bluuebunny/crossref_metadata_2025_split huggingface dataset:
1554-P: Insights Into Insulin Synthesis and Turnover Using 13C515N-Valine to Label C-Peptide in Humans
Authors: EGAN AOIFE M. , DYER ROY B. , LAURENTI MARCELLO C. , DALLA MAN CHIARA , VELLA ADRIAN 
Published: June 2024

The insulin secretory response to an intravenous, as opposed to an oral, glucose challenge is characterized by 2 phases the first of which is thought to represent preformed insulin whereas the second represents provisioning from newly synthesized insulin or newly mobilized insulin granules. The first phase is absent in people with type 2 diabetes (T2DM). Whether this reflects defects in insulin synthesis, insulin storage or both remains to be ascertained. To address these questions, we studied 10 subjects with and without T2DM. Subjects were admitted to the Clinical Research Unit at 1700 and consumed a standard meal and fasted overnight. At 2100 5g of arginine was given. An infusion of 13C515N-Valine (Prime 1.5mg/Kg body mass and then 1.5mg/Kg/hr) commenced and continued till 0600 the following morning. At 0800 subjects underwent a 75g OGTT followed by 5g I.V. arginine at 1200. Blood was sampled at frequent intervals to measure hormone and substrate concentrations as well as C-peptide labelled with 13C515N-Valine. This was purified via repetitive cycling through an affinity resin conjugated with C-peptide antibody in a custom manufactured mass spectrometric immunoassay pipette tip. Peptides were detected in positive mode with a high resolution, Q Exactive Plus Mass Spectrometer optimized by infusion of synthetic peptides. During the overnight fast, average C-peptide enrichment in the peripheral circulation was higher in T2DM (0.3 ± 0.3 vs. 2.3 ± 0.2 %, nondiabetic vs T2DM respectively, p&lt;0.01). After oral glucose, C-peptide enrichment was lower in T2DM (6.7 ± 0.5 vs. 5.0 ± 0.4 %, p=0.05). In contrast after I.V. arginine challenge, enrichment did not differ (5.6 ± 1.2 vs. 3.5 ± 0.4 %, p=0.08). These data imply that insulin turnover is higher in T2DM: a greater proportion of newly synthesized insulin is secreted during fasting. This may explain the depletion of 1st phase insulin and other easily releasable granule pools in T2DM.

Disclosure
A.M. Egan: None. R.B. Dyer: None. M.C. Laurenti: Employee; Angelini Pharma. C. Dalla Man: None. A. Vella: Research Support; Novo Nordisk. Consultant; Hanmi Pharm. Co., Ltd., Crinetics Pharmaceuticals, Inc. Advisory Panel; Rezolute, Inc.

Funding
National Institutes of Health (DK078646)
156-OR: Genome-Wide CRISPR Screen Identified That Alternative Splicing Regulates in Insulin Secretion
Authors: WEI SIYING , APARECIDA DA SILVA PEREIRA JESSICA , LEE-PAPASTAVROS ALEXANDER , YI PENG 
Published: June 2024

Pancreatic β cell functionality, especially glucose-stimulated insulin secretion (GSIS), is pivotal in diabetes pathophysiology. The impairment of GSIS is not only evident in Type 2 diabetes (T2D) but also during the early stages of Type 1 diabetes (T1D). In addition, enhancing GSIS also holds significant promise for improving the functionality of stem cell-derived β cells (SC- β cells). Addressing this limitation offers new opportunities in cell-based diabetes therapy. To uncover the intricate regulatory mechanisms of insulin secretion, we developed a unique reporter cell line carrying NPY-pHluorin transgene to visualize insulin exocytosis at single-cell level and conducted a genome-wide CRISPR screen using flow cytometry system in NIT-1 cells. Notably, our study identifies alternative splicing (AS) as a pivotal factor in insulin secretion regulation. Employing genetic Sf3b1 knockout - a key component in AS machinery in NIT-1 cells, results in a surprising outcome: increased insulin secretion and improved GSIS. Consistently, using chemical inhibitors such as FR901464 to suppress AS activities, we observed the same phenotype in β cell lines and pancreatic islets. To unravel the underlying mechanisms, we performed comprehensive RNA-seq and alternative splicing-seq analyses in Sf3b1 knockout cell lines and cells treated with AS inhibitor, revealing alterations in genes related to the insulin production machinery and synaptic/exocytic vesicle traffic pathways. Our findings emphasize the significant role of AS in modulating β cell function. This research bridges critical gaps in our current understanding and sets the stage for innovative approaches to address the intricate challenges posed by diabetes.

Disclosure
S. Wei: None. J. da Silva Pereira: None. A. Lee-Papastavros: None. P. Yi: None.
1567-P: Integrating Bioinformatics and Machine Learning to Identify Lactylation-Related Diagnostic Biomarkers and Characterize Immune Cell Infiltration in Nonalcoholic Fatty Liver Disease
Authors: ZHANG JUNYI , WU XUEYAN 
Published: June 2024

Background: This study sought to explore potential lactylation-related targets for non-alcoholic fatty liver disease (NAFLD) and examine the role of immune cell infiltration in the disease's progression.
Methods: Gene expression datasets were sourced from the Gene Expression Omnibus collection, and genomic enrichment analysis was conducted utilizing differentially expressed genes (DEGs). Overlapping lactylation and NAFLD DEGs were determined using a Venn diagram, and a protein-protein interaction (PPI) network was developed. To ascertain immune patterns, the ConsensusClusterPlus package in R was employed. Key genes were identified using three machine-learning algorithms: random forest, support vector machine-recursive feature elimination, and least absolute shrinkage and selection operator. The infiltration of immune cells was assessed by the GSVA package. The upstream transcriptional factors (TFs) and microRNAs (miRNAs) were predicted using the Regnetwork database and NetworkAnalyst.
Results: The intersection of key genes and DEGs yielded 34 genes associated with lactylation and NAFLD, comprising 22 upregulated and 12 downregulated genes. Eight genes, namely FABP5, SIRT1, TERF2, ADNP, BTF3, CDC5L, DDX17, and MNDA, were identified as diagnostic indicators with relatively high diagnostic specificity for NAFLD. The interaction among key genes were shown by PPI network. A spectrum of immune cells was found to potentially contribute to NAFLD development. The association between key genes and immune infiltration were revealed by the correlation analysis. Also, we predicted the upstream TFs and miRNAs of hub genes.
Conclusions: Our research identified 8 key genes related to lactylation and analyzed the immune cell infiltration characteristics in NAFLD patients, thereby offering additional insights into the potential mechanisms influencing NAFLD prognosis.

Disclosure
J. Zhang: None. X. Wu: None.
1568-P: Liver-specific Expression of Activated G Protein Gsα Leads to Hyperglycemia with Reduced Pancreatic α-Cell Mass and Function
Authors: GUPTA MEENAKSHI , SUN HUI , LAYTON DANIEL , ZHANG JIANHUA , LEE JI-EUN , CHEN MIN , WEINSTEIN LEE 
Published: June 2024

Gsα couples hormones to stimulate intracellular cAMP generation. We previously showed that mice with liver-specific Gsα deficiency (LGsKO) have improved glucose tolerance and enlarged pancreatic islets. Here we generated mice with liver-specific expression of an activated Gsα by breeding mice containing a Lox-STOP-Lox-GsαR201C transgene with albumin-Cre mice (LGsR201C). Compared to controls, male LGsR201C mice had normal survival but increased hepatic cAMP levels and gluconeogenetic gene expression, confirming the activation of liver Gsα/cAMP signaling in these mice. Consequently, LGsR201C mice showed elevated blood glucose, enhanced glucose response to pyruvate administration and hyperglycemic response to glucagon. LGsR201C mice had glucose intolerance and lacked insulin response to i.p. glucose injection, although having normal pancreatic β-cell mass and intact glucose-stimulated insulin secretion from isolated islets ex vivo . On the other hand, LGsR201C mice exhibited normal insulin sensitivity but lacked counterregulatory action of glucagon responding to lower blood glucose levels in vivo and had less pancreatic α-cell mass, reduced islet glucagon content and impaired arginine-stimulated glucagon secretion from isolated islets. Notably, LGsR201C mice showed significantly lower serum levels in all tested 17 amino acids. RNAseq analysis revealed that an array of genes was oppositely regulated in LGsR201C vs. LGsKO mice, including the genes of several amino acid transporters upregulated in the liver and islets of LGsR201C mice and downregulated in the liver of LGsKO mice. Thus, our data indicate that activation of Gsα/cAMP signaling in the liver triggers a liver-islet circuit that plays divergent roles in the control of pancreatic islet α- and β-cell biogenesis and functions.

Disclosure
M. Gupta: None. H. Sun: None. D. Layton: None. J. Zhang: None. J. Lee: None. M. Chen: None. L. Weinstein: None.

Funding
NIDDK Intramural Research Program
1576-P: Evaluation of Hepatic Steatosis and Adiposity in People with Type 1 Diabetes and Obesity
Authors: TRIKUDANATHAN SUBBULAXMI , AMIN KATHAN , BAMBHA KIRAN , MELHORN SUSAN J. , SEKHON MANDEEP , NAVARRO SANDI , SCHUR ELLEN A. , HIRSCH IRL B. 
Published: June 2024

Introduction: Prevalence of obesity in T1D is rising, contributing to the development of hepatic steatosis (HS) with the associated risk for metabolic dysfunction associated steatotic liver disease (MASLD). While T2 D and MASLD are reliably linked, prevalence of MASLD in T1D varies by the imaging modality employed. We therefore estimated the prevalence of HS and examined the key determinants of HS in people with T1D with BMI≥ 30 kg/m2.
Methods: In a cross-sectional study, 42 individuals with T1D on a continuous glucose monitor (CGM) with duration of diabetes &gt;10 years and BMI ≥ 30 kg/m2 were studied. MRI-Proton Density Fat Fraction (PDFF) was used to estimate HS. Visceral (VAT) and subcutaneous adiposity (SAT) measures were obtained. Linear regression models were used to evaluate the association between HS, VAT, and SAT, with outcomes of glycemic control and insulin resistance.
Results: Participants had a mean (SD) age of 47.8 (13.3) y, 59.5% were men with BMI -32.9 (3.1) kg/m2 and 30.6% had a family history of T2D. Mean A1c was 7.1 (0.77), TIR of 65% (12.5), and TAR of 32.8(12.2) and GMI of 7.2(0.45). Prevalence of HS was 9.5% in the study sample. No statistically significant associations were found between any of the parameters evaluated and HS although a trend was observed for total daily dose of insulin in units (TDDI) (p=0.06) with higher insulin doses positively associated HS. Higher A1c (p =0.004), lower TIR (p=0.02), higher TAR (p =0.009) and higher TDDI (p&lt;0.0001) were associated with higher VAT in the unadjusted models and remained significant for TDDI and VAT (p=0.01) when adjusted for age, sex, and WC.
Conclusion: Our study, which incorporates CGM metrics in a distinctive subpopulation of T1D with obesity, suggests a lower prevalence rate of HS when a sensitive imaging modality such as MRI-PDFF was used. Lack of portal hyperinsulinemia may contribute to lower HS in T1D. The results suggest that markers of insulin resistance and glycemic exposure correlate with VAT but not HS in adults with T1D and obesity.

Disclosure
S. Trikudanathan: Research Support; Insulet Corporation. K. Amin: None. K. Bambha: None. S.J. Melhorn: None. M. Sekhon: None. S. Navarro: None. E.A. Schur: Consultant; Amgen Inc. I.B. Hirsch: Advisory Panel; Abbott. Research Support; Dexcom, Inc. Advisory Panel; Roche Diabetes Care. Research Support; MannKind Corporation, Tandem Diabetes Care, Inc. Advisory Panel; embecta. Research Support; Tandem Diabetes Care, Inc. Advisory Panel; Vertex Pharmaceuticals Incorporated.

Funding
UWMDI Pilot & Feasibility Award
1579-P: Altered Serum Ceramides as Part of Dyslipidemia in Adolescents with Metabolic Dysfunction–Associated Steatotic Liver Disease
Authors: SHORT KEVIN R. , FRIEDMAN JACOB E. , WANG LIPING , SUMMERS SCOTT A. 
Published: June 2024

Introduction & Objective: Metabolic dysfunction-associated steatotic liver disease (MASLD) in adults is associated with altered plasma ceramides, and may serve as liver metabolic biomarkers. However, less is known about whether similar changes are present in adolescents. We performed a lipidomic analysis to test whether adolescents with MASLD have altered plasma ceramides.
Methods: Plasma was acquired from boys and girls, 11-20y, classified as, 1) obese with biopsy-confirmed MASLD (n=44); 2) obese without MASLD (Ob, n=28); and 3) normal weight without MASLD (NW, n=29). MASLD patients had moderate to severe liver steatosis and 91% had ≥ stage 1 liver fibrosis.
Results: MASLD and Ob had similar body fat (45±7%), and were higher (p&lt;0.01) than NW (31±6%). MASLD had higher triglycerides (p&lt;0.01) than Ob and NW (199±110, 107±48, & 86±40 mg/dl, respectively) and had lower HDL-C than Ob or NW (40±10, 47±12, & 59±12 mg/dl respectively). Of the 6 major ceramide species measured, d18:1/24:0 was ~18% lower (p&lt;0.001) in MASLD and Ob versus NW, while the others did not differ among groups. Of the 6 major dihydroceramide (dhCer) species measured, 4 were 32-185% higher (p&lt;0.001) in the MASLD group versus the Ob and NW groups. The dhCer, but not the ceramide species, were positively correlated with trunk fat (r=0.34 to 0.55), but not with insulin resistance. dhCer 18:0/18:0, the most altered species, was positively correlated with liver steatosis (r=0.36), and along with 3 other dhCer species was positively correlated with MASLD Activity Score (r=0.35 to 0.41). There were no group differences in hexosylceramides, sphingosine, sphinganine, or the major sphingomyelins or dihydrosphingomyelins.
Conclusions: Similar to adults, adolescents with MASLD have altered plasma ceramides and dyslipidemia compared to NW and Ob peers without MASLD. The patterns observed suggest that ceramide flux may be elevated and ceramide-producing cells may be damaged in pediatric MASLD patients.

Disclosure
K.R. Short: None. J.E. Friedman: None. L. Wang: Stock/Shareholder; Centaurus Therapeutics. S.A. Summers: Board Member; Centaurus Therapeutics.

Funding
NIH R01DK129656 and U54GM104938, Presbyterian Health Foundation, Harold Hamm Diabetes Center, and Children's Health Foundation
1590-P: Lian-He-Xiao-Zhi Ointment Improves Lipid Metabolism and Inflammation in MAFLD by Regulating PPARα
Authors: NIE LIJUAN , YANG XINYI , ZHOU XIQIAO 
Published: June 2024

Background and Aims: Lian-He-Xiao-Zhi ointment (LHXZO) is an empirical formula, which has a significant therapeutic effect on metabolic associated fatty liver disease (MAFLD) in clinical practice. However, the precise underlying mechanisms remain unclear. The objective of this study is to elucidate the mechanisms through which the LHXZO ameliorates MAFLD, utilizing an animal model.
Methods: C57BL/6J mice (6-8 weeks old, male) were subjected to a high-fat diet (HFD) to induce a MAFLD model. Mice received either LHXZO decoction or control solution intragastrically daily for twelve weeks. Histological and serum analyses were used to assess the liver injury and disorders of glucolipid metabolism. Total RNAseq and pathway analyses were used to identify the potential signaling pathways modulated by LHXZO in the liver. The expression levels of key genes involved in regulating lipid metabolism and inflammation were validated by qRT-PCR or Western blot analysis. The impact on gut microbiota was assessed through 16S rRNA gene sequencing.
Results: LHXZO treatment significantly improved glucolipid metabolic disorders, liver injury and liver inflammation, evidenced by decreased serum levels of fasting blood glucose, free fatty acid, TG, AST, ALT, LPS and IL-1β. HE and Oil Red O staining also showed LHXZO treatment reduced lipid deposition in the liver. Meanwhile, LHXZO significantly reduced the body weight of MAFLD mice. LHXZO mitigated inflammation by downregulating TLR4, NF-KB, NLRP3, IL-1β and IL-18. In addition, LHXZO significantly upregulated the level of PPARa. Furthermore, LHXZO significantly modulated gut microbiota.
Conclusions: LHXZO effectively improve glucolipid metabolism, liver injury and inflammation in MAFLD, indicating its potential as a therapeutic agent for MAFLD. LHXZO's improvement of liver inflammation in MAFLD mice may be achieved by regulating PPARα or gut microbiota.
Keywords: MAFLD; PPARα; inflammation; gut microbiota

Disclosure
L. Nie: None. X. Yang: None. X. Zhou: None.

Funding
the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD, 035062005002-14), the special research project of Jiangsu Administration of traditional Chinese medicine (zt202105), and the Project Funded by Jiangsu Province Hospital of Chinese Medicine (kkzx01).
1593-P: Symptoms and Correlations in Newly Diagnosed Type 2 Diabetes Mellitus (T2DM) with Nonalcoholic Fatty Liver Disease (NAFLD)
Authors: MUKHERJEE SUPRIYO 
Published: June 2024

Introduction & Objective: The study conducted in North Bihar, India, aimed to explore the prevalence of NAFLD among individuals newly diagnosed with T2DM. It investigated the potential correlations between patient symptoms and various factors such as educational background, socioeconomic class, family history of diabetes mellitus (DM), and gender.
Methods: A cross-sectional study, from December 2022 to May 2023, involved 148 newly diagnosed T2DM patients aged 18+. Exclusion criteria comprised alcohol use, pregnancy, traditional medicine, liver-related issues, and malignancy. Sociodemographic, symptom, and clinical data were collected via detailed tools, analysed using Stata 13.0 and IBM SPSS 28 for correlations and differences.
Results: Out of the 148 individuals, 73.7% presented with liver disease (NAFLD), emphasizing the high prevalence of liver involvement among newly diagnosed T2DM cases. Common symptoms included excessive thirst, polyuria, weight loss, and nocturia. Intriguingly, correlations emerged between symptom presentation and education level, socioeconomic class, family history of DM, and gender. Individuals with atypical symptoms tended to have higher education and a family history of DM. Lower-middle-class individuals exhibited a higher percentage of atypical symptoms, suggesting potential disparities in symptom recognition and healthcare-seeking behavior.
Conclusion: This study highlighted high NAFLD rates in newly diagnosed T2DM cases in North Bihar. Correlations with education, socioeconomic factors, and gender emphasize tailored interventions for early detection and management.

Disclosure
S. Mukherjee: None.
160-OR: PI3K Inhibition with Single-Dose Alpelisib as a Model of Insulin Resistance in Healthy Adults
Authors: COOK JOSHUA R. , BEDEIR NUR , SONE ZACHARY D. , WATTACHERIL JULIA , GINSBERG HENRY N. , LAFERRÈRE BLANDINE 
Published: June 2024

Introduction & Objective: Understanding the pathophysiology of insulin resistance (IR) is essential to treating type 2 diabetes, but mechanistic studies of IR in human subjects are often complicated by ancestry and phenotypic heterogeneity, variable chronicity, and difficulty in causal inference. We therefore seek to establish a safe, reliable, acute method for modeling IR in a translational setting. Our objective is to test a single dose of the phosphoinositide-3-kinase (PI3K) inhibitor alpelisib, versus placebo, for tolerability and effect on glucose and insulin metabolism.
Methods: This single-center, double-blind phase I clinical trial randomized healthy adults, ages 18-65 y with body mass index 18.0-26.9 kg/m2 and HbA1C ≤ 5.6%, to take a single oral dose of alpelisib 300 mg (n = 5) or placebo (n = 6) at bedtime. The next morning, participants had blood drawn for fasting insulin and glucose levels­ (co-primary endpoints) followed by a 3-hour, 75-g oral glucose tolerance test (OGTT). Data are analyzed based on intention to treat using an independent two-sample t test.
Results: Fasting plasma glucose trended higher with alpelisib (mean ± SEM = 93 ± 6 mg/dL) versus placebo (84 ± 2 mg/dL) (p = 0.07). Mean fasting serum insulin increased nearly fivefold: 23 ± 6 μIU/mL for alpelisib versus 5 ± 1 μIU/mL for placebo (p &lt; 0.01), resulting in Homeostasis Model Assessment of Insulin Resistance scores of 5.4 ± 1.4 vs. 1.1 ± 0.2 and Matsuda Index scores of 8.6 ± 2.2 vs. 3.0 ± 1.6, respectively. During OGTT, area under the curve (AUC) for insulin was greater with alpelisib (26.0 ± 7.5 mIU/mL × min) than placebo (5.8 ± 1.0 mIU/mL × min) (p &lt; 0.02); glucose AUC trended higher with alpelisib. Single-dose alpelisib was well tolerated without serious adverse events.
Conclusion: A single dose of alpelisib produced metabolic derangements consistent with acute induction of IR, validating its use as a safe and effective tool for studying insulin action in humans.

Disclosure
J.R. Cook: None. N. Bedeir: None. Z.D. Sone: None. J. Wattacheril: Research Support; Intercept Pharmaceuticals, Inc. Advisory Panel; GlaxoSmithKline plc. Consultant; Alphasights. Research Support; AMRA, Galectin Therapeutics. H.N. Ginsberg: None. B. Laferrère: Consultant; Up-To-Date.
1600-P: Sunflower Oil HFD Feeding in Mice Leads to Gut Dysbiosis, Mild Steatohepatitis, and Impaired Glycemic Control
Authors: SINDHU SARDAR T.A.K. , BAHMAN FATEMAH , ZUBBAIR MALIK MOHAMMAD , AREFANIAN HOSSEIN , ALRASHED FATEMA , NIZAM RASHEEBA , KOCHUMON SHIHAB P. , THOMAS REEBY S. , ALBELOUSHI SHAIMA , HASAN AMAL , ABU-FARHA MOHAMED , AL MADHOUN ASHRAF , THANGAVEL ALPHONSE T. , ALMULLA FAHD , AHMAD RASHEED 
Published: June 2024

Introduction & Objective: High-fat diets (HFDs) influence gut microbiota and promote obesity & metabolic dysregulation. Dietary sucrose is a co-player in modifying gut microbial ecosystem towards dysbiosis. Fish & sunflower dietary fats may be consumed with/without sucrose; however, gut bacterial changes from excessive intake of these fats, without the added effect of sucrose, remain unclear.
Methods: To address this objective, we compared effects of sucrose-free, sunflower high-fat diet (S-HFD) & fish (F)-HFD feeding (24 wks) on the gut microbiota, obesity, liver inflammation, and glycemic control in C57BL/6 mice.
Results: The 16S rRNA sequencing of fecal microbiome showed that gut bacterial communities were differentially expressed between two dietary groups. Firmicutes were highly-abundant in S-HFD (40%) compared to F-HFD (3%) mice, while Verrucomicrobia were abundant in F-HFD (26%) compared to S-HFD (˂1%) mice. Firmicutes/Bacteroidetes ratios in S-HFD & F-HFD were 1.32 & 2.45, respectively. MicrobiomeAnalyst & MIAOME analyses identified highly-abundant taxa in S-HFD mice, including Oscillospira, guilliermondii, Allobaculum, Yaniella, Ruminococcus, gnavus, Staphylococcus, Clostridium, Adlercreutzia, Aerococcaceae, Anaeroplasma, Mogibacteriaceae, Christensenellaceae, & RF39, having predicted associations with host genes of metabolic disorders/pathways linked to inflammation & glucose metabolism. Compared to F-HFD mice, the S-HFD mice showed slightly increased body weights (P = 0.06) but significant differences regarding fasting hyperglycemia, hepatic macrophage infiltration (F4/80 expression), macrovascular steatosis, lobular inflammation, & expression of genes associated with de novo lipogenesis (Acaca, Fasn, and Scd1) & monocyte chemotaxis (Ccl2).
Conclusion: Taken together, S-HFD feeding in mice leads to gut dysbiosis and the clinical manifestations of moderate liver inflammation, steatosis and impaired glycemic control.

Disclosure
S.T. Sindhu: None. F. Bahman: None. M. Malik: None. H. Arefanian: None. F. Alrashed: None. R. Nizam: None. S.P. Kochumon: None. R.S. Thomas: None. S. Albeloushi: None. A. Hasan: None. M. Abu-farha: None. A. Al Madhoun: None. A.T. Thangavel: None. F. Almulla: None. R. Ahmad: None.

Funding
Kuwait Foundation for the Advancement of Sciences (KFAS) (Grants #: RA 2010-003)