bluuebunny/crossref_metadata_2025_split huggingface dataset:
1511-P: Identifying How Endothelial Protease-Activated Receptors Control Insulin Signaling—Implications for Diabetes
Authors: RAJALA RAHUL , GRIFFIN COURTNEY 
Published: June 2023

Endothelial cell (EC) dysfunction occurs in diabetes. Thrombin, a circulating serine protease with increased activity in diabetics, signals through protease-activated receptors 1 and 4 (PAR1/PAR4). To determine EC-specific roles of PARs in diabetes, we generated Par1fl//fl; Par4fl/fl; iCdh5 (PAC)-CreERT2 (Par1/4dECko) mice and challenged them with streptozotocin (STZ) to induce hyperglycemia. STZ-treated Par1/4dECko mice had reduced blood glucose levels, weighed significantly more, and lost less abdominal fat than control littermates. Neither Par4ECko nor Par1ECko mice phenocopied Par1/4ECko mutants, suggesting that loss of both thrombin receptors is required to drive these phenotypes. Notably, STZ-treated Par1/4dECko mice showed comparable insulin levels to their STZ-treated littermates, suggesting mutant mice had heightened insulin sensitivity, which was verified by insulin/glucose tolerance testing. Using primary human ECs that express PAR1 but not PAR4, we found that PAR1/4 depletion increased basal insulin receptor (IR) activity as measured by the uptake of a glucose-analog. PAR1/4-depleted ECs also showed increased insulin receptor Type-A (IR-A) spliceform expression, which has high insulin-independent basal activity. Conversely, treatment of ECs with a PAR1 ligand upregulated the insulin-dependent IR Type-B (IR-B) spliceform, suggesting that endothelial PAR1/4 can modulate IR splicing/activity. Additionally, we found that ECs in many wildtype mouse organs, including skeletal muscle, predominantly express IR-B in vivo. Therefore, a switch to IR-A expression in Par1/4dECko mice may enhance endothelial IR activity. IR activity in ECs regulates insulin transport from the bloodstream to the parenchyma. We propose that loss of EC PAR1/4 elevates whole-body insulin sensitivity by increasing IR splicing/activity and subsequent transcytosis to the parenchyma. Thus, EC PAR1/4 modulation may hold promise for treating diabetes.

Disclosure
R.Rajala: None. C.Griffin: None.

Funding
American Heart Association (23PRE1014240); National Institutes of Health (HL144605)
1514-P: Insulin Sensitivity Initially Worsens but Later Improves with Aging in C57BL/6N Mice
Authors: KONDO HIROYA , ONO HIRAKU , YOKOTE KOUTARO 
Published: June 2023

Objective: Aging is believed to induce insulin resistance in humans. However, when and how insulin sensitivity changes with aging remain unclear in both humans and mice.
Methods: Groups of C57BL/6N mice at 9-15 weeks (young), 34-67 weeks (one-year-old) and 107-121 weeks (two-year-old) of age underwent hyperinsulinemic-euglycemic clamp studies with somatostatin infusion under awake and non-restrained conditions.
Results: The glucose infusion rates for maintaining euglycemia were 17.11±2.94, 5.58±1.28 and 25.30±4.42 mg/kg/min in young, one-year-old and two-year-old mice, respectively. Thus, compared with young mice, one-year-old mice exhibited the expected insulin resistance. In contrast, the insulin sensitivity of two-year-old mice showed significant improvement. These age-related changes were mainly observed in glucose uptake into adipose tissue and skeletal muscle. Epididymal fat weight and hepatic triglyceride levels were highest in one-year-old mice, while being similar in young and two-year-old mice.
Conclusions: Our observations indicate that, in mice, insulin resistance appears at one year of age but subsequently improves markedly. These alterations in insulin sensitivity are attributable to changes in visceral fat accumulations and age-related factors.

Disclosure
H.Kondo: None. H.Ono: None. K.Yokote: None.

Funding
Japan Society for the Promotion of Science (15K09398, 18K08502, 20H00524); Japan Agency for Medical Research and Development (17GM5010002H0001, 18GM5010002H0002, 19GM5010002H0003, 20GM5010002H0004)
1516-P: Free Fatty Acid and Amino Acid Predictors of Response to Metformin in Polycystic Ovary Syndrome
Authors: CHANG ALICE Y. , ASOKAN ANEESH K. , SAKRIKAR DHANANJAY , LALIA ANTIGONI , LANZA IAN R. , PETTERSON MAI , SREEKUMARAN NAIR K. 
Published: June 2023

Metformin is one of the most widely used antidiabetic drugs and is also effectively used in treating women with polycystic ovary syndrome (PCOS). Our global objective is to separate women with PCOS into subtypes by their response to metformin. This pilot study aims to determine which free fatty acids and amino acids metabolites predict response to metformin as categorized by changes in insulin sensitivity and testosterone concentrations after three months of metformin therapy. We recruited 42 participants with PCOS aged between 18-50 years. Participants were randomly categorized into metformin (n= 29; 32.41 ± 7.4 years) and placebo groups (n=13; 32.15 ± 5.65 years). The Metformin group was challenged with three months of metformin titrated to at least 1500 mg daily. Body weights decreased significantly in the metformin treatment group from 98.55 ± 22.76 to 97.27 ± 23.14 (p-value=0.01) but did not change in the placebo group. Fasting plasma glucose decreased only in the metformin group from 89.25 ± 9.12 to 86.37 ± 9.53. Testosterone levels decreased in the metformin-treated group after 12 weeks from 29.0 (18.5 - 35.5) to 23.0 (15.5 - 32.5), but no significant change was observed in the placebo group from 28.0 (20.0 - 43.0) to 27.0 (21.5 -49.0). However, we did not observe a statistically significant change in insulin sensitivity values in PCOS and placebo groups. Linear regression analysis was performed to check the predictive power of those associated metabolites with delta values of testosterone and insulin sensitivity. Our results showed that changes in insulin sensitivity and testosterone levels after metformin treatment could be significantly predicted by using concentrations of metabolites (Fig.1).

Disclosure
A.Y. Chang: None. A.K. Asokan: None. D. Sakrikar: Employee; The Binding Site. A. Lalia: None. I.R. Lanza: None. M. Petterson: None. K. Nair: None.

Funding
National Institutes of Health (UL1TR000135)
1519-P: Role of Cyr61 in Palmitic Acid-Induced Insulin Resistance and Inflammation in Hepatocyte
Authors: HEO YUJUNG , JUNG KIM DAE , JIN KIM HAE , LEE NAMI , LEE KWANWOO , CHOI SUNG E.Y. , KANG YUP Y. , JIN HAN SEUNG , YOUNG JEON JA 
Published: June 2023

Objective: Cysteine-rich protein 61 (Cyr61, also known as Cellular Communication Network Factor 1, CCN1) is involved in a variety of diseases, such as carcinoma, inflammation, and fibrosis. It was recently reported that Cyr61 was associated with nonalcoholic fatty liver disease. Here, we evaluated the effects of Cyr61 on hepatic inflammation and insulin resistance in HepG2 cells and examined the molecular mechanisms involved.
Methods: We treated HepG2 cells with palmitic acid (PA) and then evaluated insulin signaling pathway using immunoblotting. After treatment with PA, the pro-inflammatory cytokines, including IL-6, TNF-α, and IL-1β, and Cyr61 were assessed by RT-PCR and ELISA. To investigate the effects of Cyr61 on insulin resistance and inflammation, we used Cyr61 small interfering (si) RNA and anti-Cyr61 neutralizing antibodies. The Cyr61 siRNA were transiently transfected into HepG2 cells using Lipofectamine 2000.
Results: Compared to the control, PA reduced IR phosphorylation, IRS-1 (Tyr612) phosphorylation, AKT phosphorylation, and GSK-3α/β phosphorylation after insulin stimulation. PA stimulated NF-κB and JNK activation in a dose-dependent manner. PA significantly increased the expression and secretion of Cyr61 and pro-inflammatory cytokines. Interestingly, neutralization of Cyr61 reduced NF-κB and JNK signaling, which was increased by PA, and recovered insulin signaling. In addition, Cyr61-si RNA treatment reduced the mRNA expression of PA-induced IL-6, TNF-α and IL-1β, and decreased NF-κB and JNK signaling.
Conclusion: Inhibition of Cyr61 recovered insulin signaling and reduced inflammation in HepG2 cells. Cyr61 may play a role in PA-induced insulin resistance and inflammation in HepG2 cells.

Disclosure
Y.Heo: None. D.Kim: None. H.Kim: None. N.Lee: None. K.Lee: None. S.E.Y.Choi: None. Y.Y.Kang: None. S.Han: None. J.Jeon: None.

Funding
National Research Foundation of Korea (2020R1F1A1074019, 2021M3H1A104892211)
152-OR: The Infected Diabetic Foot—The MRSA Incidence, Conversion of Non-MRSA to MRSA Pathogens, and Reinfection in Soft Tissue and Bone Infections
Authors: SULUDERE MEHMET A. , KILLEEN AMANDA L. , CRISOLOGO PETER , LAVERY LAWRENCE A. 
Published: June 2023

Aim: To evaluate the incidence of MRSA infection, re-infection, and the conversion of non-MRSA bacterial pathogens to MRSA in re-infections.
Methods: This was a post hoc analysis of 233 patients that participated in an RCT for moderate and severe diabetic foot infections with 12-month outcomes. We used bone culture and/or pathology to define osteomyelitis (OM). Soft tissue infections (STI) were defined based on negative bone biopsy or negative MRI. Moderate and severe infection was based on criteria of the International. Working Group on the Diabetic Foot infection classification. All subjects required surgery. We evaluated bacterial pathogens from bone and deep soft tissue from their initial infection and when there was re-infection. We defined conversion when a subject did not have MRSA in either their first OM or STI culture and had MRSA when there was re-infection. We used χ2 for comparison of clinical events, with alpha of &lt;0.05.
Results: OM was present in 73.0% (n=170) of cases. MRSA incidence was 8.6% (n=20), with no difference between STI and OM (14.3% vs. 6.5%, p=0.06). Reinfection occurred in 29.2% (n=68), with no difference between STI and OM (36.5% vs. 26.5%, p=0.14). MRSA was seen in 29.4% (n=20) of the reinfections, with no difference between STI and OM (13.0% vs. 28.9%, p=0.16). Conversion to MRSA was seen in 14.7% (n=10), with no difference between STI and OM (4.3% vs. 20%, p=0.12). MRSA was 4.4 times more likely in re-infection compared to index infections (29.4% vs 8.6%, CI 2.2-8.9, p&lt;0.01).
Conclusion: The incidence of MRSA is 4.4-fold higher after re-infection. The incidence of MRSA for the first infection (8.6%) was low. Re-infection with MRSA (29.4%) and conversion to MRSA (14.7%) were common. There was no difference seen in the incidence rates for reinfection, MRSA, reinfection with MRSA, and conversion to MRSA between patients with STI and OM.

Disclosure
M.A.Suludere: None. A.L.Killeen: Research Support; 3M/KCI, AstraZeneca, BioTissue surgical, Hologenix, LLC, Integra LifeSciences, Kent Imaging Inc., Microbion, PolarityTE, Inc., Smith+Nephew, Stark Enterprises, LLC, Xyken, VasoActiv, ViOptix. P.Crisologo: None. L.A.Lavery: Research Support; AstraZeneca, Tissue tech, American Diabetes Association, PolarityTE, Inc., Microbion, Jarvis, Integra Lifesciences Holdings Corp, Smith+Nephew.
1520-P: Metabolic Dysregulation in Probable Alzheimer’s Disease
Authors: READING CHRISTOPHER L. , AHLEM CLARENCE , PALUMBO JOSEPH M. , TESTA MARCIA A. , SIMONSON DONALD C. 
Published: June 2023

Insulin resistance, T2D, and chronic inflammation increase risk of developing mild cognitive impairment and Alzheimer’s disease (AD). We report here baseline data from a multicenter, randomized, placebo-controlled Phase 3 study (NCT04669028) in patients with mild to moderate AD evaluating the effects of NE3107, an oral small molecule, blood-brain permeable anti-inflammatory insulin sensitizer that binds extracellular signal-regulated kinase. Inclusion criteria are diagnosis and progression of AD, supportive brain scan and Hachinski scale to exclude vascular dementia, Clinical Dementia Rating (CDR) score of 1-2, and Mini Mental State Exam (MMSE) score of 14-24. Primary outcomes are changes in cognition and function from baseline to 30 wks. Secondary outcomes include changes in neuropsychological deficits, functional performance, and regulation of glycemia. Aβ+ subjects were older and had worse MMSE, ADAS-Cog12 and AD Composite Scores than Aβ-patients, but had less evidence of inflammation (lower CRP) and less insulin resistance (lower fasting glucose, less IFG and T2D, lower fasting insulin and HOMA2-IR, and less hypertension) (Table). Metabolic dysregulation (elevated C1q, increased CGM mean and MAGE, increased cholesterol, triglycerides, and waist-hip ratio) was also observed independent of Aβ and APOEε4 status. Inflammation and insulin resistance may contribute to AD even in the absence of classical risk markers for AD.

Disclosure
C.L.Reading: Employee; BioVie, Inc. C.Ahlem: Employee; Biovie, Inc. J.M.Palumbo: Employee; BioVie, Zynerba, Other Relationship; Merck Sharp & Dohme Corp. M.A.Testa: Stock/Shareholder; Phase V Technologies, Inc. D.C.Simonson: Stock/Shareholder; Phase V Technologies, Inc., GI Windows.
1525-P: Ventromedial Hypothalamic Nucleus Subset Stimulates Tissue Thermogenesis via Preoptic Outputs
Authors: ELMENDORF ANDREW J. , MAHLER CONNOR A. , LORENTZ BETTY , FLAK JONATHAN 
Published: June 2023

Changes in energy status ignite neural circuits that maintain body weight by engaging peripheral mechanisms that control energy expenditure; obesity results for when these responses are no longer sufficient. The key neural circuits that stimulate energy expenditure have not yet been established, but previous research has identified several critical sites (e.g. preoptic area (POA) and ventromedial hypothalamus (VMN)) that could be part of an interconnected system that controls tissue thermogenesis, which is essential for body weight control. Activating SF1-cre VMN cells in male diet-induced obese mice reduces body fat by 25% in 72 hours. This function is likely mediated at least in part by release of the peptide pituitary adenylate cyclase-activating polypeptide (PACAP), since knockout of PACAP in the VMN induces obesity at thermoneutrality (30C) and suppresses temperature above both the brown and beige adipose tissue in response to feeding. Because some VMN cells densely project to the POA, we hypothesized that VMN PACAP cells activate tissue thermogenesis via preoptic outputs. We placed temperature-sensing microchips above a traditional brown (interscapular brown adipose tissue (iBAT) and beige (inguinal white adipose tissue (iWAT) adipose tissue in Adcyap1(gene for PACAP)cre mice with DIO-ChR2 in the VMN and fiber optic implants in two major VMN outputs: the POA and the bed nucleus of the stria terminalis (BST). Activating VMN fibers in the POA that contain PACAP in both male and female mice, but not the BST, increased iBAT and iWAT temperature, and the effect was most robust when the mice were fasted. In summary, these data indicate that VMN PACAP connections with the POA is important for the induction of tissue thermogenesis, particularly when feeding conditions change. VMN PACAP, as well as associated signaling targets, are critical components to energy balance by activating adipose tissue mechanisms that stimulate energy expenditure.

Disclosure
A.J.Elmendorf: Other Relationship; Eli Lilly and Company. C.A.Mahler: Stock/Shareholder; AbbVie Inc., Pfizer Inc., AstraZeneca. B.Lorentz: Research Support; Eli Lilly and Company. J.Flak: Research Support; Eli Lilly and Company.

Funding
American Diabetes Association (Pathway 17-INI-15 to J.F.); Ralph W. and Grace M. Showalter Research Trust; Indiana University; Indiana Biosciences Research Institute
1527-P: Anorexia Induced by the Central Action of Fibroblast Growth Factor 1 (FGF1) Is Dependent on Reduced GABA Receptor Activation
Authors: SCARLETT JARRAD M. , ALONGE KIMBERLY M. , PHAN BAO ANH N. , MORTON GREGORY J. , SCHWARTZ MICHAEL W. 
Published: June 2023

Previously, we reported that in multiple rodent models of type 2 diabetes, remission of hyperglycemia lasting for weeks can be induced by a single intracerebroventricular (icv) injection of FGF1. This effect is accompanied by a potent but transient anorexic response that is sufficiently robust as to hinder efforts to translate these findings towards the clinic. To understand the mechanism underlying FGF1-induced anorexia, we considered 1) our recent finding that icv FGF1 induces acute and sustained inhibition of hypothalamic AgRP neurons, and 2) prior work showing that parabrachial nucleus that express calcitonin gene-related peptide (PBNCGRP neurons) are both highly anorexigenic and tonically inhibited by GABAergic projections from AgRP neurons. We therefore hypothesized that FGF1-induced anorexia involves activation PBNCGRPneurons arising from reduced GABAergic input. Consistent with this hypothesis, we report that PBNCGRPneurons are robustly activated following icv FGF1 injection (based on c-Fos staining) via a mechanism that appears to be indirect, since the c-Fos induction is not associated with FGF receptor (FGFR) activation (detected by pERK staining in the PBN). We next asked whether the anorexic response to icv FGF1 administration can be blunted by systemic administration of a GABAA receptor agonist. Consistent with this hypothesis, we found that continuous subcutaneous infusion of the GABAA receptor agonist Bretazenil blunts the anorexic response to icv FGF1 injection by ~50% at a dose that has no effect intake in control mice. Similarly, the effect of FGF1 to induce c-Fos in the PBN was largely blocked by Bretazenil, and our preliminary findings indicate that CGRP neurons are among those in which this effect occurred. Together, these data suggest that FGF1-induced anorexia results from reduced GABAergic inhibition of PBNCGRP neurons.

Disclosure
J.M.Scarlett: None. K.M.Alonge: None. B.N.Phan: None. G.J.Morton: Research Support; Novo Nordisk A/S. M.W.Schwartz: Consultant; NodThera, Research Support; Novo Nordisk A/S.

Funding
U.S. Department of Defense (PRMRP W81XWH-21-1-0635); National Institutes of Health (DK083042)
1530-P: Retinoic Acid Signals in the Hypothalamus Regulate Energy Balance in a Sexually Dimorphic Manner
Authors: YU MENG , YANG YONGJIE , HE YANG , BEAN JONATHAN , LIU HAILAN , WANG CHUNMEI 
Published: June 2023

Retinoic acid (RA) is the active metabolite of Vitamin A (Vit-A), and both Vit-A and RA are essential micronutrients for life. Vit-A-deficient mice are leaner than Vit-A-sufficient mice fed with either chow or high-fat-diet (HFD), implying the role of RA in promoting body weight gain. Global loss of key RA synthesis enzyme retinaldehyde dehydrogenase-1 (Aldh1a1) or RA receptor retinoid X receptor gama (RXRγ) both protects mice from diet-induced obesity (DIO). However, the metabolic roles of central RA, especially the endogenously RA synthesized in the hypothalamus, are unclear. Both the RA synthesizing enzymes including Aldh1a1 and RA receptors including RXRγ and retinoic acid receptor beta (RARβ), are accumulated in the hypothalamus. We found that loss of RXRγ or RARβ in pro-opiomelanocortin (POMC)-expressing neurons in the arcuate nucleus of hypothalamus (ARH) produced lean phenotypes in a sexually dimorphic manner. In particular, loss of RXRγ in POMC neurons reduce body weight and food intake only in chow-fed female mice, but not in male mice; meanwhile, loss of RARβ in in POMC neurons reduce body weight in both sexes, with reduced food intake in male mice but not in female mice. Independent of body weight, glucose tolerance and insulin tolerance were not changed by POMC specific deficiency of either RXRγ or RARβ in either sex. Since RA can bind to both RXRγ and RARβ, and RA can be locally synthesized in the hypothalamus using Vit-A metabolites as substrates, these results support that RA acts on RXRγ and RARβ in POMC neurons to regulate energy balance in a sexually dimorphic manner. These studies will advance our understanding about the physiology of body weight/glucose balance regulated by RA and RXR or RAR complexes, which will facilitate the characterization of the current RA or RXR-based therapies and provide novel targets for anti-obesity/diabetes therapies.

Disclosure
M.Yu: None. Y.Yang: None. Y.He: None. J.Bean: None. H.Liu: None. C.Wang: None.
154-OR: Metabolomics Analysis Reveals Links between Stress Hyperglycemia and Underlying Differences in Branched-Chain Amino-Acid Metabolism and Microbiome–Host Interactions
Authors: ABRAHAM AMALIA , SMITH MATTHEW R. , PEREZ-GUZMAN MIREYA , ZAHEDI TAJRISHI FARBOD , GUERRERO ARROYO LIZDA I. , CHAKRAGIRI AMSHUMANTH , SAM VARGHESE JITHIN , PASQUEL FRANCISCO J. 
Published: June 2023

The mechanisms that lead to stress-hyperglycemia are poorly understood. We examined metabolic signatures associated with stress-hyperglycemia in patients without diabetes undergoing cardiac surgery. We enrolled patients without diabetes [BG ≤125mg/dl and HbA1c&lt;6.5%] undergoing CABG surgery and compared those with stress hyperglycemia (“High”, mean glucose &gt;140 or glucose &gt;180mg/dl) vs those with preserved glucose metabolism (“Low”). Utilizing untargeted LCMS-derived high-resolution metabolomics (HRM) analysis we examined metabolites and metabolic pathways associated with incident stress hyperglycemia. Metabolic pathways were identified from HILIC positive and Reverse Phase C18 analysis, with metabolites mapped to Kyoto Encyclopedia of Genes and Genomes (KEGG) human pathways. A total of 36 patients (age: 62.7 years, 83% male, HbA1c 5.5%) underwent CABG surgery. Nine patients developed stress hyperglycemia immediately after starting surgery and showed differences in branched-chain amino acids (BCAA, e.g., methyl-oxopentanoate), lipid metabolism (e.g., diacylglycerols), and microbiome metabolism (e.g., linoleate), figure. This analysis reveals links between stress-hyperglycemia and BCAA, and lipid metabolism, and microbiome-host interactions among nondiabetic patients undergoing cardiac stress.

Disclosure
A. Abraham: None. M.R. Smith: None. M. Perez-Guzman: None. F. Zahedi Tajrishi: None. L.I. Guerrero Arroyo: None. A. Chakragiri: None. J. Varghese: None. F.J. Pasquel: Consultant; Dexcom, Inc., Medscape. Research Support; Dexcom, Inc., Ideal Medical Technologies.

Funding
National Institutes of Health (K23GM128221)