bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effects of Cardiovascular Risk Factor Combined Antiplatelet Therapy and the Risk of Cerebrovascular Events in Patients with T2DM in an Urban Community over 96-Month Follow-Up—The Beijing Communities Diabetes Study 19
Authors: ZHANG XUE LIAN , YUAN SHEN YUAN , GROUP BCDS
Published: June 2018

Objective: We investigated the prognostic significance of metabolic risk scores and aspirin with respect to cerebrovascular events.

Methods: A total of 25 communities of diabetic patients were enrolled in Beijing Community Diabetes Study (BCDS) from 2008. 3413 patients with T2DM in BCDS have complete screening data, including blood glucose, blood pressure, lipid profiles and anti-platelet therapy, which were assigned metabolic score (MS) and add up to the total metabolic score (TMS). According to the total metabolic score (TMS), the patients were divided into four equal groups: Group 1 (24<TMS<40), Group 2 (40<TMS<47), Group 3 (47<TMS<55) and Group 4 (55<TMS<87). After 96 months, patients were followed-up to assess the long-term effects of the multifactorial interventions.

Results: During 96-months follow-up, a total of 91 cerebrovascular events occurred, including acute cerebral infarction, acute cerebral hemorrhage and transient ischemic attack (TIA). The incidence of cerebrovascular events was higher in the Group 4 than in the Group 1. In Cox multivariate analyses, there are significant differences in incidences of cerebral infarction events among the four groups during the 96-months follow-up. Cox proportional hazards analysis revealed that, HbA1c (P=0.0000), systolic pressure (P=0.0015), aspirin free treatment (P=0.0023) are independent predictor for cerebrovascular events in diabetic patients.

Conclusions: This study indicates that total metabolic score (TMS) influences the incidence of cerebrovascular events in diabetic patients. In addition to good control of blood glucose, blood pressure and lipid profiles. The patients with diabetes need long-term anti-platelet therapy, which is important for the prevention of cerebrovascular events.

Disclosure

X. Zhang: None. S. Yuan: None.
Diabetes Remission Clinical Trial (DiRECT)—Changes in Hepatic VLDL1-TG Production and Intrapancreatic Fat during Weight Maintenance Phase
Authors: AL-MRABEH AHMAD , ZHYZHNEUSKAYA SVIATLANA V. , PETERS CARL , BARNES ALISON C. , ARIBISALA BENJAMIN , HOLLINGSWORTH KIEREN G. , SATTAR NAVEED , LEAN MICHAEL E. , TAYLOR ROY
Published: June 2018

Type 2 diabetes is characterized by excess hepatic and intrapancreatic fat deposition. Liver-derived Very Low Density Lipoprotein-Triglyceride (VLDL1-TG) delivers fat to all peripheral tissues. Relative changes in VLDL1-TG production and intrapancreatic fat were investigated in a sub-group of the prospective, randomized Diabetes Remission Clinical Trial (DiRECT). Individuals with complete datasets at 12 months (n=45) were included. Detailed metabolic tests were carried out at baseline, 4 months, and 12 months after low calorie diet (825-853 kcal/day). Intra-organ fat was quantified using 3-point Dixon MRI, VLDL1-TG production was quantified using a competitive blocking non-isotopic method, and insulin secretion was measured by the Stepped Insulin Secretion Test with Arginine (SISTA). Weight loss induced major changes in liver and intrapancreatic fat with 69% remission of diabetes (responders, defined as HbA1c <6.5%). This was followed by weight regain between 4-12 months in responders (82.3±2.8 to 85.4±3.2kg, p=0.001) and non-responders (84.5±3.2 to 89.6±3.3kg, p<0.0001). However, liver fat, VLDL1-TG production, and intrapancreatic fat remained stable in responders (2.5 ±1.9 to 2.9±0.6%, p=0.20; 386.7±30.2 to 428.5±21.2 mg/kg/day, p=0.14; and 8.0±0.5 to 7.7 ±0.4%, p=0.26, respectively). In contrast, these parameters increased in non-responders (2.7±0.5 to 6.2 ±1.8%, p=0.02; 460.2±39.8 to 596.2±36.6 mg/kg/day, p=0.001; and 6.7±0.3 to 7.0±0.4%, p=0.18, respectively). The recovered first phase insulin secretion in responders continued to improve between 4-12 months (0.13±0.02 to 0.17±0.04 nmol/min/m2, p=0.17). There was no change in the non-responders (0.03±0.01 to 0.03±0.01 nmol/min/m2, p=0.98). These data consistent with VLDL1-TG being the link between liver fat and intrapancreatic fat, and a major modulator determining remission of type 2 diabetes.

Disclosure

A. Al-Mrabeh: None. S.V. Zhyzhneuskaya: None. C. Peters: None. A.C. Barnes: None. B. Aribisala: None. K.G. Hollingsworth: None. N. Sattar: Advisory Panel; Self; Boehringer Ingelheim GmbH. Speaker's Bureau; Self; Boehringer Ingelheim GmbH, Janssen Pharmaceuticals, Inc.. Advisory Panel; Self; Novo Nordisk A/S. Speaker's Bureau; Self; Novo Nordisk A/S, Eli Lilly and Company. Research Support; Self; Boehringer Ingelheim GmbH. Advisory Panel; Self; Amgen Inc.. Speaker's Bureau; Self; Amgen Inc., AstraZeneca, Mitsubishi Tanabe Pharma Corporation, Medscape, Sanofi-Aventis Deutschland GmbH. M.E. Lean: Advisory Panel; Self; Novo Nordisk Inc., Orexigen Therapeutics, Inc.. Research Support; Self; Cambridge Weight Plan. Consultant; Self; Counterweight Ltd. Stock/Shareholder; Self; Eat Balanced. R. Taylor: None.
Multinational, Multicenter, Prospective, Randomized, Double-Blinded, Placebo-Controlled Trial to Evaluate the Efficacy of Cyclical Topical Wound Oxygen Therapy (TWO2) in the Treatment of Chronic Diabetic Foot Ulcers
Authors: FRYKBERG ROBERT , FRANKS PETER J. , EDMONDS MICHAEL E. , BRANTLEY JONATHAN N. , TÉOT LUC , WILD THOMAS , GAROUFALIS MATTHEW G. , LEE ALIZA M. , THOMPSON JANETTE , REACH GERARD , DOVE CYAANDI R. , LACHGAR KARIM , GROTEMEYER DIRK
Published: June 2018

Non-healing DFUs lead to increased mortality, morbidity and health economic burden. Our RCT (NCT02326337) was undertaken to explore the efficacy of Topical Wound Oxygen (TWO2) homecare therapy in healing DFUs that had failed to heal with Standard of Care (SOC) alone. A Group Sequential Design was utilized with 2 interim analyses, requiring a significance of p<0.022 at each. All subjects meeting the incl/excl criteria were enrolled into a run-in of SOC that included gold-standard offloading and sharp debridement. Only DFUs not on a proven healing trajectory with SOC alone (<30% wound area reduction) were randomized into the active phase of the study, where they were assigned (double blind) to either active, or sham (placebo), TWO2 device treatment arms. The primary endpoint of the study was ulcers healed at 12 weeks. At the first interim analysis point of 73 subjects, the active TWO2 arm was shown to be significantly superior to the sham arm (Pearson Chi2 =7.2707, P=0.007). Multivariable analysis using logistic regression and Cox proportional hazards modelling of the secondary outcome measure of time to heal showed no other covariates achieved significance. The active TWO2 arm showed nearly 4 times the likelihood to heal DFUs in 12 weeks compared to the sham arm HR 3.88 (95% CI 1.40 to 10.71), p=0.009.

Disclosure

R. Frykberg: Research Support; Self; AOTI, Podometrics. P.J. Franks: Consultant; Self; AOTI. M.E. Edmonds: Advisory Panel; Self; Urgo Medical, Edixomed. J.N. Brantley: None. L. Téot: Advisory Panel; Self; Acelity, Integra LifeSciences. T. Wild: None. M.G. Garoufalis: None. A.M. Lee: None. J. Thompson: None. G. Reach: Other Relationship; Self; Abbott, AbbVie Inc., Janssen Pharmaceuticals, Inc., LifeScan, Inc., Eli Lilly and Company, Novo Nordisk Inc., Sanofi-Aventis, Servier, Becton, Dickinson and Company, AstraZeneca. C.R. Dove: None. K. Lachgar: None. D. Grotemeyer: Other Relationship; Self; AOTI Ltd.. Research Support; Self; Gore, Endologix.
Pro-apoptotic and Inflammatory Markers Enhance the Ability of MDA-LDL in Circulating Immune Complex to Predict Acute Myocardial Infarction in the VADT Study
Authors: LOPES-VIRELLA MARIA F.L. , HUNT KELLY J. , AGARWAL RASHI , BAKER NATHANIEL L. , VIRELLA GABRIEL , INVESTIGATORS VADT
Published: June 2018

There is considerable interest in identifying biomarkers indicative of plaque instability and acute events. Previously we reported that MDA-LDL in circulating immune complex predicted acute myocardial infarction (MI) in the VADT study. Interleukin (IL)-6, Fas Ligand, tissue inhibitor of MMP (TIMP)-1 and monocyte chemotactic protein (MCP)-1 have been identified as potentially clinically relevant biomarkers related to plaque instability. Therefore, our objective was to examine the role of these biomarkers in conjunction with MDA-LDL in circulating immune complex as predictors of acute MI in type 2 diabetes. Biomarker levels were measured in 833 VADT patients, a median of 2.0 years after study entry and participants were subsequently followed a median of 3.7 years. Biomarkers were examined individually and as composite scores created by summing z scores across biomarkers. Hazard ratios (HRs) for acute MI were calculated by Cox proportional hazard models. During follow-up, 5.2% had an MI. After adjustment for conventional risk factors, results of composite biomarkers scores were promising with a hazard ratio for acute MI of 4.65 (95% CI: 1.56, 13.9) comparing the fourth to first quartile of our summary composite score which included MDA-LDL-IC, IL-6, Fas Ligand,TIMP-1 and MCP-1 (Table).

Disclosure

M.F.L. Lopes-Virella: None. K.J. Hunt: None. R. Agarwal: None. N.L. Baker: Research Support; Self; Bristol-Myers Squibb Company. G. Virella: None.
Different Augmentation of Left Ventricular Contractility of Mice during Myocardial Infarction following Gliptin and Sulfonylurea Treatment
Authors: HAJ-YEHIA ELIAS , DIEBOLD SEBASTIAN , MOELLMANN JULIA , KAHLES FLORIAN , KLEIN THOMAS , MARK MICHAEL , MARX NIKOLAUS , LEHRKE MICHAEL
Published: June 2018

DPP-4 inhibitors are a class of antidiabetic drugs which act by augmentation of the GLP-1 system and insulin secretion. Despite similar glucose lowering efficacy within the class, only saxagliptin increased heart failure hospitalization in diabetic patients. This unexpected effect might relate to the metabolism of saxagliptin (saxa). Here, we directly compare linagliptin (lina, 3 mg/kg, p.o.), saxa (10 mg/kg, p.o.), and its metabolite 5-hydroxy-saxagliptin (OH-saxa, 10 mg/kg, p.o.) to the sulfonylurea glimepiride (glim, 2 mg/kg, p.o.) and control during acute myocardial infarction in mice. Millar catheter analysis under dobutamin stress 6h post (permanent) LAD ligation were performed. Drugs were given 3 days prior infarction. Active GLP-1 was significantly increased by Lina (1,69 fold), saxa (1,49 fold) and OH-saxa (2,27 fold) vs. control (each p < 0.05), but not by glim. Despite similar GLP-1 levels, only lina and saxa but not OH-saxa or glim increased left ventricular contractility (Lina: 11.549 ± 795 dp/dt max; p < 0.0001; n=9. Saxa: 10.121 ± 200 dp/dt max; p < 0.05; n=12. OH-saxa: 8.201 ± 567 dp/dt max; n=7. Glim: 9.947 ± 813 dp/dt max; n=6 relative to control: 8.764 ± 284 dp/dt max; n=21). We further isolated mitochondria from non-infarcted myocardial tissue and found for lina 2.89 fold (p < 0.0001; n=3), saxa 1.78 fold (p < 0.05; n=8) and OH-saxa 2.2 fold (p < 0.01; n=4) increase in mitochondrial respiration by Clark electrode, but not for glim.

In conclusion, only linagliptin and saxagliptin increased left ventricular contractility and myocardial mitochondrial respiration following LAD ligation; 5-hydroxy-saxagliptin had no effect on contractility despite increasing mitochondrial respiration. Glimepiride had no effect on contractility and mitochondrial function. Metabolism of saxagliptin might be linked to differences found in the DPP-4 class regarding heart failure.

Disclosure

E. Haj-Yehia: None. S. Diebold: None. J. Moellmann: None. F. Kahles: None. T. Klein: Employee; Self; Boehringer Ingelheim GmbH. M. Mark: Employee; Self; Boehringer Ingelheim GmbH. N. Marx: Advisory Panel; Self; Boehringer Ingelheim GmbH. Speaker's Bureau; Self; Boehringer Ingelheim GmbH. M. Lehrke: Research Support; Self; Boehringer Ingelheim GmbH. Consultant; Self; Novo Nordisk A/S. Advisory Panel; Self; Sanofi, Amgen Inc., Bayer AG, MSD K.K..
Cold Atmospheric Pressure Plasma as a Novel Treatment Modality in Diabetic Foot Ulcers—A Pilot Study
Authors: PETERS EDGAR J. , LAGRAND RIMKE , SMITS PAULIEN , PEMEN A , SOBOTA ANA , ZEPER BAS , SABELIS LOUISE W.
Published: June 2018

Introduction: Antimicrobial resistance is a growing problem in the treatment of diabetic foot ulcers. Plasma devices generate an ionized gas with a cocktail of highly reactive species, electric fields and UV light. Cold atmospheric plasma (CAP) treatment has advantages over antiseptic or antimicrobial infection prevention and control; it disinfects efficiently, painlessly and instantly, without development of antimicrobial resistance. Concurrently, plasma can stimulate human cell proliferation and migration as well as microcirculation. We studied the safety of a novel CAP device, that is simple to use and in the future can be applied at a patient’s home.

Method: We included subjects with diabetic foot ulcers (maximum depth 5 millimeters) without clinical signs of infection or exposed bone or joint in the wound base. Subjects were treated with CAP on a daily basis for ten days in a two-week period. Primary endpoint of this study was the occurrence of serious adverse events (SAE) as a result of treatment. Safety was defined as ≤ 10% of subjects experiencing SAE related to treatment other than infection, and ≤ 60% of subjects developing infection within 30 days after treatment. Standard protocols for wound treatment were deployed, including weekly debridement and offloading.

Results: Twenty subjects were enrolled. Three SAEs (infections) occurred at the site of application within one month of treatment, of which one occurred during treatment. Three SAEs unrelated to treatment occurred: pneumonia, toe amputation on the contralateral foot and a soft tissue infection of the ipsilateral leg. Transient adverse events (AE grade 1) during one or more applications were reported by 53% of subjects.

Discussion: No SAE other than infection occurred as a result of treatment and ≤ 60% of subjects developed an infection. The AEs were low graded and transient. The results of our study demonstrate that the application of CAP in diabetic foot ulcers is safe.

Disclosure

E.J. Peters: None. R. Lagrand: None. P. Smits: Employee; Self; Plasmacure BV. A. Pemen: None. A. Sobota: None. B. Zeper: Stock/Shareholder; Self; Plasmacure. L.W. Sabelis: None.
The Effect of Hypercholesterolemia on the Characteristics of Cerebral Microvasculature
Authors: TODATE YUSUKE , ISHIGAKI YASUSHI
Published: June 2018

Aim: It remains unclear whether the increase of LDL cholesterol (LDL-C) is a risk factor for cerebral vascular disease (CVD). To clarify the effects of hypercholesterolemia on cerebral small vessel disease (SVD), we investigate the characteristics of microvasculature including lenticulostriate arteries (LSA) using high resolution brain magnetic resonance imaging (MRI) in subjects with familial hypercholesterolemia (FH).

Methods: Twenty-seven patients with FH and 25 patients with type2 diabetes (DM), 25 healthy controls underwent 7Tesla (7T) brain MRI. The prevalence of SVD and LSA structural changes, vessel wall plaque images were determined in each group.

Results: The prevalence of SVD, including white matter hyperintensities and lacunar infarctions, significantly increased in DM compared with both FH and controls. In addition, the average numbers of stems of LSA were significantly smaller in DM than FH as well as controls. Interestingly, both SVD prevalence and LSA structural changes showed no difference between FH and controls. Next, we examined the vessel wall imaging analyses of middle cerebral arteries (MCA) and vertebral-basilar arteries (VA-BA). The prevalence of vessel wall plaque on both horizontal segment of MCA (FH vs. controls; 51.8% vs. 20.0%, p<0.05) and VA-BA (37.0% vs. 12.0%, p<0.05) were significantly higher in FH than controls.

Conclusion: The prevalence of SVD and LSA structural impairment definitely increased in DM, suggesting that DM was strong risk for CVD compared to hypercholesterolemia. On the other hand, the prevalence of vessel wall plaque on MCA and VA-BA were significantly high in FH compared to controls. These results suggested long-term increase of LDL-C induced the plaque formation in cerebral microvasculature. Image analyses using 7T MRI enables us to investigate the characteristics of cerebral microvasculature prior to the development of SVD, leading to earlier interventions aimed at the prevention of atherosclerotic events.

Disclosure

Y. Todate: None. Y. Ishigaki: None.
Impaired Gastric Emptying Is Associated with a Higher Incidence of Coronary Heart Disease in Subjects with Diabetes
Authors: PARK SEON-YOUNG , CHUNG JIN OOK , CHO DONG HYEOK , CHUNG DONG JIN , CHUNG MIN YOUNG
Published: June 2018

Results: Impaired gastric emptying is observed in long-standing diabetic patients. To date, there is limited information on cardiovascular events in diabetic subjects with upper gastrointestinal (UGI) symptoms according to the presence of impaired gastric emptying. The aim of this study was to identify cardiovascular events after gastric emptying study in diabetic subjects with UGI symptoms. This study collected data from 160 diabetic subjects (23 with type 1 diabetes and 137 with type 2 diabetes) with UGI symptoms who had undergone gastric emptying scintigraphy from 2001 to 2016. We gathered data on the incidence of cardiovascular events (coronary heart disease and ischemic stroke) after gastric emptying study. There were 66 diabetic subjects with normal gastric emptying and 94 diabetic subjects with impaired gastric emptying (58 with delayed gastric emptying and 36 with rapid gastric emptying). During 7.5 years of follow-up, 22.5% of subjects (36/160) developed the cardiovascular events (19 with coronary heart disease, 12 with ischemic stroke and 5 with both). The incidence of coronary heart disease was higher in the subjects with impaired gastric emptying (4.2% at 1 year, 10.6% at 3 years, and 12.7% at 5 years) than those with normal gastric emptying (2.4% at 1 year, 3.6% at 3 years and 4.8% at 5 years p=0.039). Our results showed that diabetic subjects with impaired gastric emptying had higher incidence of coronary heart disease than those with normal gastric emptying. This study may warrant a more careful cardiovascular assessment with a high index of suspicion for possible events in management of diabetic subjects with impaired gastric emptying.

Disclosure

S. Park: None. J. Chung: None. D. Cho: None. D. Chung: None. M. Chung: None.
Reducing Hypoglycemia through Increased Knowledge of an Evidence-Based Guideline
Authors: CLARK THAYER A. , BAK LEIGH
Published: June 2018

Purpose: The purpose of this project is to increase compliance with a hypoglycemia guideline to improve patient care of those experiencing hypoglycemia in the emergency department at a large academic medical center.

Methods: In order to gain an understanding of current compliance to the hypoglycemia guideline, for four weeks all hypoglycemia events were analyzed looking at eight measures (Table) pre-intervention (n=43), and this step was repeated post intervention(n=39). The intervention period included one month dedicated to educating the staff about the guideline, implementing visual aids throughout the department, and having a staff member act as a glucose champion for all blood glucose related questions.

Results: Overall there was low compliance to the protocol initially, but after the intervention there was improvement in all eight areas analyzed (Figure). Areas that showed the most improvement included: immediate blood glucose recheck (9.1% to 23.1%), 15 minute recheck (29.5% to 51.3%), follow-up with a carbohydrate (40.9% to 71.8%), and hourly recheck x3.

Conclusion: Increasing awareness of the hypoglycemia guideline through education, and visual aids led to increased compliance. Anecdotally staff found having a glucose champion to be the most helpful in adhering to the guideline and delivering best care. Going forward having at least one assigned RN as a glucose champion is recommended.

Disclosure

T.A. Clark: None. L. Bak: None.
Gemigliptin Inhibits IL-1ß–Induced Endothelial-Mesenchymal Transition via BMP2/Smad/MAPK/Runx2 Pathway
Authors: KWON HYUK-SANG , HONG OAKKEE , KANG YANGGYO , KWON HEE SUN , YOO SOON JIB
Published: June 2018

Objective: Endothelial-to-mesenchymal transition (EndMT) contributes to inflammatory conditions inducing conversion of endothelial cells (ECs) into activated fibroblasts, promoting fibrotic diseases. Pro-inflammatory cytokine interleukin (IL)-1ß is the most potent inducer of EndMT. We investigated inhibition of IL-1ß-induced EndMT by gemigliptin, a DPP-4 inhibitor.

Methods: We exposed human umbilical vein ECs (HUVECs) to 10 ng/mL IL-1ß/20 uM gemigliptin and analyzed the expression of endothelial, smooth muscle, mesenchymal, and osteoblastic markers, bone morphogenetic protein (BMP), Smad, and non-Smad signaling pathway proteins.

Results: Morphological changes showed gemigliptin blocked IL-1ß-induced EndMT, upregulated EC markers, and downregulated smooth muscle and mesenchymal markers. IL-1ß activation of HUVECs is initiated by the BMP/Smad and canonical BMP signaling pathways. Gemigliptin inhibited IL-1ß induction of BMP2 and 7, activin receptor type IA, BMP receptor type IA, and BMP receptor II. Reversal of IL-1ß-mediated inhibition of BMP-induced Smad 1/5/8, Smad2, and Smad3 phosphorylation by gemigliptin suggests involvement of the Smad pathway in gemigliptin action. In the canonical BMP pathway, gemigliptin treatment significantly increased the deactivation of ERK, p38, and JNK by IL-1ß. Gemigliptin treatment suppressed BMP-2-induced expression of key osteoblastic markers including osterix and runt-related transcription factor 2 during IL-1ß-induced EndMT.

Conclusions: We demonstrated a novel protective mechanism of gemigliptin against fibrosis by suppressing IL-1ß-induced EndMT.

Disclosure

H. Kwon: None. O. Hong: None. Y. Kang: None. H. Kwon: None. S. Yoo: None.
CRISPR-Cas9 Gene Editing Restores Beta-Cell Differentiation and Function in Patient-Specific iPSCs
Authors: MILLETTE KATELYN , VOGT KYLE R. , SALAS ANDREW , PITUKCHEEWANONT PISIT , AUSTIN JULIANA , MARTIN MARTIN , GEORGIA SENTA
Published: June 2018

Genetic knockout studies in mice concluded that the differentiation of endocrine lineages in the pancreas and intestine requires the transcription factor neurogenin3 (NGN3). However, case reports of patients with NGN3 mutations show that most patients do not develop diabetes until later in childhood, suggesting that NGN3 may not be required for human beta cell differentiation in vivo. We have identified a patient with a previously undescribed NGN3 loss-of-function mutation and generated induced pluripotent stem cells from patient fibroblasts to analyze the role of NGN3 in human beta cell differentiation. We used established protocols to differentiate patient-specific iPSCs (PS-iPSCs) into pancreatic progenitor cells and beta-like cells in parallel with the H1 hESC line. The PS- iPSCs displayed significantly lower pancreatic progenitor cell differentiation with only 10.7% of the double positive PDX1+/NKX6.1+ cell population present, whereas the control H1s hESCs produce 60% double positive cells. After further differentiation into beta-like cells, H1 cells produced multiple endocrine cell types, while the PS-iPSC line was not able to make significant numbers of endocrine cells. To assess if the loss of NGN3 was the primary cause for the loss of pancreatic progenitor cells and beta-like cell differentiation, we used CRISPR-cas9 gene editing to correct the patient’s mutation in the PS-iPSCs (cPS-iPSCs). Correcting the patient’s mutation restored the PDX1+/NKX6.1+ cell population in the pancreatic progenitor stage. Beta-like cells differentiated from cPS- iPSCs were capable of glucose responsive insulin secretion at the end of the differentiation protocol.

In conclusion, our data suggests that NGN3 may have a previously unidentified role important for the differentiation of pancreatic progenitor cells and that there may be a relationship between NKX6.1 and NGN3 that is critical for pancreatic development.

Disclosure

K. Millette: None. K.R. Vogt: None. A. Salas: None. P. Pitukcheewanont: None. J. Austin: None. M. Martin: None. S. Georgia: None.
Canagliflozin Inhibits Human Endothelial Cell Inflammation via the Induction of Heme Oxygenase-1
Authors: PEYTON KELLY J. , BEHNAMMANESH GHAZALEH , DURANTE WILLIAM
Published: June 2018

Vascular disease is the major cause of mortality in diabetes. Although many factors contribute to diabetes-associated vascular disease, inflammation of endothelial cells (ECs) plays a critical role. Sodium-glucose transporter 2 (SGLT2) inhibitors are recently approved anti-hyperglycemic drugs that cause a significant reduction in cardiovascular events in type 2 diabetic patients, but the mechanisms underlying this protection are not fully known. In the present study, we investigated the effect of SGLT2 inhibitors on the expression of the vasoprotective protein heme oxygenase-1 (HO-1) in human umbilical vein ECs. Treatment of ECs with canagliflozin, dapagliflozin, or empagliflozin stimulated the expression of HO-1 mRNA and protein; however, only clinically achievable concentrations of canagliflozin (3-10µM) increased HO-1 levels. The induction of HO-1 by canagliflozin was associated with an increase in Nrf2 activity and reactive oxygen species (ROS) formation. Canagliflozin also stimulated HO-1 promoter activity, and this was prevented by mutating the antioxidant responsive element or by overexpressing dominant-negative Nrf2. In addition, the canagliflozin-mediated induction of HO-1 was abolished by N-acetyl-L-cysteine or rotenone. Significantly, prior treatment of ECs with canagliflozin inhibited the induction of the adhesion molecules ICAM-1 and VCAM-1 in response to high concentrations of glucose (25mM) and oxidized low-density lipoprotein (50mg/L) that mimic the diabetic milieu. Significantly, the anti-inflammatory action of canagliflozin was abrogated by blocking HO-1 activity or expression.

In conclusion, this study demonstrates that canagliflozin stimulates HO-1 gene expression in the vascular endothelium via the ROS-Nrf2 pathway, and that canagliflozin mitigates inflammation in human ECs exposed to a diabetic environment via the induction of HO-1. Thus, canagliflozin may exert some of it cardiovascular benefit in diabetes by targeting HO-1.

Disclosure

K.J. Peyton: None. G. Behnammanesh: None. W. Durante: None.
Actions of Liraglutide on Vascular Endothelial Cells Play a Central Role in the Suppression of Atherosclerosis in Diabetic Mice
Authors: HIROMURA MUNENORI , MORI YUSAKU , KOSHIBU MASAKAZU , KUSHIMA HIDEKI , KOHASHI KYOKO , TERASAKI MICHISHIGE , HIRANO TSUTOMU
Published: June 2018

Background: Glucagon-like peptide-1 receptor agonists (GLP-1RAs) have been shown to exert direct anti-atherosclerotic effects through multiple actions on various cells. However, it is unclear which action is crucial. We investigated the role of the anti-atherogenic effects of liraglutide (Lira) on vascular endothelial cells (VECs) in vivo.

Methods: Streptozotocin-induced diabetic apolipoprotein E-null mice (male, 20 weeks) were randomly assigned to treatment with saline or Lira (17 nmol/kg/d) and underwent femoral artery (FA) wire injury to remove VECs. Thioglycolate-exudated peritoneal macrophages and vascular samples (aorta, FA, and right brachiocephalic artery [RBCA]) were collected 4 weeks after treatment.

Results: In the Lira group, active GLP-1 levels were significantly higher (4.7 ± 1.1 vs. 2.8 ± 0.3 pmol/L) and levels of HbA1c (8.9 ± 0.4% vs. 8.3 ± 0.5%) and total cholesterol (492 ± 8 vs. 457 ± 19 mg/dL) were significantly lower than in controls. Lira significantly reduced the atherosclerotic plaque (oil red O [ORO]-stained area) on the surface of the whole aorta by 36% and the plaque volume and intraplaque macrophage infiltration in the aortic root by 56% and 55%, respectively. Additionally, Lira suppressed the expression of inflammatory cytokines in macrophages by 40-45% (p < 0.05) and the expression of inflammatory cytokines in the RBCAs, which are prone to lesion, plaque formation, and rupture. In FA, no plaque was observed in uninjured lesions, while severe plaque formation occurred in injured lesions. In contrast, Lira failed to suppress plaque volume or neointimal thickening in FAs where the VECs were removed (ORO-stained area/vascular wall area: 0.51 ± 0.vs. 0.55 ± 0.08, neointimal area/medial area: 2.4 ± 1.2 vs. 2.6 ± 0.7), indicating that Lira cannot suppress atherosclerosis independent of VECs.

Conclusions: Of the multiple actions on various cells, VECs play a central role in the suppression of atherosclerosis mediated by Lira.

Disclosure

M. Hiromura: None. Y. Mori: None. M. Koshibu: None. H. Kushima: None. K. Kohashi: None. M. Terasaki: None. T. Hirano: Speaker's Bureau; Self; Novo Nordisk Inc., AstraZeneca.
Coronary Microvascular Smooth Muscle Cells Contribute to Heart Failure in Diabetes Mellitus by Increasing Methylglyoxal Flux
Authors: BIDASEE KESHORE
Published: June 2018

Prognosis for patients with type 1 diabetes mellitus (T1DM) who have heart failure is poor because of an incomplete understanding of molecular cues that trigger it and a lack of specific pharmacologic agents/strategies to slow/reverse its development. Supra-physiologic levels of the α-oxoaldehyde methylglyoxal (MG) have emerged as a possible cue. However, its source remains poorly characterized. Here we investigated MG levels and expression of its degrading and synthesizing enzymes glyoxalase-I (Glo-I) and vascular adhesion protein-1 (VAP-1) in ventricular tissues from rats with chronic streptozotocin-induced T1DM. We also used a gene transfer approach to prevent Glo-I downregulation in ventricular tissues to determine if it would be cardio-protective. When compared to controls, hearts of rats with chronic T1DM (10 -12 weeks) had 5-fold higher MG in ventricular tissues, 2.2-fold lower level of Glo-I in ventricular tissues and 2.5-fold higher in coronary microvascular smooth muscle cells (cSMCs). Preventing Glo-I levels downregulation in myocytes and cSMCs of rats by injecting them with an adeno-associated virus contain Glo-I driven by the endothelin promoter one week after the onset of T1DM, attenuated ventricular MG increase, with no effect in blood MG. Increasing Glo-I also blunted VAP-1 upregulation in cSMCs, attenuated microvascular leakage, inflammation, and fibrosis. These data support the notion that an increase in MG arising in part from increased expression of VAP-1 is contributing to HF development in T1DM rats and lowering ventricular MG levels T1DM rats is cardio-protective.

Disclosure

K. Bidasee: None.
Rho-Kinase Induces CTGF Expression through Actin Dynamics in Mesangial Cells
Authors: NAGAI YOSUKE , KAWANAMI DAIJI , MATOBA KEIICHIRO , TAKEDA YUSUKE , AKAMINE TOMOYO , ISHIZAWA SHO , KANAZAWA YASUSHI , YOKOTA TAMOTSU , UTSUNOMIYA KAZUNORI
Published: June 2018

Introduction: Transforming growth factor (TGF)-β promotes the development of diabetic nephropathy by enhancing excessive production of extracellular matrix such as connective tissue growth factor (CTGF) in mesangial cells. However, the mechanism by which TGF-β induces CTGF expression has not been elucidated. In this study, we aimed to investigate contribution of small GTPase Rho and Rho-kinase to TGF-β-mediated CTGF expression in mesangial cells.

Methods and Results: Exposure of mouse mesangial cells (MES-13) to TGF-β induced activation of Rho and Rho-kinase. Y-27632, a specific Rho-kinase inhibitor, reduced TGF-β-mediated CTGF protein expression, indicating that TGF-β induces CTGF expression via Rho-kinase activation. Western blot analysis demonstrated that Y-27632 attenuates TGF-β-mediated phosphorylation of Erk1/2 and JNK/SAPK and nuclear translocation of NF-κB. Rho-kinase has been shown to regulate cytoskeleton reorganization. We therefore investigated whether Rho-kinase is involved in actin dynamics-induced NF-κB activation. Immunostaining studies demonstrated that Y-27632 and Cytochalasin D, an inhibitor of actin polymerization, attenuated TGF-β-mediated nuclear translocation of NF-κB, indicating that Rho-kinase-mediated actin reorganization regulates NF-κB activation. Furthermore, siRNA-mediated knockdown of Rho-kinase isoforms demonstrated ROCK2 but not ROCK1 is involved in these observations. In the present study, we demonstrate for the first time that Rho-kinase, especially ROCK2, mediates TGF-β-induced expression of CTGF via phosphorylation of Erk1/2 and JNK/SAPK and subsequent actin reorganization-mediated nuclear translocation of NF-κB in mesangial cells.

Conclusion: Rho-kinase could be an important therapeutic target against diabetic nephropathy.

Disclosure

Y. Nagai: None. D. Kawanami: None. K. Matoba: None. Y. Takeda: None. T. Akamine: None. S. Ishizawa: None. Y. Kanazawa: None. T. Yokota: None. K. Utsunomiya: None.
Dietary Intervention for Inpatient Glycemic Control
Authors: CROMER SARA , BOGUN MAGDALENA M.
Published: June 2018

While recommendations for inpatient glycemic control suggest a conservative target of 140-180mg/dL, many struggle to achieve this goal. Glycemic control begins with a carbohydrate-controlled diet, but many inpatient diets continue to be suboptimal and can contribute to hyperglycemia. At our institution, the carbohydrate-controlled breakfast diet was changed to better reflect carbohydrate goals (e.g., replacing french toast on the “bad diet” with vegetable frittatas on the “good diet”). Pre- and post-prandial glucose values for patients with types I and II diabetes receiving a carbohydrate-controlled diet were collected by retrospective chart review during the weeks before and after these changes took effect. Patients on steroids were analyzed separately. Individual post-prandial glucose values were recorded in a binary fashion as meeting or exceeding predefined glucose targets (<180 mg/dL, <250mg/dL, and <350mg/dL). During this period, 369 patients not receiving steroids and 39 patients receiving steroids had at least one relevant glucose value. In the non-steroid group, there was no difference in proportion of patients who exceeded any of the predefined thresholds. However, in the steroid group, there was a trend toward reduction in proportion of patients exceeding blood glucose of 180mg/dL (absolute risk reduction 19%, 95% confidence interval -4% to 42%, p = 0.12) and 250mg/dL (absolute risk reduction 17%, 95% confidence interval -2% to 35%, p = 0.08) on the days after the dietary change. Although all patients with diabetes may experience hyperglycemia proportionate to their carbohydrate intake, those on steroids experience induced insulin resistance leading to especially diet-sensitive glucose excursions. Our data suggests that patients receiving steroids can likely achieve improved inpatient glucose control through dietary modification. Physician and administration attention to carbohydrate-controlled menus represents an opportunity for improved quality of care for inpatients with diabetes.

Disclosure

S. Cromer: None. M.M. Bogun: Other Relationship; Self; Mytonomy.
DNA Hydroxymethylation Regulates Beta-Cell Maturation and Expansion
Authors: GURLO TATYANA , GEORGIA SENTA , DHAWAN SANGEETA
Published: June 2018

Epigenetic mechanisms play a central role in governing functional beta-cell mass. In particular, the methylation of DNA to generate 5-methylcytosine (5mC), directed by DNA methyltransferases, has been shown to regulate the establishment and maintenance of beta-cell identity and function. While the contribution of DNA methylation to beta-cell homeostasis and diabetes pathogenesis has been studied in detail, not much is known about the role of DNA demethylation in beta-cells. Oxidative conversion of 5mC to 5-hydroxymethylcytosine (5hmC) by Ten Eleven Translocation (Tet) enzymes constitutes a key step towards active DNA demethylation. In the present study, we provide evidence that the 5hmC landscape is dynamic, and undergoes specific changes during embryonic and postnatal islet development, islet adaptation, and in diabetes. We report that the Pdx1-positive pancreatic progenitors are marked by very low 5hmC, and beta-cell differentiation and functional maturation is accompanied by a progressive increase in 5hmC. However, under conditions during adaptive beta-cell expansion as well as in diabetes 5hmC levels decline in part due to the oxidative stress. These changes in 5hmC pattern are conserved in mouse and human pancreas, and are accompanied by the changes in Tet2 levels. We show that deletion of Tet2 in the pancreatic lineage results in improved glucose tolerance and beta-cell function. Together, our data show the importance of Tet2-dependent 5hmC patterning in maintaining a differentiated beta-cell phenotype, and highlight the remodeling of beta-cell epigenetic landscape as a key component of diabetes pathogenesis.

Disclosure

T. Gurlo: None. S. Georgia: None. S. Dhawan: None.
Renin Production and Involvement of ß1 Integrin in Blood Pressure Regulation in Diabetic Nephropathy
Authors: TOYODA MASAO , SAITO NOBUMICHI , KIMURA MORITSUGU , MORIYA HITOMI , SAWADA KAICHIRO , FUKAGAWA MASAFUMI
Published: June 2018

Diabetic nephropathy (DN) and hypertension mutually exacerbate each other, but many aspects of the mechanisms of these two pathologies remain unclear. The juxtaglomerular cells (JGC) secrete renin and trigger the renin-angiotensin-aldosterone system involved in blood pressure regulation. Integrins (Itgs) are cell surface proteins that interact with the extracellular matrix and transduce intracellular signals in response to extracellular stimuli. There are several phosphorylation sites in the β chain, and phosphorylation plays an important role in the cell-binding ability and intracellular signaling functions of Itgs. Our previous immunohistochemical analysis of renal tissue sections of diabetic patients showed that β1 Itg, a cellular matrix adhesion factor, undergoes JGC-specific phosphorylation. To characterize the relationship between the phosphorylated β1 (pβ1) Itg and renin expression in DN, the following studies were conducted in rats with streptozotocin (STZ)-induced diabetes.

Immunohistochemical analysis of the prepared sections showed JGC-specific phosphorylation of β1 Itg in the kidneys of diabetic rats. Immunofluorescent staining of these kidney tissue specimens confirmed the coexpression of pβ1 Itg and renin in JGC.

The mRNA levels of pβ1 Itg decreased continuously with time in diabetic rats after the onset of diabetes, till month 2. However, the mRNA levels of pβ1 Itg increased subsequently, but ultimately showed recovery to the baseline level. In contrast, the renin mRNA ratio exhibited an inverse correlation with pβ1 Itg. Quantification by image analysis of confocal laser microscopic images confirmed the negative correlation between pβ1 Itg and renin.

Our results showed a negative correlation between pβ1 Itg and renin expression in renal JGC, suggesting a link between β1 Itg and the regulatory system that controls renin production and secretion.

Disclosure

M. Toyoda: None. N. Saito: None. M. Kimura: None. H. Moriya: None. K. Sawada: None. M. Fukagawa: None.
Klotho Prevents Epithelial-Mesenchymal Transition through Egr-1 Down-Regulation in Diabetic Kidney Disease
Authors: LI YANG , XUE MENG , HU FANG , JIA YIJIE , YANG YAN-LIN , XUE YAO-MING
Published: June 2018

Background: As a key event leading to tubulointerstitial fibrosis in diabetic kidney disease (DKD), epithelial-mesenchymal transition (EMT) has drawn increasing attention from researchers. Klotho attenuates renal fibrosis in part by inhibiting ERK1/2 signaling in DKD. Early growth response factor 1 (Egr-1), activated mainly by ERK1/2, has been shown to play an important role in EMT. However, whether Klotho prevents EMT by inhibiting ERK1/2-dependent Egr-1 expression in DKD is unclear. The aim of this study was to investigate whether Klotho prevents EMT through Egr-1 down-regulation by inhibiting the ERK1/2 signaling pathway in DKD.

Methods: Male C57BL/6J mice fed an HFD for 4 weeks received 120 mg/kg STZ. Klotho and Egr-1 expression was detected in the renal cortices of these mice upon their sacrifice at 6 and 12 w after STZ treatment. In vitro studies, we incubated HK2 cells under high-glucose (HG) or TGF-β1 conditions to mimic DKD. We then transfected the cells with an Egr-1-containing plasmid, Egr-1 siRNA, a Klotho-containing plasmid or Klotho siRNA. Klotho, Egr-1, E-cadherin, α-smooth muscle actin (α-SMA), fibronectin(FN), ERK1/2 and p-ERK1/2 were determined.

Results: Klotho expression was significantly decreased in the renal cortices of diabetes mellitus (DM) mice compared to control mice at 6 w and was even more significantly decreased at 12 w. In contrast, Egr-1 expression was significantly increased in DM mice compared to control mice only at 12 w. We also found that Klotho overexpression down-regulated Egr-1 expression and the (p-ERK1/2)/(ERK1/2) ratio in HG- or TGFβ1-treated HK2 cells. Conversely, Klotho silencing up-regulated Egr-1 expression and the (p-ERK1/2)/(ERK1/2) ratio. Moreover, the effects of si-Klotho were abolished by the ERK1/2 inhibitor PD98059.

Conclusions: Klotho prevents EMT during DKD progression, an effect that has been partially attributed to Egr-1 down-regulation mediated by ERK1/2 signaling pathway inhibition.

Disclosure

Y. Li: None. M. Xue: None. F. Hu: None. Y. Jia: None. Y. Yang: None. Y. Xue: None.
Serpin B13 Plays a Role in Beta-Cell Development and Progression to Insulin-Dependent Diabetes
Authors: KRYVALAP YURY , JIANG MATTHEW L. , KRYVALAP NADZEYA , MUELLER KYLE A. , CZYZYK JAN
Published: June 2018

Identifying natural factors of the progression to type 1 diabetes (T1D) provides a unique opportunity to develop novel preventative strategies for this disease. Our laboratory has identified serpin B13 proteinase inhibitor as a potential player, activity of which is involved in modifying the timing of T1D development. In this study we examined whether blocking of serpin B13 with a monoclonal antibody (mAb) in early life causes any changes in pancreatic islet development that may explain beneficial impact of this activity later in life. First, our animal studies showed that blocking of serpin B13 during mid-pregnancy caused robust increase in the number of Ngn3+ progenitor cells at day E16.5 and accelerated conversion of these cells to beta cells. Similar changes were observed in embryonic pancreatic buds in vitro. On the other hand, the serpin B13 protein significantly downregulated the output of Ngn3+ cells in these in vitro cultures. Importantly, at 2 to 3 months after birth, the offspring of serpin B13 mAb-treated mothers demonstrated increased islet cell mass and responded to the diabetes-inducing agent, streptozotocin, with markedly improved glucose tolerance and renal function, compared with control animals. Second, our screening of 280 children, previously enrolled in a DPT-1 clinical trial, for serpin B13 autoantibody (AA) revealed an inverse correlation of this AA with risk level for T1D, as well as a positive association with longer diabetes-free interval. These data demonstrate the significant, long-term impact of serpin B13 activity on islet biology and suggest that blocking this serpin has the potential to partially prevent, or at least slow down, the development of T1D both in the mouse and human.

Disclosure

Y. Kryvalap: None. M.L. Jiang: None. N. Kryvalap: None. K.A. Mueller: None. J. Czyzyk: None.
Klotho Prevented Epithelial-to-Mesenchymal Transition in High-Fat Diet/Streptozotocin-Induced Mice by Targeting NFkB/Microrna-21/Smad7 Signaling Pathway
Authors: XUE MENG , HU FANG , LI YANG , JIA YIJIE , XUE YAO-MING
Published: June 2018

Klotho is an antiaging hormone present in the kidney that protect renal via attenuating the nuclear factor-kB (NFkB) p65 activity. And in kidney epithelium miRNAs become active in response to injury. But little is known regarding to the role of Klotho on miRNAs in the pathogenesis of diabetic nephropathy(DN). In this study, we found that the expression of Klotho was reduced with upregulation of miR-21 and key parameters of DN in HFD/STZ-induced diabetic mice. Klotho-treatment reduced p65 nucleus expression in association with modulation of miR-21/Smad7 and amelioration of fibrosis. Furthermore, we found that Klotho expression was decreased in high glucose induced HK2 at both mRNA and protein, accompanied by the significantly increased miR-21 expression. NF-κB was demonstrated to regulate miR-21 expression by directly binding to its promotor. Additionally, exogenous addition of Klotho or inhibition of NF-kB restrained the activity of miR-21/Smad7 signaling pathways, which suppressed fibrosis in high glucose cultured HK2. This study provides a new basis to elucidate the protection mechanism of anti-aging protein Klotho in diabetic kidney. We firstly found that Klotho treatment could delay the fobrosis progression of diabetic nephropathy in type 2 diabetic mice via inhibition of NF-kB/miR-21/Smad7 signaling cascade pathways.

Disclosure

M. Xue: None. F. Hu: None. Y. Li: None. Y. Jia: None. Y. Xue: None.
Empagliflozin and Linagliptin Alleviate Glomerular Changes in a Model of Diabetic Nephropathy
Authors: KORBUT ANTON I. , KLIMONTOV VADIM , TASKAEVA IULIIA , BGATOVA NATALIYA P. , ISHCHENKO IRINA , ZAVJALOV EVGENII L.
Published: June 2018

The aim: to assess the effects of SGLT2 inhibitor empagliflozin, DPP-4 inhibitor linagliptin, and their combination on structural changes in kidneys in a model of type 2 diabetic nephropathy. Eight-week-old male db/db diabetic mice (BKS.Cg-Dock7m+/+Leprdb/J) were treated with empagliflozine (10 mg/kg), linagliptin (10 mg/kg), combination of both agents, or placebo for 8 weeks. Nondiabetic heterozygous db/+ mice were used as control. Renal structural changes were analyzed quantitatively from the light and electron microscopic images. Both empagliflozin and linagliptin, either alone or in combination, attenuated mesangial expansion estimated by mesangial volume (p=0.00for empagliflozin, p=0.03 for linagliptin and combination). The width of glomerular basement membrane was diminished in all treated groups (all p<0.01). In diabetic mice, podocytopathy was manifested by the foot process effacement (p=0.03). Under treatment, the number of podocyte foot processes increased (p=0.003 for linagliptin, p=0.03 for empagliflozin and combination) and the width of podocyte foot processes decreased in all treated groups (p=0.003 for empagliflozin and linagliptin, p=0.001 for combintion). The number of endothelial fenestrae in glomerular capillaries was increased after the treatment (all p<0.01). The interaction between empagliflozin and linagliptin was significant for the number and width of podocyte foot processes, and the number of endothelial fenestrae (all p<0.01). The effects of both agents on structural changes were not associated with hypoglycemic activity. The obtained results demonstrate that empagliflozin and linagliptin ameliorate renal fibrosis and podocyte injury in a model of type 2 diabetic nephropathy.

Disclosure

A.I. Korbut: None. V. Klimontov: None. I. Taskaeva: None. N.P. Bgatova: None. I. Ishchenko: None. E.L. Zavjalov: None.