bluuebunny/crossref_metadata_2025_split huggingface dataset:
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&lt;0.05, for each measure). AZD5904 during HFD selectively and completely restored insulin-induced microvascular sensitivity (P&lt;0.05). Furthermore, AZD5904 during the last 2 of 4 weeks of HFD feeding fully restored insulin-mediated microvascular perfusion (P&lt;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&lt;0.04) and greater renal fibrosis as compared to the corn-oil (CO) injected mice (p&lt;0.02). Kidney Injury Molecule-1 (KIM1), PAS-positive glomerulosclerosis, and serum creatinine were all increased (p&lt;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&lt;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&lt;0.02), less renal fibrosis (p&lt;0.04), and lower KIM1 expression (p&lt;0.02). T-KMCAKT mice exhibited less glomerulosclerosis and better survival (p&lt;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 &lt; .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)