bluuebunny/crossref_metadata_2025_split huggingface dataset:
387-P: N-acetyl Cysteine Does Not Prevent Impaired Awareness of Hypoglycemia
Authors: MOHEET AMIR , KUMAR ANJALI , COLES LISA , RUBIN NATHAN , EBERLY LYNN E. , SEAQUIST ELIZABETH R. 
Published: June 2019

Impaired awareness of hypoglycemia (IAH) is common in diabetes. Administration of N-acetyl cysteine (NAC) during episodes of hypoglycemia (HG) prevents the development of IAH in rodents (Fioramonti 2013). Here we tested the hypothesis that NAC will be effective in preventing IAH in healthy humans using a double blind cross over study design where subjects received either NAC or saline (S) in random order on day 1 of a 2 day protocol that involved 3 HG clamps. On day 1, 350 mg NAC or S was given IV over 5 hours starting 30 min before start of IV insulin (2 mg/kg/min). Blood glucose (BG) was targeted at 50 mg/dl for 2 hours, then at 95 mg/dl for 2 hours, then 50 mg/dl for 2 hours. After BG recovery subjects were sent home to return on day 2 for a final 2-hour HG clamp. During the 1st and 3rd HG periods, serum epinephrine (EPI) and hypoglycemic symptoms (HSx) were collected. Subjects received 25 mg diphenhydramine IV at 0 and 4 hours due to a high occurrence of drug reactions. Additional doses or odansetron were given as needed.
EPI during clamp 1 was higher during NAC compared to S but the HSx were the same. No difference was seen between subjects who did or did not have a reaction to NAC. EPI and HSx on day 2 following NAC were lower than on day 1 but no different from response measured following S on day 1. NAC elicited many adverse events, enhanced EPI during HG, and did not alter EPI or HSx during HG on day 2 compared to S. Further development of IV NAC as a therapy for IAH is unwarranted.

Disclosure
A. Moheet: None. A. Kumar: None. L. Coles: None. N. Rubin: None. L.E. Eberly: None. E.R. Seaquist: Advisory Panel; Self; Eli Lilly and Company, Zucara Therapeutics Inc. Consultant; Self; 360 Consulting, MannKind Corporation. Research Support; Self; Eli Lilly and Company, JDRF, National Institutes of Health. Other Relationship; Self; American Diabetes Association, Sanofi, WebMD.

Funding
JDRF
4-LB: Feeding-Stimulated Regulation of the Epigenome Controls Adaptive Insulin Secretion
Authors: WORTHAM MATTHEW , LIU FENFEN , FLEISCHMAN JOHANNA , WALLACE MARTINA , HARRINGTON AUSTIN R. , CHIOU JOSHUA , GAULTON KYLE , METALLO CHRISTIAN , SANDER MAIKE 
Published: June 2019

Adaptation of the insulin secretory response to fluctuating insulin demand is critical for blood glucose homeostasis. We postulated that the epigenome could link changes in metabolic state with adaptive insulin secretion. Here we show that perturbations of nutrient state lead to extensive changes to H3K27 acetylation in pancreatic islets. Genomic sites of H3K27ac deposition acutely regulated by feeding were enriched for binding by the histone demethylase Lsd1, a candidate nutrient sensor known to respond to hormonal and metabolic stimuli. Conditional deletion of Lsd1 in mouse β-cells resulted in hypoglycemia and fasting hyperinsulinemia. Lsd1-deficient β-cells exhibited changes in H3K4me1 and H3K27ac deposition at Lsd1-bound sites. These changes at the chromatin level associated with aberrant expression of nutrient signaling and metabolic genes. We further show that Lsd1 controls feeding-stimulated transcription downstream of cAMP, which has been implicated in the control of β-cell metabolism. Accordingly, Lsd1-deficient β-cells failed to attenuate glucose metabolism in the fasted state, resulting in basal insulin hypersecretion. Further supporting a role for Lsd1 in adaptive insulin secretion, islets from hyperinsulinemic db/db mice exhibited increased H3K27ac and H3K4me1 levels at Lsd1-bound sites. Moreover, genomic sites bound by LSD1 in human islets were enriched for genetic variants associated with fasting glucose and type 2 diabetes in genome-wide association studies, suggesting a conserved role of LSD1 in the regulation of insulin secretion. Combined, our findings provide evidence that nutrient state modulates adaptive insulin secretion through Lsd1-dependent changes to the epigenome.

Disclosure
M. Wortham: None. F. Liu: None. J. Fleischman: None. M. Wallace: None. A.R. Harrington: None. J. Chiou: None. K. Gaulton: None. C. Metallo: Employee; Spouse/Partner; ICON plc. Research Support; Self; Gilead Sciences, Inc., Nimbus Therapeutics. M. Sander: None.

Funding
National Institutes of Health (R01DK068471); JDRF (3-PDF-2014-193-A-N)
402-P: Higher Glucose Thresholds for Hypoglycemia Alarms on Continuous Glucose Monitoring Systems Correlate with Less Time in Hypoglycemia
Authors: LIN YU KUEI , HUNG MAN , SHARMA ANU , CHAN OWEN , VARNER MICHAEL , GROAT DANIELLE , DOPITA DANA , FISHER SIMON 
Published: June 2019

Based on the set glucose thresholds for hypoglycemia alarms, continuous glucose monitoring (CGM) systems can provide audible alerts that notify individuals for actual/impending hypoglycemic events. While CGMs have been shown to reduce hypoglycemic events in patients with type 1 diabetes (T1D), there are little data demonstrating the association between glucose thresholds for hypoglycemia alarms and clinical hypoglycemia outcomes. This cross-sectional observational study enrolled 83 T1D patients with CGM use &gt;3 months, active CGM usage time ≥70% and activated (i.e., turned “on”) hypoglycemia alarms. CGM settings and two-week CGM glucose data (total 25,683 hours) were analyzed. Glucose thresholds for hypoglycemia alarms (mean: 74 ± 8 mg/dL; range: 60-100 mg/dL) correlated inversely with time in hypoglycemia (both glucose &lt;54 and &lt;70 mg/dL) (Figure 1a). When comparing the cohort with hypoglycemia alarm threshold settings at glucose level ≤70 mg/dL (n=42), threshold settings ≥75 mg/dL (n=41) were associated with a 74% and 60% reduction in hypoglycemia time (&lt;54 and &lt;70 mg/dL, respectively) (Figure 1b). All subjects with hypoglycemia alarm set at glucose ≥75 mg/dL spent less than 3% of time with glucose &lt;54 mg/dL. The current data support the use of higher glucose thresholds on CGM hypoglycemia alarms to decrease the time in hypoglycemia in patients with T1D.

Disclosure
Y. Lin: None. M. Hung: None. A. Sharma: None. O. Chan: Research Support; Self; Zucara Therapeutics Inc. M. Varner: None. D. Groat: None. D. Dopita: None. S. Fisher: None.
406-P: Effect of Metformin on Dicarbonyl Stress in Patients with Type 2 Diabetes and Chronic Heart Failure
Authors: KOPECKY JAN , HOŠKOVÁ EVA , VELEBA JIRI , MALINSKA HANA , VELEBOVA KATERINA , MELENOVSKY VOJTECH , PELIKANOVA TEREZIE 
Published: June 2019

Background and Aims: Treatment with metformin (MET) in diabetic patients with chronic heart failure (HF) is associated with better cardiovascular outcomes. The endothelial dysfunction, atherosclerosis and macrovascular diabetic complications are associated with excessive generation of reactive dicarbonyls like methylglyoxal (MG). MET has been proposed as a scavenger of methylglyoxal. The aim of the study was to evaluate the effects of MET on substrate metabolism and dicarbonyl stress in patients with DM and HF.
Materials and Methods: A randomized, double-blind, placebo-controlled, crossover study in a total time of six months testing the effect of 3- month usage of MET (2 g/day) vs. placebo. A group of 40 treatment naive patients with type 2 DM and HF were included. At the beginning and at the end of each intervention period various metabolic tests were done including hyperinsulinemic-isoglycemic clamp, parameters of dicarbonyl stress like MG, Glutathione (GSH), glutathione disulfide (GSSG), and the activity of Glyoxalase1 (GLO-1).
Results: MET significantly reduced HbA1c compared to placebo (p˂0.01). Insulin sensitivity, measured as a metabolic clearance rate of glucose (MCR) during the clamp, improved significantly with MET (p˂0.001) but also, although less significantly, with placebo (p˂0.05), difference between MET and placebo was not significant. Compared to placebo, GHS levels increased after the intervention (baseline vs. MET vs. PL: 866 ± 231 vs. 994 ± 342 vs. 878 ± 266µmol/l, p&lt;0.05). GSH/GSSG ratio was not affected. The levels of MG decreased after MET (p&lt;0.05), decrease after placebo was not significant. The activity of GLO-1 was not affected.
Conclusions: Our results on a unique group of patients with heart failure and diabetes support the evidence that one of the cardioprotective pathways of metformin is through reducing dicarbonyl stress. As there was no change in the GLO-1 activity, it might be due to binding and deactivation of reactive dicarbonyls by MET.

Disclosure
J. Kopecky: None. E. Hošková: None. J. Veleba: None. H. Malinska: None. K. Velebova: None. V. Melenovsky: None. T. Pelikanova: None.

Funding
Agentura pro zdravotnický výzkum Ceské Republiky (AZV16-27496A)
409-P: Cardiac Biomarkers and Incident Peripheral Arterial Disease (PAD) in Patients with Type 2 Diabetes in the ADVANCE Trial
Authors: HICKS CAITLIN W. , WANG DAN , MATSUSHITA KUNIHIRO , CHALMERS JOHN , WOODWARD MARK , SELVIN ELIZABETH 
Published: June 2019

Background: High-sensitivity cardiac troponin T (hs-cTnT) and natriuretic peptide (NT-proBNP) are strongly associated with coronary heart disease, heart failure, and death in population-based studies. Little is known about the associations of these cardiac biomarkers with incident PAD in adults with type 2 diabetes.
Methods: We used Cox models to assess the associations of elevated baseline hs-cTnT (≥ 14 ng/L) and NT-proBNP (≥ 200 pg/mL) in participants in the ADVANCE trial with measured biomarkers and without PAD at baseline. We also modeled hs-cTnT and NTproBNP using restricted cubic splines.
Results: Incident PAD occurred in 417 of 6,428 ADVANCE participants during a median follow-up of 5 years. After multivariable adjustment, increasing baseline hs-cTnT and NT-proBNP were both associated with PAD (Figure). Modeled categorically, PAD risk was higher in participants with elevated hs-cTnT (HR 1.35, 95% CI 1.07-1.70) or NT-proBNP (HR 1.44, 95% CI 1.15-1.81). Participants with elevations of both biomarkers had the highest risk (HR 1.85, 95% CI 1.38-2.49).
Conclusion: Hs-cTnT and NT-proBNP were associated with incident PAD in adults with diabetes even after adjusting for cardiovascular risk factors. Cardiac biomarkers may be useful to identify persons with type 2 diabetes at risk for future PAD.

Disclosure
C.W. Hicks: None. D. Wang: None. K. Matsushita: Consultant; Self; Akebia, Kyowa Hakko Kirin Co., Ltd. Research Support; Self; Fukuda Denshi, Kyowa Hakko Kirin Co., Ltd. J. Chalmers: Research Support; Self; Idorsia. M. Woodward: Consultant; Self; Amgen Inc., Kirin. E. Selvin: None.
414-P: Insulin Secretion Capacity Relates to Human Ventricular Myocardial Mitochondrial Function
Authors: ZWECK ELRIC , SCHEIBER DANIEL , ALBERMANN SOPHIE , BORGER JULIUS , JELENIK TOMAS , PESTA DOMINIK , HORN PATRICK , BOEKEN UDO , AKHYARI PAYAM , KELM MALTE , RODEN MICHAEL , WESTENFELD RALF , SZENDROEDI JULIA 
Published: June 2019

Hyperinsulinemia and impaired insulin secretion are potential contributors to the development of diabetes-related heart failure. Interference with ventricular myocardial mitochondria might play a key pathogenic role, but human data confirming these connections are limited. We hypothesized that ventricular myocardial mitochondrial function is impaired in hyperinsulinemia but is positively related to insulin secretion capacity. We recruited 32 heart transplant recipients without heart failure or antihyperglycemic treatment (age: 53.5±13.3years; BMI: 24.8±2.9kg/m²; HbA1c: 5.9±0.7%). We measured serum insulin levels and blood glucose every 30 minutes for three hours after an oral 75g glucose uptake. Seven patients were newly diagnosed with type 2 diabetes and six had impaired glucose tolerance. High-resolution respirometry was performed on transcatheter ventricular endomyocardial biopsies to determine mitochondrial oxidative capacity, coupling efficiency (respiratory control ratio, RCR) and intrinsic uncoupling (leak control ratio, LCR). Fasting serum insulin, a marker of insulin resistance, was inversely associated with ventricular myocardial mitochondrial RCR (r=-0.39; p=0.02) and positively with LCR (r=0.42; p=0.02), but not with oxidative capacity (p=0.34). Maximum increase of insulin in proportion to fasting levels strongly related to ventricular myocardial mitochondrial RCR (r=0.62; p=0.0001) and inversely to LCR (r=-0.46; p=0.01). This insulin secretory response related to fatty acid-dependent respiration (r=0.35; p=0.05) and tended to relate to combined oxidative capacity from fatty acid and glycolytic substrates (r=0.31; p=0.09). Maximum insulin response following glucose ingestion relates to cardiac mitochondrial efficiency and oxidative capacity, while insulin resistance might affect cardiac energy metabolism. These data suggest a pathogenic role of hyperinsulinemia as well as beta cell failure in diabetes-related heart failure.

Disclosure
E. Zweck: None. D. Scheiber: None. S. Albermann: None. J. Borger: None. T. Jelenik: None. D. Pesta: None. P. Horn: None. U. Boeken: None. P. AKhyari: Research Support; Self; Abbott, Edwards Lifesciences Corporation, Medtronic. M. Kelm: None. M. Roden: Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Poxel SA, Servier. Board Member; Self; Eli Lilly and Company. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi. Speaker's Bureau; Self; Novo Nordisk Inc. R. Westenfeld: None. J. Szendroedi: None.

Funding
German Research Foundation (CRC1116); Heinrich-Heine University
419-P: LDL Particle Size Is a Predictor of Cardiovascular Events in Cardiovascular Disease Patients with Nonalcoholic Fatty Liver Disease as Well as in Those without Nonalcoholic Fatty Liver Disease
Authors: LEIHERER ANDREAS , MUENDLEIN AXEL , SAELY CHRISTOPH H. , VONBANK ALEXANDER , LARCHER BARBARA , MADER ARTHUR , DOPHEIDE JÖRN F. , BAUMGARTNER IRIS , FRAUNBERGER PETER , DREXEL HEINZ 
Published: June 2019

Nonalcoholic fatty liver disease (NAFLD) is associated with atherogenic dyslipidemia, in particular with a small LDL particle size. However, there are no data on the association between LDL particle size and cardiovascular events in cardiovascular disease patients with NAFLD. This issue is addressed in the present study. We measured LDL particle size and concentration in 298 statin-naïve cardiovascular disease patients, of whom 141 had NAFLD. In patients with NAFLD, LDL particle size was significantly lower (21.0 vs. 21.2 nm, p&lt;0.001) and particle concentration was significantly higher (1487 vs. 1336 nmol/l, p=0.006) than in those without NAFLD. Prospectively, we recorded 135 cardiovascular events during a follow-up time of 8.2±3.3 years. In multivariate Cox regression analysis LDL particle size (standardized adjusted HR 0.68 [0.54-0.85], p=0.001), but not LDL particle concentration (standardized adjusted HR 0.97 [0.79-1.19], p=0.774) was a significant and independent predictor of cardiovascular events after adjustment for age, sex, BMI, hypertension, smoking, HDL particle size and HDL particle concentration, and for the presence of T2DM and NAFLD. In subgroup analysis regarding the presence of NAFLD, LDL particle size significantly predicted cardiovascular events both in patients with NAFLD (standardized adjusted HR 0.60 [0.42-0.86], p=0.005) and in those without NAFLD (standardized adjusted HR 0.64 [0.46-0.90], p=0.010). We conclude that LDL particle size is a predictor of cardiovascular events in cardiovascular disease patients with NAFLD as well as in those without NAFLD.

Disclosure
A. Leiherer: None. A. Muendlein: None. C.H. Saely: None. A. Vonbank: None. B. Larcher: None. A. Mader: None. J.F. Dopheide: None. I. Baumgartner: None. P. Fraunberger: None. H. Drexel: None.
42-LB: A New Laminopathy Caused by an Arg133/leu Mutation in Lamin A/C and the Effects thereof on Adipocyte Differentiation and the Transcriptome
Authors: WANG ZHE , HE YINGZI , ZHENG FENPING 
Published: June 2019

Mutations in LMNA trigger changes in nuclear architecture, signal transduction, and gene regulation, associated with a group of multisystem diseases termed laminopathies. Here we report a new laminopathy in a 26-year-old female, characterized by generalized lipoatrophy, insulin-resistant diabetes, micrognathia, symptoms of premature aging, hypertension, and biopsy-proven focal segmental glomerulosclerosis. The de novo heterozygous mutation R133L lamin A/C gene (LMNA) was found in the patient and inherited to her son who has not (yet) developed a lipodystrophic phenotype. We analyzed the nuclear morphology and laminar distribution of both 3T3-L1 pre-adipocytes and mouse glomerular mesangial cells overexpressing human wild type lamin A/C (LMNA WT) or lamin A/C with the R133L mutation (LMNA R133L) that causes the new laminopathy. We found that nuclear morphology was abnormal; the nuclei evidenced hypertrophy, nuclear membrane invagination or blebbing, and a disorganized A-type lamin meshwork in both cell lines overexpressing LMNA R133L. The differentiation from 3T3-L1 pre-adipocyte into adipocytes was impaired in cells expressing LMNA R133L; the expression levels of genes associated with adipose tissue self-renewal, including adipogenesis and angiogenesis were downregulated. Furthermore, the insulin-signaling pathway was inhibited in LMNA R133L adipocytes. Microarray gene expression profiling showed that the most prominent differences between 3T3-L1 cells expressing LMNA WT and LMNA R133L were in genes implicated in metabolic pathways, the cellular response to DNA damage, DNA repair, and RNA binding. We thus expanded the clinical spectrum of laminopathy and concluded that the LMNA R133L mutation associated with lipodystrophic features, multisystem disorders and the repairation of DNA damage.

Disclosure
Z. Wang: None. Y. He: None. F. Zheng: None.

Funding
Zhejiang Provincial Natural Science Foundation of China (Y17H070002); National Natural Science Foundation of China (81471099)
42-OR: Hub Cells Orchestrate 3-Dimensional Pancreatic Beta-Cell Ca2+ Dynamics In Vivo
Authors: SALEM VICTORIA , DELGADILLO SILVA LUIS F. , SUBA KINGA , ALONSO ALDARA MARTIN , KIM WHEI-CHANG , KALOGIANNI VASILIKI , AKHTAR NADEEM , GHARAVY NEDA MOUSAVY , GEORGIADOU ELENI , LECLERC ISABELLE , BRIANT LINFORD , HODSON DAVID , DISTASO WALTER , NINOV NIKOLAY N. , RUTTER GUY A. 
Published: June 2019

Aim: In-vitro studies reveal a subpopulation of β cells which co-ordinate insulin release from pancreatic islets. This connectivity has not been established in 3 dimensions across the islet or in the living animal. We investigated this in the living zebrafish D. rario or in mouse islets engrafted in the anterior eye chamber (ACE), using the recombinant Ca2+ sensor GCaMP6 under the insulin promoter/C57Bl6, Ins1Cre:GCaM6ff/f.
Methods: Whole islet live imaging of D. rario primary islets in situ at an acquisition rate of 0.8Hz, covering 700μm, was achieved using resonant scanner technology with a Leica inverted laser scanning confocal system, using a 25X/N.A. 0.95 water lens. Fully vascularized islets in the murine ACE were imaged under isofluorane anaesthesia on a modified Nikon Ti-E spinning disc confocal microscope (20x 0.75 NA water immersion objective, 3 Hz frame rate).
Results: 3-dimensional β cell connectivity (Pearson R statistic) significantly rose to 88% across the zebrafish islet in response to glucose stimulation. Focused ablation of the co-ordinating “hub” (but not follower) β cells significantly attenuated the subsequent response to a glucose challenge. In mouse islets, high circulating glucose levels were also associated with a significant rise in 3D beta cell connectivity from 65 to 86% (n=3; p=0.02). Granger causality analysis on these islets revealed that even on prolonged (&gt;10 minute) glucose stimulation, pan-islet connectivity was sustained and the most highly connected cells were those in the region of the islet from which Ca2+ waves emanated. Binarized signal analysis (n=6) separately confirmed the presence of a highly-connected β cell subpopulation (8.7 ± 3.6% of cells).
Conclusions: These studies evidence beta cell “hubs” which are responsible for the initiation and sustained coordination of the Ca2+ response to glucose across fish and mouse islets in vivo.

Disclosure
V. Salem: None. L.F. Delgadillo Silva: None. K. Suba: None. A. Martin Alonso: None. W. Kim: None. V. Kalogianni: None. N. Akhtar: None. N. Mousavy Gharavy: None. E. Georgiadou: None. I. Leclerc: None. L. Briant: None. D. Hodson: None. W. Distaso: None. N.N. Ninov: None. G.A. Rutter: Consultant; Self; Sun Pharma.

Funding
Diabetes UK; UK Wellcome Trust
425-P: Inhibition of miR-181c Rescues Diabetes-Impaired Angiogenesis
Authors: SOLLY EMMA , HOURIGAN SAMUEL T. , MULANGALA JOCELYNE , PSALTIS PETER J. , BARTOLO BELINDA DI , NG MARTIN , NICHOLLS STEPHEN , BURSILL CHRISTINA , TAN JOANNE T.M. 
Published: June 2019

Diabetic vascular complications are characterized by impaired angiogenic responses to ischemia, resulting in delayed healing following tissue injury. MicroRNAs (miRNAs), small non-coding RNAs that target gene expression, regulate angiogenesis in a time-dependent manner. Using a miRNA array approach, we discovered that miR-181c was regulated by angiogenic stimuli but had no previous reported role in diabetes. We sought to elucidate the role of miR-181c in diabetes-impaired angiogenesis. We found that miR-181c was inhibited in endothelial cells (ECs) exposed to hypoxia but were, conversely, elevated in high glucose. To assess the importance of miR-181c in angiogenesis, ECs were transfected with mimics or antimiRs to mimic or suppress miR-181c activity respectively. Cells with increased miR-181c activity had a reduced capacity to form tubules and lower VEGFA protein levels. Conversely, suppression of miR-181c activity augmented tubule formation and VEGFA expression. This suggests that miR-181c plays an inhibitory role in angiogenesis, at least in part, through VEGFA regulation. In vivo, miR-181c was temporally tracked following ischemic induction in the hindlimb. Early post-ischemia, when angiogenesis is important, diabetic mice had elevated miR-181c levels compared to nondiabetic controls. However, at later timepoints, miR-181c levels dropped to levels similar to nondiabetic mice. This indicates that miR-181c suppression may be critical to facilitate early responses to ischemia. Diabetic mice were injected with antimiR-181c or antimiR-Neg to determine if miR-181c knockdown rescues diabetes-impaired angiogenesis. Inhibition of miR-181c completely rescued diabetes-impaired blood flow reperfusion, restoring it back to that of nondiabetic levels. Taken together, this study has identified that miR-181c plays a key role in the regulation of angiogenesis in ischemia and in diabetes. This highlights a potential new therapeutic approach for the treatment of diabetic vascular complications.

Disclosure
E. Solly: None. S.T. Hourigan: None. J. Mulangala: None. P.J. Psaltis: None. B. Di Bartolo: None. M. Ng: None. S. Nicholls: Consultant; Self; Amgen Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Cerenis, CSL Behring, Eli Lilly and Company, Merck Sharp & Dohme Corp., Novo Nordisk Inc., Omthera, Pfizer Inc., Resverlogix Corp., Roche Pharma, Sanofi, Takeda Pharmaceutical Company Limited. Research Support; Self; Amgen Inc., AstraZeneca, Athernova, Cerenis, Eli Lilly and Company, InfraReDx, Novartis Pharmaceuticals Corporation, Resverlogix Corp., Sanofi. C. Bursill: None. J.T.M. Tan: None.

Funding
Diabetes Australia