabetes Mellitus (T2DM) . Endoscopic therapies are emerging tools to treat weight regain. This study aims to assess the effectiveness of endoscopic revision for post-bariatric surgery weight regain in people with T2DM.
Methods: A retrospective review was conducted on patients who underwent endoscopic revision for post-bariatric surgery weight regain from November 20to July 2021 at an academic center. The primary outcome was weight change at 1, 3, and 6 months post-endoscopic revision.
Results: The cohort (n=33) was mean age 55 years; 82% female; 45% non-Hispanic Black, 15% Hispanic; mean BMI 39.3; 63% with gastric bypass, 33% with sleeve gastrectomy, 3% with duodenal switch; 24% with T2DM at the time of revision. The mean time from bariatric surgery to revision was 12 years with mean 33% weight regain from post-surgery nadir. Patients with T2DM experienced similar mean percent weight loss as those without T2DM at 1 (5.34% vs. 4.93%) and 3 months (4.98% vs. 6.47%) post-revision but less weight loss at 6 months (-0.28% vs. 4.07%) (Figure 1) .
Conclusion: Endoscopic revision is an effective therapy for post-bariatric surgery weight regain. However, people with T2DM may experience less long term weight loss compared to those without T2DM, warranting further investigation.

Disclosure
S.Tejani: None. S.H.Dunn: None. A.Tavakkoli: None. S.E.Messiah: None. J.Almandoz: None.
1434-P: ANGPTL8 Variant R59W Is Associated with Increased Circulatory TNF-alpha and IL7 LEVELS and Inflammation
Authors: MOHAMMAD ANWAR , ABU-FARHA MOHAMED , MADHU DHANYA , CHERIAN PREETHI T. , KHAIRI IRINA AL , HEBBAR PRASHANTHA , KAVALAKATT SINA A. , AREFANIAN HOSSEIN , ALAM-ELDIN NADA , THANGAVEL ALPHONSE T. , ALMULLA FAHD , ABUBAKER JEHAD 
Published: May 2022

Background: ANGPTL8 has been recently identified as lipid metabolism and inflammation regulator. It interacts with ANGPTL3 and, more recently, ANGPTL4 to regulate LPL activity. It was also shown to modulate NF-κB activity through its interaction with ikkα/β. A genetic variant rs2278426 (R59W) in ANGPTL8 has been associated with reduced LDL and HDL in Hispanics and increased FBG in Arabs. The objective of the study was to investigate the impact of the R59W variant on LPL and the inflammatory activity of ANGPTL8.
Methods: ANGPTL8 was genotyped in a discovery cohort of 738 Kuwaiti individuals. ANGPTL8 level, as well as inflammatory markers and LPL, were measured in plasma. Additionally, metabolite analysis, overexpression, luciferase binding assay, confocal microscopy, and cellular fractionation analyses were conducted in the HepG2 cell line to assess differences between the ANGPTL8 and its variant.
Results: The ANGPTL8 rs2278426 variant was associated with an increase in circulatory levels of TNF-α and IL7 in our cohort. Carrier versus reference genotype of the studied variant showed differences in the following metabolites: Acylcarnitines, Phosphatidylcholine, and Cholesteryl Ester. These metabolites are implicated in modulating NF-κB and TNF-α levels. Our in vitro analyses showed that the activity levels of NF-κB p65 was higher in the R59W than the wild type, and it was further elevated with TNFα stimulations. The data was validated by the luciferase activity, confocal microscopy, and cellular fractionation.
Conclusion: This study showed that ANGPTL8 variant R59W is associated with increased circulatory TNF-α and IL7 levels but not the LPL plasma levels. We have also demonstrated the pro-inflammatory role of the R59W variant in regulating the NF-κB pathway as reflected by genotyping, in vitro, metabolomics, and structural analyses. Further studies are needed to expand our understanding of other signaling pathways and possible crosstalk in the context of inflammation.

Disclosure
A.Mohammad: None. A.T.Thangavel: None. F.Almulla: None. J.Abubaker: None. M.Abu-farha: None. D.Madhu: None. P.T.Cherian: None. I.Al khairi: None. P.Hebbar: None. S.A.Kavalakatt: None. H.Arefanian: None. N.Alam-eldin: None.

Funding
Kuwait Foundation for the Advancement of Sciences
144-LB: Pediatric Type 2 Diabetes Prevalence from Real-World Data in the United States
Authors: CHAO LILY , VIDMAR ALAINA , ESPINOZA JUAN 
Published: May 2022

Background: The SEARCH for Diabetes in Youth Study has provided critical insights into the epidemiology of pediatric T2D. The 2021 report estimated an annual percent change (APC) in prevalence of 4.8% between 2009 to 2017. Multi-center longitudinal cohort studies generate robust data, but are time, resource, and effort intensive. Large real-world data repositories may offer cost-effective opportunities to validate and expand on these findings.
Hypothesis: Youth-onset T2D prevalence is rising at a rate in excess of 5% per year in the US in the previous five years.
Methods: We queried the Trinetx Research Network which includes EHR data from 58 institutions with 86,487,854 total patients. We included patients &lt;22 years of age, who had at least one incidence of an E11 ICD-10 code for T2D associated with an ambulatory, emergency room, or inpatient encounter for each calendar year from 2016 to 2021. Patients with other diabetes ICD-10 codes (E08, E09, E10, or E13) were excluded. Summary statistics of the query cohort describing sex, race and ethnicity, labs, medications, and anthopometrics were extracted from the platform. Chi-squared test for trend was used to determine statistical significance in demographic change.
Results: The mean APC in prevalence per center rose by 9.1% between 2016 to 2019 (2016: 127 cases/center, 5334 total cases; 2019: 166.2 cases/center, 7919 total cases) , but subsequently declined by 6.6% between 2019 to 2020. There was a consistent increase in the median BMI percentile over time from 2016 to 2021. The percentage of black youth diagnosed with T2D rose (from 30 to 33%, P&lt;0.0001) whereas that of Hispanic youth was unchanged (25%, P=0.73) . The proportion of male also increased over time (P&lt;0.0001) .
Conclusion: Real-world data from Trinetx demonstrates a rising incidence of youth-onset T2D that parallels an increase in median BMI percentile. The decrease in case rate in 2020 may be the result of decreased encounters during the Covid-19 pandemic. The rising prevalence of T2D in black youth and male gender warrants further analysis, to understand the driver behind this trend.

Disclosure
L. Chao: None. A. Vidmar: None. J. Espinoza: Consultant; AI Health.

Funding
National Center for Advancing Translational Science (UL1TR001855 and UL1TR000130)
1444-P: Knockdown of Etv5 Impairs Viability of Pancreatic Beta Cells by Enhancing ROS Production
Authors: YI XUANZI , BAGHNAVI FAHIMEH , DIEKMANN CEDRIC , PÄTH GÜNTER , SEUFERT JOCHEN 
Published: May 2022

Introduction: The transcription factor ETV5 regulates various biological processes such as cell proliferation and apoptosis, and activates a whole string of obesity- and diabetes-related genes. ETV5 is involved in tissue protection since mice with pancreatic loss of Etv5 show increased severity of pancreatitis and delayed tissue recovery. Here, we investigate the role of Etv5 knockdown (KD) on survival of injured endocrine pancreatic beta cells.
Methods: We generated INS-1E beta cells with doxycycline (dox) -inducible lentiviral Etv5 sh-RNA knock-down (KD) . Cellular stress was induced by streptozotocin (STZ) and palmitate (PA) . We assessed viability (MTS) , apoptosis (caspases-3/7 activation) , necrosis (assay) , ER stress (BiP and CHOP gene expression) , oxidative stress (ROS production) and gene expression of SOD1, SOD2 and Catalase.
Results: Etv5 KD significantly increases STZ/PA-induced loss of viability without changes in apoptosis but with increased necrosis. Etv5 KD did not change STZ/PA-induced gene expression of ER stress markers BiP and CHOP but significantly increased H2O2 production. This was associated with significantly elevated gene expression of mitochondrial SOD2, which converts superoxide into H2O2. Cytoplasmatic SOD1 and H2O2-decomposing catalase, PRDX1 and GPx1 remained unaffected.
Conclusion: Increased circulating free fatty acids in obesity promote oxidative stress. Loss of ETV5 increases ROS production in mitochondria during cellular stress by PA and STZ, as indicated by upregulated SOD2 gene expression. This suggests a role for ETV5 in controlling mitochondrial oxidative stress in pancreatic β-cells.

Disclosure
X. Yi: None. G. Päth: None. J. Seufert: Advisory Panel; Abbott, Sanofi-Aventis Deutschland GmbH. Research Support; Boehringer Ingelheim International GmbH. Speaker's Bureau; Abbott Diabetes, AstraZeneca, Bayer AG, Boehringer Ingelheim International GmbH, Lilly, Novo Nordisk, Sanofi-Aventis Deutschland GmbH.

Funding
XY is supported by China Scolarship Council (CSC) China,GP received Projektförderung from the German Diabetes Society (DDG)
1448-P: Partial Pancreatectomy Increased α-Cell Volume Density, but Not Compensated ß-Cell Volume in Japanese Subjects
Authors: TAKEUCHI YUKI , MIZUKAMI HIROKI , OSONOI SHO , KUDOH KAZUHIRO , SASAKI TAKANORI , YAGIHASHI SOROKU 
Published: May 2022

In type 2 diabetes, deficit of insulin secretion is often ascribed to the reduction of β-cell volume, implying that regeneration of β-cells could be one of its radical treatments. Hitherto, several models are known to elicit β-cell compensation in rodents. Among them, application of partial pancreatectomy can induce replenishment of β-cell volume in young mice, while it is unclear whether this occurs in humans. In the present study, islet pathology was evaluated using the pancreatic specimens from the patients with pancreatic cancer who underwent repeated (i.e., partial followed by total) pancreatectomy because of its local recurrence after the initial surgery. Ten subjects (5 males and 5 females) at Hirosaki University Hospital or Chiba University Hospital were evaluated. Morphometric analysis was performed on formalin fixed pancreatic sections, in which the quadruple immunostaining for each hormone and the fluorescent immunostaining for ALDH1A3 was conducted. The mean age at the first and second operations was 66.2±and 71.6±5.8 years old, respectively. The mean duration until recurrence was 3.2±2.0 years. There were no significant differences in HbA1c and BMI between first and second preoperation. In morphometrical analysis, α-cell volume in the second surgery was significantly increased compared to the first surgery (p&lt;0.04) , while β-cell volume was comparable. There was a positive correlation between the frequency of conduit glucagon positive cells and islet α-cell volume (r=0.66, p&lt;0.05) . The frequency of ALDH1A3 positive cells were positively correlated with the α-cell/ β-cell ratio in the second surgical specimen (r=0.81, p&lt;0.01) . Taken together, unlike rodents, our results indicate that partial pancreatectomy did not induce replenishment of β-cell volume, but increased α-cell volume in human subjects. As its mechanism, the neogenesis of ductal endocrine cells and/or the transdifferentiation of β-cells to α-cells in the islets may be involved.

Disclosure
Y.Takeuchi: None. H.Mizukami: None. S.Osonoi: None. K.Kudoh: None. T.Sasaki: None. S.Yagihashi: None.
1450-P: Identifying Hidden Sources of Tissue Plasticity in Pancreatic Islets with a Novel Multicolor Flow Cytometry Approach
Authors: KRYVALAP YURY , MENG SHAWN Z. , CZYZYK JAN 
Published: May 2022

Identifying pancreatic islet cells that acquire a beta-cell like phenotype is important for developing new diabetes therapies. Our studies have revealed a novel serpinB13 protease inhibitor antibody, that augments cathepsin L protease activity, resulting in Notch1 receptor cleavage and mobilization of endocrine progenitor cells to become beta cells in the embryonic pancreas. It is unclear, however, whether a similar induction of additional beta cells occurs with exposure to serpinB13 monoclonal antibody (mAb) in later life.
To explore the exact source (s) of new beta cells in response to serpinB13 mAb after birth we developed a novel, multicolor flow cytometry analysis approach to study pancreatic islets. Five-week old normal Balb/c mice were injected with serpinB13 mAb (or control IgG) , and sacrificed at 8 weeks for islet isolation. Islet cell suspensions were stained with a viability dye, and a panel of antibodies to CD31, CD45, cytokeratin 19, EpCAM, insulin (Ins) , glucagon (Glu) , somatostatin (SS) , c-peptide, and four transcription factors, Maf-A, Nkx6.1, NeuroD1, and Pdx-1.
Exposure to the serpinB13 mAb caused a significant increase in the number of hormonal-negative (e.g., Ins-Glu-SS-c-peptide-) cells (p=0.0011) , and hormonal negative, but c-peptide positive, cells (p=0.002) . At least one of four transcription factors examined were positive in 5-10% and 10-30% of these populations, respectively. At the same time, there was a trend in the drop of Glu+ and SS+ cells. The average number of pancreatic islets, and the total islet cellularity per mouse, was augmented in animals receiving serpinB13 mAb.
Although additional studies are necessary to characterize the hormonal negative cells and assess whether Glu+ and SS+ cells transdifferentiate to beta cells, these findings demonstrate that enhanced tissue plasticity and beta-cell output can be achieved in the pancreas following treatment with antibodies to protease inhibitors.

Disclosure
Y.Kryvalap: None. S.Z.Meng: None. J.Czyzyk: None.
1459-P: Human Islet Cell Transcriptome Analysis Using Single Nucleus RNA-seq Reveals New Beta-Cell Markers and Define Distinct Beta-Cell Subpopulations with Different Transcriptional Activity
Authors: KANG RANDY , LI YANSUI , ROSSELOT CAROLINA , SCOTT DONALD , GARCIA-OCANA ADOLFO , LU GEMING 
Published: May 2022

Single-cell RNA sequencing (scRNA-seq) of human islets requires cell dissociation and does not provide information on the transcriptional status of islet cells. On the other hand, single-nucleus RNA sequencing (snRNA-seq) does not require cell dissociation and provides abundant information on intronic sequences that can be used to identify actively transcribed genes. Based on this, we seek to compare scRNA-seq and snRNA-seq approaches in human islets to determine: 1) whether similar cell clusters could be detected; 2) whether new gene markers could be discovered in human islet endocrine cells using intronic reads; 3) whether human beta cell subpopulations could be identified based on INS expression dynamics; and 4) whether snRNA-seq could be used in human islet grafts transplanted in immunosuppressed mice. We performed scRNA-seq and snRNA-seq on three pairs of human islets obtained from three healthy adult human donors using exon only or exon plus intron reads, respectively. Analysis of integrated data revealed similar human islet cell clusters. In the snRNA-seq data, however, top differentially expressed genes were identified as new markers of human endocrine cells such as ZNF385D (β-cells) , PTPRT (α-cells) , LRFN5 (δ-cells) and CACNA2D3 (PP cells) . These markers also accurately define endocrine cell populations in human islet grafts. Additionally, we distinguished several beta cell sub-clusters- INS rich cluster, HNRNPA2B1 (vesicle) rich cluster and active INS transcribing cluster.
In conclusion, snRNA-seq analysis of human islet cells is a previously unrecognized tool for the identification of human islet cell types in samples where nuclear RNA processing is required. By comparing INS expression between scRNA-seq and snRNA-seq data sets, we can detect different beta cell sub-populations with distinct gene expression patterns representing different biological dynamic states.

Disclosure
R.Kang: None. Y.Li: None. C.Rosselot: None. D.Scott: None. A.Garcia-ocana: Consultant; Sun Pharmaceutical Industries Ltd. G.Lu: None.

Funding
NIH DK105015
1460-P: Tauroursodeoxycholic Acid (tudca) Improves the Morphofunction of Pancreatic Beta Cells in Postweaning Protein-Restricted Mice
Authors: SANTOS LOHANNA M. BARRETO DOS , ARAUJO THIAGO , ALVES BRUNA L. , FERREIRA SANDRA MARA , ZIMATH PRISCILA L. , VETTORAZZI JEAN F. , CARNEIRO EVERARDO M. 
Published: May 2022

Protein restriction in the postweaning diet in mice impairs glucose metabolism by reducing insulin secretion from pancreatic β cells. TUDCA in obese rodents and humans normalized insulin secretion, increased insulin clearance, furthermore recovered mass and the number of β cells per islet in animal models of diabetes. This study aimed to evaluate the effects of TUDCA on the morphology and function of pancreatic β cells in mice subjected to postweaning protein restriction. Male C57Bl/6J mice at 30 days of age received a control diet (14% protein - group C) or a protein-restricted diet (6% protein - group R) for 16 weeks. During the last 2 weeks, 50% of the animals in each group received intraperitoneal treatment with daily doses of 300 mg/kg of body weight of TUDCA (CT and RT) , and the remainder received PBS (C and R) . The R mice that showed decreased total plasma protein and serum albumin concentration had lower weight gain and feed efficiency, but increased white adipose tissue weight. There were no differences in fasting blood glucose between the groups. The R mice that have increased insulin sensitivity after treatment with TUDCA had normalized sensitivity. Isolated islets from R mice have lower insulin secretion, which has been associated with decreased morphometric parameters compared to C. In RT mice secretion was restored in response to 2.8 mM and 11.mM glucose and the pancreas of the RT showed a greater area of β cells in relation to the area of the islets and a greater number of islets compared to R. Furthermore, RT mice showed an increase in the expression of genes of the SNARE complex, genes of function, transcription and differentiation of pancreatic β cells. We conclude that TUDCA improves glycemic homeostasis, increases insulin secretion and improves the functioning of pancreatic β cells in mice subjected to protein restriction, indicating that TUDCA can be used as a therapeutic strategy to normalize insulin secretion in protein malnutrition.

Disclosure
L.M.Barreto dos santos: None. T.Araujo: None. B.L.Alves: None. S.Ferreira: None. P.L.Zimath: None. J.F.Vettorazzi: None. E.M.Carneiro: None.

Funding
Conselho Nacional de Desenvolvimento Científico e Tecnológico (134270/2019-3)