bluuebunny/crossref_metadata_2025 huggingface dataset:
400-P: Impact of Hypoglycemic Episodes (Hypos) on Type 1 Diabetes Mellitus (T1DM)
Authors: SULLIVAN EMMA D. , KEMP JESSALYN , PIERCY JAMES , HIGGINS VICTORIA
Published: June 2019

Objective: Compare functionality of T1DM patients who do or do not experience hypos and compare across hypo severities and requirement for medical help.

Methods: Data were drawn from 2016 Adelphi Disease Specific Programme, a real-world, cross-sectional study in 5EU/U.S. Physicians completed forms for the next 2 T1DM patients capturing comorbidities, demographics, disease duration, hypo details. Patients self-reported work, activity impairment, lifestyle impact. Patients were grouped based on hypo experience by 1. ever experienced vs. never, 2. mild vs. severe, 3. never needed medical attention vs. needed. Propensity score matching on gender, BMI, disease duration facilitated comparison between matched groups. p-values generated using Abadie-Imbens standard errors, and standardized mean differences <10% in matching covariates were sought.

Results: Of 1662 patients, 861 (51.8%) never experienced hypos, 637 (38.3%) experienced mild and 164 (9.9%) severe hypos. Only 46 (2.8%) experienced a hypo needing medical help. Patients with hypos were more likely ‘sometimes/greatly’ affected in feelings about the future and freedom to eat/drink (both p<=0.001), leisure activities (p<0.05) and self-confidence (p=0.056) vs. patients with no hypos. Severe hypo patients were more likely ‘sometimes/greatly’ affected when traveling (p=0.019) and were more likely to be anxious (p=0.007) or depressed (p=0.064) vs. mild hypo patients. While analysis of hypos needing medical attention was limited due to low bases, patients needing medical attention were more likely to have depression (p=0.003), anxiety (p=0.053) and increased work absenteeism (p<0.01).

Conclusion: While physicians report half of T1DM patients do not experience hypos, those who do are more impacted concerning lifestyle, confidence and outlook. More severe hypos translates to increased psychological manifestations. During the management of these patients, the broader implications of hypo experience should not be underestimated.

Disclosure

E.D. Sullivan: None. J. Kemp: None. J. Piercy: None. V. Higgins: None.
415-P: Detection of Vascular Endothelial Cell DNA in the Circulation Using Dual Amplification Refractory Mutation System PCR
Authors: KURODA AKIO , YAMADA MISUZU YAMASHITA , TAKASHI YUICHI , OHISHI MAMI , MORI HIROYASU , ISHIZU MASASHI , SUZUKI REIKO , YINHUA OTSUKA , TOMINAGA YUKARI , MATSUHISA MUNEHIDE
Published: June 2019

Background: Vascular complication of type 1 diabetes is associated with multiple factors. However, biomarkers which directly reflect vascular complication have not been elucidated. We have developed a method to detect unmethylated status of pancreatic β cell specific CpG sequences of insulin gene from cell-free DNA (cfDNA) in the circulation. Transmembrane protein ROBO4 is highly expressed in vascular endothelial cells, and it has been reported that the CpG sites in the promoter region are specifically unmethylated.

Aim: To develop a novel PCR method to specifically detect unmethylated CpG sites of ROBO4 gene promoter region in serum cfDNA and to evaluate this method in the supernatant of vascular endothelial cell culture and serum of patients with type 1 diabetes.

Material and Methods: After bisulfite conversion of cfDNA extracted from human umbilical vein endothelial cells (HUVEC) and patient serum, unmethylated ROBO4 gene was detected by the dual Amplifications Refractory Mutation Systems PCR, which amplifies only if 4 CpG sites (-185, -144, -118 and -103bp of transcription start sites of ROBO4) are all unmetylated. These methods were applied to 83 patients with type 1 diabetes (duration 16.1±13.9 years) and clinical characteristics were compared.

Results: In HUVEC culture supernatant under constant and intermittent high glucose condition, the detection of ROBO4 gene copy number increased depending on the number of dead cells and cfDNA concentration. A positive correlation (p <0.05) with the mean intima-media thickness (IMT) and MCV were observed for the ROBO4 copy numbers in patient serum cfDNA. According to the multiple regression analysis, unmethylated ROBO4 copy number (p <0.001) and age (p <0.001) were selected as the determinant factors of mean IMT.

Conclusion: Novel PCR method for ROBO4 could detect vascular endothelial cell injury and atherosclerosis quantitatively in patients with type 1 diabetes.

Disclosure

A. Kuroda: Speaker's Bureau; Self; Eli Lilly and Company, Novo Nordisk Inc., Sanofi. M.Y. Yamada: None. Y. Takashi: None. M. Ohishi: None. H. Mori: None. M. Ishizu: None. R. Suzuki: None. O. Yinhua: None. Y. Tominaga: None. M. Matsuhisa: Advisory Panel; Self; Eli Lilly and Company. Board Member; Self; Daiichi Sankyo Company, Limited. Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Mitsubishi Tanabe Pharma Corporation. Speaker's Bureau; Self; Astellas Pharma Inc., Eli Lilly and Company, Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Sanofi, Takeda Pharmaceutical Company Limited.

Funding

Japan Society for the Promotion of Science
417-P: Serum Ceramide Ratios Predict Cardiovascular Events in Patients with Type 2 Diabetes Independently from the Presence of Coronary Artery Disease
Authors: LEIHERER ANDREAS , MUENDLEIN AXEL , SAELY CHRISTOPH H. , LAAKSONEN REJO , LAAPERI MITJA , VONBANK ALEXANDER , LARCHER BARBARA , MADER ARTHUR , FRAUNBERGER PETER , DREXEL HEINZ
Published: June 2019

The ceramide ratios Cer(d18:1/16:0)/Cer(d18:1/24:0), Cer(d18:1/18:0)/Cer(d18:1/24:0) and Cer(d18:1/24:1)/Cer(d18:1/24:0) have recently been reported to be associated with cardiovascular risk. Data on their power to predict cardiovascular events in patients with type 2 diabetes (T2DM) are sparse, and it is unknown whether they prospectively predict cardiovascular events independently from the presence of coronary artery disease (CAD) at baseline. These issues are addressed in the present study. We measured the above-mentioned serum ceramide ratios in a cohort of 894 patients in whom the presence of CAD was determined angiographically. Prospectively, we recorded cardiovascular events over a mean follow-up time of 9.2±3.2 years. At baseline, the ratios Cer(d18:1/16:0)/Cer(d18:1/24:0), Cer(d18:1/18:0)/Cer(d18:1/24:0) and Cer(d18:1/24:1)/Cer(d18:1/24:0) were significantly higher in T2DM patients (n=239) than in patients without T2DM (0.11 vs. 0.10, p=0.011; 0.46 vs. 0.39, p<0.001; and 0.46 vs. 0.41, p<0.001), but did not differ significantly between patients with CAD (n=491) and those without CAD. Prospectively, vascular events occurred in 110 (47%) of the patients with T2DM and in 212 (33%) of nondiabetic patients (p<0.001). For the ratio Cer(d18:1/16:0)/Cer(d18:1/24:0) a significant association with cardiovascular events in T2DM patients could be demonstrated univariately (standardized HR 1.23 [1.07-1.41], p=0.003), and after adjustment for age, gender, BMI, smoking, LDL cholesterol, HDL cholesterol, hypertension, statin intake, and also the for the CAD status at baseline (standardized adjusted HR 1.28 [1.10-1.50], p=0.002). We conclude that ceramide ratios have the potential to predict cardiovascular events in patients with T2DM, independent from their baseline CAD status. Further research addressing the selection of optimal ceramide species for risk prediction appears warranted.

Disclosure

A. Leiherer: None. A. Muendlein: None. C.H. Saely: None. R. Laaksonen: None. M. Laaperi: None. A. Vonbank: None. B. Larcher: None. A. Mader: None. P. Fraunberger: None. H. Drexel: None.
435-P: Elevated One-Hour Post-Challenge Glucose Levels Associated with Increased Arterial Stiffness
Authors: OKADA SHUICHI , OKADA JUNICHI , YAMADA EIJIRO , SAITO TSUGUMICHI , YAMADA MASANOBU
Published: June 2019

We reported a positive correlation between brachial-ankle pulse wave velocity (baPWV) and one-hour post-challenge glucose (1-h PG). Furthermore, the subpopulation defined by 1-hour PG beyond 183 mg/dL (corresponded to Impaired glucose tolerance range of 2-h PG) in normal glucose tolerant (NGT) subjects. These findings suggested that if NGT subjects convert to higher 1-h PG levels from a normal 1-h PG range they will develop arterial stiffness. To address this issue, we initiated a clinical study that included 40 NGT subjects (23 men and 17 women, mean age 60.9 ±9.7 years), with a comprehensive medical check-up was performed on each subject at Kan-etsu Chuo Hospital every year. In addition, over the 9-year period these individuals were annually assessed by a 75-g oral glucose tolerance test (OGTT) and for atrial stiffness by baPWV. Based on the 75-g OGTT, NGT subjects alone were selected according to the 2006 WHO criteria. NGT subjects in this study were identified as individuals having a normal range of all measured parameters. In this population, there was no significant correlation between baPVW and all measured parameters. One-h post challenge glucose (1-h PG) in 28 subjects was below 154 mg/dL and their baPWV was not changed (1266.4 ±125.42 cm/sec from 1243.62 ±108.1) after nine years. On the other hand, 1-h PG in the rest of 12 subjects was significantly increased to 160.33 ±4.73 from 124.67 ±20.13 and their baPVW was also significantly elevated to 1489.33 ±41.05 from 1255.01 ±64.08 after nine years. We found that rapid blood glucose change in NGT could be a risk factor for arterial stiffness and we would like to point out the possibility for physicians to select high-risk subjects by 75g-OGTT with the evaluation of 1h-PG. Taken together, we found that rapid blood glucose change could be a risk factor for arterial stiffness.

Disclosure

S. Okada: None. J. Okada: None. E. Yamada: None. T. Saito: None. M. Yamada: None.
44-OR: Human Pseudo Islets Reveal Differential Effects of Gq Signaling in ß Cells in Low and High Glucose
Authors: WALKER JOHN T. , HALIYUR RACHANA , ARAMANDLA RADHIKA , POFFENBERGER GREG , NELSON HEATHER A. , LUCHSINGER JOSEPH , WANG PENG , GARCIA-OCAÑA ADOLFO , BRISSOVA MARCELA , POWERS ALVIN C.
Published: June 2019

Efforts to advance our understanding of human islet biology are hindered by a paucity of approaches to effectively manipulate gene expression in human islets. To address this, we developed a pseudoislet system of human islet dispersion and reaggregation via modified hanging drop with 2500 cells/drop to generate 150-200 μm diameter pseudoislets (n=9 normal donors; 26-73 years). Pseudoislet morphology, dithizone uptake, and expression of markers of α and/or β cell identity (PDX1, NKX6.1, ARX, MAFB, NKX2.2, and PAX6) were similar to native islets; in paired comparisons from the same donor, pseudoislets had slightly fewer β cells and more α cells (53% β, 44% α, 3% δ cells) than native islets (62% β, 35% α, 3% δ cells; p<0.05 β and α cells). In perifusion, pseudoislets were similar to native islets in biphasic glucose-stimulated and cAMP-potentiated insulin secretion as well as in epinephrine and low glucose stimulation of and high glucose inhibition of glucagon release. Adenoviral-transduced dispersed human islet cells (>90% β cell transduction) formed functioning pseudoislets similar to untransduced controls. Using this approach, we virally expressed hM3Dq, a Gq-coupled designer receptor exclusively activated by a designer drug (DREADD), which selectively responds to the otherwise inert ligand clozapine-n-oxide (CNO) and signals through PLC/IP3. In low glucose (2mM G + 10μM CNO), hM3Dq pseudoislets secreted more insulin than control mCherry pseudoislets (19.2 vs. 1.0 ng/100IEQ/hr; p<0.0001). In contrast, hM3Dq and control pseudoislets had similar insulin secretion in high glucose (11mM G + 10μM CNO; 24.8 vs. 23.2 ng/100IEQ/hr; p>0.05), indicating that activation of Gq signaling does not potentiate insulin secretion in high glucose. These data demonstrate how manipulation of gene expression in pseudoislets can provide insight into human islet biology and suggest that activation of Gq signaling in β cells elicits robust insulin secretion that appears to be uncoupled from glucose metabolism.

Disclosure

J.T. Walker: None. R. Haliyur: None. R. Aramandla: None. G. Poffenberger: None. H.A. Nelson: None. J. Luchsinger: None. P. Wang: None. A. Garcia-Ocaña: None. M. Brissova: None. A.C. Powers: None.
442-P: Excess Burden of Adverse In-Hospital Outcomes in Patients with Diabetes Hospitalized for Stroke
Authors: DAURIZ MARCO , TREGNAGHI ELENA , SANTI LORENZA , LUCIANER TIZIANO , ALTOMARI ANNA , RINALDI ELISABETTA , TARDIVO STEFANO , BOVO CHIARA , BONORA ENZO
Published: June 2019

Cerebrovascular accidents (CVA) represent a major complication in diabetes (DM). Real-life evidence as to whether modern management of CVA and DM have softened this relationship is currently limited. We assessed the prevalence of DM, previously known or newly-diagnosed, in all patients (N=542, males 51.5%) admitted for CVA at the Stroke Unit (SU) of Verona University Hospital from 1/1/2015 to 12/31/2016. DM was ascertained by previous diagnosis, glucose-lowering therapy at admission/discharge or admittance plasma glucose (PG) ≥11.1 mmol/L. Prevalence of DM was 21.03% (78.1% known-DM; 21.9% new-DM). In-hospital death rate was 10.5%. Compared to non-DM, patients with DM showed an increased risk of death (15.8 vs. 9.1%; OR 1.87 95% CI, 1.03-3.41) and complications (48.2 vs. 33.3%; 1.87, 1.22-2.86) with similar duration of hospitalization (mean±SEM, 9.14±1.06 vs. 8.80±0.39 days, p=0.72). All deaths occurred in the SU. The “survivors” with DM transferred to non-intensive ward had an out-of-SU hospital stay twice as longer than non-DM (2.32±1.02 vs. 0.98±0.19 days, p=0.038). After multivariable adjustment, DM remained predictor of complications (adj-OR 1.87, 1.22-2.86) but not mortality (1.79, 0.92-3.46). Patients in the highest PG quartile (≥6.55 mmol/L) displayed the highest risk of death (7.89, 2.73-22.79; PG<4.77 mmol/L, ref.) and complications (2.30, 1.24-4.23), independent of diabetes status, glucose-lowering medications, CVA treatment and established CVA risk factors. Individuals in the older age tertile (≥81 years) were at increased risk of death (3.25, 1.40-7.54; age <72 years, ref.), while risk of infectious/cardiorespiratory complications was higher at younger age (3.13, 1.42-6.9). No difference was observed in new-DM vs. known-DM.

In conclusion, these data highlight that DM frequently occurs in patients admitted for CVA and adds an excess burden of adverse clinical outcomes that urgently calls for strategies to anticipate DM diagnosis in high-risk individuals.

Disclosure

M. Dauriz: None. E. Tregnaghi: None. L. Santi: None. T. Lucianer: None. A. Altomari: None. E. Rinaldi: None. S. Tardivo: None. C. Bovo: None. E. Bonora: None.
447-P: Low Muscle Mass Is Associated with Carotid Atherosclerosis in Korean Men with Type 2 Diabetes
Authors: NAM MOON-SUK , SEO DA HEA , JUNG MIHYE , AHN SEONG HEE , HONG SEONGBIN , LEE YONG-HO , CHOI YOUNGJU , HUH BYOUNG WOOK , HUH KAP BUM , KIM SO HUN
Published: June 2019

Background: Sarcopenia, an age-related decline in skeletal muscle mass, leads to metabolic and vascular abnormalities. However, little is known regarding the independent relationship between skeletal muscle mass and atherosclerosis in patients with type 2 diabetes mellitus (T2DM). The purpose of this study was to evaluate the association of skeletal muscle mass with markers of carotid atherosclerosis, carotid intima-media thickness (CIMT) and carotid artery plaque, in men and women with T2DM.

Methods: In this cross-sectional study, a total of 8,918 patients (4,520 men and 4,398 women) with T2DM were recruited from the outpatient clinic of Huh’s Diabetes Center from 2003 to 2016. Skeletal muscle mass was estimated from bioimpedance analysis measurements and skeletal muscle mass index (SMI, %) was defined as skeletal muscle mass (kg)/total body weight (kg) × 100. Carotid ultrasonography was performed to measure mean CIMT of both common carotid arteries. Carotid artery plaque was defined as a focal structure protruding into the lumen of the vessel of ≥50% than the surrounding area.

Results: In both men and women, the presence of carotid artery plaque was higher with decreasing SMI quartiles. In men, this association remained significant after adjustment for additional risk factors (ptrend <0.001). Odds ratio for having carotid artery plaque was 1.76 (95% confidence interval [CI] 1.34-2.33; p <0.001) in men with lowest SMI quartile compared with highest quartile. However, in women, the association was attenuated after adjustment for additional risk factors. There was no significant association between SMI quartiles and CIMT in both men and women.

Conclusion: Low muscle mass is associated with the presence of carotid artery plaque in men with T2DM.

Disclosure

M. Nam: None. D. Seo: None. M. Jung: None. S. Ahn: None. S. Hong: None. Y. Lee: None. Y. Choi: None. B. Huh: None. K. Huh: None. S. Kim: None.

Funding

National Research Foundation of Korea (2017R1D-1A1B03034581)
448-P: Predictors and Their Impact on Coronary Artery Disease (CAD) According to Glucose Tolerance Status (GTS) and Prior CAD: Historical Cohort Study in Japan
Authors: KITAZAWA MASARU , FUJIHARA KAZUYA , OSAWA TAEKO , YAMAMOTO MASAHIKO , HARADA MAYUKO , KANEKO MASANORI , MATSUBAYASHI YASUHIRO , YAMADA TAKAHO , YAMANAKA NAUTA , SEIDA HIROYASU , OGAWA WATARU , SONE HIROHITO
Published: June 2019

Although GTS, prior CAD and other traditional risk factors independently affect incidence of subsequent CAD, their impact in the same cohort has been scantly studied. We analyzed a nationwide claims-based database of 138,162 men aged 18-72 y in 2008-14. Participants were classified as normal glucose tolerance (NGT, n=78,230), prediabetes (PreDM, n=45,610) or diabetes mellitus (DM, n=14,322) based on blood tests and prescriptions and with prior (CAD+) or without prior CAD (CAD-) based on ICD-10 codes and questionnaires. Among CAD-, management of traditional risks differed from those with and without CAD events, while such differences were weaker in preDM and DM with CAD+. Cox regression model showed that PreDM and DM confer approximately 1.2- and 2.8-fold excess risk of CAD, respectively, compared with CVD- with NGT. CAD+ confers approximately a 5- to 8-fold increased risk. Most traditional risks were independently associated with subsequent CAD in the CAD- group regardless of GTS. Conversely, in the CAD+ group, their impact was weakened and only HDL-C was a significant risk factor for future CAD in those with DM and CAD+ (Table). Thus, the impact of known risks on subsequent CAD differs markedly according to GTS and CAD+. Those with CAD+ and glucose abnormality, especially DM, need treatment residual risk factors as well as traditional risk factors.

Disclosure

M. Kitazawa: None. K. Fujihara: None. T. Osawa: None. M. Yamamoto: None. M. Harada: None. M. Kaneko: None. Y. Matsubayashi: None. T. Yamada: None. N. Yamanaka: None. H. Seida: None. W. Ogawa: Consultant; Self; Abbott. Research Support; Self; Astellas Pharma Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Daiichi Sankyo Company, Limited, Mitsubishi Tanabe Pharma Corporation, Takeda Pharmaceutical Company Limited. Speaker's Bureau; Self; Lilly Diabetes, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi, Sumitomo Dainippon Pharma Co., Ltd. H. Sone: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Kowa Pharmaceutical Europe Co. Ltd., Kyowa Hakko Kirin Co., Ltd., Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Teijin Pharma Limited.
458-P: 1H-MRS Lipid Spectra of Interventricular Septum (SEP): A Novel Marker of Cardiovascular (CV) Events in Hyperglycemic (HG) Patients
Authors: CODELLA ROBERTO , LEO GIOVANNI DI , ZANARDO MORENO , SCHIAFFINO SIMONE , FABIANO SALVATORE DIONIGI , MONTI C. B. , CODARI MARINA , LUZI LIVIO , SARDANELLI FRANCESCO
Published: June 2019

Proton magnetic resonance spectroscopy (1H-MRS) can detect and quantify lipid accumulation in specific myocardial or peri-myocardial districts. Thirty-three patients (20M/13F; BMI 26±4 kg/m2; 64±10 years) with suspect of CV diseases underwent 1.5-T 1H-MRS using a water-suppressed chemical-shift imaging sequence to quantify the lipid profile of SEP, epicardial (EAT), pericardial (PAT), and subcutaneous (SAT) adipose tissues. The integral of the lipid peaks at 0.9, 1.3, 2.0, 5.3 ppm was measured as percentage over the total amount of lipids (Table). A subgroup of 15 subjects were identified as HG (12 impaired-fasting-glucose and 3 diabetics: 121±26 mg/dL). Bulk methylene (1.3 ppm) of SEP was higher in HG vs. other patients (P=0.047). In HG patients, SEP was positively correlated to glucose, visceral fat, C-reactive protein (Spearman ρ>0.63, P<0.018); EAT was positively correlated to visceral fat, age, body weight, and inversely to muscular-, fat free-, bone mass (Pearson ρ<|0.53|, P>0.039). In all subjects, the main lipid peaks correlated with metabolic parameters.1H-MRS allowed to measure lipid peaks in different adipose compartments with cardiometabolic relevance. SEP resulted a myocardial site associated with biomarkers of secondary heart events in pre/diabetic patients at higher risk of CV episodes. High SEP fat may be proposed as a marker of future CV events.

Disclosure

R. Codella: None. G. Di Leo: None. M. Zanardo: None. S. Schiaffino: Speaker's Bureau; Self; General Electric. S.D. Fabiano: None. C.B. Monti: None. M. Codari: None. L. Luzi: Consultant; Self; McKinsey & Company. Research Support; Self; Allergan, Gelesis, SunStar Inc. Speaker's Bureau; Self; A. Menarini Diagnostics, AstraZeneca, Eli Lilly and Company, GlaxoSmithKline plc., Menarini Group, Novartis AG, Novo Nordisk Inc., Sunstar Foundation. F. Sardanelli: None.
459-P: Risk for Type 2 Diabetes and Cardiovascular Health in Children Treated with Second-Generation Antipsychotics
Authors: HENDERSON AMANDA M. , ISLAM NAZRUL , SANDOR GEORGE GABOR , PANAGIOTOPOULOS CONSTADINA , DEVLIN ANGELA M.
Published: June 2019

Background: Second-generation antipsychotics (SGAs) are used to treat mental illness in children but have side effects including weight gain, dyslipidemia, hypertension, and risk for type 2 diabetes. These side effects are well-known risk factors for cardiovascular disease. However, little is known about the cardiovascular health of SGA-treated children.

Methods: Cross-sectional study of children (aged 6-18 years) with mental illness that were SGA-treated (n=47) or SGA-naïve (n=37), and control children (n=83, no mental illness). Anthropometrics and a fasting blood sample were collected. Subclinical indicators of atherosclerosis were assessed. Central pulse-wave velocity (PWV) by standard M-mode echocardiography and Doppler tracing, an indicator of arterial stiffness, and carotid intima-media thickness (cIMT) were assessed.

Results: SGA-treated children had greater BMI z-scores (p<0.001) and higher (p<0.001) prevalence of overweight/obesity (BMI ≥85th percentile for age and sex) than SGA-naïve and control children. SGA-treated children had higher fasting plasma insulin (p=0.016) and 21.7% had hyperinsulinemia (fasting insulin ≥100pmol/L) compared to 12.9% of SGA-naïve children. No differences in fasting glucose or circulating lipids were observed. Hypertension was observed in 14.9% of SGA-treated children, compared to 5.4% of SGA-naïve and 6.0% of control children. SGA-treated (p=0.002) and SGA-naive children (p<0.001) had higher PWV compared to controls (adjusted for age, sex, BMI, systolic blood pressure). No effect of SGAs or mental health diagnosis on cIMT was observed.

Conclusion: Children with mental illness have greater arterial stiffness than control children.

Disclosure

A.M. Henderson: None. N. Islam: None. G.G. Sandor: None. C. Panagiotopoulos: Advisory Panel; Self; Dexcom, Inc. A.M. Devlin: None.

Funding

British Columbia Mental Health & Substance Use Services
461-P: Prothrombotic State Is Associated with Impaired Arterial Wall Elastic Properties in Patients with Type 2 Diabetes Mellitus
Authors: SIASOS GERASIMOS , STAMPOULOGLOU PANAGIOTA K. , BLETSA EVANTHIA , BATZIAS KONSTANTINOS , PASCHOU STAVROULA A. , ANTONOPOULOS ALEXIOS , TSIGKOU VASILIKI , GOULIOPOULOS NIKOLAOS , ZAROMYTIDOU MARINA , MAZARIS SAVVAS , OIKONOMOU EVANGELOS , TSALAMANDRIS SOTIRIS , VOGIATZI GEORGIA , THANOPOULOU ANASTASIA , VRYONIDOU ANDROMACHI , POLITOU MARIANNA , TOUSOULIS DIMITRIS , TENTOLOURIS NIKOLAS
Published: June 2019

Background: Arterial stiffness is a prominent macrovascular complication in patients with type 2 diabetes mellitus (DM2). Antiplatelet therapy is the cornerstone of CVD therapy. There is little evidence about the influence of antiglycemic agents on arterial wall properties and prothrombotic state.

Methods: We enrolled 80 consecutive patients (males=64.4%), aged 64.19 ± 8.82 years receiving either metformin plus another antiglycemic agent such as sulphonylureas, DPP-4i, GLP-1 agonists, insulin, or metformin alone (n=16 per group). Applanation tonometry was used to assess aortic pulse wave velocity (PWV) as a measure of arterial stiffness. Platelet reactivity was measured with PFA-200, collagen/epinephrine (CEPI) and PFA-200 collagen/ADP closure time (CADP).

Results: There was no difference between the study groups regarding gender, age, hypertension, dyslipidemia, smoking, PWV, CADP or CEPI (p=NS for all). When conducting a separate analysis between patients receiving aspirin (AS), dual antiplatelet therapy (DAPT) and those without antiplatelet therapy (nAPT), we observed that nAPT patients exerted significantly lower CEPI values (134.42 ± 40 s for nAPT vs. 170.36 ± 64 s for AS vs. 178.90 ± 74 s for DAPT, p=0.03). PWV was 10.96 ± 2.54 m/s for nAPT vs. 11.20 ± 2.88 m/s for AS vs. 10.82 ± 2.76 m/s for DAPT (p=0.9). Within-group analysis showed that CEPI values were significantly reduced in nAPT under sulphonylureas (121 s vs. 243 s vs. 262 s for nAPT, AS and DAPT respectively, p=0.02) or GLP-1 (138 s vs. 203 s vs. 168 s for nAPT, AS and DAPT respectively, p=0.05). Additionally, we found an inverse, linear association between CEPI and PWV (rho= -0.45, p=0.02) and CADP and PWV (rho= -0.04, p=0.05) in nAPT patients alone.

Conclusion: Increased prothrombotic state is associated with impaired arterial wall elastic properties. Antidiabetic and antiplatelet treatment interactions may regulate the relationship between arterial stiffness and thrombosis in patients with DM2.

Disclosure

G. Siasos: None. P.K. Stampouloglou: None. K. Batzias: None. S.A. Paschou: None. A. Antonopoulos: None. V. Tsigkou: None. N. Gouliopoulos: None. M. Zaromytidou: None. S. Mazaris: None. E. Oikonomou: None. S. Tsalamandris: None. G. Vogiatzi: None. A. Thanopoulou: None. A. Vryonidou: None. M. Politou: None. D. Tousoulis: None. N. Tentolouris: None.
475-P: Exendin-4 Stimulates eNOS Activation by Upregulation of Scavenger Receptor Type B Class I in Human Endothelial Cells
Authors: LYU JINGYA , IMACHI HITOMI , FUKUNAGA KENSAKU , SATO SEISUKE , DONG TAO , IBATA TOMOHIRO , KOBAYASHI TOSHIHIRO , MATSUMOTO MARI , MURAO KOJI
Published: June 2019

Exendin-4 (Exenatide), a long-acting agonist of the glucagon-like peptide 1 receptor (GLP-1R), has been confirmed to protect the cardiovascular system from atherosclerosis in type 2 diabetes. However, the detailed mechanism of this process is not clear. Activation of endothelial nitric oxide synthase (eNOS) to produce nitric oxide (NO) plays an important role in endothelial homeostasis. Previous study shows that HDL activates eNOS via its receptor scavenger receptor type B class I (SR-BI). In this study, we checked the effect of exendin-4 on HDL/SR-BI-mediated eNOS activation in HUVECs. Firstly, we demonstrated the expression of GLP-1R in HUVECs and exendin-4 significantly induced HDL-mediated eNOS activation. Next, we found exendin-4 increased the expression and promoter activity of SR-BI, and then blocking of PKA or AMPK by its specific inhibitor cancelled this effect. Consistently, cAMP enhanced SR-BI promoter activity while domain negative AMPK inhibited the effect of exendin-4 on SR-BI promoter activity. Blocking of GLP-1R by exendin 9-39 cancelled exendin-4-induced SR-BI promoter activity and AMPK phosphorylation. As a downstream of AMPK, a transcription factor FoxO1 was confirmed to directly bind to the SR-BI promoter region by Chromatin Immunoprecipitation (ChIP) assay and overexpression of FoxO1 enhanced SR-BI promoter activity. Moreover, exendin-4 markedly increased the expression and nuclear recruitment of FoxO1 by decreasing its phosphorylation. As well as, mutation of FoxO1-binding site in the SR-BI promoter region or silence of FoxO1 cancelled exendin-4-induced SR-BI expression.

In summary, our results show that exendin-4 upregulates the expression and transcription of SR-BI via PKA/AMPK/FoxO1 pathway, which contributes to HDL-mediated eNOS activation and NO generation in human endothelial cells.

Disclosure

J. Lyu: None. H. Imachi: None. K. Fukunaga: None. S. Sato: None. T. Dong: None. T. Ibata: None. T. Kobayashi: None. M. Matsumoto: None. K. Murao: None.
477-P: Inhibiting Myeloperoxidase (MPO) Prevents Onset and Reverses Established High-Fat Diet (HFD)-Induced Microvascular Insulin Resistance (IR)
Authors: BARRETT EUGENE , CHAI WEIDONG , AYLOR KEVIN W. , GAN LI-MING , MICHAËLSSON ERIK
Published: June 2019

HFD rapidly recruits neutrophils to insulin target tissues and induces microvascular IR. MPO is abundant in neutrophils, inhibits nitric oxide mediated vaso-relaxation, and associates with increased cardiovascular risk. AZD5904 (AstraZeneca) irreversibly inhibits MPO and is in clinical trials. MPO knockout, or chemical inhibition, improves glucose regulation in HFD-fed mice. Whether MPO affects muscle microvascular or metabolic IR in vivo is unknown. To test whether AZD5904 can prevent onset of HFD-induced vascular and metabolic IR, Sprague-Dawley rats were fed chow or a HFD (60% calories) for 2 weeks. HFD-fed rats received vehicle (n=8) or AZD5904 subcutaneously (n=10) via Alzet pump. Chow-fed rats (n=8) received vehicle alone. On day 15, we measured muscle microvascular perfusion in all rats by contrast-enhanced ultrasound, at 0, 60 and 120 minutes of a 2-hour euglycemic insulin clamp (3 mU/kg/minutes) and glucose disposal was determined. We next tested whether AZD5904 can reverse established vascular IR. Two groups of rats (n = 10, each) were fed the HFD for 2 weeks to induce vascular and metabolic IR, HFD was continued for 2 additional weeks while giving vehicle or AZD5904. Microvascular and metabolic insulin action was measured again using the insulin clamp. HFD for 2 weeks completely blocked insulin-mediated muscle capillary recruitment, decreased glucose utilization by 35%, and inhibited muscle Akt phosphorylation (P<0.05, for each measure). AZD5904 during HFD selectively and completely restored insulin-induced microvascular sensitivity (P<0.05). Furthermore, AZD5904 during the last 2 of 4 weeks of HFD feeding fully restored insulin-mediated microvascular perfusion (P<0.05) again without affecting glucose disposal. We conclude that inhibiting MPO selectively corrects vascular IR. This effect may connote a therapeutic potential for MPO inhibition in the prevention of vascular disease/dysfunction seen in IR humans.

Disclosure

E. Barrett: Research Support; Self; AstraZeneca. W. Chai: None. K.W. Aylor: None. L. Gan: Employee; Self; AstraZeneca. E. Michaëlsson: Employee; Self; AstraZeneca.

Funding

University of Virginia; AstraZeneca
489-P: Loss of Neuropilin 1 in Podocytes Aggravates Diabetic Nephropathy
Authors: CHENG RUI , HE XUEMIN , MA JIAN-XING
Published: June 2019

Podocyte injury is a crucial pathological event in diabetic nephropathy. Neuropilin 1 (NRP1) is a multi-functional transmembrane protein and functions as a co-receptor of extracellular ligands, such as VEGF. Previous studies have shown that NRP1 levels are significantly decreased in the kidney of diabetic patients, and advanced glycation end products down-regulate NRP1 in a mouse podocyte cell line. The function of NRP1 in podocytes has not been clearly understood. We found that NRP1 levels are significantly decreased in the glomeruli of db/db mice, STZ-induced diabetic mice and OVE26 mice. A podocyte-specific NRP1 knockout mouse strain (NRP1-CKO) is generated by crossbreeding NRP1flox/flox mice with Nphs2-Cre transgenic mice. Compared to NRP1flox/flox mice, the NRP1-CKO mice in the absence of diabetes showed increased 24h urine albumin excretion in the age of 2 months and a reduced creatinine clearance at the age of 4 months. Enlarged foot processes of podocytes and flattened podocyte body was found in NRP1-CKO mice, compared to age-matched NRP1flox/flox mice. NRP1-CKO mice showed significantly decreased podocyte numbers per glomerulus and increased apoptosis. We generated diabetic NRP1-CKO mice (NRP1-CKO-OVE26) by crossbreeding the NRP1-CKO mice with OVE26 diabetic mice. NRP1-CKO/OVE26 diabetic mice developed more severe proteinuria, compared to the age- and genetic background-matched NRP1flox/flox/OVE26 mice. As shown by PAS staining, the glomerular structure was disturbed, and glomerular mesangial expansion was more severe in NRP1-CKO/OVE26 mice. Phosphorylation of MAPKP38 was significantly elevated in the NRP1-deficient glomeruli, compared to age-matched NRP1flox/flox glomeruli, suggesting that NRP1 deficiency induces activation of P38 signaling, further confirming podocyte stress and injury in NRP1-CKO mice. These results suggest that NRP1 ablation impairs renal function and exacerbates diabetic nephropathy and NRP1 is essential in the regulation of podocyte function and survival.

Disclosure

R. Cheng: None. X. He: None. J. Ma: None.

Funding

Oklahoma Center for the Advancement of Science and Technology; National Institutes of Health
495-P: Kidney May Regulate SGLT2-Dependent Glucose Reabsorption through Sweet Taste Receptors: Implications for Diabetes
Authors: ZHOU LUPING , HUANG WEI , LIU NAN , MA XIU M. , GUO MAN , CHEN QING , GAO CHENLIN , XU YONG
Published: June 2019

Sodium-glucose cotransporter 2 (SGLT2) mediates 90% glucose reabsorption in kidney, and SGLT2 inhibitors have emerged as a novel oral antihyperglycaemic agents by selectively reducing renal reabsorption of glucose and sodium. Recent research has suggested that sweet taste receptors (STRs) are important participants of diabetes. However, it is unclear whether STRs regulate reabsorption of glucose via SGLT2 in the pathogenesis of diabetes. Therefore, we evaluated the role and mechanism of STRs in reabsorption of glucose via streptozotocin (STZ)-induced diabetes mouse in vivo and in high glucose-induced mouse glomerular mesangial cells (GMCs) and human proximal renal tubular epithelia cells (HK-2) in vitro. Our results showed that the expression of STRs were obviously downregulated while SGLT2 was upregulated under the condition of diabetes in vivo and in vitro. We hypothesized that high glucose-induced downregulation of STRs might modulate reabsorption of glucose. STRs inhibitor lactisole or siRNA-STRs was used as an intervention reagent respectively, distribution of STRs in vivo was detected by PET/CT via intravenous injection of 18F-labeled lactisole, glucose level, renal pathology and SGLT2 expression in vivo and fluorescence labeled glucose uptake rate in vitro were assayed. The effect of high glucose on STRs and SGLT2 expression as well as the glucose absorption were obviously mitigated by lactisole or siRNA-STRs. Overall, this is the first observation of the potential relationship between STRs and SGLT2, suggesting that kidney may regulate SGLT2-depended glucose reabsorption through STRs, which may act as new therapeutic targets of diabetes.

Disclosure

L. Zhou: None. W. Huang: None. N. Liu: None. X.M. Ma: None. M. Guo: None. Q. Chen: None. C. Gao: None. Y. Xu: None.

Funding

National Natural Science Foundation of China (81800741)
498-P: Involvement of Phosphorylation of Beta-1 Integrin in Renin Production and Blood Pressure Regulation in Diabetic Nephropathy
Authors: SAWADA KAICHIRO , TOYODA MASAO , SAITO NOBUMICHI , KIMURA MORITSUGU , MORIYA HITOMI , FUKAGAWA MASAFUMI
Published: June 2019

Diabetic nephropathy (DN) and hypertension exacerbate each other, but many aspects of the mechanisms between these two pathologies remain unclear. The juxtaglomerular cells (JGC) secrete renin and trigger the renin-angiotensin-aldosterone system involved in blood pressure regulation. Our previous immunohistochemical analysis showed JGC-specific phosphorylation of β1 integrin (Itg), a cellular matrix adhesion factor. Quantitative analysis indicated the negative correlation between the phosphorylated β1 (pβ1) Itg and renin expression in JGC. To analyze the details of this relationship, As4.1, a cell-line of JGC was used. As protein kinase C (PKC) subtype epsilon is a putative kinase of the β1 Itg in JGC, PKC activator (phorbol 12-myristate 13-acetate, PMA) and PKC inhibitor (bisindolylmaleimide I, BIM-1) as well as TGF β1 which is known to stimulate a pathway to phosphorylate β1 Itg in JGC were applied to the culture of As4.1 cells. After 24 hours, expression of the mRNA levels of renin was gained by BIM-1 but suppressed by PMA or TGF β1. In the course of the cell culture with PMA, the mRNA levels of renin was increased till 90 minutes after application. However, it decreased subsequently, ultimately to 10% of the baseline level. Additional application of PMA at 6, 12 and 24 hours after the first application of PMA did not reproduce the temporal increase of renin expression. Time course of the cell culture with TGF β1 showed continuous decrease of the mRNA levels of renin. As PMA has broad spectrum for PKC subtypes, it was estimated that some activated PKC subtypes promoted temporal increase of renin expression in the early phase, however, later changes of cell state produced by pβ1 Itg suppressed the renin expression irreversibly. Our results confirmed a negative correlation between pβ1 Itg and renin expression in JGC cell-line, suggesting a link between β1 Itg and the regulatory system that controls renin production and secretion.

Disclosure

K. Sawada: None. M. Toyoda: Speaker's Bureau; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Eli Lilly and Company, Kyowa Hakko Kirin Co., Ltd., Medtronic Japan Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Novo Nordisk Inc., Sanofi, Sumitomo Dainippon Pharma Co., Ltd., Takeda Pharmaceutical Company Limited. N. Saito: None. M. Kimura: None. H. Moriya: None. M. Fukagawa: Research Support; Self; Kyowa Hakko Kirin Co., Ltd. Speaker's Bureau; Self; Bayer Yakuhin, Ltd., Kissei Pharmaceutical Co., Ltd., Ono Pharmaceutical Co., Ltd., Torii Pharmaceutical Co., Ltd.
499-P: The Relative Roles of Pro-oxidant Enzymes Nox4 and Nox5 in Diabetic Kidney Disease
Authors: JHA JAY C. , COOPER MARK E. , KENNEDY CHRIS , JANDELEIT-DAHM KARIN
Published: June 2019

Background: Renal oxidative stress plays an important role in mediating kidney injury in diabetes. There is increasing evidences that recently discovered pro-oxidant enzyme, Nox5 plays a significant role in human diabetic kidney disease (DKD). We examined the role of Nox5 in human DKD, in human renal cell populations as well as in a high fat fed and alloxan rabbit model of DKD. In addition, we also examined the contribution of Nox5 in the in DKD in the absence of Nox4 expression using renal cell specific Nox5Tg mice.

Results: Expression of Nox5 was increased in both glomerular and tubular compartments of kidney biopsies obtained from diabetic individuals. Nox5 is the Nox isoform which shows the highest upregulation in response to high glucose. Silencing of Nox5 reduces ROS formation and expression of proinflammatory and profibrotic cytokines and growth factors as well as putative elements that are implicated in DKD. Our data also suggest that Nox5 is upstream of Nox4 and that Nox5 inhibition downregulates Nox4, but not vice versa. High fat feeding and alloxan induced diabetes rabbits showed upregulation of Nox5 and increased ROS in association with increased mesangial area, ECM protein accumulation and upregulation of a range of pro-inflammatory (MCP-1) and pro-fibrotic genes (CTGF) as analysed by RNAseq and confirmed by RT-PCR. In addition, over expression of Nox5 in mesangial cells of Nox4 KO diabetic mice demonstrated 30% increase in albuminuria, mesangial expansion, increased renal injury and inflammation via enhanced ROS production.

Conclusions: These studies will for the first time explore in a comprehensive manner the role of Nox5 in recently generated Nox5KO rabbit models in the context of type 1 and type 2 diabetes as well as delineate the relative role of Nox4 versus Nox5 in diabetic complications. These studies will provide evidence if Nox5 blockade alone or combined with Nox4/Nox5 blockade is the optimal approach to treat and prevent diabetic complications, in particular nephropathy.

Disclosure

J.C. Jha: None. M.E. Cooper: Advisory Panel; Self; AstraZeneca, Lilly Diabetes. Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Servier. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Novo Nordisk A/S. Speaker's Bureau; Self; Bayer AG, Merck Sharp & Dohme Corp., Novartis AG. C. Kennedy: Research Support; Self; Prometic Life Sciences Inc. K. Jandeleit-Dahm: Advisory Panel; Self; Australian Diabetes Society, Genkyotex. Consultant; Self; Metavention.
515-P: Mitochondrial AKT1 in Renal Tubules Is a Novel Modulator for the Development of Glomerulosclerosis and Subsequent Renal Failure
Authors: YOU-HSIEN LIN HUGO , CHEN YU-HAN , TA ALBERT , LEE HSIAOCHEN , CHEN YUMAY , WANG PING H.
Published: June 2019

Diabetes increases the risk of renal ischemia/reperfusion injury (IRI). Recent evidence suggests that tubular dysfunction is associated with glomerular impairment, but its causal relationship is unclear. Renal tubules have abundant mitochondria; Akt is translocated into mitochondria upon IRI. To test the hypothesis that tubular mitochondrial Akt1 (mito-Akt) plays a protective role during IRI and subsequent renal failure, we have generated novel renal tubule-specific transgenic mice harboring inducible mitochondria-targeting dominant negative Akt1 (KMDAKT) or constitutively active Akt1 (KMCAKT) with the Cre-lox system. After KMDAKT mice were induced with tamoxifen (T) and subjected to IRI, histological analysis showed increased tubule injury index Jablonski score (p<0.04) and greater renal fibrosis as compared to the corn-oil (CO) injected mice (p<0.02). Kidney Injury Molecule-1 (KIM1), PAS-positive glomerulosclerosis, and serum creatinine were all increased (p<0.01) in the T-KMDAKT mice when compared to the CO mice 45-days post IRI. Renal inflammatory markers, including IL-1B, were also increased, suggesting inflammation played a role. Kaplan-Meier analysis showed much lower survival in the T-KAMDAKT mice post IRI than the CO mice (p<0.01). To study whether augmentation of tubular mito-Akt can protect against renal IRI, T-KMCAKT and CO-KMCAKT were subjected to renal IRI.T-KMCAKT mice showed an improved Jablonski score (p<0.02), less renal fibrosis (p<0.04), and lower KIM1 expression (p<0.02). T-KMCAKT mice exhibited less glomerulosclerosis and better survival (p<0.01).

Summary: 1] tubular mito-Aktcritically modulated the outcomes of kidney IRI beyond renal tubules, 2] tubular mitochondria dysfunction causally triggered development of glomerulosclerosis during acute and chronic kidney failure, and 3] renal tubular mito-Akt, which is activated by insulin, may represent a novel therapeutic target for diabetic nephropathy.

Disclosure

H.Y. Lin: None. Y. Chen: None. A. Ta: None. H. Lee: None. Y. Chen: None. P.H. Wang: None.
551-P: The Impact of Type 1 Diabetes on Motor Dynamics in the Brain
Authors: EMBURY CHRISTINE M. , WERE VEGA A. , LORD GRACE H. , DRINCIC ANDJELA , DESOUZA CYRUS , WILSON TONY W.
Published: June 2019

Type 1 diabetes affects the structure and function of the brain, with psychomotor speed as the most commonly reported deficit. The current study sought to uncover the dynamics underlying motor processing in the brain of adults with type 1 diabetes using an arrow-based Flanker task. Adults (ages 19-35, N = 39) with type 1 diabetes and no major complications and a matched control group (N = 40) underwent magnetoencephalography (MEG) while performing the task. During the task, participants were presented with a row of five arrows facing right or left, and participants were instructed to respond by button press to the direction of the middle arrow. Analyses focused on the time period around movement execution to examine motor-related dynamics. Time series analyses were performed on the peak beta and gamma responses from the motor cortices. An ANOVA model and follow-up regression analyses were used to delineate the predictive value of each neural response in driving behavioral outcomes. Subsequent t-tests between frequent (3+ episodes per week) and infrequent (0-1 episodes per week) hypoglycemic groups were performed to examine response differences within the adults with type 1 diabetes. Responses driving behavior differed between adults with and without type 1 diabetes. In adults with type 1 diabetes, beta frequency responses were significant predictors of reaction time, where greater ipsilateral recruitment led to faster reaction times (b = .58, t = 3.64, p = .001). In contrast, gamma drove behavioral outcomes in the control group (b = -.43, t = -2.96, p = .005). Further, within group analyses revealed that more frequent episodes of hypoglycemia were associated with alterations specific to beta responses during the pre-movement baseline period (p < .05). These results show frequency specific alterations in motor dynamics in adults with type 1 diabetes, with an added effect of increased hypoglycemia. These findings strongly indicate that diabetes directly impacts motor control components in the brain, regulating behavioral outcomes.

Disclosure

C.M. Embury: None. V.A. Were: None. G.H. Lord: None. A. Drincic: Board Member; Self; Corcept Therapeutics. C. Desouza: Consultant; Self; Novo Nordisk A/S, Sanofi US. T.W. Wilson: None.

Funding

National Institutes of Health (R01MH103220, R01MH116782)