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COMPLICATIONS—HYPOGLYCEMIA an activating role of SAMSN1, L-triiodothyronine, IFNA4, JAK1 and mTORC1, and an inhibitory action of BDNF, POR, ESR1, CTNNB1 and ERG on the gene networks identified in our samples. Moderated Poster Discussion: Hypoglycemia—Novel Concepts Our study for the first time characterizes the transcriptional responses (Posters: 381-P to 386-P), see page 19. of the BBB compartment to recurrent hypoglycemia exposure and may help identify novel therapeutic targets to restore the impaired responses against 381‑P hypoglycemia in patients with type 1 . & Supported By: National Institutes of Health; JDRF Hypoglycemia-Associated Autonomic Failure Is Associated with POSTERS Complications

Coordinated miRNA-mRNA Network Changes in the Ventromedial Acute and Chronic & 383‑P RAHUL AGRAWAL, CASEY TAYLOR, ADRIANA VIEIRA-DE-ABREU, SIMON J. KATP Channel Opening Increases Severe Hypoglycemia-Induced FISHER, Salt Lake City, UT Cardiac Arrhythmias We tested the hypothesis that the maladaptive response to recurrent CANDACE M. RENO, JUSTIN BAYLES, ALLIE SKINNER, SIMON J. FISHER, Salt hypoglycemia (RH; i.e., HAAF) is mediated by the coordinated changes in Lake City, UT microRNA-mRNA expression in ventromedial hypothalamus (VMH). The mechanisms of how severe hypoglycemia leads to fatal cardiac induced (2-1.2 U/kg, sc) RH (30-50 mg/dl x 3 days, n=10) or recurrent arrhythmias are unknown. It is hypothesized that ATP sensitive potassium (RS, n=10) Sprague-Dawley rats were subjected to either, 1) hyperinsulinemic (KATP) channels regulate arrhythmias during severe hypoglycemia. To test (20 mU/kg/min) hypoglycemic (40-50 mg/dl) clamps, or 2) were euthanized to the role of KATP channels in severe hypoglycemia-induced arrhythmias, obtain RNA from the VMH for genome-wide microRNAome and transcrip- nondiabetic Sprague Dawley rats were infused with either vehicle (VEH, tome profiling. As expected, the epinephrine response to hypoglycemia was 18% ethanol, n=6), KATP blocker (, GLIB, 5 mg/kg/hr n=5), or blunted by 55% in RH rats. A total of 205 miRNAs and 1013 mRNAs were KATP opener (diazoxide, DIAZ, 5 mg/kg/hr, n=4) during a hyperinsulinemic differentially expressed (false discovery rate; FDR<0.05) between groups. A (0.2 U/kg/min) severe hypoglycemic (10-15 mg/dl) clamp with electrocardio- miRNA target filtering analysis using Ingenuity Pathway Analysis (IPA) soft- gram recordings for 3 hours. The infusion rates to maintain severe ware demonstrated coordinated changes in miRNA-mRNA with 82 miRNA hypoglycemia were not different among the groups indicating the drugs did pairing to 402 mRNA (FDR <0.05). miRNA-mRNA network analysis of RH not affect hypoglycemic counterregulation. Various types of cardiac arrhyth- induced genomic changes identified miR-23a-3p and miR-7a-5p based on mias were noted. Compared to VEH, 2nd degree heart block was not differ- their large predicted network and association with mRNA target changes, ent in GLIB, but DIAZ significantly increased 2nd degree heart block 10-fold specifically Clcn3 and Mknk2 (gene products involved in GABAergic synaptic (p<0.05; Figure). Third degree heart block was reduced by 22% in GLIB and vesicles release and pancreatic β-cells function, respectively). increased 19% in DIAZ compared to VEH. However, severe hypoglycemia- In conclusion, RH induced changes in VMH levels of miR-23a-3p and miR- induced mortality was not different among the groups. 7a-5p might be potential mediators or regulators of HAAF. In summary, during severe hypoglycemia, KATP blockade tended to reduce arrhythmias whereas KATP opening increased cardiac arrhythmias. Figure. In conclusion, acute modulation of KATP channel activity during severe hypoglycemia can affect the incidence of potentially fatal cardiac arrhythmias. Figure.

Supported By: JDRF & 382‑P -Brain-Barrier Transcriptome Response to Recurrent Hypo‑ glycemia Exposure in a Rodent Model of Hypoglycemia Unaware‑ ness ZEJIAN LIU, YUYAN DING, RAIMUND I. HERZOG, New Haven, CT Recurrent hypoglycemia (RH) results in hypoglycemia unawareness, which substantially increases the risk of severe hypoglycemic episodes. Pre- Supported By: National Institutes of Health vious studies suggested that changes of energy substrate transporters and other regulators at the level of the Blood-Brain-Barrier (BBB) may contribute to this problem. To understand response of the BBB to antecedent RH, we & 384‑P isolated cortical brain microvessels (MVs), immediately after an episode Hypoglycemia Effects on Neurocognitive Function in Type 1 Diabe‑ of acute hypoglycemia, from our rodent model, in which rats were either tes Mellitus Measured by Neuroimaging pretreated with 3 consecutive days of recurrent hypoglycemia (3dRH) or KAITLIN G. BRAU, ALEX I. WIESMAN, TIMOTHY J. MCDERMOTT, ANDJELA T. left untreated (n=15 per group). We then analyzed extracted RNA by whole DRINCIC, TONY W. WILSON, CYRUS V. DESOUZA, Omaha, NE exome sequencing to identify transcriptional differences. Hypoglycemia has been associated with diabetes-related cognitive dys- Verification of MVs purity confirmed a 50-fold enrichment of the endothe- function. We aimed to better understand neuronal changes that precipitate lial compartment in comparison to whole brain cortical tissue. Out of 13,951 the development and progression of this cognitive dysfunction in type 1 genes detected, we identified a signature of 50 genes that were character- diabetes mellitus (T1DM) by using a noninvasive functional neuroimaging istic for exposure to 3dRH, with 19 being upregulated and 31 being down- tool that directly evaluates neural activity, known as magnetoencephalogra- regulated. Manual annotation of the 50 genes suggests that RH attenuates phy (MEG). We hypothesized that MEG brain function parameters would be inflammatory responses, ameliorates vascular permeability and inhibits abnormal in patients with T1DM with more frequent hypoglycemia. Thirty- cycle progression. Ingenuity Pathway Analysis (IPA) revealed key signaling three patients with T1DM completed a somatosensory task during MEG. cascades affected by RH, including , ER stress/unfolded During this task, electrical paired pulse stimulation was applied to a par- response (UPR), NF-κB, STAT3 and LXR/RXR signaling. Candidate ticipant’s right median nerve. All MEG data was processed following stan- pathways were validated by qPCR and/or Western blot in independently dard protocols, and the resulting images of functional brain activity were generated confirmatory MV samples. Upstream regulator analysis predicted averaged across all patients. The time series of activation in the peak voxel (cube of tissue) was then evaluated statistically for group differences. Data

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A101 COMPLICATIONS—HYPOGLYCEMIA regarding outpatient frequency of hypoglycemia was collected. Patients & 386‑P were divided into two groups, 1-or-less and 3-or-more episodes of blood An Aptamer-Based Platform for Platelet Proteomics following glucose < 70 mg/dL per week. Patients with more frequent hypoglycemia Induced Hypoglycaemia in Diabetes and Control Subjects had significantly stronger neural activity during the baseline period (before HASSAN KAHAL, ERIC S. KILPATRICK, THOZHUKAT SATHYAPALAN, ADITYA M. stimulation onset) relative to peers with less frequent episodes (p < 0.05). BHAGWAT, JOHANNES GRAUMANN, STEPHEN L. ATKIN, Coventry, United King- Patients with more episodes also had stronger absolute responses to the dom, Doha, Qatar, Hull, United Kingdom paired stimulations (p < 0.05), along with significantly reduced somatosen- Aims and Hypothesis: Strict glycaemic control has been associated with sory gating (i.e., equal responses to both stimulations; p < 0.05). Patients

POSTERS an increased mortality rate in subjects with (T2DM), the Complications with more frequent hypoglycemic episodes had substantially elevated cause of which is unclear. Acute and Chronic spontaneous neural activity in somatosensory regions (i.e., brain “noise”) Methods: Using hyperinsulinaemic clamps, blood glucose levels were stabi- and abnormal responses to the stimulation. Such brain noise has been asso- lised at 90mg/dl for 1 hour (1 h), then reduced over 1 h to 50mg/dl for 1 h. Blood ciated with advancing age, which may suggest increased neural aging in samples were collected at times 0 (baseline), 2 h (euglycaemia), 4 h (hypoglycae- patients with more frequent hypoglycemic episodes. Such premature aging mia) and at 24 h after the clamp studies. Plasma was analysed using SOMAscan, could contribute to the cognitive decline that has been associated with dia- an aptamer technology to comparatively quantify a platelet panel of 114 . betes. Results: 10 subjects with T2DM and 8 controls were recruited. In sub- jects with T2DM, there were no acute platelet function changes but at 24 h & 385‑P there was increased coagulability with a decreased response to prostacy- Prevention of Hypoglycemia-Induced Neural Damage by Verapamil clin (1nM). No significant changes in platelet function were noted in controls DAVID A. JACKSON, TREVIN MICHAEL, ADRIANA VIEIRA DE ABREU, RAHUL either acutely or after 24 h. Significant changes in 13/114 platelet associated AGRAWAL, SIMON J. FISHER, Salt Lake City, UT proteins were see at hypoglycaemia (Figure) with significant though weak Type 1 diabetic patients at an increased risk of severe hypoglycemia differences for clusterin, thrombin and fractalkine at 24 h. induced brain damage. Rapid correction of hypoglycemia with glucose may Conclusions/Interpretation: Significant differences in platelet associated worsen brain damage due to rapid calcium influx/cytotoxicity. It was hypoth- proteins are induced by hypoglycaemia in comparison between normal and esized that a calcium channel blocker, Verapamil (VER), would significantly subjects with T2DM that are not reflected in altered platelet function, but reduce brain damage caused by insulin-induced severe hypoglycemia. Male residual protein effects may still be evident after 24 h when there is an Sprague-Dawley rats (4 per treatment group) were subjected to either, 1) increase in coagulability. -1 -1 a control hyperinsulinemic (200mU.kg .min ) euglycemic (80-100mg/dL) Figure. clamp, 2) hyperinsulinemic hypoglycemic (≤15mg/dL) clamp, or 3) hyperinsu- linemic hypoglycemic clamp followed by VER. All clamps were 90 minutes. The clamps were followed by a recovery period with concurrent treatment of glucose infusion and an IP injection of either 0.9% saline or VER (20mg/kg). Seven days after this one-time treatment, neural damage was quantified in euthanized rats by counting Fluoro-Jade B positive (+) cells (evaluated by a blind observer). Severe hypoglycemia markedly increased neural damage in the hippocampus (p<0.01) and cortex (p<0.01) compared to euglycemia. VER treatment immediately following severe hypoglycemia decreased neural damage by 85% in the hippocampus (p<0.05; Figure) and 83% in the cortex (p=0.055). In summary, VER attenuates neural damage caused by severe hypoglyce- mia. For insulin treated people with diabetes, VER may be a useful drug to reduce the risk of hypoglycemia induced brain damage. Figure.

Moderated Poster Discussion: Risk Factors and Prediction of Hypogly- cemia (Posters: 387-P to 392-P), see page 21. & 387‑P Predicting Inpatient Hypoglycaemia Risk with Cardiff Hypoglycae‑ mia Risk Score (CHRiS) VINAY S. ELIGAR, PETER TAYLOR, ARSHIYA TABASUM, PREETHI NALLA, ANGE- LIKI GIANNOPOULOU, SAM RICE, KOFI OBUOBIE, ALED ROBERTS, ANIL KUMAR PURA NARAYANASWAMY, NEERA AGARWAL, JULIA PLATTS, Cardiff, United Kingdom, Llanelli, United Kingdom, Newport, United Kingdom Introduction: Up to 22% of inpatients with diabetes can develop hypo- glycaemia during their hospital stay, which increases the length of stay and health care costs. We have developed a predictive scoring system to prevent inpatient hypoglycaemia. Methods: We analysed the risk factors for developing hypoglycaemia in 142 diabetes inpatients. The risk score was generated based on 10 criteria’s including type of diabetes, length of diabetes, , hepatic/renal , recurrent hypoglycaemia, hypoglycaemic unawareness and other medical conditions. Higher scores were associated with more severe hypo- Supported By: National Institutes of Health; University of Utah glycaemia (p=0.003) and more frequent hypoglycaemic episodes (p<0.0001). Those with the highest score had higher odds of having more than 2 hypos OR=6.16 (95% CI 2.39, 15.9) p<0.001. Exploratory analysis revealed that recurrent hypoglycaemia, , type 2 diabetes on insulin, dura- tion of diabetes of >10 years and hypoglycaemia unawareness were the key

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A102 COMPLICATIONS—HYPOGLYCEMIA drivers in the risk score. Using these 5 risk factors alone predicted severity rapidly increased plasma glucose (PG) by ≥20 mg/dL (9-14 min) and to PG of hypoglycaemia (p=0.0001) and having more than 2 hypoglycaemic epi- ≥70 mg/dL (within 6-10 min), similar to GLUCA (Table). All patients on DASI sodes (p=0.0001). reached these endpoints within 30 min (predefined success criteria). Both Conclusion: Our data strongly suggest that 5 simple questions on admis- treatments were well tolerated. was the most frequent adverse sion can reliably predict and differentiate patients at increased risk of hypo- event occurring at a similar rate of 44-56% patients. glycaemia. Appropriate changes to medications and enhanced monitoring of In conclusion, DASI was well tolerated and showed similar early PD at risk patients will improve morbidity and mortality outcomes. response to GLUCA at equivalent doses suggesting they have comparable clinical effects. may be an effective and reliable emergency Table. Cardiff Hypoglycaemia Risk Score (CHRiS). Score 1 for Each of POSTERS the Risk Factor if Present. treatment for severe hypoglycemia in a ready-to-use pen. Complications Table. Acute and Chronic Type 1 Diabetes — Dasiglucagon* GlucaGen® Type 2 Diabetes using insulin — Dose 0.1 mg 0.3 mg 0.6 mg 1.0 mg 0.5 mg 1.0 mg Duration of Diabetes >10 years — N 5 16 17 16 15 31 Recurrent Hypoglycaemia (Do you have frequent Hypoglycaemia’s) — Hypoglycaemia Unawareness (Can you sense your blood glucose is Time to reach plasma glucose levels ≥70 mg/dL [min] low or not) — Median (min/max) 10.0 (2.0-17.0) 6.0 (0-13.0) 6.0 (0-9.0) 6.0 (0-9.0) 6.0 (0-9.0) 7.0 (0-10.0) Total Score — Time to increase in plasma glucose levels ≥20 mg/dL [min] Score ≥3 Consider reducing insulin dose, closer monitoring and Median (min/max) 14.0 (11.0-27.0) 10.0 (7.0-20.0) 9.0 (6.0-16.0) 9.0 (7.0-15.0) 10.0 (6.0-13.0) 10.0 (5.0-15.0) Diabetes team Consult at earliest — *Proposed int. non-proprietary name.

388‑P & 390‑P Evaluation of a Scoring System to Determine Impaired Hypoglycaemia Awareness WITHDRAWN ANDREW M. SOLOMON, OLUWASEUN ANYIAM, PETER WINOCOUR, Stevenage, United Kingdom, Welwyn Garden City, United Kingdom Introduction: In those with type 1 diabetes, there is a need for precise evaluation of altered hypoglycaemic awareness to reduce the risk of poten- tially life threatening events. In 2015, new guidelines for type 1 diabetes in the UK mandated inclusion of a hypoglycaemia awareness scoring system into routine practice. Our clinical database already enabled reporting of unexpected and/or nocturnal episodes. In order to assess the impact of the guideline, a new mechanism was added into our diabetes database using a modification of the Clarke score. The setting was a specialist diabetes service, incorporating 50 accredited diabetes professionals covering a UK region with a population of 500,000. Methodology: First, we determined the qualitative level of hypoglycaemic awareness reported using the established methods. Second, we inserted the scoring system into the clinical database. Impaired awareness was defined as a score≥ 4 out of 5. Results: In the period preceding introduction of the new scoring system analysis of qualitative data demonstrated a 17% rate of patients reporting at least 1 episode of hypoglycaemia causing loss of consciousness without warning. In addition, at least 27% described nocturnal episodes and 29% the finding of glucose <2mmol/l. Over a 6 month period, the 5-point modi- fied Clarke score was completed in 223 patients. Of those just 5% scored ≥4 out of 5. Discussion: In our extensive outpatient diabetes service, the ability for diabetes professionals to detect and act upon reports of altered hypogly- caemic awareness was more comprehensive before the introduction of a modified Clarke score. The regimented need to ask 5 questions in sequence and/or the unfamiliarity of the professionals using it may have influenced the result showing much lower prevalence of relevant features than a self- reported qualitative record. Clinical trials carefully combining qualitative and quantitative scoring systems for hypoglycaemia awareness are urgently needed. & 391‑P & 389‑P Higher Glycemic Variability Is Associated with Increased Risk of Dasiglucagon, a Novel Soluble Analog, Successfully Hypoglycemia in Well or Poorly Controlled Type 1 or Type 2 Diabetes Restores Blood Glucose Levels after Hypoglycemia in People with Patients Type 1 Diabetes Mellitus (T1DM) LESZEK CZUPRYNIAK, ANNA BORKOWSKA, ELEKTRA SZYMANSKA-GARBACZ, TIM HEISE, BRITTA VÆVER BYSTED, ULRIK MOURITZEN, ULRIKE HÖVELMANN, Warsaw, Poland, Lodz, Poland DANIELA LAMERS, DANIÉL VEGA MØLLER, Neuss, Germany, Glostrup, Denmark Avoiding hypoglycemia is an important treatment goal in diabetes. We Currently available glucagon formulations for rescue treatment of severe assessed the relationship between blood glucose (BG) variability in well or hypoglycemia need to be reconstituted before use which delays its use and poorly controlled type 1 (DM1) or type 2 diabetes (DM2). The patients with may lead to underutilization. In this single center, randomized, double-blind HbA1c ~7% (i.e., 6-8%) and ~10% (i.e., 9-12%) were enrolled into the study trial we investigated the pharmacodynamic (PD) and pharmacokinetic (PK) and formed 4 study groups: DM1 7% (n=29); DM1 10% (n=30); DM2 7% characteristics of dasiglucagon (DASI), a novel stable soluble glucagon ana- ® (n=28); and DM2 10% (n=35). Their mean age, duration of diabetes, BMI and log, in comparison to GlucaGen (GLUCA). Fifty-eight patients with T1DM HbA1c were 42±13, 41±14, 62±11, 64±8 years; 14±11, 13±7, 11±6, 16±8 years; received subcutaneous injections of 0.1, 0.3, 0.6 or 1 mg DASI or 0.5 or 1 mg 23.1±3.2, 26.1±4.0, 30.7±5.3, 33.0±5.1 kg/m2; 7.1±0.6, 10.1±0.9, 7.2±0.6, GLUCA after induction of hypoglycemia (55 mg/dL) through intravenous insu- 9.6±0.8%, respectively. Each patient underwent blinded continuous glucose lin infusion. DASI demonstrated a dose-dependent and rapid increase in PK monitoring (CGMS; iPro2, Medtronic) for 5.8±0.8 days. In each subject dura- reaching maximum levels at ~35 min with a half-life of ~0.5 hours. DASI

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tion of the time spent at BG <70 mg/dl (hypoglycemia threshold) was deter- 393‑P mined and expressed as a percentage of total CGMS use time (T hypo). BG High One-Year Mortality Rate following Admissions with Severe variability was assessed with coefficient of variation (CV) calculated as the Hypoglycemia to the Hospital—Experience from Tertiary Care ratio of the standard deviation to the mean of daily glucose measurements Acute Hospital in Asia obtained from CGMS. There was a statistically significant and highly posi- WEIYING LIM, EDMUND TICK CHIA CHAN, ANDREW XIA HUANG TAN, ZON- tive correlation between glucose variability and duration of hypoglycemia GWEN WEE, LI CHANG ANG, XIAOHUI XIN, YONG MONG BEE, SU YEN GOH, in all studied groups (Figure 1). The results of the study may suggest that MING MING TEH, Singapore, Singapore decreasing BG variability may be an effective way to reduce the risk of hypo-

POSTERS Aim: Self-reporting of severe hypoglycemia is associated with increased

Complications glycemia in diabetes patients with various degree of metabolic control. mortality. However, less is known regarding the short term mortality follow- Acute and Chronic ing admission with severe hypoglycemia to the acute care hospital. The aim of the study is to investigate one year mortality following admis- sion with severe hypoglycemia to Singapore General Hospital (SGH), which is the largest tertiary acute care hospital in Singapore. Method: Clinical, biochemical and 1-year mortality data from diabetes patients who were admitted with severe hypoglycemia to SGH in the year of 2014 admission were extracted from institutional medical record. Com- parison was made between the patients who survived, with those who are deceased within a year of admission. Bivariate analysis was done to explore factors which influence the 1-year mortality rate. Results: 312 eligible patients (185F: 127M) were identified and the mean capillary blood glucose on admission was 2.3+0.7 mmol/l (41.1+ 12.6mg/dl). Mean age, (HbA1C) and Charlson’s comorbidity index (CCI) of the cohort are 70.5 +11.3 year old, 6.9+1.4% and 4.5+2.1 respectively. 22% of the cohort died within a year of admission. Those deceased are older (69.2 vs. 73.3 year old, p<0.05) with higher Charlson’s comorbidity index (CCI) (4.1 vs. 6.1, p<0.05) and longer length of stay (5.0 vs. 10.0 days, p<0.05). However, there was no significant difference in glycemic control between the 2 groups (HbA1C 7.0 vs. 6.7%, p>0.05). Age is associated with increased Supported By: Medical University of Lodz 1-year mortality risk ratio 1.06 (95% confidence interval (CI):1.03-1.09) with p <0.001. CCI of at least 6 is associated with minimum 1-year mortality risk & 392‑P ratio of 9.46 (95% CI: 1.96-45.62) with P = 0.01. Is Frailty a Risk Factor for Severe Hypoglycemia in Older Adults? Conclusions: There is high 1-year mortality rate of 20% following admis- The Risk in Communities (ARIC) Study sions with severe hypoglycemia. Therefore, admissions with severe hypo- ALEXANDRA K. LEE, A. RICHEY SHARRETT, BEVERLY G. WINDHAM, ELBERT S. glycemia may have important prognostic implications. HUANG, KAREN BANDEEN-ROCHE, ELIZABETH SELVIN, Baltimore, MD, Jackson, MS, Chicago, IL 394‑P Many common risk factors for hypoglycemia, such as , malnutri- Day-to-Day Variability of Self-Measured Plasma Glucose tion, and disability, overlap substantially with frailty, which reflects diminished (SMPG) Correlates with Risk of Hypoglycemia in Adults with Type 1 physiologic reserve. We therefor hypothesized that frailty would be associ- (T1D) or Type 2 Diabetes (T2D) ated with hypoglycemia in a cross-sectional study of 1826 ARIC participants TIMOTHY S. BAILEY, ANUJ BHARGAVA, J. HANS DE VRIES, GREGG GERETY, with diabetes at Visit 5 (2011-13, mean age 75 years, 45% male, 29% black). JANUSZ GUMPRECHT, WENDY S. LANE, CAROL H. WYSHAM, BRITTA ANKER Prior episodes of severe hypoglycemia requiring treatment by hospitalization, BAK, ELISE HACHMANN-NIELSEN, ATHENA PHILIS-TSIMIKAS, Escondido, CA, emergency department, or ambulance were identified from CMS claims from Des Moines, IA, Amsterdam, Netherlands, Albany, NY, Zabrze, Poland, Asheville, NC, 1991-2013 using ICD-9 codes. Fifty participants (3%) had a history of severe Spokane, WA, Søborg, Denmark, Frederiksberg, Denmark, San Diego, CA hypoglycemia at Visit 5. Frailty was defined as >3 of the following components: The relationship between hypoglycemia and day-to-day variability of weight loss, low physical activity, slow walking speed, exhaustion, or weak grip glycemic control has not been well established. A post-hoc analysis was strength. Pre-frailty was defined as 1 or 2 components. The prevalence of prior performed correlating day-to-day variability of fasting SMPG with hypogly- hypoglycemia in frail, pre-frail, and robust persons was 6.0%, 3.2%, and 1.2%, cemia in two double-blind, treat-to-target, crossover trials that compared respectively (Figure). In an adjusted Poisson model, the prevalence of prior once daily (OD) with U100 OD in adults with hypoglycemia was over three times greater in frail compared to robust persons. T1D (SWITCH 1, n=501) or insulin-experienced adults with T2D (SWITCH 2, When examining each frailty component individually, walking speed was most n=721). Available SMPG measurements were used to determine a weekly strongly associated with a history of severe hypoglycemia. To understand the variance for each patient, using the log SMPG values to allow for relative directionality of these findings, prospective studies are needed to determine comparisons. For each patient and treatment, the geometric mean of the if baseline walking speed predicts future risk of hypoglycemia in older adults. weekly variance was calculated and these values were categorized into low, medium and high tertiles as a measure for day-to-day variability. The effect of having low or high variability compared with medium variability was ana- lyzed in relation to overall symptomatic (severe or blood glucose [<56 mg/ dL] confirmed), nocturnal symptomatic (00:01-05:59), and severe (requiring third-party assistance and confirmed by a blinded adjudication committee) hypoglycemia. Day-to-day SMPG variability was a significant predictor for the risk of overall and nocturnal hypoglycemia in T1D and T2D, and severe hypoglycemia in T1D (Table). In conclusion, day-to-day glycemic variability relates to hypoglycemia risk.

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Table. Effect of Fasting SMPG Variability on Hypoglycemia in SWITCH 1 and Figure. 2: Low and High Tertiles Compared with Medium Tertile. POSTERS Complications Acute and Chronic

Supported By: Novo Nordisk

395‑P Effects of Chronic on Neurocognitive Function in Supported By: National Institutes of Health Type 1 Diabetes Mellitus: Evidence from Neuroimaging KAITLIN G. BRAU, TIMOTHY J. MCDERMOTT, ALEX I. WIESMAN, CYRUS V. 397‑P DESOUZA, ANDJELA T. DRINCIC, TONY W. WILSON, Omaha, NE The Relationship between Hypoglycemia Severity and Health- Chronic hyperglycemia in type 1 diabetes mellitus (T1DM) is known to Related Quality of Life among U.S. Patients with Type 2 Diabetes play a role in diabetes-related cognitive dysfunction. In this study, we used MANJIRI PAWASKAR, KRISTY IGLAY, EDWARD A. WITT, SWAPNIL RAJPATHAK, an advanced noninvasive functional brain imaging tool known as magne- SAMUEL ENGEL, L. KEOKI WILLIAMS, Kenilworth, NJ, New York, NY, Detroit, MI toencephalography (MEG) to identify the neuronal alterations associated Although studies have shown that hypoglycemia impacts morbidity and with such cognitive dysfunction and to determine the impact of glycemic mortality in type 2 diabetes (T2D), most of these studies have focused on dysregulation as measured by A1C. Thirty-three patients with T1DM with- severe hypoglycemic events (SE). This study examined the relationship out complications completed two tasks during MEG. Task one was a visual between hypoglycemia severity and health related quality of life (HRQoL) in attention task where participants viewed a screen showing five congruent adults with T2D. The study sample included 2013 U.S. National Health and or incongruent arrows and responded with a button press depending on Wellness Survey respondents with diagnosis of T2D and treated with an the orientation of the middle arrow. In task two, electrical stimulation was antihyperglycemic agent. Participant recall of a single hypoglycemia event applied to the right median nerve of participants to stimulate the somato- in the preceding 3 months was used to categorize each individual into one sensory system. All MEG data was processed following standard protocols, of 3 groups: no event (NE), non-severe event (NSE), or SE. HRQoL was mea- and the resulting images of functional brain activity were examined statis- sured using validated instrument, the Medical Outcomes Study Short-Form tically. Reaction time and accuracy were also collected for all behavioral (SF-36). Generalized linear regression models were used to derive outcome tests. Data on A1C was also collected and a median split of this parameter means adjusted for potential confounders. Of the 3,630 participants who was performed, allowing comparison of the resulting two groups (midpoint met study criteria, 52.3% reported at least one hypoglycemic event and 4.7% excluded). In the visual attention task, patients with higher A1C had reduced reported a SE. Hypoglycemia severity was inversely associated with HRQoL activity in the prefrontal cortex relative to those with lower A1C (p<0.05). (measured as health utility scores), including both the mental and physical Reaction time during this task was also significantly correlated with A1C val- components (Table). Similar trends were observed for all subdomains of the ues, and this indicated slower responses in those with higher A1C (p<0.05). SF-36, even after accounting for potential confounders. This study suggests In the somatosensory stimulation task, patients with higher A1C had sig- that increasing severity of hypoglycemia is associated with a decrease in nificantly weaker responses in somatosensory cortices relative to those HRQoL, and includes reductions in both physical and mental health. with lower A1C (p< 0.05). These results support the hypothesis that we can Table. Estimated Means for Health Related Quality of Life (SF-36) Utility identify abnormal cognitive and neural function in patients with T1D who are Scores, Summary Scores, and Subdomain Scores by the Level of Hypoglyce- virtually free of current health complications. Poor glycemic control acceler- mia Severity in Type 2 Diabetes Patients. ates cognitive decline, and chronic hyperglycemia, represented by elevated A1C, leads to aberrant function in visual and somatosensory systems.

396‑P Does a History of Hospitalization for Severe Hypoglycemia (HSH) Predict All-Cause Mortality in Long-Duration Type 1 Diabetes (T1D)? GEORGIA PAMBIANCO, TREVOR J. ORCHARD, Pittsburgh, PA A past history of severe hypoglycemia (HSH) has been shown to be associ- ated with increased mortality in type 2 diabetes and in T1D where HSH with coma or seizure independently predicted mortality (DCCT/EDIC). Data from the Pittsburgh Complications (EDC) study of childhood onset T1D (mean age 28 and duration 19 years, n=658) were thus examined to determine if HSH hospitalization within the 5 years prior to baseline (1986- 1988, n=658) is associated with total mortality after 22 years of follow-up. 398‑P Participant hospitalizations were categorized as: 1) no hospitalization for SH Clinical and Psychological Characteristics of People with Type 1 (n=594) 2) one SH hospitalization (n=52) 3) two or more SH hospitalizations Diabetes and a High Risk of Hypoglycemic Events (n=12). was confirmed by death certificate and classified according to LUTZ HEINEMANN, GABRIELE FABER HEINEMANN, NORBERT HERMANNS, Diabetes Epidemiology Research International (DERI protocol) by a physician DOMINIC EHRMANN, PETER WINTERGERST, GUIDO FRECKMANN, Düsseldorf, committee. In a Cox proportional hazards model, adjusting for baseline diabe- Germany, Bad Mergentheim, Germany, Ulm, Germany tes duration (p=.0001), HbA1c (p=.0001), (p=.0001), estimated glomer- For the HypoDE study, people with type 1 diabetes treated by MDI with ular filtration rate (eGFR, p=.006), and albumin excretion rate (AER, p=.0001), a high risk of hypoglycemic events were recruited. Baseline data of 126 the hazard rate for 2 or more SH hospitalizations was 3.49 (1.3-9.6). Gender, study participants were analyzed (age 46.5±11.6 yrs, 36.5% female, HbA1c and non-hospitalized SH were available but not predictive. A his- 7.5±1.0%). They reported 4.5±9.2 episodes of severe hypoglycemia per year tory of multiple SH hospitalizations was independently related to all-cause (third party assistance for recovery required) and 1.0±2.4 episodes of hypo- mortality and those affected merit special attention. glycemic episodes with coma or seizure per year prior to study participation.

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COMPLICATIONS—HYPOGLYCEMIA

This corresponds to 700%, respectively 600% more hypoglycemic episodes atic delta cells. Because neuronostatin enhances mRNA expres- than observed in the DCCT (0.64, respectively 0.16 episodes per year). sion and glucagon release from isolated islets and immortalized alpha cells, Blinded CGM recordings for 28 days during the run-in phase of the study we hypothesize that neuronostatin is an important component of the counter- revealed that the participants had 12.7±11.8 hypoglycemic events per 28 regulatory response. In support of this hypothesis, we have found that plasma days (= glucose readings ≤55 mg/dl for at least 20 min). They spent 109 min levels of neuronostatin increase following a hypoglycemic challenge, suggest- per day at glucose levels ≤70 mg/dl and 34 min per day ≤50 mg/dl. This cor- ing that hypoglycemia is the physiologic stimulus for neuronostatin release. responds to 32.5%, respectively 55% more time in this range that the adult Furthermore, while neuronostatin enhances glucagon release from the alpha participants in the JDRF CGM-trial. The hypoglycemia unawareness score of cell, the appears to inhibit the release of insulin from isolated islets POSTERS

Complications the participants was 5.0±1.1 (out of a maximum score of 7); 95.2% yielded a and in vivo. In addition, infusion of exogenous neuronostatin increases basal Acute and Chronic score ≥4, which is used as a cut-off score for hypoglycemia unawareness. plasma glucose levels in rats. Experiments are underway to confirm the role In the Hypoglycemia Fear Survey, HypoDE participants achieved a score of of neuronostatin in the counterregulatory response and to evaluate the role of 32.3±15.5 and a Diabetes Distress Scale mean item score of 2.5±1.2. Both neuronostatin in intra-islet communication during hypoglycemia. Because of scores were higher than those which could be expected in a sample of people neuronostatin’s effect on glucagon release, we propose that the peptide could with type 1 diabetes, not specifically selected by hypoglycemia problems. serve as a novel therapeutic target for the treatment of iatrogenic hypoglyce- In summary, these data suggest that HypoDE participants represent mia and hypoglycemia unawareness. patients with a high risk of clinical as well as biochemical hypoglycemic Supported By: National Institutes of Health events. Compared to the “typical” patient with type 1 diabetes, these sub- jects reported a high amount of hypoglycemia worries and diabetes-related 401‑P distress. Progress of Diabetes Severity Associated with Higher Risk of Supported By: Dexcom, Inc. Severe Hypoglycemia EDY KORNELIUS, YI SUN YANG, CHIUNG HUEI PENG, SHIH CHANG LO, YUNG 399‑P RUNG LAI, JENG YUAN CHIOU, CHIEN NING HUANG, Taichung, Taiwan Hypoglycemia Management through the Eyes of the Significant Background: The association between the progression of diabetes sever- Other: Highlights from the InHypo-DM Study (Canada) ity and risk of severe hypoglycemia is unknown. We aimed to evaluate the SONJA M. REICHERT, ALEXANDRIA RATZKI-LEEWING, BRIDGET L. RYAN, association between the progression of diabetes severity and severe hypo- SELAM MEQUANINT, SUSAN WEBSTER-BOGAERT, JUDITH B. BROWN, STEW- glycemia in diabetic patients. ART B. HARRIS, London, ON, Canada Methods: We conducted an 11-year population-based retrospective cohort While the risk of hypoglycemia can detrimentally affect people with both study of new diabetic patients. Diabetic progression was evaluated by the type 1 and type 2 diabetes (DM), it can also impose an immense burden on adapted Diabetes and Severity Index (aDCSI) from index date the lives of their significant others (SO). It is important to understand how to end of follow-up. We classified the progression of diabetes severity into 3 hypoglycemia impacts the lives of SO when they are required to help man- categories; slow, moderate, and rapid. We further compared those 3 catego- age hypoglycemia in people with DM. To date, this important population has ries and evaluate the risk of first hospitalization due to severe hypoglycemia. been largely understudied. Results: A total of 353,767 new-onset diabetic patients were recruited. To address this research gap, our team developed and administered a Mean age of diabetic patients in this study was 56.8 years old, and mean theory-driven, 63-item questionnaire (InHypo-DMSOQ). It was disseminated follow-up duration was 9.3 years. Mean initial aDCSI was 0.7, while mean online, in French and English, to a nationwide panel managed by Ipsos Inter- aDCSI at the event date or end date was 2.9. A rapid progression of aDCSI active Services Ltd., Canada’s largest research polling firm. was associated with higher risk of severe hypoglycemia compared with A total of 552 SOs of people with DM (≥17 years old) completed the slower progression of aDCSI. Compared with slow progression of diabetes questionnaire (female: 56%, mean age: 48 (SD 16.3)). The majority of SOs severity, the hazard ratios of patients with moderate and rapid progression identified as either a spouse (24%) or a parent (22%) of a person with DM. increased the risk of severe hypoglycemia in proportion to diabetic progres- Just over half (52%) of SOs reported caring for a person with type 2 DM; sion, with HRs of 1.10 (95% CI 1.04-1.16) and 5.23 (95% CI 5.00-5.47), respec- 16% did not know the type of DM. Overall, 87% of SOs of persons with tively. The results were unchanged after adjustment for age, gender, disease DM responded that they would forgo other things in their lives to help man- comorbidities, and prescriptions for antidiabetic drugs. age hypoglycemia, yet over half (56%) reported struggling to make such Conclusions: This study demonstrated that rapid progression of diabetes life changes. Sixty-two percent of SOs were involved in making decisions complications was associated with higher risk of severe hypoglycemia. It is regarding hypoglycemia management; yet, 71% of respondents expressed imperative that the treating physician identify patients with acute worsening feelings of inadequacy around helping to prevent or treat an event. Ninety- of diabetic severity and provide proper hypoglycemia education and prevention. two percent of all respondents felt committed to helping manage hypogly- cemia, and for the vast majority (86%), doing so, was seen as an unavoidable 402‑P part of life. Predicting Severe Hypoglycaemia with Self-Monitoring of Blood The InHypo-DMSOQ, the largest survey of its kind to date, underscores Glucose in Type 1 Diabetes the significant involvement of SOs of people with DM in the management LOUIS POTIER, ABDUL MOUTAIROU, BERNARD CHARBONNEL, KAMEL MOHAM- of hypoglycemia. Our study revealed a strong sense of altruism and com- MEDI, ABDOULEYE LEYE, BRUNO DETOURNAY, RONAN ROUSSEL, Paris, France, mitment amongst SOs to help individuals with DM manage their hypoglyce- Nantes, France, Dakar, Senegal, Bourg-la-Reine, France mia. Healthcare professionals should focus on partnering with SOs in their Severe hypoglycemia (SH) is the most important adverse effect of insu- efforts to optimize glycemic care, equipping them with the necessary edu- lin therapy in patients with type 1 diabetes. To avoid SH, identification of cational support and appropriate resources. risk factors is essential. Our aim was to study the association between mild Supported By: Sanofi Canada hypoglycemia detected by self-monitoring blood glucose (SMBG) and SH in the 1441 participants of the Diabetes Control and Complications Trial. One 400‑P SMBG profile (7 measurements/24 h) was collected at each visit. We calcu- Neuronostatin Enhances Glucagon Secretion: A Potential Target for lated a score (hypo-score) by dividing the number of SMBG values <70 mg/dl Hypoglycemia Unawareness by the total number of SMBG measurements. Associations between hypo- STEPHEN M. GROTE, WILLIS K. SAMSON, GINA L.C. YOSTEN, St. Louis, MO score and SH were assessed in unadjusted and adjusted logistic regression Patients with type 1 diabetes whose blood glucose levels are tightly regu- models. Mean number of SMBGs during the follow-up was 248±16. Mean lated with exogenous insulin are at significant risk of iatrogenic hypoglyce- hypo-score was 9.3±7.0. The hypo-score was higher in the intensive arm mia and its serious consequences, including coma and death. Under normal compared with the standard one (13.3±6.5 vs. 5.5±4.9, p<0.001). The inci- conditions, the counterregulatory response to low glucose includes endocrine dence of SH by quartile of hypo-score was 17±7.9% (Q1), 38±5.1% (Q2), (glucagon and epinephrine release) and behavioral (increased food-seeking 58±5.2% (Q3), and 71±4.8% (Q4) (p<0.001). Risk of SH was linearly associ- behavior) mechanisms to increase plasma glucose levels. However, repeated ated with hypo-score either in unadjusted or adjusted model (Figure). Similar exposure to hypoglycemia blunts these normal responses, causing “hypogly- analyses were conducted with the SMBGs collected only during the first cemia unawareness,” which can lead to an inability to recognize the signs of year of follow-up, and SH episodes which occurred after the first year. Con- hypoglycemia during future hypoglycemic events. We recently identified a sistently, the first-year hypo-score was associated with the risk of future novel peptide, neuronostatin, which is encoded by the prepro- SH. These results underline that patients with type 1 diabetes and frequent hormone and is produced in somatostatin-expressing cells, including pancre- unremarkable hypoglycemia have a higher risk of SH.

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Figure. analysis showed that an increase in within-day variability has a significant correlation with an increasing risk of overall and nocturnal hypoglycemia (Table). However, no correlation was found for severe hypoglycemia in this dataset. In conclusion, within-day glycemic variability is associated with a risk of overall and nocturnal hypoglycemia. Table. Effect of Within-Day Variability (9-Point Profile) on Hypoglycemia in

SWITCH 1 and 2: Low and High Tertiles Compared with Medium Tertile. POSTERS Complications Acute and Chronic

403‑P Clinical Correlates of Predictors of Hypoglycemia over Four Years in 2,729 People with Type 2 Diabetes Starting Insulin PHILIP D. HOME, LAWRENCE BLONDE, FRANCOISE CALVI-GRIES, VALERIE PILOR- GET, JOSEPH BERLINGIERI, MARTIN PFOHL, NICK FREEMANTLE, Newcastle upon Tyne, United Kingdom, New Orleans, LA, Reze, France, Paris, France, Hamilton, ON, Canada, Duisburg, Germany, London, United Kingdom We aimed to identify factors associated with documented symptomatic hypoglycemia (DH), and severe hypoglycemia (SH), in people starting insu- Supported By: Novo Nordisk A/S lin therapy in the CREDIT study, an international prospective observational study. Data was collected from routine care semi-annually over 4 years; 2,729 and 2,271 people had data from the last 6 months of years 1 and 4. 405‑P Accurate Assessment of Hypoglycemia Risk with U500 Insulin Multivariable logistic regression selected factors associated with DH - the in Markedly Insulin Resistant Diabetes (MIRD) from Direct Meter models were then applied to SH. Participants with 1 DH event (excluding ≥ Downloads via Home Telehealth (HT) SH) were 18.5% and 16.6% in years 1 and 4, 24.6% and 18.3% if achieving DINESH EDEM, MICHAEL GRIMES, RONALD CODARIO, R. HARSHA RAO, HbA1c <7.0% (SH 3.0% and 4.6%), and 16.5% and 16.2% if not (SH 1.5% Pittsburgh, PA and 1.1%). On multivariable analysis, baseline variables associated with Introduction: Five-fold concentrated (U500) insulin is a viable therapeutic DH at year 1 were lower , more physical activity, and no option in MIRD (Total daily dose of insulin 200 units/d), with well-docu- oral agents vs. , with lower HbA1c, higher total insulin dose, ≥ mented glycemic efficacy but uncertain hypoglycemia risk. Studies report and sulfonylurea use at year end. In year 4, baseline variables were lower conflicting data, with both higher and unchanged hypoglycemia risk, but the body mass index, higher diastolic (BP), lower HbA1c, mac- methods used to assess hypoglycemia, such as diagnosis codes, question- rovascular disease, and more physical activity, and explanatory variables naires, and blood glucose [BG] logs, are unreliable and known to under-report higher systolic BP, lower HbA1c, and higher insulin dose. Insulin regimen was hypoglycemia. a univariate associate, weaker at year 4 than year 1, and not retained on Objective: To assess the true risk of hypoglycemia in MIRD patients taking multivariate analysis. The models developed for DH predicted SH. Thus the U500 insulin. results confirm that thinner, active people, taking oral agents and having Methods: Retrospective chart reviews of MIRD patients being treated better glucose control with higher insulin doses have more DH in the years with either U500 (n=221) or U100 insulin (n=199), to determine the fraction after starting insulin. Predictor and explanatory variables developed for DH of BG values <70 mg/dl and number of hypoglycemic episodes over a year, can be extended to SH. The suggestion from intensive control studies that using BG data derived from meter downloads via HT or self-documented (SD) DH/SH is more of a problem at higher HbA1c is not supported in routine patient BG logs. care. Hypoglycaemia (DH and SH) occur at significant but not high rates after Results: The U500 cohort was more likely to check BG (199/221 [90%] starting insulin therapy, but with active management rates are stable over 4 vs. U-100 113/199 [57%], p<0.001), use HT (99/221 [45%] vs. 47/199 [24%], years despite increased insulin dosage. The association with insulin regimen p<0.001), and maintain lower A1c (8.1±0.1% vs. 8.7±0.1%, p<0.01). U-500 and with oral agent use declines from year 1 to year 4 with active clinical patients also documented more BG values <70mg/dl (1362/50013 [3%] vs. management. 456/23714 [2%], p<0.001), and more experienced hypoglycemic episodes Supported By: Sanofi (143/199 [72%] vs. 62/113 [55%], p<0.01). However, the increase was limited to mild (BG 60-69 mg/dl; 128/199 [64%] vs. 52/113 [46%], p<0.01) or moder- 404‑P ate hypoglycemia (BG 40-59 mg/dl; 107/199 [54%] vs. 45/113 [40%], p<0.05), Within-Day Variability Based on 9-Point Profiles Correlates with not severe hypoglycemia (BG <40mg/dl; 20/199 [20%], vs. 7/113 [6%], p=0.3), Risk of Overall and Nocturnal Hypoglycemia in Adults with Type 1 and only evident by HT (948/31021 [3%] vs. 205/12518 [2%], p<0.001), not SD (T1D) and Type 2 Diabetes (T2D) (414/18992 [2%] vs. 251/11196 [2%], p=0.7). TIMOTHY S. BAILEY, ANUJ BHARGAVA, J. HANS DEVRIES, GREGG GERETY, Conclusions: U500 insulin therapy improves glycemic control, but JANUSZ GUMPRECHT, WENDY S. LANE, CAROL H. WYSHAM, BRITTA ANKER increases the risk of mild and moderate, but not severe hypoglycemia. How- BAK, CHARLOTTE THIM HANSEN, ATHENA PHILIS-TSIMIKAS, Escondido, CA, Des ever, higher hypoglycemic risk is seen only in verified BG data from direct Moines, IA, Amsterdam, Netherlands, Albany, NY, Katowice, Poland, Asheville, NC, meter downloads by HT, not in self-documented BG logs, indicating a signifi- Spokane, WA, Søborg, Denmark, San Diego, CA cant recall bias in reporting hypoglycemia. Higher glycemic variability has previously been linked to an increased risk of hypoglycemia. The correlation between within-day variability, based on 9-point profiles, and hypoglycemia was investigated in two double-blind, 406‑P Hypoglycemia Rates in Type 2 Diabetes Mellitus: Results of the treat-to-target, crossover trials comparing insulin degludec once daily (OD) Population-Based InHypo-DM Study (Canada) with insulin glargine U100 OD in adults with T1D (SWITCH 1, n=501) or insu- ALEXANDRIA RATZKI-LEEWING, STEWART B. HARRIS, BRIDGET L. RYAN, lin-experienced adults with T2D (SWITCH 2, n=721). Within-day glycemic SONJA M. REICHERT, SELAM MEQUANINT, SUSAN WEBSTER-BOGAERT, variability was calculated as the relative fluctuation of the 9-point profile, JUDITH B. BROWN, London, ON, Canada defined through the integrated absolute distance from the mean within-day A thorough understanding of -related hypoglycemia incidence variability. Variabilities were subsequently categorized into low, medium is important to the therapeutic optimization of type 2 diabetes (T2DM). This and high tertiles based on the geometric mean. Hypoglycemia was defined study aimed to substantiate the population-based effect of insulin and/or as overall symptomatic (severe or blood glucose [<56 mg/dL] confirmed), sulphonylureas (SUs) on the rate of hypoglycemia. nocturnal symptomatic (00:01-05:59) and severe (requiring third-party All data were obtained from the InHypo-DM cross-sectional cohort study, assistance and confirmed by a blinded adjudication committee) events. This Canada’s largest hypoglycemia-related survey to date. Negative binomial

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regression (NBR) was used to model the effect of insulin, SUs, or insulin plus 408‑P SUs on the retrospective incidence of non-severe daytime (DH), non-severe Patients’ Knowledge of Hypoglycaemia Management in Outpatient nocturnal (NH), and severe hypoglycemia (SH) events. A directed acyclic Settings graph was constructed to ascertain the minimally sufficient adjustment set. ATENA GOGOKHIA, PREETHI NALLA, LAXMI P. PATEL, LAKDASA D. PREMAWAR­ Results of the NBR were reported as incidence rate ratios. DHANA, MOHAMED A. ADLAN, Ystrad Mynach, United Kingdom A total of 458 people with T2DM (male: 53%, mean age: 51.5 (SD: 15.3) Introduction: Hypoglycaemia (Hypo) is an acute complication in patients years) completed the InHypo-DM survey; 47% were on SUs alone, 35% were with diabetes especially when treated with insulin and . It on insulin alone, and 18% were on insulin and SUs. Annualized crude inci- increases morbidity and mortality, hospital admissions and is an expensive POSTERS

Complications dence rates of hypoglycemia were highest among individuals on combina- complication to manage. We surveyed patients with diabetes attending sec-

Acute and Chronic tion therapy as compared to insulin or SUs alone, especially for SH (3.66 ondary care clinics to assess patients’ knowledge of Hypos and its manage- events/person-year). Based on the NBR analysis, adjusting for all covariates, ment. individuals on combination therapy vs. SUs alone experienced 2.22 (95% CI: Aims: Evaluate patients’ knowledge about Hypo and its management. 1.44-3.45) and 2.10 (95% CI: 1.04-4.27) times the rate of DH and SH, respec- Methods: A questionnaire based on the British Diabetes Societ- tively. Moreover, people on insulin vs. SUs alone experienced a 1.56 (95% ies guidelines on Hypo identification and management was distributed to CI: 1.05-2.33) factor increase in the expected rate of DH and were also 3.27 patients attending secondary care diabetes clinics. A total of 200 patients (95% CI: 1.67-6.42) times as likely to develop SH. Similar trends, though sta- completed the questionnaire. tistically non-significant, were observed for NH. Results: The majority of patients were over 40 years old (89%), had type 2 This large Canadian-based study aimed to assess the causal link between diabetes (68%) and were insulin-treated (73%). Blood sugars were checked antidiabetic therapy and hypoglycemia frequency in T2DM. Reported event at least twice a day by 47% of patients, where as 10% never did (the median rates were remarkably high, particularly among individuals taking insulin in frequency of monitoring blood sugars was 2-4 times a day). A significant combination with SUs. Prescribers should remain vigilant to differences in proportion of patients had never experienced Hypos (39%), but around 10% drug-induced hypoglycemia across classes of treatment agents. of patients had more than 10 Hypos over the past 12 months (the median Supported By: Sanofi Canada number of Hypos was 1-2). The majority of patients (70%) had self-managed Hypos and only 11% required help from ambulance or hospital staff. The 407‑P patients’ awareness of Hypos was poor as 44% stated that they never or Improving Detection of Hypoglycemia with Standardized Blood rarely were aware of having Hypos. However, only 44% of patients had Glucose Checks after Administration of Insulin for Hyperkalemia sufficient knowledge of Hypo symptoms. A lack of knowledge of Hypo risk Treatment factors was also reported in 43% of the patients. Interestingly, 73% of the HEIDEMARIE W. MACMASTER, BRADLEY MONASH, ALLEN TRAN, ROBERT J. patients felt confident about self-managing Hypos. 63% stated that they had RUSHAKOFF, San Francisco, CA received education about Hypos and the diabetic specialist nurse was their main educator (30% of cases). The majority (44%) opted for printed informa- Acute Hyperkalemia (K ≥ 5.1 mEq/L) is often treated with a bolus of IV insulin. Nationally, this treatment results in a 20% rate of hypoglycemia and tion leaflets as their preferred method of education. a 7% rate of severe hypoglycemia. At UCSF, a 12-month baseline review Conclusions: This survey identified significant deficiencies in knowledge prior to a December 2015 hyperkalemia orderset revision found a 12% preva- about the symptoms and risk factors for hypoglycaemia and its management lence of hypoglycemia amongst those treated with insulin for hyperkalemia in patients attending secondary care diabetes clinics locally. but > 40% of patients had no POCT within 6 hours post insulin administra- tion. Utilizing a new Hyperkalemia order set that requires blood glucose 409‑P POCT pre-insulin administration and post-insulin administration (1, 2, 4 and New Tools for the Evaluation of Adherence in Type 1 Diabetes: Use‑ 6 hours post insulin, respectively) we sought to determine the rate for post- fulness and Relationship with Glycemic Control treatment hypoglycemia and also Identify risk factors which predispose REBECA REYES-GARCIA, PEDRO MEZQUITA-RAYA, OLGA GONZALEZ-FEIJOO, adult inpatients receiving insulin for hyperkalemia to hypoglycemia. Of 100 FUENSANTA MARTINEZ-MINGOT, INES TALAVERA-LOPEZ, ALEJANDRA DE patients treated with insulin for hyperkalemia, 23% had glucose <70 mg/dl TORRES-SANCHEZ, PEDRO ROZAS-MORENO, Almeria, Spain, Ciudad Real, Spain and 5% <40 mg/dl. 9%, 61% and 22% of these occurred 1,2 and 3 hours, Aims: Hypoglycemia is related to poor adherence to treatment in type 1 respectively, post insulin. The risk factors for hypoglycemia are in Table. diabetes mellitus (T1DM). The use of simple tools for the evaluation of hypo- Table. Risk Factors for Hypoglycemia. glycemia and adherence may improve patient care. Our objectives were: to evaluate the frequency of hypoglycemia in T1DM patients attended at endo- Identified Risk Factors: crinology, and to evaluate the usefulness of different scales of adherence. • Pre-Insulin Blood Glucose <140mg/dL Also we tested the utility of a new measure of adherence to treatment, Scale • No Diabetes Diagnosis Treatments • No Concomitant Albuterol Treatments with Post-Insulin of Evaluation of Adherence to Treatment in T1DM (T1DM-SEAT). This scale • No Concomitant without Post-Insulin Hypoglycemia has 5 simple questions, which indicates poor adherence with higher scores. • Administration of >0.11 units/kg Insulin Hypoglycemia (<70 mg/dL) Patients and Methods: Cross-sectional study including 262 T1DM patients • Serum > 1.22 mg/dL n = 77 n = 23 attended in . For the evaluation of adherence we used 3 scales: No Disposing Factors 0 0 Measurement of Adherence to Treatment (MAT) insulin, Morisky Green test, and T1DM-SEAT. We also recorded the frequency of auto-reported hypogly- 1 Disposing Factor 0 0 cemia and Clarke test. 2 Disposing Factors 2 (3%) 0 Results: The auto-reported frequency of hypoglycemia was: 1-6 episodes/ 3 Disposing Factors 24 (31%) 1 (4%) month in 44%, 2-3 every week in 26%, 4-5 episodes/week in 13.5%, one or 4 Disposing Factors 27 (35%) 9 (39%) more hypoglycemia every day in 9.5% of subjects. 28% of patients had hypo- glycemia unawareness. The scales of adherence showed a good correlation 5 Disposing Factors 16 (21%) 8 (35%) between them. There was no relationship between Morisky-Green test, the 6 Disposing Factors 8 (10%) 5 (22%) frequency of hypoglycemia or HbA1c. MAT scale was not related to the fre- As most hypoglycemic events occur within 3 hours post-insulin admin- quency of hypoglycemia, but it was related to HbA1c (Rho 0.179, p = 0.004). istration in the treatment of hyperkalemia, implementing scheduled POCT We observed a positive correlation between the frequency of hypoglycemia blood within the first 6 hours may provider earlier detection of and T1DM-SEAT score (Rho 0.174, p = 0.004), and an inverse relationship these events. This early detection may improve inpatient safety and prevent between a high score in T1DM-SEAT, indicative of poor adherence, and a the progression of symptoms. worse glycemic control (Rho -0.248, p < 0.001). Conclusions: T1DM patients have a high frequency of auto-reported hypo- glycemia, and a high risk of hypoglycemia unawareness. A simple scale of five questions may allow to identify patient with poor adherence, in whom the actions for reduce the hypoglycemia may be of greater utility.

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410‑P Table. The Association between Frequency of Hypoglycemic Events and With Symptoms No Symptoms P-value Proportion of Asymptomatic Hypoglycemia in Type 1 Diabetes Assessed by Continuous Glucose Monitoring Number of patients, n (%) 138 (55) 112 (45) MARIE MOTH HENRIKSEN, HENRIK ULLITS ANDERSEN, BIRGER THORSTEINS- Age, years 55±13 59±11 0.003 SON, ULRIK PEDERSEN-BJERGAARD, Hillerød, Denmark, Gentofte, Denmark Female, n (%) 72 (52%) 41 (37%) 0.014 Recurrent hypoglycemia promotes impaired awareness resulting in an Male, n (%) 66(48%) 71(63%) 0.014

increased risk of asymptomatic hypoglycemia. However, there are no firm POSTERS data on the frequency of daily life hypoglycemia needed to initiate this HbA1c, % 9.13±2.92 8.19±2.61 0.006 Complications vicious circle. To investigate this we performed 6 days of blinded continu- BG on admission, mg/dL 248±183 221±160 0.431 Acute and Chronic ous glucose monitoring (CGM iPro2, Medtronic Inc.) in 153 patients with Previous day mean BG, mg/dL 156±66 152±65 0.57 type 1 diabetes (T1DM). The patients were instructed to record symptoms Risk Factor, n (%) of hypoglycemia during the period of monitoring. At entry, the patients filled in a questionnaire about self-estimated state of hypoglycemia awareness - Reduced oral intake (NPO status) 49 (39%) 48 (48%) 0.154 - GFR <60 mL/min 69 (51%) 62 (56%) 0.42 (Clarke, Gold and Hillerød methods) and HbA1c and C-peptide were measured. CGM readings were reviewed for hypoglycemic events, defined by glucose Mean BG during hypo event, mg/dl 53.8±11 59.9±8 0.0001 values continuously <4.0 mmol/l for ≥20 minutes and classified according - BG 60 - 69, mg/dL, n (%) 58 (42%) 73 (65%) to presence of warning symptoms or not. The patients were grouped by number of events during the recording period (20 patients had no events): - BG 40 - 59, mg/dL, n (%) 63 (46%) 37 (33%) group 1: 1 event, group 2: 2-3 events, group 3: 4-6 events, group 4: ≥7 events - BG <40, mg/dL, n (%) 17 (12%) 2 (2%) and fractions of asymptomatic events were calculated. The overall rate of Neurological symptoms, n (%) hypoglycemia was 5.8 events/patient-week corresponding to 10.1 hours - Confusion/low concentration 70 (52%) — with hypoglycemia/patient-week. Asymptomatic events comprised 74% of all recorded events (4.3 events/patient-week). In the 4 groups, the fraction - Blurred vision 53 (40%) — of asymptomatic hypoglycemia increased from 57% in group 1 to 61%, 65%, - Slurred speech 39 (30%) — and 80% in group 2, 3, and 4, respectively (p<0.001). Group 4 consisted of Adrenergic symptoms, n (%) patients with longer diabetes duration (group 1-3 vs. group 4; 19 years ± 14 - Sweating 79 (61%) — (mean ± SD) vs. 28 ± 14, p=0.001), lower HbA1c (66 mmol/mol ± 13 vs. 60 ± 10, p=0.003) and more frequent impaired awareness of hypoglycemia (p=0.03). - Anxiety 70 (52%) — Patients with T1DM with hypoglycemic rates corresponding to daily expo- - Trembling 72 (55%) — sure to hypoglycemia had an increased fraction of asymptomatic events and - Dry mouth 57 (44%) — were characterized by the presence of multiple risk factors. Such patients deserve particular attention in clinical practice. Supported By: Nordsjællands Hospital; Danish Medical Research Grant; Toyota- 412‑P Fonden, Denmark Determinants of Severe Hypoglycaemia in Type 1 Diabetes JEAN-BAPTISTE JULLA, PAUL DEBOISSIEU, TIPHAINE VIDAL TRECAN, CLARA 411‑P BOUCHE, JEAN-PHILIPPE KEVORKIAN, MARIE LALOI, FLORINE FERON, LAU- Asymptomatic Hypoglycemia in Non-ICU Patients with Diabetes RENCE SALLE, VANESSA JUDDOO, TULIO SARRON, HERVÉ LEBLANC, JEAN- J. SONYA HAW, PATRICIA GOMEZ, ISABEL ANZOLA, CLEMENTINA RAMOS, FRANÇOIS GAUTIER, JEAN-PIERRE RIVELINE, Paris, France MARRIA URRUTIA, SAUMETH CARDONA, PRIYATHAMA VELLANKI, FRANCISCO Low HbA1c is considered as a severe hypoglycemia risk factor in patients PASQUEL, MAYA FAYFMAN, HEQIONG WANG, GUILLERMO E. UMPIERREZ, with type 1 diabetes (T1D). Considering recent novelties, our aim was to Atlanta, GA, Decatur, GA assess if today, low HbA1c was still a risk factor of SH. Inpatient hypoglycemia occurs in up to 32% of insulin treated patients Data from 1221 patients with T1D followed in consultation during the in non-ICU settings. The prevalence of asymptomatic hypoglycemia is last 2 years had been collect. SH for the last 6 months, age, gender, BMI, unknown, but a recent study with continuous glucose monitoring reported diabetes duration, number of self-monitoring of blood glucose (SMBG) by ~half of patients with BG<70mg/dl were asymptomatic. This prospective day, number of mild hypoglycemia by week (MHBW), insulin treatment and study determined the frequency of asymptomatic events in non-ICU settings diabetes complications have been analyzed. Glycemic liability scores have in 250 consecutive patients with diabetes and BG values <70mg/dl. Data been calculated from CGM: standard deviation (SD) of glucose levels, Mean was collected on symptoms and signs of hypoglycemia, risk factors, compli- Amplitude of Glycemic Excursions score (MAGE) and the Low Blood Glucose cations, and insulin regimen using a standardized questionnaire. Index (LBGI). A logistic regression model was used for assess SH risk factors. Results: Asymptomatic hypoglycemia was reported in 45% of patients. Data are presented as mean (+/- SD). Asymptomatic events were more common in males, older patients with Information about SH were available for 625 patients: age 41±15 years old, lower admission HbA1c. Age and gender were associated with asymptom- men 56%, diabetes duration 20.3 ±12.6 years, HBA1c 7.87 +/- 3.7. 16,8% had atic hypoglycemia independently of blood glucose at the time of index event. at least one SH for the last 6 months (105/625). Univariate analysis shows The odds of asymptomatic events were 2 times higher in men (OR 2.05, 95% no correlation between SH and HbA1c (split in quartile <6.5%; 6.5-7.4%; 7.5- CI: 1.15, 3.64). There were no differences in BMI, duration of diabetes, insulin 8.4% and >8.5%), insulin treatment, or presence of any diabetes complica- regimen, length of stay, vs. services, or hospital complica- tion. Diabetes duration and high number of MHBW were associated with tions between patients with or without symptoms. increased SH risk. Multivariate analysis showed a significant increase of SH Conclusion: Asymptomatic hypoglycemia is common in insulin treated number with more than 10 years of diabetes duration (10-30 years: OR 3.03 non-ICU patients with diabetes. Younger females with high HbA1c had more [1.14-8.07] p= 0.03; >30 ans: OR 6.64 [2.29-19.20] p=0.001) and having 3 or symptoms compared to older males with lower HbA1c who had more asymp- more MHBW (MHBW=0: ref; MHBW =3-4: OR 3,95 [1,27-12,22]; MHBW≥5: tomatic events. More research is needed to understand factors leading to OR 11,45 [3,67-35,76]). Among 61 patients with CGM, univariate analyzes asymptomatic hypoglycemia in hospitalized patients. found a relationship between SH and SD (p = 0.03), but this relationship disappeared in mutivariate analysis. Surprisingly, risk of SH remains high but is not related to low HbA1c neither treatment modality in adult T1D. Diabetes duration and high occurrence of mild hypoglycemia are positively correlated to SH. A correlation was found between SD and SH. Knowing the determi- nants of SH in this population makes it possible target prevention.

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COMPLICATIONS—MACROVASCULAR— with T2DM, 24hBPV was higher with use, however there was ATHEROSCLEROTIC CARDIOVASCULAR DISEASE AND a trend towards reduced 24hBPV with NaCl supplementation. Of the anti- HUMAN DIABETES hypertensive classes, ARBs may not be as effective at lowering 24hBPV as calcium channel blockers or .

Moderated Poster Discussion: Estimation and Management of Cardio- & 415‑P vascular Risk (Posters: 413-P to 418-P), see page 16. Risk of 30-Day Myocardial Infarction and Mortality after Vascular

POSTERS Surgery Complications KHUMARA HUSEYNOVA, BAQIYYAH CONWAY, Morgantown, WV Acute and Chronic & 413‑P Impact of Beta-Blockers on Peripheral Disease Outcome in There is high prevalence of peripheral vascular disease among patients Type 2 Diabetes Mellitus Patients with diabetes mellitus (DM) as well as increased perioperative complication YU WEN SU, YUNG TAI CHEN, CHIEN YI HSU, PO HSUN HUANG, Taipei, Taiwan rates. We undertook a study to evaluate 30-day myocardial infarction (MI) Beta-blockers theoretically reduce peripheral perfusion. The use of beta- and mortality rates among patients undergoing open vascular surgery strati- blockers in PAD is therefore controversial. According to the European soci- fied by DM status using the American College of Surgeons National Surgical ety of cardiology guideline for PAD, beta-blockers are not contraindicated. Quality Improvement Program (NSQIP) database. However, there is few data regarding the safety of beta-blocking agents use Data on 130,698 patients (30.7%, n=40,149 with diabetes) who underwent in diabetes mellitus (DM) patients with PAD. We conducted a retrospective open vascular surgery from April 1, 2011 to March 31, 2013 were reviewed. registry analysis with Taiwan’s National Health Insurance Research Database Cox proportional hazards model was used to assess the relationship of DM to analyze the impact of beta-blockers use on limb outcome in patients with with MI and death within 30 days of surgery. Covariates included were age, type 2 DM and PAD. A total of 20,125 propensity score-matched pairs of beta- sex, diabetes (insulin treated, noninsulin treated, no diabetes), race, history blocker users and nonusers with type 2 DM and known PAD were examined in of congestive heart failure, MI, smoking, alcohol, preoperative blood urea the period 2009 to 2011. A total of 365 beta-blocker users and 434 non-users nitrogen (BUN), creatinine and total length of hospital stay. were amputated. Compared with nonusers, beta-blocker users were associ- 30-day MI rates were 2.3%, 1.6%, and 1.1% among those with insulin ated with a lower risk of for PAD (hazard ratio 0.83; 95% confi- treated DM, noninsulin treated DM, and no DM. Respective mortality rates dence interval, 0.72-0.96). Beta-blocker users also had a lower risk of all-cause were 3.4%, 2.1%, and 2.7%. Compared to no DM, the adjusted hazard of MI mortality (HR 0.94; 95% CI, 0.91-0.98). Risks of in-hospital cardiovascular was 1.7 (95% CI 1.5; 1.9) in patients with insulin treated DM and 1.3 (95% CI death, myocardial infarction, and ischemic were not significantly differ- 1.1; 1.5) in patients with noninsulin treated DM. ent. This large-scale nationwide population-based cohort study demonstrated Among those with DM, African American race was protective of MI (HR that treatment with beta-blockers is associated with lower risk of all-cause 0.6, 95% CI 0.5; 0.8). No difference was observed based on sex. Among mortality and amputation in type 2 DM patients with PAD. those without DM, no difference was observed based on sex or race. Adjusted hazard of mortality was 1.2 (95% CI 1.1; 1.4) for those with insulin Table. Incidence and Risks of ______After Propensity treated DM and 0.72 (95% CI 0.6; 0.8) for those with noninsulin treated DM Score Matching. compared to those without DM. Beta blocker user Beta blocker non-user Crude Both 30-day MI and mortality rates are highest among patient with insulin No. of Incidence No. of Incidence Hazard Ratio p treated DM. Efforts should be directed to optimization of blood sugar and Events Rate Events Rate (95% CI) reducing immediate complication rates I this patient subgroup. All-cause mortality 4332 34.82 4536 36.8 0.94(0.91-0.98) 0.006 In hospital 1159 9.32 1209 9.81 0.95(0.87-1.03) 0.178 & 416‑P cardiovascular death Does Risk-Factor Control Influence ’s Cardiovascular Myocardial infarction 733 5.98 717 5.89 1.01(0.91-1.12) 0.797 Benefits after Stroke? SILVIO E. INZUCCHI, CATHERINE M. VISCOLI, LAWRENCE H. YOUNG, KAREN L. Ischemic stroke 2070 17.56 2101 18.03 0.97(0.92-1.04) 0.401 FURIE, MARK GORMAN, ANNE M. LOVEJOY, GREGORY G. SCHWARTZ, WALTER Amputation 365 2.98 434 3.55 0.83(083-0.96) 0.010 N. KERNAN, IRIS TRIAL INVESTIGATORS, New Haven, CT, Providence, RI, Portland, ME, Denver, CO In the Intervention after Stroke (IRIS) trial, the insulin & 414‑P sensitizing , pioglitazone (45 mg daily), reduced the primary Effect of Angiotensin II Receptor Blocker (ARB) and Sodium Chlo‑ outcome of myocardial infarction (MI) or recurrent stroke compared with pla- ride (NaCl) Supplementation on Blood Pressure Variability in cebo in 3876 insulin-resistant, non-DM patients with recent stroke or tran- Patients with Type 2 Diabetes (T2DM) sient ischemic attack (TIA.) Standard secondary prevention strategies after ANGELA X. CHEN, JOHN L. MORAN, SARA BAQAR, RENATA LIBIANTO, CHRISTO- stroke include anti-platelet and/or anti-thrombotic therapy (APT/ATT), blood PHER O’CALLAGHAN, RICHARD J. MACISAAC, GEORGE JERUMS, ELIF I. EKINCI, pressure (BP) and lipid control, and smoking cessation. Here, we determined Melbourne, Australia, Adelaide, Australia, Fitzroy, Australia, Heidelberg, Australia if the incremental benefit of pioglitazone on the primary outcome depended Raised blood pressure variability (BPV) increases cardiovascular risk, and upon achievement of standard secondary prevention goals. is affected by many factors. However, the effect of renin angiotensin system The interaction of risk factor control and randomization to pioglitazone on blockade and NaCl supplementation on BPV in T2DM is poorly understood. the primary outcome was determined for APT/ATT use, BP (< vs. ≥140/90), We aimed to determine the effect of NaCl supplementation on 24 hour mean and LDL-C (< vs. ≥100), and non-smoking status (yes/no) at randomization. arterial BPV (24hBPV) in the setting of ARB (telmisartan) use and the effects Cox model hazard ratios (HRs; pioglitazone vs. placebo) and Kaplan-Meier of age, sex, plasma renin activity (PRA) and serum aldosterone on 24hBPV. cumulative outcome-free rates were calculated. In a randomised, double-blind, crossover study, patients with T2DM (n=28), Primary outcome HRs did not differ significantly for those achieving vs. treated with telmisartan 40mg for four weeks received NaCl (100mmol/24 not achieving any of the 4 secondary prevention goals: APT/ATT, 0.76 vs. hr) or placebo capsules in the final two weeks of telmisartan use. Following a 0.84 (interaction p=0.96); BP, 0.67 vs. 0.94 (p=0.09); LDL-C, 0.78 vs. 0.73 six week washout, the protocol was repeated in reverse. 24hBPV was evalu- (p=0.77); and non-smoking status, 0.73 vs. 0.86 (p=0.49). Results were not ated as a co-efficient of variation (CV(%)= [mean/standard deviation] x100). substantially affected by adjustment for baseline risk features, or in sensi- 24 hour urinary sodium excretion (24 hUNa), ambulatory blood pressure and tivity analyses for LDL-C < vs. ≥70 or for prevalent statin therapy (yes/no). biochemical tests were performed at each phase. Results were analysed Finally, HRs were not significantly different in those patients meeting all using a linear mixed model. 4 vs. ≤3 of the goals (0.69 vs. 0.81, p=0.47) as well as in those meeting 3-4 24hBPV was higher with telmisartan vs. predicted baseline (p=0.01), with vs. 0-2 of the goals (0.74 vs. 0.87, p=0.51). a trend towards reduced 24hBPV with NaCl supplementation (predicted Among insulin-resistant, non-DM patients with recent stroke or TIA, pio- BPV telmisartan and placebo 11.69% vs. predicted BPV telmisartan and glitazone decreased the risk of MI and recurrent stroke irrespective of base- NaCl 10.56%, p=0.052). 24hBPV was lower in females (females vs. males line APT/ATT, BP and LDL-C control, and smoking status. Thus, the benefits -0.537%, p=0.017) with increasing age (-0.020% per year age, 95% CI -0.31 of pioglitazone in this vulnerable population appear to be incremental to and to -0.009, p=0.001) and PRA (-0.015% per unit PRA, 95% CI-0.265 to -0.004, not attenuated by standard evidence-based therapies. p=0.011). There was no significant correlation between systolic blood pres- Supported By: National Institute of Neurological Disorders and Stroke sure and BPV during telmisartan and/or salt supplementation. In patients (U01NS044876); Takeda Pharmaceutical Company Limited

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A110 COMPLICATIONS—MACROVASCULAR—ATHEROSCLEROTIC CARDIOVASCULAR DISEASE AND HUMAN DIABETES & 417‑P Methods: We confirmed the supervention of macroangiopathy from medi- Relationship between Arterial Stiffness and Left Ventricular Dia‑ cal histories of 748 type 2 diabetes patients. The vascular age was deter- stolic Function in Patients with Type 2 Diabetes mined from CAVI/ABI measured with a vascular screening system VaSera SOICHI KURIOKA, Neyagawa, Japan VS-1500AE (Fukuda Denshi, Tokyo). Relation of ARFs and the data of CAVI/ Increased arterial stiffness and left ventricular (LV) diastolic dysfunction ABI with macroangiopathy development was analyzed by the multivariate are common in patients with type 2 diabetes mellitus (T2DM). However, it logistic regression (MLR). The stepwise procedure was used for the vari- remains unclear whether there is an association between the increased able selection method. C statistics in which some independent variables are arterial stiffness and LV diastolic dysfunction (LVDD). The aim of this study combined were evaluated. Relation of the vascular age and CAVI/ABI data POSTERS was to investigate the relationship between arterial stiffness and indices of with the existence of any of the complication and ARFs was examined by Complications LV diastolic function. One-hundred and twenty consecutive asymptomatic Student t-test. Acute and Chronic patients with T2DM [63% men; age 66±10 years (mean±SD); diabetic dura- Results: The MLR analysis confirmed the following independent variables: tion 11±9 years; HbA1c 7.6±1.8%; systolic blood pressure (SBP) 130±14mmHg] brachial systolic blood pressure (SBP), CAVI, and brachial diastolic blood were enrolled in this study. Subjects with , severe pressure (DBP) for cardiac disorders, CAVI, ABI, and HT for CS, and ABI and valvulopathy, atrial fibrillation, or peripheral arterial disease were excluded CAVI for ASO. The odds ratios were larger in order of sex, CAVI, and SBP for from the study. Arterial stiffness was evaluated by measuring the brachial- cardiac disorders, ABI, HT, and CAVI for CS, and ABI and CAVI for ASO. The ankle pulse wave velocity (baPWV). LV diastolic function was assessed with C statistics were larger in the order of CD, CS, and ASO, and were of 0.71, echocardiographic examination. Doppler echocardiographic indices includ- 0.72, and 0.93, respectively. A difference of mean vascular age between two ing E, A, and e’. Mean baPWV, E/A ratio, e’, and E/e’ ratio were 1760±373 groups of patients with or without CD, CS, ASO, and the present existence cm/s, 0.85±0.36, 6.0±1.6 cm/s and 12±3, respectively. of any of complication was 8.5 (P<0.001), 7.7 (P<0.001), 9.6 (P=0.0015), and baPWV correlated with age (r=0.529, p<0.0001), diabetic duration 9.4 (P<0.001), respectively. (r=0.293, p=0.001), SBP (r=0.243, p=0.007), E/A ratio (r=-0.248, p=0.002), Conclusion: The present results indicate that concomitant development e’ (r=-0.473, p<0.0001), E/e’ ratio (r=0.247, p=0.007). In multiple regres- of macroangiopathy in type 2 diabetes patients is probably predicted based sion analysis, age (β=0.365, p<0.001), SBP (β=0.231, p=0.005), and e’ on the ARFs and the measurement data provided by a CAVI/ABI. The past (β=-0.259, p=0.026) correlated independently with baPWV, however, E/A ratio and present existence of complications of CD, cerebral sclerosis, ASO, and (β=-0.055, p=0.547) and E/e’ ratio (β=-0.119, p=0.294) did not. In this study, any of the complication, ARFs, and CAVI/ABI data are useful predictors of increased arterial stiffness was associated with LVDD. Among the indices of macroangiopathy development. LV diastolic function, e’ correlated most significantly with baPWV. 420‑P & 418‑P Prevalence of Asymptomatic and Associ‑ WITHDRAWN ated Factors among Type 2 Diabetes Mellitus Patients in Havana, Cuba YORDANKA PINA RIVERA, LIZ ODELMIS CRUZ BENITEZ, Gaborone, Botswana, Havana, Cuba Introduction: The most frequent echocardiographic finding in patients with asymptomatic diabetes mellitus is Left Ventricle (LV) diastolic dysfunc- tion with preserved LV ejection. The objective was to determine the preva- lence of diabetic cardiomyopathy in patients with type 2 diabetes mellitus and its correlation with different demographic and biochemical parameters. Methods: We conducted a cross-sectional study to identify the preva- lence of Diabetic cardiomyopathy (DC) among asymptomatic type 2 diabetic patients attending Dr. Carlos J. Finlay Hospital in Havana, Cuba from May 2010 to May 2012. A resting M-mode Echocardiography was performed in 79 type 2 diabetic patients without history of Hypertension, kidneys or heart . For the purpose of this study, DC was defined as having diastolic dysfunction with preserved left systolic function. A comparison was made between patients with DC and those without DC to determine associating demographic and biochemical factors. A p-value < 0.05 was considered sta- tistically significant. Results: Diabetic cardiomyopathy was found in 41/79 (51.9%); and it was significantly associated with male gender (p < 0.02), elderly age (p < 0.0004), and longer duration of diabetes (p < 0.04). Unlike previous studies; body mass index, fasting blood sugar and HbA1c were not statistically associated with DC. Hypercholesterolemia and was more frequent in patients with DC with p= 0.002 and p < 0.0001 respectively. The risk for DC was 4.2 and 11.4 times higher in patients with hypercholesterolemia and hypertriglyceridemia respectively (OR = 4.2; CI 95% 1.6-10.9 and OR = 11.4; CI 95% 2.9-43.4). Conclusion: Diabetic cardiomyopathy was very common in the study popu- lation and it appeared to correlate broadly to the high burden of dyslipid- 421‑P emia. There is a need to conduct multicentric studies to verify the findings. Low Serum Bilirubin Concentration Is Associated with High Preva‑ Furthermore aggressive cholesterol reduction protocols are advocated in lence of and Macrovascular Diseases in Type 2 Dia‑ this setting. betic Patients MASAMI TANAKA, TAKESHI NISHIMURA, SHU MEGURO, JUNICHIRO IRIE, 419‑P YOSHIFUMI SAISHO, MASANORI MITSUISHI, YOSHINAGA KAWANO, HIROSHI Can We Quantitatively Assess Macroangiopathy Development in ITOH, Tokyo, Japan Patients with Type 2 Diabetes? Objective: Low serum total bilirubin concentration (TBC) is a risk factor for HIROKATSU NIWA, MIDORI DANNOURA, KAZUNO OOMORI, KIYOHIKO TAKA- and macrovascular diseases (Macro) (stroke, ischemic HASHI, ARINA MIYOSHI, Sapporo, Japan heart disease, or peripheral arterial disease) in diabetic patients. In addition, Background and Aims: This study aims to elucidate if any association it is reported that diabetic retinopathy is a risk factor for Macro. Therefore, exists between the development of macroangiopathy in type 2 diabetes we investigated the relationships among TBC, retinopathy, and Macro. patients, and arteriosclerotic risk factors (ARF) and the data of cardio ankle Methods: Six hundred and forty four patients (425 men) with type 2 dia- vascular index (CAVI) and ankle brachial pressure index (ABI). betes were investigated in this cross-sectional study. The proportion of the patients with Macro was compared between the patients with and with-

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out diabetic retinopathy by a chi-square test. TBC was compared between Figure 1. Incidence Rates of HF Hospitalization and Death in Inpatients with the patients with and without retinopathy, and with and without Macro by T2D Participating in HF Trials of Different HF Interventions and Their Hazard unpaired t-test. Logistic regression analyses were performed to find explan- Ratios (95% Confidence Interval). Abbreviations: NR-Not Reported. (*Com- atory factors for retinopathy and Macro. posite Outcome Comprises HF Hospitalization and CV Death). Results: The age, diabetes duration, HbA1c, and TBC of 644 patients were 64.6 + 14.0 years, 13.9 + 10.9 years, 9.1 + 2.2%, and 0.68 + 0.24 mg/ dl, respectively. Two hundred and seventy-two (42%) patients had retinopa- thy. Two hundred and eighteen (34%) patients had Macro. The proportion of POSTERS

Complications the patients with Macro was significantly higher in those with retinopathy

Acute and Chronic than in those without retinopathy (p<0.001). TBC was significantly lower in patients with retinopathy than in those without retinopathy (p<0.001). Simi- larly, TBC was lower in patients with Macro than in those without Macro (p=0.02). TBC, age, diabetes duration, systolic blood pressure, HbA1c, and eGFR were shown to be independent explanatory factors for retinopathy. Age, diabetes duration, BMI, smoking, and LDL-C were independent factors for Macro. Conclusions: Diabetic patients with retinopathy showed high prevalence of Macro. Although low TBC was related to both retinopathy and Macro, the Figure 2. Proportion of Patients with Heart Failure Hospitalization in Car- effect of bilirubin to retinopathy might be stronger than that to Macro. The diovascular Outcome Trials of Different Glucose-Lowering Drugs and Their factors associated with retinopathy and Macro were quite different. Hazard Ratios (95% Confidence Interval). 422‑P The Ratio of Visceral to Subcutaneous Area Predicts Cardiovas‑ cular Events in Patients with Type 2 Diabetes TATSUYA FUKUDA, RYOTARO BOUCHI, TAKATO TAKEUCHI, ISAO MINAMI, HAJIME IZUMIYAMA, KOSHI HASHIMOTO, TAKANOBU YOSHIMOTO, YOSHI- HIRO OGAWA, Tokyo, Japan Recent investigations have demonstrated that the ratio of visceral fat area (VFA) to subcutaneous fat area (SFA) (V/S ratio) is a better predictor of atherosclerosis than SFA or VFA; however, it has still been unclear whether the V/S ratio could be predictive of incident cardiovascular disease (CVD) in patients with diabetes. We therefore conducted a prospective observational study to examine the relationships between the V/S ratio and incidence of CVD in Japanese patients with type 2 diabetes. In this study, 682 Japanese patients with type 2 diabetes (mean age 64 ± 13 years; 41% female) were enrolled. VFA (cm2) and SFA (cm2) were assessed by dual bioelectrical imped- ance analyzer (BIA). The patients were divided into four groups according to the quartiles of V/S ratio. Over a median follow-up of 2.50 years (range: 0.02-4.18), incident CVD occurred in 21 participants. The 3-year cumulative incidence of the endpoint were 0, 13.2, 15.2, 28.6% according to the quar- tiles of V/S ratio from Q1 to Q4, respectively (Log-rank p value = 0.047). In Cox proportional hazards regression analysis, baseline V/S ratio was signifi- 424‑P cantly associated with incident CVD after adjustment for the confounding Factors Associated with Statin Initiation for Primary Coronary variables including brain-type (BNP), use of antiplatelet Artery Disease (CAD) Prevention in Type 1 Diabetes (T1D) agents, HbA1c [hazard ratio (HR) 31.58, 95% CI 1.68-594.02, p = 0.021]. In RACHEL G. MILLER, TREVOR J. ORCHARD, Pittsburgh, PA contrast, neither VFA (HR 1.01, 95% CI 1.00-1.02, p = 0.066) nor SFA (HR There is little information regarding the use and efficacy of statin therapy 1.00, 95% CI 0.99-1.01, p = 0.802) were significantly associated with incident in T1D. We thus examined the characteristics of those with T1D who ini- CVD. Addition of V/S ratio to age, eGFR, BNP, antiplatelet agents, and HbA1c tiated statin therapy for primary CAD prevention and determined whether significantly improved classification performance for incident CVD using net statin use was associated with reduced CAD risk after accounting for these reclassification improvement (NRI 0.60, 95% CI 0.21-1.00) and the integrated factors. The Pittsburgh Epidemiology of Diabetes Complications (EDC) Study discrimination improvement (IDI 0.02, 95% CI 0.00-0.05). This study provides is an ongoing prospective cohort study of childhood-onset (<17 yrs) T1D diag- evidence that V/S ratio measured by dual BIA is an independent predictor of nosed 1950-1980 (n=658, 49% women, mean age 27, T1D duration 18 yr at incident CVD in patients with type 2 diabetes. baseline). Baseline exams took place 1986-88 and biennially thereafter for Supported By: Japan Ministry of Education, Culture, Sports, Science and Tech- the first 10 yrs and at 18 and 25 yr. Statin use was ascertained through bien- nology nial questionnaires throughout follow-up. Most recent risk factor data prior to statin initiation were used. CAD incidence was defined as CAD death, 423‑P myocardial infarction, revascularization or coronary stenosis ≥50%. Preva- Concomitant Heart Failure (HF) and Type 2 Diabetes (T2D): A Deadly lent CAD cases at baseline were excluded (n=44). Duo 34% (218) initiated statin therapy prior to first CAD event. Mean age and ANNE PERNILLE OFSTAD, DAN ATAR, LARS GULLESTAD, GISLE LANGSLET, ODD T1D duration at statin initiation were 43 and 35 yr respectively. Those initiat- ERIK JOHANSEN, Rud, Norway, Oslo, Norway ing statins had significantly higher non-HDL cholesterol (nonHDLc) (p=0.001), We explored the incremental effect on mortality/morbidity of co-existing lower white cell count (WBC) (p=0.02), higher estimated glomerular filtration HF and T2D. The review included HF trials of ACEi, digoxin, β blocker, ARB, rate (eGFR) (p=0.0001), and were more likely to have >/=Bachelor’s degree (p=0.02). In a multivariable model lower HbA1c (OR 0.8, p=0.005), higher If-blocker, MRA, and ARNI, that included patients (n=38600) with preva- lent T2D, and CV outcome trials (CVOTs) in T2D with or without prevalent nonHDLc (OR 1.01, p<.001), higher eGFR (OR 1.01, p<.001), and lower WBC HF (n=64209). In all HF trials, HF outcomes by prevalent T2D were reported (OR 0.9, p=0.04) were associated with greater odds of statin initiation. After with an incremental risk of HF and death confessed by prevalent T2D and adjusting for these factors, statin use was not significantly associated with a treatment effect similar to those without T2D (Figure 1). All T2D CVOTs CAD risk (HR 1.1, p=0.6). drugs reported on HF outcomes (Figure 2) with heterogeneity between tri- These results suggest that apart from nonHDLc those who initiated statin als with one reporting benefits () and two reporting increased therapy for primary CAD prevention were generally healthier and had a risk (, pioglitazone). In vulnerable T2D patients with concomitant higher level of education than those not initiating statins. More research is HF, guideline recommended HF drugs are effective. When choosing glucose needed to determine whether the highest risk T1D patients are being appro- lowering therapy, outcomes from available CVOT should be considered. priately targeted for statin therapy. Supported By: National Institutes of Health

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A112 COMPLICATIONS—MACROVASCULAR—ATHEROSCLEROTIC CARDIOVASCULAR DISEASE AND HUMAN DIABETES

425‑P 427‑P Type 2 Diabetes in Young Females Results in Increased Serum Amy‑ All-Cause Mortality in Randomized Controlled Trials of Type 2 Dia‑ loid A and Changes to High Density Lipoprotein Function in Both betes: An Updated HDL2 and HDL3 EBRAHIM BARKOUDAH, BRIAN L. CLAGGETT, MARC A. PFEFFER, Boston, MA KAYLEIGH GRIFFITHS, AGNIESZKA PAZDERSKA, MOHAMMED AHMED, ANNE With the recent regulatory guidance to evaluate cardiovascular risk MCGOWAN, ALEXANDER P. MAXWELL, JANE MCENENY, JAMES GIBNEY, of new antidiabetic agents the number of major randomized clinical out- GARETH J. MCKAY, Belfast, United Kingdom, Dublin, Ireland come trials (RCTs) of in subjects with type 2 diabetes mellitus (T2DM) has

Persons with type 2 diabetes mellitus (T2DM) are at increased risk of expanded. We examined RCTs which randomized over 1,000 adults with POSTERS developing atherosclerosis. High density lipoproteins (HDL) transport cho- T2DM and followed for a minimum of 1 year. Excluded were RCTs of subjects Complications lesterol from cells and tissues back to the providing protection against within 3 months of an acute coronary syndrome event or receiving dialysis. Acute and Chronic the development of cardiovascular disease (CVD), which may be reduced We related the RCT inclusion and exclusion criteria to the annualized mortal- by the release of serum amyloid A (SAA). In a case-control study of young ity rates observed in the trials. RCTs were categorized according to mortality females, blood samples were compared between T2DM (n=42) and con- rates of: ≤1; >1 - ≤2; >2 - ≤3; or >3 per 100 patient-years. trols without T2DM (n=42). HDL2 and HDL3 subfractions were isolated by Included were 33 RCTs with 175,310 patients and 693,665 patient-years rapid ultracentrifugation, and SAA and apolipoprotein AI (apoAI) concentra- of follow-up. The overall mortality rate in this population was 1.97 per 100 tions, paraoxonase 1 (PON-1), cholesteryl ester transfer protein (CETP) and patient-years, and varied widely by RCT from 0.28 to 8.24 (Figure 1). The lecithin-cholesterol acyltransferase (LCAT) activities were measured in the mortality rate categories were characterized for the most part by patterns serum and/or subfractions. SAA concentrations in serum, HDL2 and HDL3 of inclusion and exclusion: ≤1 only hypertension was permitted, and mild were higher in individuals with T2DM compared to controls: serum [29573 chronic disease (CKD) and established cardiovascular disease (CVD) μg/L (17211, 67664) vs. 14911 μg/L (7037, 36008); p=0.002], HDL2 [956.5 μg/L were excluded; 1- ≤2 higher concentrations of CVD risk factors were mostly (553.0, 2239.8) vs. 386.7 μg/L (188.3, 722.1); p<0.001], and HDL3, [13083 μg/L required; >2 - ≤3 CVD had to be established; and >3 advanced CKD was an (7802, 28615) vs. 5809 μg/L (3147, 13314); p<0.001]. Serum PON-1 activity inclusion. was significantly lower in participants with T2DM compared to controls Across the RCTs of subjects with T2DM, rates of all-cause mortality vary [38245 U/L (7025) vs. 41109 U/L (5690); p=0.043]. CETP activity was sig- widely (almost 30 folds) with requirement of advanced CKD associated with nificantly higher in individuals with T2DM vs. controls in HDL2 [232.6 μmol/L the highest mortality rates. (14.1) vs. 217.1 μmol/L (25.1); p=0.001] and HDL [279.5 μmol/L (17.7) vs. 245.2 3 Figure 1. All-Cause Mortality in Randomized Controlled Trials of Type 2 Dia- μmol/L (41.2); p<0.001]. These results suggest individuals with T2DM have betes Subjects. increased SAA-related and biochemical features of pro-ath- erogenic HDL. Consequently, SAA may prove a useful biomarker in subjects with T2DM given its influence on increased CVD risk.

426‑P The Effect of Time Window on Measurements of Postprandial Hypo‑ tension JENNIFER K. FERRIS, KENNETH M. MADDEN, Vancouver, BC, Canada Postprandial hypotension (PPH) is a common complication of diabetes mellitus (DM), and is associated with increased falls risk and mortality in older adults with DM. PPH is defined as a drop in systolic blood pres- sure (SBP) of ≥20mmHg after a meal; however, optimal SPB measurement parameters have yet to be empirically determined. We examine the impact of the time window of SBP data collection in assessing PPH, comparing 2 commonly employed outcome measures: total number of PPH episodes, and mean change in SBP. 428‑P We recruited 15 adults aged 65 and older with a diagnosis of type 2 DM Factors that May Account for CV Risk Reduction Associated with (mean age: 76; age range: 67-85). 2 testing sessions were performed for DPP-4 Inhibitor Treatment in Young T2D Patients each individual. After a 20-minute baseline measure, participants were MARC EVANS, PLAMEN KOZLOVSKI, PÄIVI M. PALDÁNIUS, JAMES E. FOLEY, given a standardized meal, and beat-beat SBP was continuously measured VAISHALI BHOSEKAR, CARMEN SERBAN, ANGELO AVOGARO, Cardiff, United with a finometer for 90 minutes after the meal. SBP data were binned into Kingdom, Basel, Switzerland, East Hanover, NJ, Hyderabad, India, Padua, Italy different time windows for analysis (30s, 1m, 2m, 3m, 5m, 6m, 9m, 10m, 15m, In a CV meta-analysis from a safety pool of 17,446 patients, there was a 18m, 30m, 45m, 90m). Number of PPH episodes was defined as the total significant 37% relative risk reduction in prospectively adjudicated MACE number of events with a SBP change of 20mmhg. Mean change in SPB ≥ (non-fatal MI, non-fatal stroke, CV death) with (VILDA) vs. com- was defined at the average difference of SPB post-meal relative to baseline parators (COMP) in patients <65 years, without an increased risk of MACE measures. Repeated-measures ANOVAs were used to assess the impact of in patients 65 years. We present an explorative analysis of factors which time interval on PPH episodes or mean change in SBP. ≥ may have contributed to the observed CV risk reduction. In pre-defined 14 participants had at least one PPH episode in the post-meal assess- subgroups <65 and >65 years, on-treatment differences (VILDA vs. COMP) ment. Number of PPH episodes differed based on the time window of data for the change from baseline (BL) in CV risk factors, were analyzed using analysis (F = 13.329, p < 0.0001). 12 participants showed a mean SPB 12,29 ANCOVA with BL value for each variable of interest, treatment and study as change of 20mmHg in at least one assessment time window. Mean change ≥ covariates. Additional adjustment for antihypertensive and statin use at BL in SBP did not differ based on time window of data analysis (F = 0.589, 12,29 was performed for BP and lipids, respectively. At BL patients <65 had slightly p = 0.851). higher HbA1c than patients 65 (8.3% vs. 8.0%). More patients 65 had The measurement of number of PPH events is highly influenced by the ≥ ≥ hypertension (73.1% vs. 51.3%), (53.3% vs. 43.9%) and history time frame of data collection, and is therefore difficult to compare between of CV events (32.2% vs. 12.9%). There were small, statistically significant different SBP analysis protocols. Mean change of SPB is a more robust mea- differences in the change in HbA1c and total cholesterol in favor of VILDA, sure that is likely to have good reliability between different study protocols, similar in both subgroups. In the <65 subgroup, but not in the 65 subgroup, and may have higher validity as an assessment of PPH. Future research on ≥ significant on-treatment differences were observed in the reduction in PPH should use mean change in SBP as a preferred outcome measure. SBP, LDL-cholesterol and weight. VILDA’s effect on CV risk factors like SBP, Supported By: Canadian Diabetes Association (OG-3-13-4157) LDL-cholesterol and weight could explain the observed CV risk reduction in patients <65 years.

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Table. Adjusted Mean Change in Parameters from Baseline to Endpoint in Methods: We enrolled 1,240 asymptomatic subjects who underwent repeated Age Stratified Subgroups. CAC score measurement during routine health examinations. CAC score progres- sion was defined as either incident CAC in a population free of CAC at baseline, or an increase by ≥2.5 units between the baseline and final square root of CAC scores in participants with detectable CAC at baseline. Subjects were stratified by body mass index (cut-off, 25.0 kg/m2) and metabolic health state using Adult Treatment Panel-III criteria. FLD was assessed via ultrasonography. Results: Over 2.9 years of follow-up, 25.2% of total subjects exhibited CAC POSTERS

Complications score progression. The MHO phenotype was not significantly associated

Acute and Chronic with CAC score progression (multivariate adjusted-odds ratio [OR], 1.45; 95% confidence interval [CI], 0.93-2.25), as compared to the metabolically healthy nonobese (MHNO) phenotype. However, subgroup analysis indicated that the MHO/FLD phenotype was significantly associated with CAC score progres- sion (multivariate adjusted-OR, 2.37; 95% CI, 1.34-4.16), as compared to the MHNO/no FLD phenotype, whereas the MHO/no FLD phenotype was not (mul- tivariate adjusted OR, 1.25; 95% CI, 0.71-2.24). Conclusions: Obese individuals with FLD have an increased risk of athero­ sclerosis progression, despite their healthy metabolic profile. Preventive interventions targeting cardiometabolic risk factors should be considered in such individuals, regardless of the weight status.

431‑P WITHDRAWN

Supported By: Novartis

429‑P Group-Based Trajectories of Two-Year Visit-to-Visit Glucose Vari‑ ability in Outpatients with Diabetes Were Associated with All- Cause and Cardiovascular Mortality in the Next Four Years CHIA LIN LEE, JUN SING WANG, YU FEN LI, Taichung, Taiwan Background: Visit-to-visit glucose variability has been associated with micro- and macro-vascular complications. However, the effects of glucose variability changes over time remained unclear. The aim of this study was to investigate 1. glucose variability changes over time, and 2. the impact of glucose variability changes on all-cause and cardiovascular mortality. Method: Diabetes outpatients from 2009 to 2014 were enrolled. Coeffi- cient of variation of fasting plasma glucose (glu_CV) within each 3 months was defined as glucose variability. Changes of glucose variability was defined as glu_CV changes over first 2 years after enrollment. We used a group based modeling approach to identify glu_CV trajectories overtime and compared the mortality between groups in the next 4 years. Results: Among the 3569 subjects (mean age 69 years old, male 55%), five distinct trajectories of glucose variability were identified: low glucose vari- ability (N=3025, set as reference group), increasing glucose variability (N=110), fluctuated glucose variability (N=248), decreasing glucose variability (N=119) and high glucose variability (N=67). The multivariable adjusted hazard ratio for death from any cause was 2.05 (95% confidence interval 0.83-5.02), 2.63 432‑P (1.40- 4.93), 2.78 (1.33-5.80) and 4.44 (1.78-11.06) for increasing glucose vari- Combined and Eye Screening: A Feasibility Study ability, fluctuated glucose variability, decreasing glucose variability and high JANE E.A. LEWIS, KERI HUTCHINSON, Cardiff, United Kingdom, Pontypridd, United glucose variability group, respectively. After considering competing death, the Kingdom corresponding adjusted hazard ratios of cardiovascular-specific death were The majority of non-traumatic are most often caused by PAD, 3.34(0.61-18.29), 4.92(1.30-18.65), 6.58(1.52-28.57) and 15.98(2.92-87.39). followed by diabetes or a combination of both. People with diabetes are 6 Conclusions: Compared with low glucose variability, other trajectories of times more likely to have an amputation than those without (up to 20/day glucose variabilities within first 2 years were associated with all-cause and in the UK) with both high monetary and human costs. The increased risk cardiovascular mortality in the next 4 years. for cardiovascular morbidity, such as myocardial infarction and stroke, and increased risk for mortality is observed in both symptomatic and asymp- 430‑P tomatic patients. A diagnosis of PAD presents the opportunity to initiate Fatty Liver Disease Determines the Progression of Coronary Artery secondary prevention by instituting atherosclerosis risk factor modification. Calcification in a Metabolically Healthy Obese Population This study aimed to explore the feasibility of combining foot and eye CHANG HEE JUNG, YU MI KANG, YUN KYUNG CHO, SEUNG EUN LEE, MIN JUNG screening within one appointment. It also aimed to evidence the prevalence LEE, JENIE YOONOO HWANG, EUN HEE KIM, JOONG YEOL PARK, WOO JE LEE, of asymptomatic PAD, uptake of the additional screening and whether the HONG KYU KIM, Seoul, Republic of Korea patient experience welcomed a combined service in the future. Potential Background: This study aimed to investigate whether metabolically healthy participants were recruited whilst attending their retinopathy screening obese (MHO) phenotype is associated with progression of atherosclerotic activ- appointments 1 day a week over an 8 week period. Those consenting had ity, reflected as the changes in coronary artery calcification (CAC) over time. If so, a lower limb arterial assessment using an automated device measuring ABI we sought to determine the role of fatty liver disease (FLD) in this progression. and pulse volume negating the need to rest the patient, and plantar light

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A114 COMPLICATIONS—MACROVASCULAR—ATHEROSCLEROTIC CARDIOVASCULAR DISEASE AND HUMAN DIABETES touch sensation checked, during the waiting time required for pupil dilation anion gap metabolic acidosis with hyperglycemia is diabetic . to occur prior to retinal screening. High clinical suspicion and prompt treatment can save a life. 360 participants were invited to take part. 89% (n=321) accepted the PAD screening and met the inclusion criteria. 12% (n=38) were found to have 435‑P neuropathy; 24.6% (n=79) to have previously undiagnosed asymptomatic mRNA Expression of Platelet Activating Factor Receptor in Periph‑ PAD with a 100% positive patient experience. The combined screening was eral Blood Mononuclear Cells Is Associated with and well-received and prevalence of undiagnosed asymptomatic PAD in this Macroangiopathy in Patients with Type 2 Diabetes study would indicate targeted screening could allow earlier identification MIHOKO KURANO, SAHAR GHAVIDEL DARESTANI, ATUSHI SHINNAKASU, KIYO- POSTERS and intervention of potentially treatable limb and life threatening disease. AKI YAMAMOTO, YUKARI DOCHI, KAYO UEMURA, YUKO IKEDA, AKIRA KIKUCHI, Complications HIROSHI HASHIGUCHI, TAKAHISA DEGUCHI, YOSHIHIKO NISHIO, Kagoshima, Acute and Chronic 433‑P Japan Uric Acid Is a Strong Predictor of Myocardial Infarction in Men with High urinary albumin to creatinine ratio (ACR) and low estimated glomeru- Decreased Renal Function lar filtration rate (eGFR) are known as independent risk factors for cardio- SPOMENKA LJUBIC, IVANA ANTAL, ANAMARIJA JAZBEC, MARTINA TOMIC, vascular events. Several studies showed that changes in genes expression LEA SMIRCIC-DUVNJAK, Zagreb, Croatia in peripheral leukocyte play a role in the pathogenesis of atherosclerosis in Prediction of myocardial infarction (MI) in relation to previous patient patients with diabetes. We have hypothesized that any molecular changes or family history was studied. Metabolic parameters were determined in in blood cells in the process of developing nephropathy cause renal dysfunc- 383 patients assigned into groups according to MI (no:0, with:1, albumin/ tion and through impaired endothelial function in creatinine ratio (ACR) (<3.0, ≥3.0) and the presence of glucose intolerance type 2 diabetes (T2D). We recruited 95 patients with T2D and classified them (GI), type 1 (DM1), type 2 (DM2) diabetes, and control group (CG). In all sta- into three groups according to the level of ACR: 42 with normoalbuminuria, tistical tests: T-test, Mann Whitney test, ANOVA and Tukey post-hoc test, 26 with , 27 with macroalbuminuria. We also determined logistic regression, α=0.05 was considered statistically significant. Patients their eGFR, flow-mediated dilatation of brachial artery (FMD), and mRNA with MI had significantly higher values of fibrinogen (FIB) and uric acid (UA), expression in peripheral blood mononuclear cells (PBMCs) by real-time whereas apolipoprotein A1 (Apo A1), glomerular filtration rate (GFR) and reverse transcription polymerase chain reaction. The mRNA expression of diastolic blood pressure (DBP) were decreased. Women had significantly platelet activating factor receptor (PAFR) showed the most prominent dif- higher values of FIB and Apo A1 compared to men, whereas UA was higher in ferences among three groups, and it was significantly higher in macroalbu- men. ANOVA revealed significant differences in UA, FIB and Apo A1 among minuric group compared with the other two groups. Furthermore, Spearman groups according to DM or GI presence. UA was increased in DM2 and GI, correlation analysis revealed that the mRNA expression of PAFR significantly and FIB in DM2 and CG compared to DM1. Apo A1 and GFR were increased in correlated with log transformed ACR (r = 0.424, p < 0.001), however, it did not DM1 and CG compared to DM2. GFR, UA and DBP significantly differ among significantly associate with eGFR (r = -0.14, p = 0.177). It also had a signifi- groups according to MI and ACR (MI:0 + ACR<3.0, MI:0 + ACR≥3.0, MI:1 + cant negative correlation with FMD (r = -0.373, p < 0.001). Stepwise multiple ACR<3.0, MI:1 + ACR≥3.0). UA was significantly increased in MI:1+ACR<3.0 logistic regression analysis showed that duration of diabetes and mRNA and GFR decreased in MI:1 + ACR<3.0 and MI:1+ACR≥3.0 compared to MI:0 + expression of PAFR were independent explanatory variables for prevalence ACR<3.0 group. DBP was decreased in MI:1 + ACR≥3.0 group compared to of cerebrovascular disease (duration: OR = 1.127, p = 0.010; PAFR mRNA: MI:0 + ACR<≥3.0 group. Univariate logistic regression for MI as a dependent OR =2.581, p = 0.004). These results indicate that the increased expression variable point out UA, as statistically significant predictors, both in men, of PAFR in PBMCs is possibly may link to macroangi- OR=1.014, 95% CI (1.004,1.023) and women OR=1.019, 95% CI (1.003,1.036). opathy through the impaired endothelial function in patients with T2D. Significant difference in FIB, GFR and DBP was observed in siblings according to the presence of MI in parents. Patients, particularly DM2, with increased 436‑P UA and decreased GFR and DBP may be at increased risk for MI, especially Visceral Fat, Not Subcutaneous Fat, Impairs Endothelial Function in men related to UA behavior. Apo A1 could be protective in women. GFR more Patients with Type 2 Diabetes than ACR seem to be connected with MI. Insight in UA, DBP, Apo A1 and HARUKA TAMURA, YOSHINOBU KONDO, SHINOBU SATOH, YASUO TERAUCHI, renal function could identify subjects with increased MI risk. Kanagawa, Japan, Yokohama, Japan Aim: Coronary heart diseases (CHD) occur more frequently in patients 434‑P with diabetes than in those without diabetes. The onset rate of myocardial DKA or Not DKA? infarction in patients with diabetes having no history of CHD was equal to SAPNA RAGHUNATHAN, SHAILEJA PAMNANI, Derby, CT the relapse rate in those without diabetes having a history of CHD. Insu- A 56-year old Caucasian with PMH of DMT2, HLD, depression, alcohol lin resistance was caused by visceral fat accumulation, and observed to dependence, benzodiazepine dependence (h/o intentional overdose), presented be related to endothelial function. Endothelial dysfunction is a predictor of to the ED with severe back pain after mechanical fall. In the ED, labs WNL, long-term arteriosclerosis and the incidence of cardiovascular event. That except for blood glucose (201 mg/dL). He was given NSAID’s for pain and dis- was noninvasively assessed by flow-mediated dilation (FMD). We examined charged. Few hours later, found to be confused, incoherent and nauseous and the effects of visceral fat on endothelial function in patients with diabetes was brought to the ED. Vitals: Temp 96.3 F; HR 80-100 beats/min; BP 90/60 mm by using FMD and abdominal CT. of Hg, RR 32 breaths/min. Labs revealed WBC 24.3, 6 bands; blood glucose level Methods: The study included patients hospitalized for type 2 diabetes. FMD 601 mg/dL; potassium 5.5 mmol/L; anion gap 41; BUN 10 mg/dL; Cr 3.4 mg/dL; and abdominal CT were evaluated in all subjects. The fat area at the navel bicarbonate <5 mmol/L; lactic acid 26.3 mmol/L. ABG showed pH 6.90; PCO2 11; level was measured using images obtained with the SYNAPSE VINCENT. HCO3 2 mEQ/L. Utox positive for benzodiazepine, negative for acetone. Started Results: We enrolled 70 subjects (36 males and 34 females; age, 65.2 ± on an insulin drip for DKA. Bicarbonate drip added for low bicarbonate levels. 12.9 years; disease duration, 9.3 ± 10.4 years; BMI, 25.3 ± 5.9 kg/m2; HbA1c, Anion gap was elevated despite insulin and bicarbonate drips. Other causes 10.4 ± 2.5%; FMD, 5.7 ± 3.0%). BMI was significantly correlated with FMD for elevated anion gap metabolic acidosis, i.e., toxic alcohol ingestion, sepsis (P = 0.03). If BMI increased by 1 kg/m2, FMD decreased by 0.13%. Multiple were investigated. Patient became hemodynamically unstable, was started on linear regression analysis revealed that BMI was an independent predictor pressors and broad spectrum . Hyperglycemia resolved but low urine of reduced FMD (β = −0.25; SE, 0.06; 95% CI, −0.23 to −0.005; P = 0.04). Dis- output and persistent metabolic acidosis continued. History from the daugh- ease duration, sex, HbA1c, hypertension, and smoking history were included ter revealed the patient had made threats about overdosing on . As in this model. Subcutaneous and visceral fat areas were 163.4 ± 92.6 cm2 creatinine worsened, severe metabolic acidosis persisted and hyperkalemia and 141.3 ± 68.6 cm2, respectively. If subcutaneous fat area increased by 10 (6.8 mmol/L) developed, he was intubated and dialyzed emergently. Metformin cm2, FMD decreased by 0.06%. Thus, subcutaneous fat was not correlated levels were obtained before dialysis. Toxic alcohol levels were negative. Post with FMD. In contrast, if visceral fat increased by 10 cm2, FMD decreased by dialysis, pH, acidosis, anion gap and urine output improved significantly. Anion 0.15%. Thus, visceral fat was significantly correlated with FMD (P = 0.003). gap normalized after 2nd dialysis. Pressor was discontinued on day 2 and extu- Conclusions: was correlated to FMD. Visceral fat accumulation bated on day 5. Creatinine went back to the baseline (1.0 mg/dL) at discharge. in obesity decreased FMD, whereas subcutaneous fat accumulation in Metformin levels were 28 mcg/ml (normal: 1-2 mcg/ml). obesity did not. Visceral fat may deteriorate the progression of endothe- Conclusion: This case illustrates the potential for severe lactic acidosis lial dysfunction. with the overdose of metformin. It is important to understand that not all

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437‑P status was ascertained using the Michigan Neuropathy Screening Instru- Baroreflex Failure Plays an Important Role in the Development of ment clinical examination (MNSIE). MNSIE score ≥ 2.5 defined the presence Diabetic Cardiomyopathy via Oxidative Stress in Type 2 Diabetic of DPN. CIMT was measured using high-resolution B-mode ultrasonography Rats and the maximal CIMT value was measured. Of the 114 subjects, 44 (39%) HIROYUKI IUCHI, MASAYA SAKAMOTO, DAISUKE MATSUTANI, HIROFUMI had DPN. Subjects with and without DSP had similar age (58.0 ±8.5 vs. 57.0 ± SUZUKI, KAZUNORI UTSUNOMIYA, Tokyo, Japan 7.8 years), diabetes duration (10.8 ± 7.4 vs. 8.2 ± 5.6 years), body mass index Increased blood pressure variability caused by baroreflex failure is (24.8 ± 3.1 vs. 24.6 ± 3.2 kg/m2), eGFR (76.3 ± 15.4 vs. 81.3 ± 12.8 ml/min/1.73 m2), and not poor glycemic control (7.8 ± 1.5 vs. 7.4 ± 1.6%). However, sub-

POSTERS recently thought to be a novel risk factor for cardiovascular disease in type

Complications 2 diabetic patients. In addition, it is also demonstrated that baroreflex func- jects with DPN had higher CIMT than those without DPN (1.80 ± 0.92 vs. Acute and Chronic tion tends to be impaired in diabetic patients. However, it remains unclear 1.31 ± 0.52 mm, P=0.048). Linear regression analysis showed that CIMT how blood pressure variability caused by baroreflex failure affects cardiac was associated with MNSIE score (R2 = 0.1126, P = 0.016). In asymptomatic function in type 2 diabetes. Here we show for the first time that baroreflex type 2 diabetic subjects, CIMT is significantly associated with MNSIE score. failure aggravates diabetic cardiomyopathy via oxidative stress in nonobese Subjects with DPN are at high risk of cardiovascular disease and careful type 2 diabetic Goto-Kakizaki (GK) rats. We found that myocardial fibrosis assessment of the combined risks and intensive interventions are needed. was dramatically worsened in GK rats with sino-aortic denervation (SAD), which is the model of baroreflex failure and only increases blood pressure 440‑P variability but not blood pressure level. Also, SAD highly induced left ven- Prospective Associations of Apolipoprotein-Defined Lipoprotein tricular hypertrophy. Interestingly, SAD-induced cardiac dysfunction was Subclasses and Serum Apolipoproteins with Carotid Intima-Media strongly observed in GK rats until one year, compared with Wistar rats. Thickness in Type 1 Diabetes Also, oxidative stress, such as 4-hydroxynonenal and mRNA of NADPH oxi- ARPITA BASU, ALICIA JENKINS, JULIE STONER, YING ZHANG, RICHARD KLEIN, dase NOX2, was markedly upregulated in the heart by SAD. Furthermore, we MARIA F. LOPES-VIRELLA, W. TIMOTHY GARVEY, DAVID S. SCHADE, JAMIE R. found that the administration of metformin attenuated cardiac dysfunction WOOD, PETAR ALAUPOVIC, TIMOTHY J. LYONS, THE DCCT/EDIC RESEARCH and myocardial fibrosis in SAD-operated GK rats. On the other hand, glicla- GROUP, Stillwater, OK, Oklahoma City, OK, Charleston, SC, Birmingham, AL, zide and insulin did not prevent cardiac dysfunction and myocardial fibrosis. Albuquerque, NM, Los Angeles, CA Oxidative stress markers were reduced in the response to only metformin. Background and Aims: Type 1 diabetes (T1DM) is associated with dyslipo- Our results demonstrate that baroreflex failure-induced blood pressure vari- proteinemia and with accelerated atherosclerosis. Apolipoprotein-defined ability is involved in the development of cardiac remodeling via oxidative lipoprotein subclasses (ADLS) and serum apolipoprotein concentrations pre- stress in type 2 diabetic rats, and that metformin is effective to prevent dict vascular events in the general and type 2 diabetic populations, but data SAD-induced cardiac remodeling. As our results reflect clinical settings well, on their associations with atherosclerosis in T1DM are limited. we anticipate that baroreflex function is a next therapeutic target to prevent Methods: ADLS, serum apolipoproteins and conventional lipids were mea- diabetic cardiomyopathy and diabetic heart failure. sured in a subset (n=417) of the Diabetes Control and Complications Trial Supported By: Japan Society for the Promotion of Science (15J11642) (DCCT)/Epidemiology of Diabetes Interventions and Complications (EDIC) cohort, using samples collected in 1997-2000. The results were compared to 438‑P carotid IMT at approximately the same time, EDIC ‘year 6’ (1998-2000), and six years later at EDIC ‘year 12’). Associations were defined using multiple WITHDRAWN linear regression stratified by gender and following adjustment for concur- rent HbA1c, diabetes duration, BMI, albuminuria, DCCT treatment random- ization, smoking, statins, and ultrasound devices. Results: In men, cross-sectional associations were observed between Lp-B, Lp-B:C, Apo-B, Apo-C-III HP (heparin precipitate, i.e., Apo-C-III in VLDL) and Apo-E and common and/or internal carotid IMT. In conventional lipid pro- files, total-, LDL-, and non-HDL-cholesterol were also associated. In women, cross-sectional associations were observed between Apo-C-III (total and HP) and internal carotid IMT. Prospective associations were observed in men between Lp-B, Apo-B, Apo-C-III HP, and Apo-E and subsequent common or internal carotid IMT at EDIC year 12. In women, prospective inverse associa- tions were observed between Lp-A-I:A-II and Apo-A-1 and internal carotid IMT: there were no associations with conventional lipid profiles. Conclusions: ADLS analysis and detailed serum apolipoprotein concentra- tions may provide gender-specific biomarkers and mechanisms for vascular complications in men and women with T1DM. Supported By: National Institutes of Health

441‑P Correlation of Periodontal Disease Status with Glycemic Control, Duration of Diabetes, and Dosage of Insulin in Adolescent and Adult Subjects with Type 1 Diabetes Mellitus CHANDNI RADHAKRISHNAN, ROSAMMA J. VADAKKEKUTTICAL, INDU VENU- GOPALAN, JOSERAJ MG, Kozhikode, India Recent studies suggest that inflammatory mediators have a broader role in type 1 diabetes mellitus (T1DM), but the evidence for the correlation of periodontal status with glycemic control, duration of diabetes and dosage of 439‑P insulin in T1DM is scarce. This hospital based cross sectional study assessed Diabetes and Carotid Atherosclerosis in the proportion and severity of periodontal disease in T1DM as well as the Asymptomatic Type 2 Diabetic Subjects correlation between the duration of diabetes, dosage of Insulin, glycemic TAE SEO SOHN, HANNAH SEOK, HYUN SHIK SON, Uijeongbu, Republic of Korea control and periodontal disease severity. Study comprised of 64 T1DM and Diabetic peripheral neuropathy (DPN) is the most prevalent chronic com- 64 systemically healthy subjects with an age range of 14 to 59. The subjects plications of diabetes with high mortality rates and carotid intima-media were assessed for periodontal parameters {Modified gingival index, Plaque thickness (CIMT) has been shown to predict cardiovascular risk. However, index, OHI-S index, DMFT index, Probing pocket depth, and CAL (Clini- little is known about the association between DPN and atherosclerotic cal attachment loss)} and systemic parameters (FPG, FLP, and HbA1c). The vascular changes. The aim of this study is to investigate the association groups were comparable in terms of mean age and socio demographic char- between DPN and CIMT in asymptomatic type 2 diabetic subjects. The pres- acteristics. T1DM group had higher proportion of periodontitis (71.9%) com- ent study included 114 subjects with T2DM (age 57.4 ± 8.0, diabetes duration pared to control group (28.1%). Periodontal disease severity was significant 9.2 ± 6.4 years, HbA1c 7.5 ± 1.6%, eGFR 79.3 ± 1.6 ml/min/1.73 m2). DPN between T1DM group (39.1%) and control (l7.8%), (p = 0.000). There was no

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A116 COMPLICATIONS—MACROVASCULAR—ATHEROSCLEROTIC CARDIOVASCULAR DISEASE AND HUMAN DIABETES statistically significant difference in the disease severity among T1DM sub- In our study, cFLC was associated with carotid atherosclerosis, measured jects when stratified based on dosage of insulin (< 30, 30-39, 40-49, >50 IU). by the B-score in subjects with T2DM. However, further larger trials need A negative correlation was observed between dosage of insulin and mean to confirm this association and determine whether cFLC could be used as CAL among T1DM subjects (Pearson Correlation coefficient -0.039, p=0.757). a potential biomarker for atherosclerosis and future cardiovascular risk in The periodontal disease severity was significantly higher for T1DM patients patients with T2DM. with ≥12 years of duration of diabetes (p value=0.000). A positive correlation was observed between duration of diabetes and mean CAL (r =0.530, p value 444‑P 0.000). A positive correlation was found between HbA1c level and mean CAL Stress Hyperglycemia Ratio, an Index of Relative Hyperglycemia, POSTERS among the study subjects (r=0.351, p value 0.000). This study showed that as a Predictor of Clinical Outcomes after Percutaneous Coronary Complications the proportion and severity of periodontal disease is higher among T1DM Intervention Acute and Chronic patients. YEOREE YANG, TAE HOON KIM, EUN YOUNG LEE, JAE HYOUNG CHO, KUN HO YOON, BONG YUN CHA, KIYUK CHANG, SEUNG HWAN LEE, Seoul, Republic of 442‑P Korea Oxidative and Advanced Glycation End Products Predict Cardiovas‑ Objective: We aimed to investigate the outcome-predicting value of cular Disease in Type 1 Diabetes a novel index of stress hyperglycemia in coronary artery disease (CAD) PAUL J. BEISSWENGER, IONUT BEBU, SCOTT HOWELL, HELEN PAN, JOHN M. patients who underwent percutaneous coronary intervention (PCI). LACHIN, THE DCCT/EDIC RESEARCH GROUP, Lebanon, NH, Rockville, MD Research Design and Methods: Stress hyperglycemia ratio (SHR) was cal- Background: People with type 1 diabetes have a 10-fold increased risk of culated by dividing the first-measured random serum glucose at admission cardiovascular disease (CVD) relative to those without. New biomarkers are with estimated average glucose derived from HbA1c. 4362 subjects from the needed for early detection. We investigated the association and predictive CathOlic university of Korea percutAneous Coronary inTervention (COACT) value of Oxidative Products and Advanced Glycation End products (OP and registry were used to estimate the risk of major adverse cardiovascular and AGEs) for CVD events over 30 years in the Diabetes Control and Compli- cerebrovascular events (MACCE) defined as a composite of all-cause death, cations Trial and its follow-up Epidemiology of Diabetes Interventions and non-fatal myocardial infarction (MI) and non-fatal stroke. Complications (DCCT/EDIC). Results: Over a median follow-up of 900 days, 344 (7.9%), 43 (1.0%), and Methods: Using a case-control design, we measured the plasma OP 89 (2.0%) cases of death, non-fatal MI, and non-fatal stroke occurred. Com- sulfoxide (METHSO) and AGEs carboxyethyllysine (CEL) and 3 pared with the subjects in the lower three quartiles of SHR, the HR (95% CI) deoxyglucosone hydroimidazolone (3DGH) by LC/mass spectrometry in 93 of the highest SHR quartile (Q4) group for MACCE was 1.31 (1.05, 1.64) in CVD cases and 366 controls at baseline, year 1, DCCT closeout, and year 1-2 total population and 1.45 (1.02, 2.06) in nondiabetic population, after adjust- of EDIC. CVD included nonfatal MI and stroke, CVD death, confirmed angina, ing potential covariates. The risk of MACCE in SHR Q4 group was signifi- congestive heart failure, and coronary revascularization. Cox models looked cantly higher in patients presenting as ST-elevation MI (STEMI), but not in at associations between biomarkers as fixed (baseline and early EDIC) or patients presenting as other types of CAD. The prognostic impact of SHR time-dependent (all 4 time points) covariates and CVD. To assess prediction was more prominent for 30 days MACCE. Similar results were observed in improvement, C-statistic was computed for biomarkers that were signifi- another cohort consisting of patients presenting as acute MI only. cantly associated with the outcomes in fully adjusted models. Conclusions: SHR is a useful predictive marker of MACCE after PCI, espe- Results: Baseline CEL was associated with CVD (p=0.03, HR 1.15) when cially in nondiabetic patients with STEMI. It could be utilized for identifying fully adjusted for all covariates (Age, A1c, BMI, HDL, LDL, Sex, SBP, and DBP), the patients who are at higher risk of adverse outcomes. while METHSO at EDIC beginning and in the time dependent analysis was strongly inversely associated with CVD (p < 0.001 and HR 0.58 and 0.60, 445‑P respectively). Adding METHSO also significantly increased the predictive Increased Internal Carotid Flow Resistivity Index in Patients with c-statistic from 0.711 to 0.783 (p<0.001) when added to the other covariates. Long-Term Type 1 Diabetes: A Reflection of Cerebral Microvascular Conclusions: High MetSO, a product of the free oxygen radical scavenger ? methionine, is associated with lower CVD incidence during DCCT/EDIC, and FANG FANG, MARNI GREIG, LEANNE HUNT, DINESH SELVARAJAH, SOLOMON significantly improves predictive value over traditional risk factors. Baseline TESFAYE, IAIN D. WILKINSON, Shanghai, China, Sheffield, United Kingdom levels of the AGE CEL may also predict later CVD. Early identification of CVD Aims: Although disease duration is the main risk factor for microvascular risk in type 1 diabetes may facilitate early preventive therapies. complications, few studies have focused on brain small vessel disease (SVD) Supported By: National Institute of Diabetes and Digestive and Kidney Diseases in type 1 diabetes (T1DM). We investigated brain SVD and internal carotid (1R44DK101226-01A1) artery (ICA) flow resistivity index (RI) in patients with varying durations of T1DM. 443‑P Methods: 66 subjects (49 T1DM and 17 HV) with no history of cardio- Polyclonal Serum Free Light Chains Levels Are Associated with vascular disease/stroke were assessed and grouped by diabetes duration Carotid Atherosclerosis in Subjects with Type 2 Diabetes Mellitus [Short≤15yrs, Medium=15-30yrs; Long≥30yrs]. Lacunes, white matter FELIX ABERER, NORBERT J. TRIPOLT, MICHAELA EDER, HUBERT SCHARNAGL, WMH and were rated on T2 MRI. Quantitative ICA MR angiography TATJANA STOJAKOVIC, HARALD SOURIJ, Graz, Austria assessed carotid RI. Patients with type 2 diabetes mellitus (T2DM) face a 2-4 fold increased Results: There were significant differences in group mean ICA RI (HV= risk for cardiovascular events compared to nondiabetic counterparts. Chronic 0.54±0.05, Short=0.56±0.08, Medium=0.63±0.06, Long term= 0.65±0.06; inflammation and activation of the immune system plays an important role P<0.001). Post-hoc analysis indicated higher RI in Long and Medium-term in atherosclerosis and plaque formation. Recent investigations suggest com- compared to both Short-term (P<0.005; P<0.05) and HV (P<0.001; P<0.001) bined serum free light chains (cFLC) as a potential biomarker for cardiovas- groups. There were no significant differences in group prevalence of imag- cular events. The aim of this analysis was to investigate the association of ing abnormalities (χ2=7.064, P=0.07). In cases of no imaging abnormali- cFLC with carotid atherosclerosis in subjects with type 2 diabetes. ties, ICA RI varied with retinopathy stage (none=0.53±0.05, pre-prolifera- We performed a cross-sectional analysis of data from a prospective single tive=0.56±0.08, proliferative=0.63±0.07; P<0.05). ICA RI correlated with center 2-year study of 97 patients with T2DM and insufficiently controlled duration of diabetes (r=0.6, P<0.001), prevalence of retinopathy (r=0.6, cardiovascular risk factors. Complete data on cFLC, high sensitive C-reactive P<0.001), peripheral neuropathy (r=0.3, P<0.05) and higher SVD score (r=0.3, protein (hsCRP) were available of 75 subjects. Carotid intima media thick- P<0.04), adjusting for age. ness (CIMT) and a B-score to quantify carotid atherosclerotic plaque burden Discussion: Changes in ICA RI paralleled diabetes duration. Prevalence were measured with ultrasound. The data were compared using explorative of diabetic retinopathy, polyneuropathy and higher severity of cerebral SVD statistical analysis. (all microangiopathic complications) were associated with ICA RI. Even in We analyzed data of 26 female and 49 male subjects, aged 59 ± 8 years. patients without evidence of cerebral SVD on standard MRI, ICA RI was Their mean BMI was 31.6 ± 4.4 kg/m2, the diabetes duration was 8 ± 6 years, higher in those with retinopathy than those without. This data infers ele- the median B-score of the carotid were 2 (IQR 0-3). Significant corre- vated ICA RI occurs prior to abnormalities on standard imaging, suggesting lations between cFLC and the B-score (r=0.32; p=0.007) as well as cFLC and RI is an early marker for cerebrovascular involvement. Such an index may aid hsCRP (r=0.35; p=0.002) were observed, after adjustment for age, sex and initiation of prophylactic therapeutic regimens. renal function. The correlation between cIMT and cFLC (r=0.22; p= 0.058) did Supported By: European Foundation for the Study of Diabetes; Wellcome Trust not reach statistical significance. UK

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446‑P 448‑P Postprandial Microvascular Reactivity Is Significantly Modified by Effect of Acute and Postprandial State on the Endogenous Insulin in Recently Diagnosed Type 2 Diabetic Patients Endothelial Function in Patients with Type 2 Diabetes and Metaboli‑ MARTIN PRÁZNÝ, EVA HOROVÁ, JAN ŠKRHA, JR., JAN ŠOUPAL, MARTIN cally Healthy Volunteers with or Obesity VEJRAŽKA, JAN ŠKRHA, Prague, Czech Republic JIRI VELEBA, LENKA BELINOVA, HANA MALINSKA, HANA KAHLEOVA, KAT- Aim: To analyze factors modifying microvascular reactivity (MVR) in post- ERINA VELEBOVA, EVA STOLARIKOVA, TEREZIE PELIKANOVA, Prague, Czech prandial state in early type 2 diabetes (T2DM). Republic

POSTERS Methods: Twenty patients with T2DM (58±6 yrs, T2DM 2.3±1.3 yrs, met- Background and Aims: Endothelial dysfunction has been linked to the

Complications formin only) and 14 control subjects (CON) were examined at fasting and 60 pathogenesis of cardiovascular disease. Mechanisms involved in the regula- Acute and Chronic (T60) and 180 (T180) minutes after standard breakfast. MVR was measured tion of vascular reactivity are not fully understood. The aim of the study was by laser Doppler flowmetry during post-occlusive hyperemia (PORH) at fin- to test the effects of isolated acute hyperinsulinemia and postprandial state gertip and thermal hyperemia (TH) in the forearm. Lipids (total, LDL- and HDL- on the vascular reactivity. cholesterol, ), HbA1c, glucose and insulin levels were evaluated. Materials and Methods: We enrolled 30 patients with type diabetes (T2D) Results: Postprandial glucose was higher in T2DM vs. CON. Insulin was on metformin treatment and 30 volunteers (C) with overweight or obesity non-significantly higher in T2DM vs. CON. Negative correlations were found and with normal glucose . All subjects underwent the hyperinsu- at fasting between HbA1c, glucose and the perfusion at forearm. A weak linemic euglycemic clamp combined with indirect calorimetry and meal test positive correlation between MVR at fingertip and insulin led us to further using standard breakfast. Parameters of the endothelial function - Augmen- analysis. We divided the patients into tertiles based on their insulinemia in tation Index (AI) and Reactive Hyperemia Index (RHI) were measured by fin- T60. The insulin in the low tertile (LOW) was 68±7 mU/L, in medium (MED) ger pulse plethysmography before and during the clamp and before and after 103±21 mU/L and in the high (HIGH) 219±53 mU/L (p<0.003). Basal fingertip the meal test. Repeated-measures ANOVA models, multivariate regression perfusion at T60 and T180 was higher in HIGH vs. LOW tertile (181±78 vs. using the method of orthogonal projections to latent structure (OPLS) and 72±57 PU, p=0.014 and 134±46 vs. 76±50 PU, p=0.048). PORH at T60 and Pearson’s correlation coefficient were used for statistical analysis. TH at T180 was higher in the HIGH vs. LOW tertile (255±59 vs. 156±93 PU, Results: AI did not change in response to hyperinsulinemia in either group. p=0.030 and 158±52 vs. 85±33 PU, p=0.012). TH at T180 was higher also in RHI tended to decrease in C (∆RHIC: -0.112±0.71, p>0.05), while there was a HIGH vs. MED tertile (158±52 vs. 88±33 PU, p=0.015). No differences in MVR trend toward increase in T2D (∆RHIT2D: 0.084±0.062, p>0.05). There was a were found between LOW and MED. Although insulin was higher in HIGH statistically significant difference between the groups over time (Gxt p<0.05). tertile compared to CON (219±53 vs. 118±58 mU/L, p<0.001), PORH after AI decreased in both groups in response to meal test (∆AIT2D: -10.5±2.781, meal still remained significantly lower in HIGH tertile vs. CON. Oppositely, p<0.001; ∆AIC: -7.631±2.03, p<0.01), whereas RHI did not change. RHI was TH at T180 was higher in the HIGH vs. CON (158±52 vs. 109±48 PU, p=0.045). one of the independent predictors of type 2 diabetes (p<0.01), along with Conclusions: Low endogenous insulin levels were associated with impaired HBA1c, FPG, REE, and fasting oxidation of . endothelium dependent MVR in recent T2DM. Further research should clar- Conclusion: Postprandial state decreases arterial stiffness during meal ify whether insulin treatment restores MVR which would support early use test. RHI is strong predictor of type 2 diabetes. of insulin treatment in the clinical practice. Supported By: Ministry of Health of the Czech Republic (NCT01946347) Supported By: Agency for Healthcare Research of the Czech Republic (P25/ LF1/2, 15-26705A) 449‑P Evaluation of Cerebral Vessel Disease by 7-T MRI in Type 2 Diabetes 447‑P SATOSHI YASHIRO, KAN NAGASAWA, MAKOTO SASAKI, YASUSHI ISHIGAKI, The Serum Levels of Trimethylamine-N-Oxide (TMAO) in Patients Morioka, Japan with Type 2 Diabetes/ and Chronic Heart Failure: The Objective: Type 2 diabetes (T2DM) is known to be a major risk of ischemic Effect of Metformin stroke. However, it is still controversial whether the cerebral small vessel KATERINA VELEBOVA, JIRI VELEBA, EVA STOLARIKOVA, HANA KAHLEOVA, disease (SVD), including white matter hyperintensities, lacunar infarctions JAN KOPECKY, JR., ONDREJ KUDA, MARKETA SEGETOVA, JAN KOPECKY, SR., and microbleeds, occur with greater frequency in the subjects with T2DM. VOJTECH MELENOVSKY, TEREZIE PELIKANOVA, Prague, Czech Republic To determine cerebral microvascular changes as the base of SVD, stenotic Introduction and Aims: TMAO is a product of gut microbiota. Elevated lesions of the lenticulostriate arteries (LSA) were analyzed in T2DM using circulating levels are associated with the progression of atherosclerosis ultra-high resolution 7-T MRI. and increased risk of fatal cardiovascular events. The influence of pharma- Methods: Twenty-five subjects with T2DM (mean age 57.2 years) and 25 cological intervention on TMAO levels was not investigated till now. The nondiabetic control subjects (mean age 60.1 years) underwent 7-T brain MRI aims of the study were a) to compare the serum levels of TMAO in patients scan, in evaluating the incidence of SVD. The stenotic changes of LSA were with type 2 diabetes/prediabetes and chronic heart failure (DM+HF) and in examined by MR angiography, following the slab maximum intensity projec- healthy individuals (H), b) to evaluate the effect of metformin in DM+HF and tion technique. c) to explore possible association of the TMAO levels with metabolic risk Results: The occurrence of LSA stenosis was significantly high in T2DM factors, insulin resistance and diabetes control. compared with control subjects (68.0% vs, 16.0%, p < 0.01). Intriguingly, the Methods: 26 DM + HF and 21 H were enrolled. DM+HF underwent the six- incidence of SVD had no significant difference between T2DM and control month randomized, interventional, double-blind, placebo-controlled, cross- (60.0% vs, 44.0%). In the analysis limited to T2DM, the occurrence of LSA over study, testing the effect of 3-month metformin intake (2g/day) vs. stenosis was associated with low ratio of both eicosapentaenoic acid to placebo on selected metabolic parameters, including TMAO levels. At the arachidonic acid (AA) and docosahexaenoic acid to AA. baseline and at the end of each intervention period the panel of metabolic Conclusion: Current study revealed the occurrence of LSA stenosis was and cardiovascular tests was done. It included the routine clinical laboratory markedly higher in the subjects with T2DM, while the incidence of SVD had tests, measurement of the serum levels of TMAO and the hyperinsulinemic less frequent. This result indicated that the pathological process of SVD euglycemic clamp (3 hrs) for assessment of insulin sensitivity. development in T2DM could be visualized by using ultra-high resolution MRI Results: The TMAO levels were higher in DM+HF compared to H (0.81±0.56 and newly developed imaging program. In addition, high serum concentra- vs. 0.5±0.35 ug/ml; p<0.04) and positively correlated with age (r=0.52; tion of omega-3 polyunsaturated might associated with reduced p<0.001) and fasting plasma glucose (r=0.43; p<0.001) in H. However, in risk of cerebral SVD. Taken together, LSA stenosis is potentially early change DM+HF the serum levels of TMAO were not related to any of the metabolic prior to SVD in T2DM and this technique provides important findings for the variable (HbA1c, serum lipids, BMI or MCR) and did not significantly change minute changes in cerebral microvessels. after metformin. Conclusions: The TMAO levels are higher in DM+HF compared to H. The 450‑P metformin treatment does not significantly influence the TMAO levels. Our Does Strenuous Activity Affect the Impact of Nonsevere Hypoglyce‑ results do not support the relationship between the TMAO levels and the mia on Coronary Artery Calcification? metabolic control of diabetes or insulin resistance in patients with DM+HF. ELKE R. FAHRMANN, LAURA ADKINS, HENRY K. DRISCOLL, Huntington, WV Supported By: Ministry of Health of the Czech Republic (AZV16-27496A) Research has indicated that strenuous and hypoglycemia inde- pendently activate inflammatory pathways, which may result in a predis- position to atherosclerosis. However, it is unknown whether an interaction between very hard activity and nonsevere hypoglycemia (NSH) in coronary

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A118 COMPLICATIONS—MACROVASCULAR—ATHEROSCLEROTIC CARDIOVASCULAR DISEASE AND HUMAN DIABETES artery calcification (CAC) exists. We hypothesize that time spent on strenu- vascular resistance and cardiac workload (pulse pressure (PP), mean arte- ous activity alters the relationship between NSH and CAC. rial pressure (MAP) and the double product (DP)) were assessed across We analyzed Diabetes Control and Complications Trial/Epidemiology of subgroups of age, sex, and SBP, using a mixed model repeated measures Diabetes Intervention and Complications (DCCT/EDIC) data of type 1 dia- analysis in randomized pts who received ≥1 dose of study drug using all betes patients. Log-binomial models (adjusted for relevant baseline differ- measurements obtained until study end. 2333, 2345 and 2342 pts received ences) were utilized to evaluate associations between variables of interest PBO, EMPA 10 mg or 25 mg and mean±SE baseline SBP was 135.79±0.36, and CAC score >100 Agatston units (CAC100) and validated by sensitivity 134.91±0.35, 135.65±0.35 mmHg, respectively whereas mean±SE heart analyses. rate was 70.74±0.23, 70.96±0.22 and 70.52±0.22 beats per minute. There POSTERS

During DCCT: 1173 patients reported time spent on strenuous activity. were significantly greater reductions in markers of arterial stiffness, vas- Complications

Mean NSH was 1.8 ±1.4 events/week, and mean time of activity was 35 ± cular resistance and cardiac workload among pts treated with either EMPA Acute and Chronic 84 min/week (max mean: 864 min/week). NSH and strenuous activity were dose vs. PBO across age, sex, and SBP groups (Table). EMPA consistently weakly correlated (rho=0.08, p<0.01). Length of time spent on strenuous improved cardiac and vascular hemodynamic markers, irrespective of age, activity modified the association between NSH and CAC100; with more pro- sex, or baseline SBP. Further analyses will determine the potential contribu- longed strenuous activity, NSH became a more obvious atherosclerosis fac- tion of these changes to the CV mortality benefit with EMPA. tor. Significance threshold was 190 min/week on strenuous activity. Strati- Table. fication by gender revealed a significant interaction between time spent on strenuous activity and NSH, albeit for males only (p<0.001). During EDIC: 1177 patients reported time spent on strenuous activity. Mean NSH was 2.3 ±1.6 events/week, and mean time of exercise was 18 ± 55 min/week (max mean: 627 min/week). In contrast to DCCT, strenuous activity and HbA1c were correlated (rho=0.11, p<0.001). The interaction between strenuous activity and NSH was not significant; however, interac- tion among strenuous activity, NSH, and prior treatment was. If confirmed, attention should be given to NSH frequency, strenuous activ- ity and time spent on it, and patients should be managed accordingly.

451‑P Severe Cardiac Pathology Correlates with Marked Islet Amyloid Deposition in Patients with Acute Myocardial Infarction and Type 2 Diabetes KAZUHISA TAKAHASHI, HIROKI MIZUKAMI, WATARU INABA, SOROKU YAGI- HASHI, Hirosaki, Japan Acute myocardial infarction (AMI) is the most frequent cause of death in patients with type 2 diabetes (T2DM). Recent studies suggested that such macrovascular diseases may themselves be implicated in the high preva- lence of T2DM in part related to impaired b-cell function. It still remains unclear, however, whether the structural changes of the islets in T2DM have something to do with the pathology of AMI. We attempted to character- ize the pathological alterations of islets and their relations to the cardiac changes in patients with AMI subjects complicated with T2DM. 15 non- Supported By: Boehringer Ingelheim diabetic cases (ND), 14 nondiabetic cases with AMI (AMI), 31 cases with T2DM and AMI (A-DM), and 13 cases with T2DM but without AMI (DM) 453‑P were investigated. Age, duration of diabetes, BMI and average HbA1c were Pulse Wave Velocity and Autonomic Function in Type 2 Diabetes matched among the groups for comparison. Amyloid deposition in the islets ANASTASIOS TENTOLOURIS, IOANNA ELEFTHERIADOU, PINELOPI GRIGORO- was evaluated with double immunostaining with thioflavin T and chromo- POULOU, STAMATIA CHOREPSIMA, GIORGOS KONSTANTONIS, ATHANASIOS granin A (CgA). Islet microvasculature was evaluated on immunostained sec- PROTOGEROU, NIKOLAOS TENTOLOURIS, Athens, Greece tions for CgA and CD31. Size of cardiac cells was also evaluated. As results, Increased carotid-femoral (cf) pulse wave velocity (PWV) reflects arte- cardiac weight was increased 1.2 fold in AMI compared to ND (p<0.05). It rial stiffness and is associated with cardiovascular risk. Cardiac autonomic was further increased 1.2 fold in A-DM compared to AMI (p<0.05). Islet dysfunction (AD) is a complication of diabetes and altered heart rate vari- amyloid volume density (Va) was increased 9 times in A-DM compared to ability (HRV) is one of its earliest manifestations. This study investigated DM (p<0.05). Average size of cardiomyocytes was significantly increased the association of arterial stiffness with AD in diabetes. A total of 290 in A-DM compared to AMI. Va in A-DM correlated well with cardiac weight patients with type 2 diabetes (T2DM) were included. cfPWV was measured (r=0.4, p<0.05) and size of cardiomyocytes (r=0.5, p<0.01). There was a 2-fold at the carotid-femoral segment with applanation tonometry. Participants increase in intra-islet microvessel area in AMI without an increase in the were classified as having normal (n=193) or abnormal (n=97) cfPWV values number of vessels compared to other groups (p<0.01). In contrast, periph- using age-corrected values. Cardiac autonomic nervous system activity eral-islet micro vessel area and number of microvessels was significantly was determined by measurement of parameters of HRV, while baroreflex increased in A-DM compared to other groups (p<0.01). Taken together, we sensitivity (BRS) was measured using the spontaneous sequence method. conclude that the presence of T2DM in cases with AMI is associated with Participants with abnormal cfPWV had higher arterial blood pressure (BP) augmented cardiac pathology with severe amyloid deposition and microves- and HR, higher total cholesterol and more often peripheral neuropathy as sel changes in the islet. compared to those with normal cfPWV. BRS did not differ between the 2 groups. Participants with abnormal cfPWV had significantly lower values 452‑P of high frequency (HF) power, total power of HRV, square root of the mean Effects of Empagliflozin on Cardiac and Vascular Hemodynamic of the squares of successive differences between adjacent NN intervals Markers by Subgroups of Age, Sex, and Hypertension in Patients (r-MSDD)]. Univariate logistic regression analysis showed that age, systolic, with T2DM and High CV Risk: EMPA-REG OUTCOME diastolic and central mean BP, HR, total cholesterol, triglycerides, treatment ROBERT J. CHILTON, LARS GULLESTAD, DAVID FITCHETT, SILVIO E. INZUCCHI, with statins, peripheral neuropathy and several parameters of HRV were MICHAELA MATTHEUS, HANS JUERGEN WOERLE, ODD ERIK JOHANSEN, associated with abnormal cfPWV. Multivariate analysis, after controlling for San Antonio, TX, Oslo, Norway, Toronto, ON, Canada, New Haven, CT, Ingelheim, age, HR, treatment with statins and peripheral neuropathy, demonstrated Germany, Asker, Norway that abnormal cfPWV was associated independently with central mean BP Empagliflozin (EMPA) significantly reduced the risk of CV death in the (OR=1.052, 95% CI 1.016-1.088, p=0.004), Lg of total power (OR=0.490, 95% EMPA-REG OUTCOME trial. We explored for differential effects on car- CI 0.258-0.932, p=0.030), Lg of HF power (OR=0.548, 95% CI 0.302-0.991, diac and vascular hemodynamic markers by age, sex, and systolic BP (SBP) p=0.047), and Lg r-MSDD (OR=0.565, 95% CI 0.317-1.008, p=0.053). Cardiac subgroups. Pts with T2D and high CV risk were randomized to receive pla- AD is strongly associated with increased cfPWV in people with T2DM and cebo (PBO) or 2 doses of EMPA. Changes in indices of arterial stiffness, should be monitored for cardiovascular risk stratification.

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454‑P 456‑P Pulse Wave Velocity Rather than Atherosclerotic Markers Describe Postprandial Increase of Serum -19 (FGF- Vascular Status of Patients with Type 2 Diabetes Mellitus 19) Levels after Fat Tolerance Test Was Significantly Lower and BERND STRATMANN, MAGDALENE RYDZKOWSKI, DIETHELM TSCHOEPE, Bad Delayed in Peak-Time in Subjects with Prediabetes and Diabetes Oeynhausen, Germany than Normal Controls Type 2 diabetes mellitus (T2DM) is associated with high risk of atheroscle- SEO YOUNG LEE, SUNG HEE CHOI, TAE JUNG OH, KYOUNG MIN KIM, JAE HOON rosis. Common risk factors for atherosclerosis are compared to the vascular MOON, SOO LIM, HAK CHUL JANG, Gyeonggi, Republic of Korea

POSTERS status determined by non-invasive arteriography. Patients were grouped: We aimed to investigate the clinical implication of fasting and postpran-

Complications T2DM without coronary artery disease (CAD) (T2DM, N=50), T2DM with dial FGF-19 in human. Especially, the association between postprandial Acute and Chronic CAD (T2DMCAD, N=44). Pulse wave velocity (PWV) was measured using excursion of FGF-19 after fat-meal challenge test and cardiovascular risk TL1 TensioClinic™ (Tensiomed, Bietigheim-Bissingen) and patients were markers. We compared postprandial FGF-19 response in participants with categorized in patients with abnormal or normal PWV results. Markers of IGT (n=5) or with DM (n=5) and in normal healthy participants (n=15) after atherosclerotic risk were assessed in plasma or serum using commercially standardized fat-rich meal and performed comprehensive analysis between available ELISA-kits. Patients groups were age-, BMI-, and sex-matched; FGF-19 and other cardiometablic risk factors, such as intima medial thickness HbA1c, systolic blood pressure, LDL and HDL-cholesterol were comparable. (IMT). Twenty six men, ages 23-47 (35.5±6.8), with a BMI range of 20.7-31.0 Diabetes duration was longer in T2DMCAD (15.4±8.4 years) than in T2DM kg/m2 (25.1±2.6 kg/m2) finished the standardized fat tolerance test. Median (10.9±8.0 years, p<0.05). 38% of patients with T2DM without CAD pres- value of maximum plasma FGF-19 showed a decrease tendency from healthy ent with abnormal high PWV [11.4±0.2m/s (T2DM+, N=31) vs. 8.1±0.2m/s control group to IGT group to T2DM group (p for trend = 0.09), although fast- (T2DM-, N=19), p<0.001], comparable to those with CAD [12.1±0.3m/s ing plasma FGF-19 did not (p for trend=0.180). The postprandial pattern of (T2DMCAD+, N=30) vs. 8.8±0.2m/s (T2DMCAD-, N=14), p<0.001]. BNP was serum FGF-19 was similar to postprandial pattern of plasma TG after fat significantly increased in T2DMCAD+ if compared to T2DM+ or T2DM- loading diet. Postprandial FGF-19 levels showed a maximum increase after (74.6±16.5 pg/ml vs. 34.1±8.5 pg/ml vs. 27.8±3.4 pg/ml; p<0.05) whereas 2 hours in healthy control group and after 4 hours in subjects with IGT or Lp(a), , , and hsCRP did not differ between the groups. T2DM. Postprandial FGF-19 levels returned to baseline value after 4 hours in Regarding markers of turnover and fibrosis, no differ- healthy control group, whereas it returned to baseline value after much later ences were detected within the groups for MMP-2, MMP-9, TIMP-2, and (6 hours) in IGT or T2DM group. When participants divided to 2 groups who Galectin-3, but TIMP-1 levels were significantly elevated in T2DM+ if com- were the peak time of postprandial FGF-19 below 2 hours (early responder) pared to T2DM- (210.6±10.6ng/ml vs. 176.2±7.5ng/ml, p<0.05). Pulse wave and above 2 hours (late responder), the late responder had definite athero- velocity is a powerful tool to assess the vascular status of T2DM patients. sclerotic change on carotid artery (maximum IMT of CCA, 0.72±0.09 mm in Lowering of established cardiovascular risk factors (lipid parameters, blood late responder vs. 0.60±0.10 mm in early responder, p=0.004). The maximum pressure) may contribute to the normalisation of laboratory markers that level of FGF-19 after fat-rich meal was much lower level and showed delayed correlate to the atherosclerotic risk. The high prevalence of vascular disease peak time (4 hours) in IGT or T2DM group than healthy control groups (2 in T2DM has to be taken into account when evaluating the vascular function hours). Also, the maximum carotid IMT was thicker significantly in patients using laboratory markers. with the peak time of FGF-19 level above 2 hours.

455‑P 457‑P Hemoglobin Glycation Index Is Independently Associated with Car‑ Evaluation of the Impact of Pioglitazone on Stroke Risk and Stroke diovascular Diseases in People with Impaired Glucose Metabolism: Outcomes: A Retrospective, Observational Study Analysis of a Prospective Patient Registry CHRISTOPHER L. MORGAN, SARA JENKINS-JONES, JORGE PUELLES, CRAIG J. CHANG HO AHN, SE HEE MIN, DONG WHA LEE, TAE JUNG OH, KYOUNG MIN CURRIE, SILVIO E. INZUCCHI, Cardiff, United Kingdom, London, United Kingdom, KIM, JAE HOON MOON, SUNG HEE CHOI, KYONG SOO PARK, HAK CHUL JANG, New Haven, CT JOON HA, ARTHUR SHERMAN, SOO LIM, Seoul, Republic of Korea, Seongnam, Introduction: RCT data from the PROactive trial demonstrated a potent Republic of Korea, Bethesda, MD effect of pioglitazone in reducing recurrent stroke in patients with T2DM. There is interindividual variation in the association between glycated More recently, the IRIS trial showed pioglitazone decreased the risk of vas- hemoglobin (HbA1c) and plasma glucose concentrations. The impact of this cular events following stroke or TIA in those with insulin resistance but with- variation on vascular complications of diabetes is controversial. We aimed out diabetes. The purpose of this study was to investigate this important to examine the association between disproportionately high HbA1c lev- pleiotropic effect in people exposed to pioglitazone as a glucose-lowering els compare to plasma glucose concentrations, which is estimated as the therapy. hemoglobin glycation index (HGI), and cardiovascular diseases in people Methods: Routine data from UK NHS sources for people with T2DM were with impaired glucose metabolism. We analyzed a patient registry of 1248 extracted from January 2000 to December 2012, with follow-up to 2016. treatment-naïve subjects with prediabetes or diabetes. HGI was defined as Cases were selected if exposed to pioglitazone >=90 days. The index date the measured HbA1c minus predicted HbA1c which was calculated from the was first prescription for pioglitazone. Patients with prior history of heart linear relationship between HbA1c and fasting plasma glucose. The preva- failure were excluded. Non-exposed controls were matched on age, sex, lence of cardiovascular diseases (CVD) including coronary artery disease HbA1c, diabetes duration, stroke history, co-morbidities, and prior glucose- (CAD), stroke and peripheral artery disease (PAD) were assessed according lowering regimen. Outcomes were risk of incident stroke, risk of fatal stroke, to the tertile of HGI. The overall prevalence of composite CVD, individual stroke morbidity and stroke recurrence. Time to event was evaluated using CAD, stroke, and PAD were 10.3% and 5.7%, 5.1%, and 1.3%, respectively. the Cox proportional hazards model adjusted for potentially confounding All of these prevalences significantly increased from the 1st to 3rd HGI factors, including atrial fibrillation. tertile. In multivariate analysis, the highest HGI tertile was independently Results: 4,484 patients were included in each arm of the study; 106 (2.4%) associated with composite CVD (odds ratio, [95% confidence interval]: 2.81 in each arm had prior history of stroke. For patients with no prior history [1.59-4.98]), CAD (2.30 [1.12-4.73]), stroke (3.40 [1.50-7.73]), and PAD (6.37 of stroke, there were 42 (4.1 per 1,000 patient years [pkpy]) stroke events [1.18-34.33]) after adjustment for traditional CVD risk factors and HbA1c lev- whilst exposed to pioglitazone compared with 71 (8.2 pkpy) in the control els. Two consecutive measurements of HGI obtained on different days were arm. Respective figures for the entire follow-up period were 140 (5.7 pkpy) in good correlation (r = 0.651, P <0.001) and high concordance in tertile clas- vs. 184 (8.5 pkpy). After adjustment the hazard ratio (HR) relative to controls sification (69.1%). The highest HGI tertile in the two consecutive measure- was 0.489 (0.329-0.727) on treatment and 0.661 (0.524-0.833) for the entire ments was also associated composite CVD, CAD, stroke, and PAD than the follow-up period. For patients with prior history of stroke, the respective HRs lower tertiles. High HGI was independently associated with macrovascular were 0.607 (0.277-1.331) and 0.597 (0.354-1.007). complications of diabetes and can be used as an indicator of vascular health Discussion: In support of evidence from two randomized trials, these in patients with prediabetes or diabetes. observational data show that pioglitazone has an apparent potent effect in reducing stroke events in people with T2DM. Supported By: Takeda Pharmaceutical Company Limited

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A120 COMPLICATIONS—MACROVASCULAR—ATHEROSCLEROTIC CARDIOVASCULAR DISEASE AND HUMAN DIABETES

458‑P 460‑P Adverse Outcomes in Subgroups of Type 2 Diabetes in the ACCORD Associations between Pulse Wave Velocity and Lifestyle Factors In Clinical Trial Elderly Subjects with T2D—The Oulu 45 Cohort DANIEL M. ROTROFF, SKYLAR W. MARVEL, JOHN R. JACK, TAMMY M. ERJA METSÄMARTTILA, JARI JOKELAINEN, SAULI HERRALA, JUHANI LEP- HAVENER, ALESSANDRO DORIA, HETAL S. SHAH, MARIO-LUCA MORIERI, HE PÄLUOTO, SIRKKA KEINÄNEN-KIUKAANNIEMI, ENRIQUE RODILLA, KARL-HEINZ GAO, JOSYF C. MYCHALECKYJ, HOWARD L. MCLEOD, JOHN B. BUSE, MICHAEL HERZIG, Oulu, Finland, Valencia, Spain J. WAGNER, ALISON A. MOTSINGER-REIF, THE ACCORD/ACCORDION INVESTI- Pulse wave velocity (PWV) is a measure of arterial stiffness, predicting

GATORS, Raleigh, NC, Chapel Hill, NC, Boston, MA, Charlottesville, VA, Tampa, FL cardiovascular disease (CVD). We examined, how various life style factors POSTERS

Comorbidities of type 2 diabetes (T2D) are risk factors for multiple and impaired glucose metabolism (IGM) and T2D affect PWV in elderly sub- Complications adverse outcomes, with T2D related mortality occurring most frequently jects born in 1945 (Oulu 45 cohort). The cross-sectional study consisted of Acute and Chronic due to cardiovascular disease (CVD). T2D heterogeneity in outcomes and 570 subjects of which 59% were female). Subjects with body mass index drug response stresses the need to identify underlying etiologies, and over 40 or irregular heart rate were excluded. PWV was determined by a recent evidence suggests that T2D may manifest from multiple etiologies. non-invasive applanation tonometry (AtCor Medical, Australia). Oral glu- Identifying subgroups of T2D may create opportunities for more precise cose tolerance test was performed and HbA1c, LDL and HDL cholesterol interventions, and the Action to Control Cardiovascular Risk in Diabetes were analyzed. Habitual physical activity was objectively registered by a (ACCORD) clinical trial is ideal for defining subtypes because it has a large wrist-worn accelerometer (Polar Electro, Finland) during wakeful time for cohort (n=10,251) with detailed drug response and adverse outcome data in two weeks. Information of life style factors was collected by question- a controlled clinical setting. A previously published study identified three naires. PWV values in IGM and T2D subjects were higher than in normal subtypes of T2D based on clinical variables from electronic medical records. glycemic subjects (P<0.001). Fasting and 2 h glucose and HbA1c correlated Here, we performed clustering of 34 analogous clinical variables in ACCORD significantly with PWV (P<0.001). PWV had a negative correlation with HDL (e.g., concomitant medications, serum creatinine, glomerular filtration rate) (P<0.05) but not with LDL cholesterol. PWV was significantly reduced as using Euclidean distance and Ward’s method clustering, which also resulted daily steps exceeded 10,000. In the multivariate analysis presence of T2D or in three groups of T2D in the ACCORD data. Subjects were assigned to their alcohol consumption in males had independent effects on PWV but presence respective group: A, B, and C, and each group was tested against the other of IGM, gender, obesity, physical activity, smoking had not. Presence of T2D groups for associations with adverse outcomes (i.e., myocardial infarction, but not that of IGM as an independent factor demonstrates that duration stroke, and CVD death) using a logistic regression model while correcting of diabetes is also a predictor of arterial stiffness. Alcohol consumption in for gender and trial arm. A significant increase in CVD death was observed males exceeded 3 g/d on average and led to increased PWV. Lower amounts in Group A compared to Groups B+C (OR=1.65 95% CI [1.29, 2.12]), whereas, of physical activity had not effect on PWV; the habitual physical activity of group B was significantly decreased for CVD death compared to Groups our subjects was evidently too low to improve arterial structures. A+C (OR=0.69 95% CI [0.53, 0.89]). These results demonstrate the prom- In conclusion, T2D significantly accelerated PWV. Interestingly, lipids did ise of this approach to provide new insight into subtypes of T2D that are not have an additional effect on PWV. High physical activity (over 10,000 at increased risk of adverse outcomes or are likely to experience better or steps per day) alleviated the aorta from stiffening. poorer response to medications. Supported By: National Institutes of Health (R01HL110380) 461‑P Relationship of Glutathione Peroxidase Activity (GPx) to Sex- 459‑P Specific Differences in Vascular Stiffness and Central Obesity in The Relationship between Circulating PCSK9 Concentration and Patients with Type 2 Diabetes Coronary Damage Severity in Patients with Cardiovascular Disease MIA STEYN, KARIMA ZITOUNI, FRANK KELLY, KENNETH A. EARLE, London, YEON KYUNG CHOI, KWI HYUN BAE, MI JIN KIM, JUNG BEOM SEO, HYE JIN United Kingdom HAM, KEUN GYU PARK, Daegu, Republic of Korea Vascular stiffness is an emerging risk factor for cardiovascular disease Proprotein convertase subtilisin/kexin type 9 (PCSK9), a circulating pro- which we previously reported to be associated with central obesity in tein that promotes degradation of the low density lipoprotein (LDL) receptor, women. We hypothesized that GPx activity would be affected differently by has been emerged as a novel therapeutic target for the dyslipidemia. It is oxidative loads from visceral fat according to gender. We studied a cohort well known that the pathogenesis of cardiovascular disease (CVD) involves of 171 patients with T2DM with characteristics in Table below. A vascular lipid metabolism alteration, but the predictive value of circulating PCSK9 stiffness index (SI) was computed using infra-red finger plethysmography. levels concerning coronary disease severity is largely unknown. The main Total body fat was assessed using bio-impedance and eGFR from the CKD- objective of this study is to determine whether circulating PCSK9 concentra- EPI equation. tion is linked to coronary damage severity in patients with acute coronary Table. syndrome (ACS). We studied 137 patients with clinically suspected ACS who underwent coronary angiography. The study population was divided into Females Males p-value (n = 85) (n = 86) two groups depending on the presence of coronary artery lesion (lesion (+): n=112, lesion (-): n=25). Baseline characteristics and PCSK9 levels were mea- Caucasian (%) 33 49 sured and coronary lesions were evaluated using the SYNTAX scoring sys- Age (years) 61.58 [6.96] 59.87 [8.31] 0.148 tem. After adjustment for established CVD risk factors including age, body Waist Circumference (cm) 101.37 [14.02] 102.84 [11.72] 0.464 mass index, total cholesterol and LDL, ACS patients with coronary artery Body Mass Index (kg/m2) 31.02 [6.9] 29.87 [4.88] 0.208 lesion have significantly higher PCSK9 levels than patients without lesion (178.26±63.9 ng/mL vs. 223.0±76.1 ng/mL, p=0.026). Spearman’s correla- Sitting Systolic Blood 138.77 [17.11] 141.30 [15.36] 0.313 tions revealed that serum PCSK9 levels were positively associated with the Pressure (mmHg) number of involved coronary artery (Pearson coefficient, 0.034; P=0.01) and Duaration of Diabetes 9.46 [6.70] 11.06 [8.04] 0.186 the global registry of acute coronary events (GRACE) risk score which is a (years) risk prediction tool applicable for ACS patients (Pearson coefficient, 0.209; HbA1c (mmol/mol) 59.38 [18.43] 55.89 [18.26] 0.219 P=0.015). Furthermore, we found that plasma PCSK9 levels were positively eGFR - CKD EPI (mls/ 89.98 [16.24] 88.76 [17.95] 0.642 correlated with SYNTAX score (Spearman’s R=0.115, P=0.048). In the pres- min/1.73m2) ent study, we show that serum PCSK9 concentration is associated with angi- Bio-impedence (%) 40.44 [8.02] 28.99 [7.63] 0.000 ographic severity of ACS and GRACE score after adjustments for established CVD risk factors. Further studies are needed to confirm this observation. Vascular Stiffness Index 9.12 [6.66] 9.99 [4.55] 0.337 (m/sec) Vitamin E, selenium and the activity of plasma GPx were assessed as markers of anti-oxidant defense. GPx activity was significantly higher in women vs. men (382.75 [99.8] vs. 343.37 [128.01], p-value 0.029) but Vitamin E and Selenium levels were similar (8.76 [2.54] vs. 8.95 [3.03], p-value 0.664 and 1.27 [0.23] vs. 1.25 [0.28], p-value 0.583 respectively). A linear regression analysis with SI as the dependant variable in women showed a relationship with waist circumference (0.150; p = 0.008) and with

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A121 COMPLICATIONS—MACROVASCULAR—CELLULAR MECHANISMS OF ATHEROGENESIS IN DIABETES

eGFR (- 0.150; p 0.006), but not with GPx. In non-Caucasian men we found 464‑P a significant positive relationship with GPx (0.02; p = 0.007) and Vitamin E Differential Effect of Weight Loss and Glucose Improvement on Lung (0.670; p = 0.69) with SI. Function in Patients with Type 2 Diabetes (Sweet Breath Study) Low GPx activity in these men is associated with increased SI where in LILIANA GUTIÉRREZ, ENRIC SÁNCHEZ, FERRÁN BARBÉ, ASUNCIÓN SEMINARIO, women higher levels could be linked to greater oxidative stress loads in rela- CECILIA TURINO, CAROLINA LÓPEZ, MARTA HERNANDEZ, FERRAN RIUS, M. tion to central obesity. DOLORES SANTOS, CRISTINA HERNANDEZ, MARTA SANCHEZ, RAFAEL SIMÓ, ALBERT LECUBE, Lleida, Spain, Barcelona, Spain

POSTERS 462‑P Objective: Type 2 diabetes (T2D) exerts a deleterious effect on lung func- Complications Autofluorescence and Pulmonary Function in Type 2 Diabetes tion, with higher prevalence of restrictive pattern. However, there are still Acute and Chronic ENRIC SÁNCHEZ, ÁNGELS BETRIU, LILIANA GUTIÉRREZ-CARRASQUILLA, no data about whether improvements in glycemic control positively influ- MARTA ELÍAS, FERRAN BARBÉ, MANEL PORTERO, FRANCESC PURROY, MIQUEL ence pulmonary function. Our aim was to examine prospectively the effect BUTI, ELVIRA FERNÁNDEZ, CAROLINA LÓPEZ-CANO, ALBERT LECUBE, Lleida, of glycemic control on lung function parameters. Spain Methods: A prospective interventional study with 55 patients with T2D Tissues that are rich in and , such as the lung, can be (HbA1c: 9.6±1.6% and BMI 31.1±6.9 kg/m2) and without known pulmonary target of non-enzymatic glycation. Therefore, the production of advanced disease. All patients underwent a titration treatment over a 3-months glycation end-products (AGEs) would collaborate in the deterioration of period. The lung function was evaluated at baseline and 3-months after of lung function previously described in type 2 diabetes (T2D). Our aim was pharmacological intervention. to access the relation between AGEs and pulmonary function because little Results: HbA1c decreased to 7.3±1.2% (p<0.001), BMI to 30.6±6.4 kg/ information exits in these patients. m2 (p=0.085). The initial 30.9% of patients with a restrictive respiratory Methods: Cross-sectional study including 747 subjects (58.1±6.4 years; pattern decreased to 14.5% (p=0.001). The absolute decrease in BMI sig- 51.3% men; 6.3% with T2D) and without known pulmonary disease under- nificantly correlated with changes in forced expiratory volume in 1 second went a baseline spirometry (Datospir©). Restrictive pattern was defined by (FEV1; r=-0.312, p=0.020) and forced vital capacity (FVC; r=-0.266, p=0.049), GOLD’s guidelines pattern as FEV1/FVC>70% and FVC<80. Skin autofluores- whereas the absolute decrease in HbA1c correlated with changes in forced cence (AF), a non-invasive assessment of subcutaneous AGEs accumulation, expiratory flow between 25% and 75% of the FVC (FEF25-75; r=-0.395, was measured (AGE Reader; DiagnOptics Technologies, The Netherlands). p=0.003) and the FEV1/FVC ratio (r=-0.457, p<0.001); both changes in BMI Results: Patients with T2D showed less forced vital capacity [FVC: 77.0 and HbA1c correlated with increments in peak expiratory flow (PEF). Multi- (38.0-114.5) vs. 91.1% predicted (31.0-154.0), p<0.001], forced expiratory variate analysis showed that the BMI reduction was independently associ- volume in the first second [FEV1: 80.1 (28.0 to 124.5) vs. 90.1 (28.0 to 143)% ated with the amelioration of FEV1 (R2=0.462) and PEF (R2=0.392), whereas predicted, p=0.001], and a higher percentage of restrictive pattern (44.7% the HbA1c reduction was independently related with the improvement of vs. 19.5%, p<0.001) compared with nondiabetic subjects. In the entire popu- FEF25-75 (R2=0.395) and FEV1/FVC ratio (R2=0.442); however, age, gender, lation, subjects with a restrictive pattern showed an increased skin AF com- former smoker and evolution of T2D did not influence final results. pared with subjects with normal lung function (2.3±0.6 vs. 2.0±0.5 arbitrary Conclusions: Glycemic control improvement significantly ameliorates lung units, p<0.001). This difference persisted when only patients with T2D were function pattern in patients with T2D. Although the mechanisms are not yet evaluated (3.0±0.6 vs. 2.6±0.7AU, p=0.048). Univariate analysis showed fully understood, this effect is independent of weight reduction. that skin AF negatively correlated with FVC (r=-0.233, p<0.001) and FEV1 Supported By: PI12/00803, PI15/00260; CIBERDEM (r=-0.213, p<0.001). A multivariate regression analysis showed that skin AF (together with age, gender, BMI and packs of cigarettes per year) indepen- 2 2 dently predicted FVC (R =0.174) and FEV1 (R =0.183). COMPLICATIONS—MACROVASCULAR—CELLULAR Conclusions: Skin AF is correlated with impaired pulmonary function. This MECHANISMS OF ATHEROGENESIS IN DIABETES is the first clinical evidence that demonstrates the role of AGEs as a mecha- nism to explain the deleterious effect of T2D in lung function. Supported By: PI12/00803, PI15/00260, PI14/00008 Moderated Poster Discussion: Evolving Mechanisms of Atherogenesis (Posters: 465-P to 470-P), see page 17. 463‑P Cardiac Lipid Metabolism and Contractile Function in Type 2 Berar‑ & 465‑P dinelli-Seip Congenital (BSCL2) Disease Effects of -Rich Lipoproteins on Monocyte Phenotypes HONGYI ZHOU, JIE LI, HUABO SU, WEIQIN CHEN, Augusta, GA ZEQIN LIAN, APARNA MUKHERJEE, XIAO YUAN D. PERRARD, JERRY L. PER- Berardinelli-Seip Congenital Lipodystrophy (BSCL) is a rare metabolic RARD, ANUM SAEED, CHRISTIE M. BALLANTYNE, HUAIZHU WU, Houston, TX disorder characterized by severe generalized lipodystrophy at birth or Hypertriglyceridemia increases risk for atherosclerotic cardiovascular infancy accompanied with early-onset insulin resistance and diabetes. BSCL disease, but the mechanisms remain incompletely understood. We hypoth- patients commonly develop hypertrophic cardiomyopathy, a leading cause esize that hypertriglyceridemia with elevated levels of triglyceride-rich of premature death. However, the pathological mechanisms underlying the lipoproteins (TRLs) contributes to atherosclerosis by changing monocyte development of hypertrophic cardiomyopathy in lipodystrophy has never phenotypes. Monocytes in human subjects with hypertriglyceridemia and been addressed. By utilizing global BSCL2 knockout (gKO) mice which reca- showed increased lipid accumulation and enhanced pitulate human type 2 BSCL (BSCL2) disease, we for the first time identified inflammatory phenotypes. Here, we examined in vitro the effects of TRLs that BSCL2 gKO mice develop early onset cardiac hypertrophy along with on monocyte phenotypes. TRLs were isolated from subjects at 5 hours after an age-dependent impairment of cardiac contractile function. Dysregula- a high- meal. Treatment of THP1 monocytes (without differ- tion of myocardial triglyceride (TG) metabolism has been associated with entiation into ) with postprandial TRLs for 4-48 hours caused cardiac dysfunction in various diseases. Interestingly, BSCL2 gKO hearts monocyte lipid accumulation indicated by increased granularity and posi- have reduced myocardial TG content and increased content. Fur- tive nile red staining examined by flow cytometry, and increased expression ther analysis of cardiac substrate utilization demonstrated an elevation in of inflammatory markers (IL-6, MCP-1, and IL-1β), CD11c (a β2 ), and cardiac fatty acid oxidation (FAO) but not glucose oxidation in BSCL2 gKO CD36 (a scavenger receptor) at mRNA and protein levels examined by quan- hearts. Importantly, loss of BSCL2 increases the protein expression of car- titative RT-PCR or flow cytometry. Upregulation of monocyte CD36 by TRLs diac adipose triglyceride lipase (ATGL), a critical enzyme that catalyzes the (mean fluorescent intensity [MFI]: 4.5 ± 0.9 with TRL treatment vs. 1.8 ± initial and rate-limiting step of intracellular TG hydrolysis. Thus, our data 0.2 with PBS, P<0.05) was associated with enhanced monocyte uptake of suggest BSCL2 may play an important role in the maintenance of normal DiI-labeled cholesterol ester-rich lipoproteins (CRLs), the major lipopro- cardiac growth and substrate metabolism, and dysregulation of myocardial teins causing foam cells in atherosclerotic lesions. MFI of DiI-CRL uptake TG metabolism in the absence of BSCL2 may contribute to the development by monocytes was 6.9 ± 0.5 with TRL pretreatment vs. 4.4 ± 0.2 with PBS of cardiac dysfunction in BSCL2 disease. pretreatment (P<0.01). Treatment with TRLs also increased interferon regu- Supported By: National Heart, Lung, and Blood Institute latory factor 1 (IRF1) in monocytes. Silencing IRF1 in monocytes by siRNA significantly reduced TRL-induced proinflammatory markers and CD36 and attenuated TRL-enhanced DiI-CRL uptake by monocytes. In summary, monocytes took up TRLs and underwent phenotypic changes, with increased expression of inflammatory markers and CD36, which was

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A122 COMPLICATIONS—MACROVASCULAR—CELLULAR MECHANISMS OF ATHEROGENESIS IN DIABETES associated with enhanced monocyte uptake of CRLs and foam cell forma- & 468‑P tion. IRF1 may play an essential role in TRL regulation of monocyte pheno- Evidence of the Role of -3-Kinase in the Glycation Gap types. and in Protection against Complications in Patients with Diabetes Supported By: American Diabetes Association (1-17-IBS-082 to H.W.); National SIMON J. DUNMORE, AMR S. AL-DERAWI, ARUNA NARSHI, FAKHRA NASEEM, Institutes of Health (R01HL098839); American Heart Association (30410012) JAMES E.P. BROWN, ALAN NEVILL, ANANTH U. NAYAK, BALDEV M. SINGH, Wolverhampton, United Kingdom, Birmingham, United Kingdom, Walsall, United 466‑P Kingdom, Stoke-on-Trent, United Kingdom & The glycation gap (GGap), discrepant average glycaemia calculated using Inhibition of HDAC3 Prevents Diabetic Cardiomyopathy in OVE26 POSTERS Mice via Epigenetic Regulation of DUSP5-ERK1/2 Pathway HbA1c vs. fructosamine, is a consistent phenomenon in our diabetic patients Complications ZHENG XU, JIAN SUN, LU CAI, Louisville, KY, Changchun, China associated with greater incidence of complications in patients with +ve cf Acute and Chronic Inhibition of total histone deacetylases (HDACs) was phenomenally asso- -ve GGaps. One factor in GGap and incidence of complications could be vari- ciated with the prevention of diabetic cardiomyopathy (DCM); however, ation in the level/activity of deglycating enzymes, especially Fructosamine- which specific HDAC plays the key role remains unclear. The present study 3-Kinase (FN3K), regulating AGE. explored the prevention of DCM by specific inhibition of HDAC3 and associ- Explored relationship of erythrocyte FN3K to GGap and prothombotic and ated mechanism. Type 1 diabetes OVE26 and age-matched wild type mice endothelial dysfunction markers in patients with +ve/-ve GGaps. Assayed were given the selective HDAC3 inhibitor RGFP966 or vehicle for 3 months. FN3K protein (ELISA) and FN3K activity in erythrocytes from sub-groups of These mice were then sacrificed immediately (3 months) or 3 months later cohort with highest +ve (>+0.5%, n=43) or -ve (<-0.5%, n=55) GGaps. Immu- (6 months) for cardiac function and pathological examination. HDAC3 activ- noassayed plasma FN3K protein, AGE, E-selectin, thrombomodulin, PAI-1, ity was significantly increased in the heart of diabetic mice. Administration /adiponectin (LAR) ratio. of RGFP966 significantly inhibited HDAC3 activity and prevented DCM, evi- +ve GGap group erythrocytes: lower concentrations of FN3K protein (68% denced by improved diabetes-induced cardiac dysfunction, hypertrophy and vs.-ve, p<0.01), much lower FN3K activity (20% vs.-ve, p<0.001). Plasma: fibrosis, along with diminished cardiac oxidative stress and inflammation, +ve cf -ve GGap - AGE, PAI and LAR significantly higher (25%, p<0.05;547%, not only at 3 months but also at 6 months. This may suggest a memory of p<0.001; 45%, p<0.005 respectively) but not FN3K, E-selectin or thrombo- the cardiac protection at the 6 months from the first three-month treatment. modulin. Univariate analysis: only GGap group, not BMI, HbA1c, uACR were Mechanistically, phosphorylated ERK1/2, a well-known initiator of cardiac significant independent variables affecting any SDs for FN3K concentration hypertrophy, was significantly increased, while Dual Specificity Phospha- and activity, AGE, PAI, LAR. ANCOVA analysis-erythrocyte FN3K activity vs. tase 5 (DUSP5), an ERK1/2 nuclear phosphatase to consequently dephos- protein: log erythrocyte FN3K activity vs. FN3K protein: sig diff in activity phorylate ERK1/2, was substantially decreased in diabetic hearts. Both of -ve GGap group cf +ve GGap group, controlled for log concentration (r2=0.66, these changes were prevented by RGFP966. Chromatin immunoprecipitation F=90.68, p<0.001), significant slope parameter near unity (B=0.90, t=9.219, assay showed that HDAC3 inhibition elevated histone H3 acetylation on the p<0.001). Separate regression lines fitted each group, significantly displaced DUSP5 gene promoter at both 3 and 6 months. These findings suggest that from each other (F=61.79, p<0.001), representing 326% greater FN3K activ- diabetes-activated HDAC3 inhibits DUSP5 expression to de-repress ERK1/2 ity in -ve cf +ve GGap. through deacetylating histone H3 on the primer region of DUSP5 gene, lead- This is novel evidence supporting a role for FN3K in deglycation of gly- ing to the initiation of DCM. This study suggests the potential application of cated haemoglobin (may affect treatment strategy), and possibly in deglyca- HDAC3 inhibitor for the prevention of DCM. tion of glycated proteins associated with production of AGE and diabetic Supported By: American Diabetes Association (1-15-BS-018 to L.C.) complications. Supported By: Wolverhampton Diabetes Trust & 467‑P Caveolin-1, a Binding Protein of CD26, Is Essential for the Suppres‑ & 469‑P sion of Inflammation in Macrophages by DPP-4 Inhibitors Advanced Glycation End Products Induce Brain-Derived Neuro‑ MUNENORI HIROMURA, HIDEO KATAOKA, KYOKO NOHTOMI, YUSAKU MORI, trophic Factor Release from Human Platelets through the Src Family TOMOYASU FUKUI, HIROTAKA KUWATA, TSUTOMU HIRANO, Tokyo, Japan Kinases Activation We reported that inhibition of dipeptidyl peptidase-4 (DPP-4), the cata- KAZUO FURUKAWA, ICHIRO FUSE, YURIKO IWAKURA, HIDEKAZU SOTOYAMA, lytic site of CD26, suppresses atherosclerosis in animal models through its OSAMU HANYU, HIROYUKI NAWA, NOBUYUKI TAKEI, HIROHITO SONE, Niigata, anti-inflammatory properties; however, the mechanism has not been fully Japan elucidated. Caveolin-1 (Cav-1), a major structural protein of caveolae on the Brain-derived neurotrophic factor (BDNF) exerts beneficial effects not cell surface, has been reported to modulate inflammatory pathways by bind- only on but also on cardiovascular injury. There is argu- ing to CD26 in macrophages. We investigated the role of Cav-1 in the sup- ment regarding the levels of serum BDNF in patients with diabetes mellitus pression of inflammation by DPP-4 inhibitors using human monocyte U937 (DM). Because BDNF in peripheral blood is abundant in platelets, this may cells and macrophages taken from Cav-1 knockout (Cav-1-/-) mice. represent dysregulation of BDNF release from platelets. Here we focused The DPP-4 inhibitor (1-10 nM) suppressed pro-inflammatory on advanced glycation end products (AGEs), which are elevated in patients expression induced by lipopolysaccharide (LPS), such as IL-6 and with DM and have adverse effects on cardiovascular functions. The aim of TNF-α in U937 cells. Gene knockdown by siRNA and antibody administration this study is to elucidate the role of AGEs in the regulation of BDNF release against CD26 or Cav-1 showed similar effects. from human platelets. Platelets collected from peripheral blood of healthy Immunohistochemistry of murine peritoneal macrophages showed co- volunteers was incubated with various concentrations of AGE glycated- expression of CD26 and Cav-1. In flow cytometric analysis, approximately BSA) at 37°C for 5 min with or without BAPTA-AM, a cell permeable Ca2+ 10% of the macrophages were CD26+, and neither LPS nor teneligliptin chelator, or PP2, a potent inhibitor of Src family kinases (SFKs). Released changed the ratio of CD26+ cells. In peritoneal macrophages taken from and cellular BDNF were measured by ELISA and calculated. Phosphorylation Cav-1-/- mice, LPS-induced inflammatory cytokine expression was 50-60% of Src and Syk, a downstream kinase of SFKs, in stimulated platelets was lower than in Cav-1+/+ mice. Teneligliptin (10 nM) suppressed LPS-stimulated examined by Western blotting and immunoprecipitation. As a result, AGE- gene expression of IL-6 and TNF-α in Cav-1+/+ mice. In contrast, in Cav-/- mice, induced BDNF release from human platelets in a dose-dependent manner, teneligliptin did not reduce pro-inflammatory cytokine expression. which was dependent on intracellular Ca2+ and SFKs. We found that AGE Next, we investigated whether teneligliptin could affect binding between induced phosphorylation of Src and Syk. Cav-1 and CD26 using the Biacore™ system (biophysical interaction analy- In conclusion, AGE induces BDNF release from human platelets through sis). Both human soluble CD26 protein (sCD26) alone and teneligliptin alone the activation of the Src-Syk-(possibly phospholipase C)-Ca2+ pathway. Con- bound to the Cav-1 protein, and co-administration of teneligliptin and sCD26 sidering the toxic action of AGEs and the protective roles of BDNF, it can be additionally bound to Cav-1, suggesting that teneligliptin itself may directly hypothesized that AGE-induced BDNF release is a biological defense system interact with Cav-1. in the early phase of diabetes. Chronic elevation of AGEs may induce deple- These results indicated that Cav-1 is a key molecule regulating the anti- tion or down regulation of BDNF in platelets during the progression of DM. inflammatory properties of DPP-4 inhibitors in LPS-stimulated macrophages.

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A123 COMPLICATIONS—MACROVASCULAR—CELLULAR MECHANISMS OF ATHEROGENESIS IN DIABETES & 470‑P 471‑P A Role for DPP-4 Expression via oxLDL/TLR4/TRIF/CD36 Pathways Polymerase Delta-Interacting Protein 2 Regulates Collagen Accu‑ in Human Obesity and Atherosclerosis mulation via Activation of the AKT/mTOR Pathway in Vascular JIXIN ZHONG, SANJAY RAJAGOPALAN, XIAOQUAN RAO, Cleveland, OH Smooth Muscle Cells Dipeptidyl peptidase-4 (DPP-4) is well-known for its regulatory role in MASAKAZU FUJII, YASUHIRO SAKO, KATHY GRIENDLING, Fukuoka, Japan, Iizuka, incretin biology. Although prior studies have suggested a role for DPP-4 in Japan, Atlanta, GA atherosclerosis, the regulation of DPP-4 in diabetes and atherosclerosis has Objectives: Polymerase delta interacting protein 2 (Poldip2) has previously

POSTERS not been investigated. In this study, we examined the relationship of DPP-4 been implicated in migration, proliferation and extracellular matrix (ECM) Complications with human atherosclerosis and its role in obesity/insulin resistance. In production in vascular smooth muscle cells. Thereby, regulation of Poldip2 Acute and Chronic atherosclerosis, both cleaved and membrane bound DPP-4 were increased. protein expression is suggested to prevent the development and progres- DPP-4 expression on monocytes positively correlated with atherosclerotic sion of diabetic macroangiopathy. To better understand the role of Poldip2 plaque volume assessed by high-resolution MRI (P=0.0043) with expres- in ECM regulation, we investigated the mechanism responsible for collagen sion positively correlating with non-HDL cholesterol level (P=0.0242) but I accumulation in Poldip2+/- mouse aortic smooth muscle cells (MASMs). not fasting blood glucose or insulin. Oxidized low-density lipoprotein Approach and Results: Protein degradation and protein synthesis path- (oxLDL) up-regulated DPP-4 expression on macrophages with a preferential ways were investigated. Depletion of Poldip2 caused a significant reduction increase in CD36+ cells. OxLDL mediated increase in DPP-4 was abolished in autophagic flux. However, the rate of collagen I degradation was increased by siRNA-mediated Toll-like receptor 4 (TLR4) and considerably diminished in Poldip2+/- vs. Poldip2+/+ MASMs. Conversely, activation of the PI3K/Akt/ by CD36 knock-down. TRIF deficiency but not MyD88 deficiency attenuated mTOR signaling pathway, involved in regulation of protein synthesis, was DPP-4 increase with oxLDL. Our study suggests DPP-4 on monocytes cor- significantly elevated in Poldip2+/- MASMs as wasβ 1-Integrin expression. relates with plaque volume with a key role for oxLDL in regulating DPP-4 Surprisingly, immunocytochemistry showed immature focal adhesions expression via CD36/TLR4/TRIF pathways. Increased DPP-4 in response in Podip2+/- MASMs. Suppressing mTOR signaling using Akt inhibitor or to oxidized lipids may represent an integrated mechanism by which post- Rapamycin and reducing β1-Integrin expression using siRNA prevented the prandial glucose metabolism is linked to lipoprotein abnormalities potenti- increase in collagen I production. Immunocytochemistry showed immatured ates atherosclerosis. focal adhesion in Podip2+/- MASMs. While collagen I and fibronectin were increased in Poldip2+/- MASMs, overall protein synthesis was not different Figures. from that in Poldip2+/- MASMs, suggesting selectivity of Poldip2 for ECM proteins. Conclusions: Poldip2+/- MASMs exhibit higher β1-Integrin expression and activity of the PI3K/Akt/mTOR signaling pathway, leading to increased ECM protein synthesis. The cause of increasing β1-Integrin expression in Poldip2+/- MASMs is suggested as a compensation against immatured focal adhesion. These findings have important implications for atherosclerotic vascular diseases such as diabetic macroangiopathy in which ECM accu- mulation plays a role.

472‑P Accumulation of Endogenous Inhibitor Is a Major Contributor to Erectile Dysfunction in Type 2 Diabetic Rats YAN XIONG, YAN PING LEI, CHENG HUANG, YUAN LIN, XIAO MEI LI, ZHI MIN HE, Guangzhou, China Erectile dysfunction (ED) is one of the most common complications of dia- betes. However, the underlying mechanism is not clear. Emerging evidence shows that accumulation of endogenous nitric oxide (NO) synthase (NOS) inhibitor asymmetric dimethylarginine (ADMA) is associated with diabetic cardiovascular complications. This study was to determine whether ADMA accumulation contributes to ED of type 2 diabetic rats and elucidate poten- tial mechanisms. Corpus cavernosums were isolated for the examination of acetylcholine (ACh)-induced relaxation to reflect erectile function. NOS activity, ADMA, NO and cGMP contents were measured in corpus caver- nosum. The relaxation function of corpus cavernosum response to ACh was significantly impaired and associated with the elevation of serum ADMA level in diabetic rats compared to control rats. Furthermore, reductions of NOS activity, NO and cGMP contents, increase of oxidative stress and dis- order of ADMA signal protein expression were observed in corpus caverno- sums of diabetic rats. Logistics regression analysis shows that serum ADMA concentration was negatively correlated with NO and cGMP contents and relaxation function of corpus cavernosum. Treatment with NO precur- sor L- (100 mg/kg in gavage) for 8w could reverse above changes in diabetic rats. Similar relaxation dysfunction of corpus cavernosum and decreased NO and cGMP contents observed in diabetic rats were also found in either exogenous NOS inhibitor NG-Nitro-L-arginine-treated rats or iso- lated corpus cavernosum from normal rats incubated with ADMA in vitro. These results indicate that ADMA accumulation is a major contributor to Supported By: National Institutes of Health (K01DK105108); National Natural erectile dysfunction of diabetic rats. The underlying mechanism is related to Science Foundation of China (81670431) reduction of NO production and increase of oxidative stress. Supported By: Guangzhou Municipal Bureau of Science and Information (14J4100067); National Natural Science Foundation of China (81570751)

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473‑P of EMPA on in vivo cardiac energy status in diabetic db/db mice using 31P Single Nucleotide Polymorphisms of Metabolism-Related magnetic resonance spectroscopy (MRS). Gene DHCR7 Are Associated with Vascular Risk Db/db mice (10 weeks old) were fasted for 6 hours prior to 31P MRS. After 4 RINKOO DALAN, HUILING LIEW, LIUH LING GOH, SAFIYYA ALI MOHAMAD, hours of fasting, the mice received either vehicle (N=10) or EMPA (30 mg/kg DANIEL E.K. CHEW, BERNHARD O. BOEHM, Singapore, Singapore body weight; N=10) in 0.5% Natrosol (10 ml/kg body weight) by oral gavage. Vitamin D deficiency is associated with diabetes, obesity and atheroscle- Plasma parameters were measured after another 2 hours of fasting. 31P MRS rosis. The GG genotype of 7-dehydrocholesterol reductase (DHCR7) is known was then performed in a voxel covering the left ventricle (~5x5x5 mm3; Fig- ure 1A). The PCr/γ-ATP ratio was quantified as a measure of cardiac energy to influence vitamin D serum concentrations and atherosclerosis. We aim to POSTERS explore the association between DHCR7 genotypes and ethnicity, BMI, and status (Figure 1B). Complications vascular function. Plasma glucose levels were lower in the EMPA group compared with Acute and Chronic Subjects: 253 subjects (Total 190 T2DM, 140 Chinese, 23 Malays and 90 vehicle (12.1 ± 1.9 mM vs. 17.3 ± 5.3 mM; P=0.017), while plasma ketone Indians; 63 Controls). levels were higher (1.7 ± 0.8 mM vs. 0.6 ± 0.2 mM; P=0.003). The EMPA group Measurements: Demographics, BMI, waist circumference, blood pres- showed a higher cardiac PCr/ATP ratio compared with vehicle (0.80 ± 0.15 vs. sure, HbA1c, glucose and lipids, CIMT, hsCRP, oxidative stress index (OSI; 0.56 ± 0.13; P=0.001; Figure 1C). Diacron, Italy), DHCR7 SNP rs12785878, rs3829251 genotyping (RT-PCR). A single dose of EMPA acutely improves cardiac energy status in db/db The association between GG/non-GG genotypes with all the variables was mice, which may explain the improved cardiovascular outcome in the EMPA- assessed using logistic regression in univariate analysis and multivariate REG study. We hypothesize that this may be related to a shift in cardiac fuel analysis model using all predictor variables (STATA version 13). utilization; however, this warrants further investigations. Prevalence of GG Genotype: rs12785878: Indians: 74%; Malays: 65%; Chi- nese 34%; rs3829251: Indians: 52%; Chinese: 48%; Malays: 40% (p<0.001). Allelic frequencies were in Hardy-Weinberg equilibrium. In univariate analysis, DHCR7 rs12785878 GG was associated with OSI (p=0.001), hsCRP (p=0.013) and waist circumference (p=0.016) and DHCR7 rs3829251 GG was associated with CIMT (p=0.046). In multivariate analysis, rs3829251 GG was associated with BMI (p=0.044) and non-HDL cholesterol (p=0.038). The GG genotypes of the DHCR7 (rs12785878, rs3829251) are more preva- lent in Indians and correlates with BMI and vascular risk markers of non-HDL cholesterol, OSI, hsCRP and CIMT. These associations need to be replicated in larger cohorts and possible mechanisms elucidated.

474‑P Regulation of Endothelial Function by Dietary Potassium: Roles of Gut Microbiota and Oxidative Stress YOUNG TAEK OH, JOYCE M. RICHEY, JANG H. YOUN, Los Angeles, CA Dietary potassium (K+) intake improves cardiovascular (CV) health, but the underlying mechanisms are unclear. Endothelial function is a major deter- minant of CV health. We examined the effects of dietary K+ on endothe- lial function in rat aorta. Male Wistar rats were fed either a K+-rich (2%) or K+-deficient (0%) diet for 5 days, and endothelial function was assessed in isolated aortas by measuring acetylcholine-induced relaxation. Aortic relax- ation was enhanced in rats fed the K+-rich diet, compared to those fed the + + K -deficient diet; dietary K decreased EC50, reflecting increased sensitivity, from 0.21±0.04 to 0.12±0.01 µM (P < 0.05; n = 5 each). Dietary K+ intake increases intestinal K+ levels, which may be an important factor for gut bac- terial growth and function. We next examined whether gut may play a role in the regulation of endothelial function by dietary K+. Rats were treated with antibiotics (Abs; vancomycin, neomycin, and ampicillin) in their drinking water to remove gut bacteria. Fecal DNA content decreased to 3% of control after a week of the Ab treatment (n = 4), indicating successful removal of gut bacteria. After a week of Ab treatment, animals were fed the 0% or 2% K+ diet for 5 days, while the Abs were continued. Ab treatment prevented the improvement of endothelial function observed with dietary K+; + EC50 was 0.19±0.06 and 0.26±0.08 µM with the 0% and 2% K diet, respec- tively (P > 0.05; n = 4 each). These data suggest that gut bacteria are required for dietary K+ effects to improve endothelial function. To further elucidate underlying mechanisms, we performed an oxylipin analysis of aorta samples from rats fed with the 0% or 2% K+ diet (n = 7 each). We found significant decreases in the oxidative stress markers 9-HODE and 13-HODE with the K+-rich diet (P < 0.05), suggesting that dietary K+ decreased oxidative stress 476‑P in the aorta. Taken together, our data suggest that dietary K+ improves endo- Plasma Omentin Levels Are Associated with Vascular Endothelial thelial function by interacting with the gut microbiota and decreasing oxida- Function in Patients with Type 2 Diabetes and Renal Dysfunction tive stress. MARIKO SENDA, TOMOAKI MORIOKA, MARIKO ASADA, YUKO YAMAZAKI, Supported By: American Diabetes Association (1-16-IBS-332 to J.H.Y.) YOSHINORI KAKUTANI, REINA KAWARABAYASHI, KOKA MOTOYAMA, KAT- SUHITO MORI, SHINYA FUKUMOTO, ATSUSHI SHIOI, TETSUO SHOJI, MASA- 475‑P NORI EMOTO, MASAAKI INABA, Osaka, Japan Omentin is an adipokine predominantly expressed in visceral adipose tis- The Effect of a Single Dose of Empagliflozin on Cardiac Energy Sta‑ sue and has insulin-sensitizing and vasculoprotective effects, like adiopo- tus in Diabetic Mice nectin, in rodents. Although circulating omentin levels are inversely related DESIREE ABDURRACHIM, EMMY MANDERS, KLAAS NICOLAY, JEANINE J. to obesity and metabolic risk factors in humans, the association between PROMPERS, ERIC MAYOUX, Eindhoven, Netherlands, Biberach, Germany omenin and subclinical atherosclerosis remains unclear. In this study, we In the EMPA-REG OUTCOME trial, the sodium glucose cotransporter 2 investigated the association between plasma omentin levels and vascu- inhibitor empagliflozin (EMPA) showed impressive benefits on cardiovascu- lar endothelial function in patients with type 2 diabetes. We included 428 lar mortality in type 2 diabetes (T2D) patients. T2D is associated with a low patients with type 2 diabetes in this study. Fasting plasma omentin levels cardiac energy status. Our aim was to investigate the effect of a single dose and total adiponectin levels were measured by ELISA, and the endothelium-

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dependent, flow-mediated dilatation (FMD) of the brachial artery was mea- (P < 0.01) while standard deviations of blood glucose were unchanged sured by ultrasonography. The median of plasma omentin level, adiponectin (2.1±1.4 mM vs. 1.6±0.65 mM, P=0.24). T10, an indicator of thrombus for- level, and FMD were 572 ng/mL, 6.1 µg/mL, and 5.7%, respectively. Plasma mation, was significantly improved after glycemic control (338±65 sec vs. omentin levels were positively correlated with adiponectin and HDL-choles- 425±117 sec, P < 0.01). The improvement of T10 were significantly correlated terol, and negatively correlated with BMI, IRI, HOMA-R, triglycerides, and with the changes in the mean blood glucose levels after the treatment eGFR. Multivariate analysis showed that plasma omentin levels were not (r = 0.718, P < 0.05) but not with those in standard deviation of blood glu- significantly associated with FMD after adjustment for adiponectin levels cose. On the other hand, the platelet aggregation assessed with the same and traditional cardiovascular risk factors. However, subgroup analysis microchip flow-chamber system using a different chip was not changed after POSTERS

Complications revealed that plasma omentin level was an independent determinant of FMD the treatment. Thus, we found that short term reduction of mean blood glu- Acute and Chronic (β = 0.209, p = 0.021) in subjects with chronic (CKD, eGFR cose reduced the thrombus formation without affecting platelet function in <60mL/min/1.73m2), but not in those without CKD. Interaction analysis indi- patients with type 2 diabetes. cated a potential effect modification by eGFR on the association of plasma omentin levels with FMD (p = 0.102 for interaction). 479‑P In conclusion, plasma omentin levels are associated with vascular endo- Inhibition of A-FABP Attenuates Cerebral Injury via Alle‑ thelial function, independently of adiponectin and traditional cardiovascular viating MMP-9-Mediated Blood-Brain Barrier Disruption risk factors, in patients with type 2 diabetes and CKD. This study indicates BOYA LIAO, LEILUO GENG, KAREN S. LAM, AIMIN XU, RUBY L.C. HOO, SOOKJA that omentin plays a protective role in vascular function in patients with K. CHUNG, Hong Kong, China type 2 diabetes and CKD at high risk of cardiovascular disease. Ischemic stroke is one of the major causes of cerebral injury and perma- nent disability in the worldwide; however, the underlying molecular mecha- 477‑P nisms are poorly understood. Clinical studies demonstrate that serum levels Increases HDL-Mediated Cholesterol Efflux In of -fatty acid binding protein (A-FABP), an adipokine mainly derived Macrophages from and macrophages, are significantly increased in patients KOTA MATSUKI, YUKI KIMURA, KOKI MATSUMURA, HIROSHI MURAKAMI, with ischemic stroke. Here we investigated the role of A-FABP in ischemic MAKI YAMASHITA, JUTARO TANABE, HIROSHI MURAKAMI, MAKOTO DAIMON, brain injury. To address this question, A-FABP knockout (KO) mice and their Hirosaki, Japan wild type (WT) littermates were subjected to middle cerebral artery occlu- Both diabetic macroangiopathy and atherosclerosis are characterized by sion surgery. We found that A-FABP KO mice exhibited reduced ischemic increased arterial stiffness and hyaluronic acid (HA) deposition in the large brain injury as indicated by smaller brain infarct volume and decreased cell elastic arteries. HA is a large non-sulfated present in apoptosis comparing to their WT littermates. Meanwhile, pharmacological the extracellular matrix. Extensive research on the HA system in cardio- inhibition of A-FABP with BMS309403 (BMS) treatment alleviated ischemic vascular pathology has contributed to an understanding of the physiologi- cerebral injury in WT mice. Blood-brain barrier (BBB) disruption was signifi- cal and pathophysiological role of HA in vitro and in vivo. It is noteworthy cantly alleviated in both A-FABP KO mice and BMS-treated mice as indicated that systemic inhibition of hyaluronan synthesis by 4-methylumbelliferone by reduced expression of the tight junction proteins ZO-1 and Occludin. This interferes with the protective function of the endothelial glycocalyx, thereby was also accompanied by a lower expression levels of matrix metallopepti- facilitating leukocyte adhesion, subsequent inflammation, and progression dase 9 (MMP-9), the key proteinase to degrade tight-junction protein in BBB. of atherosclerosis. High density lipoprotein (HDL) mediates cholesterol These data suggest that inhibition of A-FABP protects mice against cerebral efflux, which is the initial and rate-limiting step of reverse cholesterol trans- ischemia injury which may via its suppression on the expression of MMP9 port preventing from atherosclerosis. The aim of the present study was to thus prevent BBB disruption. A-FABP inhibition may be a potential strategy investigate for the first time the relationship between HA and cholesterol for ischemic cerebral injury therapy. efflux in macrophages. Human monocyte derived THP-1 cells were devel- Supported By: Health and Medical Research Fund; Shenzhen Basic Research oped to macrophages and labeled with 3H-cholesterol (0.6 μCi/ml). After the Fund incubation adding HDL into the medium for 24 hours, the rate of labeled cholesterol pulled out from the macrophages was measured as cho- 480‑P lesterol efflux. The effect of HA on the cholesterol efflux was studied, add- MicroRNA-32 Promotes Calcification by Targeting PTEN in Vascu‑ ing that into the medium before the analysis of the cholesterol efflux. Under lar Smooth Muscle Cells the existence of HA (100 μg/ml), the HDL mediated cholesterol efflux sig- JIANGHUA LIU, YINGYING SHEN, LING CHEN, QINGHAI ZHANG, XUYU ZU, nificantly increased by 9.4% (n=4, p<0.0001). We also researched the rela- Hengyang, China tionship between HA and ABCG1, because expression of ABCG1 enhances Cardiovascular calcification often causes significant morbidity and mor- cholesterol efflux to HDL. Predictably, HA significantly increased ABCG1 tality. However, the precise mechanisms underlying vascular calcification mRNA expression in macrophages (n=4, p<0.01), determined by Real-time remain largely unknown. Increasing evidence has suggested Quantitative RT-PCR. We showed the possibility that HA increases HDL- might play an important role in vascular smooth muscle cells (VSMCs) calci- mediated cholesterol efflux via ABCG1 in macrophages, and it might be one fication. Here, we have identified several key miRNAs which participate in of the important mechanisms to prevent progression of atherosclerosis and vascular calcification in vivo and in vitro, and further characterized the role of diabetic macroangiopathy. microRNA-32 (miR-32) in vascular calcification. Microarray data showed that microRNA-125b (miR-125b), microRNA-30a (miR-30a) and miR-32 expres- 478‑P sion were elevated, while and the expression of microRNA-29a (miR-29a), Reduction of Thrombus Formation Assessed by an Automated microRNA-210 (miR-210) and microRNA-320 (miR-320) were downregulated Microchip Flow-Chamber System after Glycemic Control during vascular calcification progression in vivo and in vitro. Furthermore, KIYOAKI YAMAMOTO, TAKASHI ITO, TOMOKA NAGASATO, SAHAR GHAVIDEL gain-of-function and loss-of-function studies of miR-32 revealed that miR-32 DARESTANI, ATSUSHI SHINNAKASU, MIHOKO KURANO, AIKO ARIMURA, could promote mouse VSMC calcification through inducing bone morphoge- HIROSHI ARIMURA, HIROSHI HASHIGUCHI, TAKAHISA DEGUCHI, IKURO netic protein-2 (BMP-2), runt-related transcription factor 2 (Runx2), Osteo- MARUYAMA, YOSHIHIKO NISHIO, Kagoshima, Japan pontin (OPN) and matrix GLA protein (MGP) expression in vitro. Moreover, Thrombus formation is one of the important factors affecting the risks and miR-32 modulated vascular calcification progression through activating PI3K prognosis of cardiovascular events. Several studies have shown that coagu- signaling and increasing the expression and phosphorylation of Runx2 by lability, platelets aggregation or markers of thrombus formation increase in directly targeting PTEN 3’-UTR in mouse VSMCs. Finally, miR-32 level was patients with diabetes. However, it is not clear whether glycemic control higher in plasma of coronary artery disease (CAD) patients with coronary reduces thrombus formation. Therefore, we investigated the effect of short- artery calcification (CAC) than those in the non-CAC patients (P=0.016). term glycemic control on thrombus formation using an automated microchip In summary, our data disclose a dynamic miRNA expression profile during flow-chamber system (T-TAS, Fujimori Kogyo, Japan). Ten (5 male, 5 female) vascular calcification progression in vivo and in vitro, and suggest miR-32 patients with type 2 diabetes were recruited (Age 47.5±16.6 years, HbA1c could be a critical modulator and potential diagnostic marker for CAC. 10.5±3.1%). Their blood glucose levels were controlled with insulin or oral hypoglycemic agents. Before and after 2 weeks of treatment, blood glucose was analyzed with continuous glucose monitoring system and the throm- bus formation was analyzed quantitatively with the T-TAS. As a result, the level of mean blood glucose was reduced from 10.1±2.6 mM to 6.9±0.97 mM

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481‑P 483‑P Circulating Endothelial Progenitor Cells (EPCs) and Endothelial Potential Role of Circulating miR-21 in the Onset of Prediabetes Microparticles (EdMPs) across Progression of Type 2 Diabetes and T2DM: Possible Connection to Mitochondrial Dysfunction in In DANIELA LUCCHESI, LAURA GIUSTI, MONIA GAROFOLO, VERONICA SANCHO Vitro Analysis of Glucose Variability BORNEZ, GIUSEPPE DANIELE, ANGELA DARDANO, ROBERTO MICCOLI, STEFANO LUCIA LA SALA, ANNA CHIARA UCCELLATORE, GAIA SPINETTI, ELENA SAN- DEL PRATO, GIUSEPPE PENNO, Pisa, Italy GALLI, MONICA CATTANEO, SIMONA MRAKIC-SPOSTA, VALERIA DE NIGRIS, The balance between Endothelium derived Microparticles (EdMPs) FRANCESCO PRATTICHIZZO, STEFANO GENOVESE, ANTONIO CERIELLO, Milan,

and Endothelial Progenitor Cells (EPCs) reflects concomitant processes of Italy, Barcelona, Spain POSTERS endothelial injury and repair. We aimed to track changes in EPCs, EdMPs Disorders of glucose metabolism, IGT and undiagnosed T2DM are risk Complications and EdMP/EPC ratio inT2D progression. Circulating EdMPs (CD42-/CD31+) factors for diabetic complications (DC). Identifying new molecular markers Acute and Chronic and EPCs (CD34+/CD133+/KDR+) were quantified (M±SD) by FACS analy- might be beneficial to reduce or delay diabetes progression, by ameliorat- sis in: 196 T2D (age 62±9 yrs, DD 28±12 yrs, BMI 30.8±6.1 kg/m2, HbA1c ing lifestyle or developing new therapeutic targets. MicroRNAs (miRs) 7.3±1.2%); 30 newly-diagnosed T2D (nT2D; 55±10 yrs, BMI 29.5±5.2 kg/m2, are emerged as epigenetic molecules able to regulate gene expression. HbA1c 11.1±2.3%); 20 with prediabetes (pT2D; 54±9 yrs, BMI 29.7±7.7 Recently, miR-21 has been associated with diabetes, but its function is kg/m2, HbA1c 5.9±0.2%) and 20 controls (C, age 52±9 yrs, BMI 24.1±1.8 kg/m2, unclear. Here, we evaluated the potential role of circulating miR-21 (c-miR- HbA1c 5.5±0.3%). Compared to C (Krukal-Wallis test), stem cells (CD34+ and 21) in the onset of T2DM in Diabetes Prediction And Screening Observational CD34+CD133+: 2193±707 and 967±380 cells/ml) were marginally reduced in Study (DIAPASON) selected cohort (n=78) on the basis of FINDRISC ≥9. qPCR pT2D (1742±517 and 841±488 cells/ml) and nT2D (1686±979 and 796±496; was used for c-miR-21 detection in 26 controls (CTRs, 40-72 yrs), 26 predia- p=0.028 and p=0.070 vs. T2D, respectively), but not in T2D (2212±1262 and betics (PRE, 44-71 yrs), and 26 subjects with new diagnosed T2DMs drug 1067±797). On the contrary, EPCs (CD34+KDR+ and CD34+CD133+KDR+) pro- naïve (DIA, 45-74 yrs), screened on 2hPG ADA criteria. ANOVA, ROC analysis gressively dropped from C (879±354 and 435±227 cells/ml) to pT2D (590±277 and Spearman’s ρ were calculated. c-miR-21 expressions were elevated in and 302±191), nT2D (496±487 and 263±282) and T2D (408±338 and 222±207; PRE (p=0.01) and DIA (p=0.001) compare to CTRs. c-miR-21 correlates with p<0.001 and p=0.003, respectively). EdMP increased, not significantly, glycaemic parameters, FPG (ρ=0.32, p=0.004), 2hPG (ρ=0.4, p=0.0008) and from C (7903±2501 n/ml), to pT2D (11664±8050), T2D (11261±15979 ) and HbA1c (ρ=0.22, p=0.05). In addition, ROC curve demonstrated c-miR-21 diag- nT2D (14843±27238), while EdMP/EPC ratio increased from C (21.6±13.2) nostic accuracy (CTR vs. DIA, AUC=0.8, p<0.0001; CTR vs. PRE, AUC=0.78, to pT2D (55.0±46.9), nT2D (56.9±50.5) and T2D (85.2±148.5; p=0.016 vs. C). p=0.0005). We also evaluated miR-21 mechanistic value using glucose vari- By logistic regression ORs for CD34+CD133+KDR+ cells below the median ability (GV) model of HUVECs, mimicking pre- and diabetic state. We found covariated for gender, age, BMI, BP, lipids, blood glucose, HbA1c, A/C ratio c-miR-21 level was significantly consistent with intracellular miR-21 (i-miR- and CKD-EPI eGFR were 9.8 (p=0.048) for nT2D and 9.0 (p=0.040) for T2D; 21) alterations found in our cellular model. We found significant i-miR-21 for EdMP/EPC above the median, covariated ORs were 3.1 (ns) and 6.9 up-regulation coupled to ROS over production measured by EPR (Electron (p=0.029), respectively. Covariates showed no independent effects even Paramagnetic Resonance), to mitochondrial dysfunction and to dysregulated when subjects category was removed from the regression. Alteration in SOD2 protein (miR-21’s indirect target, by luciferase assay), via AKT-inhib- EPCs, EdMPs and EdMP/EPC worsen with disease progression irrespective ited pathway. of the effects of the conventional CV risk factors. In conclusion, this phenomenon could occur in pre- and diabetic subjects Supported By: Regione Toscana (D55E11002680005) and could be associated with oxidative stress and early mitochondrial dys- function. 482‑P Supported By: Fondazione Invernizzi Milano, Italy Effects of Saturated, Mono- and Poly-unsaturated Fatty Acids and Protective Effects of the GLP-1 Analog Exendin-4 on Survival of 484‑P Human Cardiospheres and Human Cardiac Progenitor Cells Elevating CXCR7 Improves Angiogenic Function of EPCs via Akt/ LUIGI LAVIOLA, ROSSELLA D’ORIA, MARIA ANGELA INCALZA, CRISTINA CAC- GSK-3β/Fyn-Mediated Nrf2 Activation in Diabetic Limb Ischemia CIOPPOLI, VALENTINA ANDRULLI BUCCHERI, ANNALISA NATALICCHIO, SEBAS- XIAOZHEN DAI, JUN ZENG, XIAOQING YAN, JING CHEN, KUPPER A. WINTERG- TIO PERRINI, FRANCESCO GIORGINO, Bari, Italy ERST, LU CAI, YI TAN, Chengdu, China, Louisville, KY Mono- and poly-unsaturated fatty acids exert beneficial effects on the Recently, dipeptidyl peptidase-4 (DPP-4) inhibitor , a major anti- heart, whereas saturated fatty acids are associated with higher cardiovas- hyperglycemic agent, has received substantial attention as a therapeutic cular risk. The vitality of human cardiac progenitor cells (hCPC) and human target for cardiovascular diseases via enhancing the number of circulating cardiospheres (hCS) is essential for healthy myocardial tissue endothelial progenitor cell (EPC) in diabetes. But the direct effects of sita- and can be impaired in diabetes. In this study, we investigated the effects of gliptin on EPC function and the underlying mechanism remain elusive. We palmitate, oleate or eicosapentaenoic acid (EPA) and the protective effects evaluated the pro-angiogenic effects of sitagliptin in a diabetic hind limb of the GLP-1 analog exendin-4 on the viability of hCS and hCPC, isolated from ischemia (HLI) model in vivo and in an EPC culture system in vitro. db/db mice human right auricle biopsies. Apoptosis was assessed by caspase-3 cleav- were chosen to establish HLI model, followed by treatment with sitagliptin age and ELISA, and autophagy by autophagosome labeling and LC3-II and (Januvia®, 25mg/kg per day) or vehicle for 1 week before HLI surgery and beclin1 immunoblotting. accumulation was evidenced by immuno- for additional 4 weeks after surgery via gavage administration. Sitagliptin fluorescence. Palmitate, but not oleate or EPA, induced apoptosis in hCPC. treatment efficiently enhanced ischemic and blood perfusion, Exposure to palmitate was also associated with impaired hCS isolation and which was accompanied by significantly increases in circulating EPC number. increased hCPC autophagy, as well as ceramide accumulation and increased EPCs derived from bone marrow of normal mice were treated with high glu- expression of ceramide synthase-5 (CerS5), a key enzyme in de novo synthe- cose (HG) to mimic hyperglycemic conditions in type 2 diabetes. We found sis of ceramide, in hCPC. However, both CerS5 inhibition with fumonisin-B1 that HG treatment induced EPC apoptosis and impairment in tube formation, and CerS5 knockdown reduced palmitate-induced apoptosis and autophagy. which were significantly prevented by sitagliptin pretreatment. Mechanistic Noteworthy, pretreatment with exendin-4 preserved hCS isolation and study revealed that HG impaired the basal autophagy of EPCs, while sita- reduced the palmitate-induced CerS5 expression, ceramide biosynthesis gliptin pretreatment restored the autophagy of EPCs, and autophagy inhibi- and hCPC apoptosis and autophagy. Co-incubation with the GLP-1 receptor tor abolished the protective effects of sitagliptin on EPCs. The protection of (GLP-1R) antagonist exendin (9-39), or inhibition of GLP-1R signaling with the sitagliptin against HG was accompanied by sitagliptin activation of AMPK/ PKA inhibitor H89 or by selective knockdown of the GLP-1R fully abolished ULK-1/mTOR autophagic signal pathway. Altogether, these results indicate the ability of exendin-4 to prevent apoptosis and autophagy. that sitagliptin preservation of EPC angiogenic function results in improve- In conclusion, exendin-4 prevents the palmitate-induced abnormalities of ment of diabetic ischemia angiogenesis and blood perfusion, which are most hCS and hCPC by counteracting ceramide generation. Hence, GLP-1 and its likely mediated by sitagliptin augmenting EPCs autophagy activity to prevent analogues may limit the lipotoxic damage in the human heart by preserving diabetic hyperglycemia induced apoptosis. the viability of myocardial progenitor cells. Supported By: American Diabetes Association (1-13-JF-53 to Y.T.); JDRF (1-INO-2014-122-A-N); National Natural Science Foundation of China (81573435, 81200917, 81200239, 81370917); Key New Drug Development Grant (2012ZX09103- 301-016); Health and Family Planning Commission of Sichuan Province (130297); China Scholarship Council

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A127 COMPLICATIONS—MACROVASCULAR—CELLULAR MECHANISMS OF ATHEROGENESIS IN DIABETES

485‑P Association of Imaging Markers of Diffuse Myocardial Fibrosis by CMR T1ρ Mapping and T1 Mapping with Myocardial Diastolic Dysfunction in Rhesus Monkeys with Spontaneous Type 2 Diabetes Mellitus YU ZHANG, FABAO GAO, JIE ZHENG, Chengdu, China, St. Louis, MO Aim: To investigate the ability of T1ρ mapping in diffuse myocardial fibro-

POSTERS sis assessment compared with T1 mapping; to explore the association of Supported By: National Natural Science Foundation of China (81520108014)

Complications imaging markers of diffuse myocardial fibrosis and diastolic dysfunction (DD) Acute and Chronic in nonhuman-primate diabetic model. 486‑P Method and Result: 14 monkeys with spontaneous type 2 diabetes Role of Lactate Dehydrogenase A (LDH-A) in Diabetic Vascular mellitus (T2DM) and 5 matched controls were studied. Blood biochemical Complication examination was tested during four-week acclimation. CMR protocol: cine, KWI HYUN BAE, JI HYUN KIM, JAE HAN JEON, HYE JIN HAM, YEON KYUNG T1ρ mapping, T1 mapping, and LGE. Pre-T1, post-T1, ECV, myocardial fibrosis CHOI, IN KYU LEE, MI JIN KIM, HYUN AE SEO, KEUN GYU PARK, SUNG WOO index (mFI) based on the dispersion characteristics of T1ρ, and T1ρ were KIM, Daegu, Republic of Korea, Gumi, Republic of Korea calculated. Before CMR, ultrasound was performed. One heart from dia- Pathological proliferation and migration of vascular smooth muscle cells betic monkey was harvested for HE and Masson staining. Ultrasound shows (VSMCs) has been implicated in the pathogenesis of accelerated athero- all monkeys in T2DM have DD whereas 5 controls were normal. The basic sclerosis in patients with diabetes mellitus. Increased aerobic glycolysis characteristics, imaging parameters, relationship between DD and imaging is a key feature of cellular phenotypes including cancer and immune cells. markers are demonstrated in Figure 1-A. CMR parametric maps shows in Fig- However, it remains largely unknown about the role of aerobic glycoly- ure 1-B. There was no evidence of regional LGE in all animals. Histopathol- sis in atherogenic phenotype of VSMCs. Here, we investigated the role ogy showed myocardial extracellular space widened with diffuse fibrosis. of lactate dehydrogenase-A (LDH-A), which is a key enzyme for glycoly- Conclusion: T1ρ mapping has potential to assess diffuse myocardial fibro- sis, on proliferation and migration of VSMCs. Activation of VSMCs with sis. The association of myocardial DD, mFI, and ECV suggest a link between platelet-derived growth factor (PDGF) or fetal bovine serum (FBS) resulted them in diabetic cardiomyopathy. in cellular proliferation and migration and increased glycolytic activ- ity accompanied by the increased expressions of 1 (GLUT1), hexokinase (HK) 2 and LDH-A in primary rat VSMCs. Through healing assay, actin stress fiber staining and transwell migration assay, we observed that both pharmacological inhibition (oxamate) and siRNA-mediated knockdown of LDH-A (siLDH-A) effectively inhibited cel- lular migration. Oxamate reduced PDGF stimulated glucose uptake, lactate production, ATP production and NAD/nicotinamide adenine dinucleotide (NADH) ratio. Nicotinamide mononucleotide (NMN), nucleotide precur- sor of NAD+, partially but significantly recovered oxamate treatment or siLDH-A induced inhibition of VSMC proliferation and migration, suggest- ing NAD+ involvement in LDH-A mediated VSMC proliferation and migra- tion. Taken together, this study shows that enhanced aerobic glycolysis in PDGF- and FBS-stimulated VSMCs plays an important role in their prolifer- ation and migration and suggests that LDHA can be a potential therapeutic target for the prevention of diabetic vascular complications.

487‑P Serum Levels of Sfrp5 Are Associated with a Favourable Cardio‑ metabolic Risk Profile: Results from the KORA F4 Study MAREN CARSTENSEN-KIRBERG, JULIA M. KANNENBERG, CORNELIA HUTH, CHRISTA MEISINGER, WOLFGANG KOENIG, ANNETTE PETERS, WOLFGANG RATHMANN, MICHAEL RODEN, CHRISTIAN HERDER, BARBARA THORAND, Düs- seldorf, Germany, Munich, Germany Secreted frizzled-related protein 5 (Sfrp5) has been described as an anti-inflammatory and insulin-sensitizing protein in mice. In contrast, findings from human studies on the role of Sfrp5 in the development of type 2 diabetes (T2D) and cardiovascular disease are currently controver- sial. Therefore, it remains unclear to what extent endogenous Sfrp5 levels are associated with cardiometabolic risk factors. The aim of this study is to identify correlates of Sfrp5 serum levels within a population-based cohort. Serum levels of Sfrp5 were measured by ELISA in 1097 participants aged 62-81 years of the Cooperative Health Research in the Region of Augs- burg (KORA) F4 study. All participants without known T2D were assigned to receive a standard 75-g oral . Individuals with known T2D (n=156) and those using lipid-lowering drugs (n=268) were excluded from the analyses of associations with glycaemia and serum lipids, respectively. In the age- and sex-adjusted model, serum Sfrp5 was lower in patients with prediabetes or T2D compared to individuals with normal glucose tolerance. Sfrp5 was inversely associated with fasting glucose, 2-h glucose, HbA1c, BMI, triglycerides, blood pressure, inter- leukin (IL)-6 and IL-1 receptor antagonist (all P<0.05). In contrast, Sfrp5 was positively associated with HDL cholesterol, adiponectin and omentin (P<0.05). After additional adjustment for lifestyle factors (smoking, alco- hol consumption, physical activity), Sfrp5 remained inversely associated with BMI, fasting glucose, HbA1c, blood pressure and IL-6, and positively with adiponectin and omentin. Thus, serum levels of Sfrp5 showed inverse associations with many cardiometabolic risk factors and positive associa- tions with factors implicated in lower risk of cardiometabolic diseases. Our

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A128 COMPLICATIONS—MACROVASCULAR—CELLULAR MECHANISMS OF ATHEROGENESIS IN DIABETES data support the hypothesis that Sfrp5 may also have a protective role in 490‑P the development of cardiometabolic diseases in humans. High Glucose With and Without High Palmitate Represses β-klotho Supported By: German Research Foundation; German Federal Ministry of Expression and Impairs FGF-21 in Mouse Heart Health; German Ministry of Innovation, Science, Research and Technology, North YONGSOO PARK, MINGGUANG CUI, HYUNOK KIM, JUNLIAN GU, LU CAI, Louis- Rhine-Westphalia; German Center for Diabetes Research; German Federal Minis- ville, KY, Seoul, Republic of Korea try of Education and Research; State of Bavaria The heart expresses FGF-21, the action of which in the heart under normal and diabetic conditions are not fully understood. This study investigated the

488‑P effects of high glucose with and without high palmitate on heart FGF-21 POSTERS A Plant-Derived Drug Improves the Cardiac Function of Monkeys actions by investigating responses of diet-induced obesity and db/db mouse Complications with Heart Failure and Type 2 Diabetes to exogenous FGF-21. The direct effects of glucose and fatty acid on FGF-21 Acute and Chronic ZHIGANG LIANG, YINAN LIANG, ZUNYUAN YANG, YUBO SHEN, ZUNWEI YAO, signaling in vitro, and the involvement of Sirt-1/PPARγ in FGF-21 pathway YUANHAI CHEN, LI GONG, WEN ZENG, BARBARA C. HANSEN, Chengdu, China, activation were also sought. Similar to what is found in the liver, the expres- Gulfport, FL sion of FGF-21 in the diabetic heart is also very high. HFD treated B6 mouse Heart failure (HF), an end stage cardiovascular complication of type 2 hearts, aged db/db mouse hearts and normal cardiomyocytes treated with diabetes (T2DM), is associated with significant morbidity and mortality, high glucose with and without palmitate in vitro displayed reduced β-klotho while safe and effective treatments for systolic HF with diastolic dysfunc- expression and suppressed responses to FGF-21 (resistance to FGF-21), and tion are lacking. To assess the efficacy and safety of a plant-derived drug decreased Sirt-1/PPARγ expression culminating in severe cardiomyocyte (HTL002) and compared to Entresto (LCZ696) as a positive control (PC) in impairment. Antioxidants similar to FGF-21 ameliorated and reversed cardiac diabetic HF monkeys, a total of 12 male monkeys (Age 13.3±3.3 years, BW FGF-21 resistance, and increased AMPK phosphorylation, Sirt-1 and PPARγ 11.6±1.5 kg, FPG 93±15.6 mg/dl, and diabetes duration 3.6±1.3 years) all with expression in the heart and thereby improved cardiomyocyte impairment. T2DM and reduced ejection fraction (EF) (>10% reduction from normal EF of Sirt-1 siRNA and nicotinamide blocked AMPK phosphorylation as well as monkey) were randomly divided into 4 groups (3 animals/group and ≥ 2 ani- Sirt-1 expression, whereas reservatrol treatment rescued Sirt-1 blockage mals combined with diastolic dysfunction in each group): HTL002 low-dose and reversed FGF-21 resistance. , an antidiabetic PPARγ ligand, group (LD)(65 to 200 mg/kg), HTL002 high-dose group (HD)(130 to 250 mg/ also ameliorated cardiac FGF-21 resistance. Our data indicate that high glu- kg), a positive control group (PC)(Entresto [ratio / doses] cose with and without high palmitate in type 2 diabetes may lead to FGF-21 from 0.80/0.86 to 6.48/6.85) and Vehicle control group (VC). Monkeys were resistance in the heart, probably through reduction of Sirt-1/ PPARγ expres- dosed (q.d. and i.g.) for 13 weeks, and blood chemistry and echocardiography sion, which provides a novel mechanism for diabetes related cardiomyocyte were obtained at week -1, 1, 3, 6, 9 and 13. Systolic parameter EF increased dysfunction. 15.2%, 20.1%, 23.5% and zero in four groups respectively; shortening frac- tion increased 14.6%, 29.1%, 30.4% and zero, and stroke volume increased 491‑P 27.1%, 40.7%, 40.1% and zero. Significant increases in end diastolic volume Inhibitory Factor 1 Knockout Protects the Hypertrophied Heart by and decreases in end systolic volume were seen in LD, HD, and PC, while Maintaining Mitochondrial Membrane Potential and Facilitating all parameters remained stable in VC. Significant improvements in diastolic Autophagy parameters (Mitral E/A ratio, Peak e’ velocity, e’/a’ ratio and Mitral E/e’ ratio) KEVIN YANG, QINQIANG LONG, FENGYUAN HUANG, LUFANG ZHOU, JUNJI were also observed in monkeys combined with diastolic dysfunction in HD NAKAMURA, MASASUKE YOSHIDA, QINGLIN YANG, Birmingham, AL, Wuhan, and PC. No side effects or deaths were observed. With safety and effective- China, Tsukuba, Japan, Kyoto, Japan ness, the plant-derived drug, HTL002, was safe and effective, and showed ATP synthase is pivotal to energy production and metabolism in the heart. similar efficacy to Entresto, indicating its potential for the improvement of In the pathological heart, ATP synthase can also maintain the mitochondrial cardiac function of HF patients with T2DM. membrane potential by hydrolyzing ATP and replenishing protons in the intermembrane space. Inhibitory factor 1 (ATPIF1 or IF1) is an ATP synthase- 489‑P interacting protein that suppresses the hydrolytic activity and reserves ATP. Global and Liver-Specific Bmal1-Deficient Mice Regulate Intestinal As a result, IF1 mediates a trade-off between preserving mitochondrial Glucose Absorption integrity and ATP supply. Our earlier study indicated that inhibition of IF1 in XIAOYUE PAN, MOHAMMED ULLAH, MAHMOOD M. HUSSAIN, Mineola, NY mice is protective against pressure-overload cardiac hypertrophy induced Clock and Bmal1 are critical clock genes that positively regulate circadian by transverse aortic constriction. Similarly, cell viability was improved in rhythms. Circadian rhythms controlled by clock genes affect plasma glucose. IF1 knockdown cells and exacerbated in IF1 overexpressed cells compared Bmal1 has been shown to regulate plasma glucose by regulating insulin to WT. To examine if the loss of IF1 would help maintain the mitochondrial secretion by the . We have shown that Clock plays an important membrane potential, we performed JC1 assay on mitochondria isolated role in lipid and absorption, and hypothesized that disruptions from the hypertrophied hearts of mice and revealed that mitochondrial in circadian expression of intestinal transporters may contribute to hyperlip- membrane potential in IF1 knockout was improved compared to WT. Consis- idemia and hyperglycemia. Here, we explored the role of Bmal1 on intestinal tently, JC1 assay showed improved membrane potential in IF1 knockdown, glucose absorption. In this study, we used global and liver-specific Bmal1 but increased depolarization in IF1 overexpressed H9C2 cells treated with deficient C57Bl6J (Bmal1−/− and L-Bmal1−/−) and Apoe−/− (Bmal1−/−Apoe−/− and isoproterenol compared to that of WT. As expected, bioenergetic study L-Bmal1−/−Apoe−/−) mice fed a chow diet ad libitum to clarify the role of Bmal1 revealed that the IF1 deficient H9C2 cells showed reduced mitochondrial- in the intestinal glucose absorption. Oral gavage of radiolabeled a-meth- dependent ATP production. Consequently, AMPK activity was upregulated ylglucose resulted in increased absorption of glucose in Bmal1−/− mice but as indicated by Western blot analysis in IF1 KO mice. To determine if the glucose absorption was reduced in L-Bmal1−/− mice. To explain differences in AMPK activation would facilitate autophagic flux, we assessed LC3 and p62 glucose absorption in global and liver specific Bmal1 deficient mice, we stud- levels using Western blot analyses. LC3 II levels were increased and p62 lev- ied glucose uptake in isolated enterocytes. Enterocytes isolated from both els were decreased in the hypertrophied IF1 KO mice, indicating an increased Bmal1−/− and L-Bmal1-/- mice took up more glucose than control mice. Further, autophagic flux in the IF1 KO heart. we quantified mRNA levels of three glucose transporters; SGLT1, Glut2 and In conclusion, IF1 plays a key role in maintaining mitochondrial function Glut5. Expression levels of SGLT1 and Glut2 were increased in Bmal1−/− mice. and integrity, thus, serving as a therapeutic target for pathological cardiac Whereas, SGLT1 mRNA levels were increased in the intestine of L-Bmal1−/− hypertrophy with metabolic disorders, such as diabetic cardiomyopathy. mice, but Glut2 and Glut 5 mRNA levels were unchanged in L-Bmal1−/− mice. In addition, Bmal1-/- and L-Bmal1-/- mice in Apoe-/- background shown simi- larly effect in glucose absorption. These studies show that global and liver- specific ablation of Bmal1 has similar effect on the expression of intestinal SGLT1, has no effect on Glut2 and Glut5 expression. Global deficiency of Bmal1 increases intestinal glucose up take by increasing the expression of SGLT1 and Glut2. These data suggest that intestinal carbohydrate absorp- tion is regulated by Bmal1 involving different mechanisms. Supported By: American Heart Association

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A129 COMPLICATIONS—NEPHROPATHY—BASIC AND EXPERIMENTAL SCIENCE

COMPLICATIONS—NEPHROPATHY—BASIC AND glucosuria in NHE3-KO may reflect enhanced renal to nor- EXPERIMENTAL SCIENCE malize ammonium secretion. Supported By: National Institutes of Health; Boehringer Ingelheim GmbH Moderated Poster Discussion: Targets and Mechanisms in Experimen- & 494‑P tal Diabetic Nephropathy (Posters: 492-P to 497-P), see page 19. Do Insulin or IGF1 Receptors Contribute to Glomerular Dysfunctions Induced by Diabetes?

POSTERS 492‑P ATSUSHI ISHIKADO, QIAN LI, KYOUNGMIN PARK, TAKANORI SHINJO, YAS- Complications & UTAKA MAEDA, HISASHI YOKOMIZO, WEIER , RONALD ST-LOUIS, GEORGE L.

Acute and Chronic Renoprotective Effects of and Empagliflozin in a Rat Model of Early Diabetic Nephropathy KING, Boston, MA ANGUS GILL, STEPHEN GRAY, ANNA WATSON, MARK E. COOPER, KARIN A.M. Mesangial cells (MCs) have receptors for and respond to insulin and IGF1 JANDELEIT-DAHM, Melbourne, Australia with biological actions, yet both the deficiency and excess of their actions Linagliptin (L) a DPP-4 inhibitor, and Empagliflozin (E), a SGLT2 inhibitor, are have been associated with diabetic nephropathy (DN). To determine their recent antihyperglycemic interventions involved in the treatment of type 2 effects individually, receptors for insulin (IR) or IGF1’s (IGF1R) were selec- diabetes mellitus. However, the glucose-independent mechanisms of reno- tively deleted in mesangial cells by cross-breeding IR or IGF1R (flox/flox) mice protection afforded by these interventions are largely unknown. with SM22-cre or MYH11-cre mice to generate either SMIRKO or SMIGF1RKO Methods: 6 wk old male Sprague-Dawley rats were randomised into 6 mice. In MCs cultured from wild type mice (WT), SMIRKO and SMIGF1RKO mice, insulin and IGF1 activated PI3 kinase/pAkt and pErk pathways and control (C) or diabetic (D) treatment groups (10-15/gp), Untreated (H2O vehi- cle control), L treated (3mg/kg/day) and E treated (10mg/kg/day) per gavage. induced mRNA and protein expressions of anti-oxidative enzyme heme oxy- Diabetic rats were rendered diabetic via STZ injection (55mg/kg) and culled genase-1 (HO-1), matrix enzyme, hyaluronan synthase HAS2, inflammatory after 12 wks. proteins VCAM-1 and MCP-1 in a dose-dependent manner (1-100nM), which Results: Doses of drugs were chosen to be associated with no statistically paralleled with increased adhesion of monocytes (THP-1) cells to MCs by significant change in HbA1c (control 3.1±0.2%, diabetic 7.0±0.8%, p<0.05, L 2-fold. At physiological levels, only IGF1 induced HAS2, VCAM-1 and MCP-1 6.3±1.2%, E 6.0±0.8%). Both interventions significantly attenuated albumin- expressions which were inhibited by MEK/MAPK inhibitor, PD98059 or PKC uria (untreated diabetic 12.3±3.0 mg/24 hours, L 7.5±2.1 and E 4.8± 0.9 mg/24 inhibitor, GFX109203x significantly. In MCs from SMIRKO mice, insulin and hours, both p <0.05) and reduced hyperfiltration significantly in diabetic IGF1 induced phosphorylation of Akt (pAkt) and their receptors (pTyr IR or rats. Renal VEGF protein was significantly attenuated by L (150±10 vs. 210± pTyr IGF1R) were inhibited, respectively. Interestingly, the deletion of IGF1R 8 ug/mg in untreated diabetic rats, p<0.05, and by E 140±20 ug/mg, p<0.05). in MCs increased insulin activation of its receptors and pAkt by 2-3 fold Diabetes increased gene expression of NOX2 and NOX4 which was signifi- but decreased IGF1’s signaling in parallel with decrease in HAS2 but not cantly attenuated by L and E. Nitrotyrosine was significantly reduced by E HO-1 expressions. In vivo studies comparing WT and SMIRKO mice with (10±2 ug/mg vs. 15±2 ug/mg in untreated diabetic rats) but not by L treat- STZ-diabetes or control, no differences were observed in insulin sensitivity, ment. Renal MCP-1 protein, a pro-inflammatory chemokine, was reduced in weight, albuminuria or glomerular after 4 months of diabetes. In E treated diabetic rats (0.18±0.50 ug/mg vs. 0.35±0.10 ug/mg in untreated contrast, STZ-diabetic SMIGF1KO mice exhibited significant less albuminuria diabetic rats, p<0.05). There was no significant attenuation of mesangial than STZ-diabetic WT mice after 2-3 months of disease. Thus, in MCs, IGF1/ area in treated diabetic animals. receptors and not insulin have physiological effects on glomerular basement Conclusion: This study demonstrates glucose-independent reductions in membranes and expressions of inflammatory which contribute to pro-inflammatory and oxidative stress markers by L and E. These findings the development of diabetic glomerular pathologies. provide a potential reason for some of the renoprotective actions of these Supported By: National Institutes of Health agents seen in clinical trials and also provide a potential rationale for these agents in type 1 diabetic patients with diabetic nephropathy. & 495‑P Supported By: Boehringer Ingelheim GmbH Oral Treatment with PBI-4547, a Novel Antidiabetic and Antifibrotic Compound: A Potential Therapy For Diabetes, Diabetic Nephropa‑ & 493‑P thy, and Obesity Effect of Renal Tubule-Specific Knockdown of Na+/H+ Exchanger BRIGITTE GROUIX, MARIE-PIER CLOUTIER, ALEXANDRA FELTON, JONATHAN NHE3 in Akita Diabetic Mice RICHARD, ALEXANDRE LAVERDURE, LIETTE GERVAIS, PIERRE LAURIN, LYNE AKIRA ONISHI, YILING FU, WINNIE HUANG, PANAI SONG, ROHIT PATEL, VOLKER GAGNON, Laval, QC, Canada VALLON, San Diego, CA Introduction and Aims: Type 2 diabetes is a major health problem world- NHE3 contributes to sodium/bicarbonate reabsorption and ammonium wide. Adiponectin has been shown to play important roles in the regulation secretion in renal tubules. However, its role in the diabetic kidney remains of energy homeostasis and insulin sensitivity, and its low level is predic- incompletely understood. We generated mice with NHE3 knock-down in the tive of future development of diabetes. PBI-4547, a novel first-in-class orally tubular system (NHE3-KO; using Pax8-Cre) and crossed with type 1 diabetic active compound, displays anti-fibrotic activities via a novel mechanism of Akita mice (n=15-27/group). Systolic blood pressure (SBP) was measured action. This study examined the effect of PBI-4547 in leptin-deficient ob/ob by automated tail cuff, and glomerular filtration rate (GFR) by FITC-sinistrin mice, a model of type 2 diabetes and obesity. kinetics, both in conscious mice at 17-20 weeks of age. In the absence of dia- Methods: ob/ob mice (6 weeks old) were treated with vehicle or PBI- - 4547 (10 and 50 mg/kg, oral once a day) from day 1 through 105. Blood glu- betes, NHE3-KO showed increased urine pH and HCO3 to creatinine ratios vs. wild type (WT) (means±SEM: 7.9±0.1 vs. 6.9±0.1; 41±5 vs. 2±1 mol/mol, cose, white (WAT) histology, pro-inflammatory/fibrotic gene P<0.01), while blood pH and urine ammonium to creatinine ratios (AmCR) expression in kidney and liver, as well as serum adiponectin levels were were maintained. This was associated with lower GFR and SBP in NHE3-KO examined. vs. WT (263±9 vs. 325±11 µl/min; 118±1 vs. 122±1 mmHg; P<0.05). In WT and Results: In oral glucose tolerance test, PBI-4547 increased glucose metabo­ NHE3-KO, diabetes increased blood glucose (~550 mg/dl) and food and fluid lism. Serum cholesterol and triglyceride levels were also reduced by PBI- intake, and reduced body weight to similar levels. Diabetes also lowered 4547. Serum level of adiponectin was reduced in ob/ob non-treated mice and blood and urine pH and increased urine AmCR ratios to the same extent in strongly increased in PBI-4547-treated mice. Histomorphometry analysis NHE3-KO and WT; this was associated with greater urine glucose to cre- was performed on WAT and PBI-4547 treatment reduced fibrosis, inflamma- atinine ratios in NHE3-KO vs. WT (1256±77 vs. 748±53 mg/mg, P<0.01). In tory cell infiltration and adipocyte size. To further characterize the activity of NHE3-KO and WT, diabetes increased GFR (to 443±17 and 531±19 µl/min) and PBI-4547, quantitative RT-PCR analysis of profibrotic markers demonstrated SBP (to 124±2 and 126±1 mmHg) with GFR levels remaining lower in diabetic that PBI-4547 reduced α-SMA, fibronectin, CTGF, and MMP-2 expression in NHE3-KO vs. WT (P<0.05). Diabetes increased urine albumin to creatinine kidney, as well as collagen type I and III, MMP-2, TIMP1, TNF-α and PPAR-γ ratios in WT (62±16 vs. 16±2 µg/mg) but not in NHE3-KO (28±6 vs. 23±3 expression in liver. µg/mg). We conclude: 1) In the absence of diabetes, tubular KO of NHE3 Conclusions: Taken together, these results suggest that PBI-4547 offers - the potential as a novel therapy for diabetes, diabetic nephropathy, liver induced urine HCO3 loss, associated with reduced GFR and SBP, and com- pensated blood acid base status. 2) Type 1 diabetes induced a modest acido- fibrosis and obesity by reducing blood glucose levels, reducing pro-inflam- sis and increased GFR, SBP and albuminuria. 3) Tubular NHE3 determines the matory and profibrotic markers in kidney and liver and by increasing serum absolute levels of GFR and albuminuria in diabetes. 4) The effect of diabetes adiponectin to regulate energy homeostasis. on blood pH and AmCR was similar in the absence of NHE3. Increases in

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A130 COMPLICATIONS—NEPHROPATHY—BASIC AND EXPERIMENTAL SCIENCE & 496‑P factor-β;1 (TGF-β;1). Since mesangial cells are known to secret TGF-β;1 from Dipeptidyl Peptidase Inhibitor Linagliptin- and Angiotensin II early-stage DN, we expect that this culture system could simulate the early Receptor Blocker Telmisartan-Induced Effects on Renal and Uri‑ responses of in DN. Upregulated may participate in the nary Exosomal miRNA Expression in Rats with 5/6 Nephrectomy enhanced in vitro migration and attachment observed in HSMP treated with DENIS DELIC, KATHRIN SONTHEIMER, OLEG TSUPRYKOV, AHMED A. HASAN, TGF-β;1. Quantitative RT-PCR revealed that changes of expression levels BERTHOLD HOCHER, PATRICK BAUM, ANDREAS KOHLER, FLORIAN GANTNER, of several genes which were involved with proliferation, apoptosis, adhe- MICHAEL MARK, THOMAS KLEIN, Biberach, Germany, Potsdam, Germany sion, and signal transduction were occurred in TGF-β;1 treated HSMP. The Dipeptidyl peptidase (DPP)-4 inhibitors attenuate chronic kidney dis- symptom observed in early-stage DN may be caused by synergistic effects POSTERS ease progression in experimental diabetic nephropathy (DN) in a glucose- of such genes accelerated/suppressed by TGF-β;1, but not a simple down- Complications independent manner, but the exact molecular mechanisms remain unclear. regulation of integrin. Acute and Chronic MicroRNAs (miRNAs) are short, non-coding RNA that are important Supported By: Japan Diabetic Nephropathy Study Group post-transcriptional regulators of gene expression and play an important role in the pathogenesis of DN. miRNAs are present in urine in a remarkably 499‑P stable form, packaged in extracellular vesicles (predominantly exosomes). In Upregulation of Semicarbazide-Sensitive Amine Oxidase (SSAO) the present study, renal miRNA profiling was assessed using the Nanostring in Endothelial Cells Is the Mechanism of Oxidative Vascular Injury nCounter technology and mRNA profiling using RNA sequencing from the in Diabetes: Blockade by UD-014, a Novel SSAO Selective Inhibitor following animal groups: sham operated plus placebo; 5/6 nephrectomy TETSUO KAWAGUCHI, SHINPEI NONOUCHI, MASARU SHINOHARA, KOJI ITOH, plus placebo; 5/6 nephrectomy plus telmisartan; and 5/6 nephrectomy plus SHIGERU USHIYAMA, Ube, Japan linagliptin. To evaluate which of the deregulated miRNAs in the kidney can Semicarbazide-sensitive amine oxidase (SSAO)/vascular adhesion pro- be reflected in urinary exosomes, TaqMan Array miRNA Cards were used. tein-1 (VAP-1) expressed on the endothelium catalyzes deamination of In kidneys from 5/6 nephrectomized rats, the expression of 13 miRNAs primary monoamines such as methylamine and aminoacetone and gener- was significantly increased (>1.5-fold, P<0.05), whereas the expression ates cytotoxic molecules, H2O2, aldehydes and ammonia. The plasma SSAO of 7 miRNAs was significantly decreased (>1.5-fold, P<0.05). Most of the activity is known to be elevated in diabetes mellitus. Thus, SSAO has been deregulated miRNA species are implicated in TGF-β induced endothelial-to- considered to be one of the causative factors to induce microvascular injury mesenchymal transition processes. The ameliorative and restoring effects in diabetes. Our recent study found that UD-014, a novel, SSAO selective on miRNA expression were more strongly pronounced in animals treated inhibitor, ameliorated albuminuria in the streptozotocin-induced diabetic with 5/6 nephrectomy plus telmisartan compared with 5/6 nephrectomy plus nephropathy model in rats. However, its mechanism remains unclear. The linagliptin. Both telmisartan and linagliptin suppressed the induction of pro- aim of this study is to clarify the mechanism of action for UD-014 using glo- fibrotic miRNAs, such as miR-199a-3p, and restored levels of anti-fibrotic merular microvascular endothelial cells (GMVECs) focusing on the vascular miR-29c. In particular, linagliptin-induced restoring effects on miR-29c injury by oxidative stress. expression could also be reflected in urinary exosomes, which might be used GMVECs treated with 1 mM methylamine in 10% FBS culture medium as a biomarker for disease progression and monitoring of treatment effects. caused cell death in 2 day-culture. Unexpectedly, SSAO mRNA was upregu- Supported By: Boehringer Ingelheim lated by more than 50-fold prior to the cell death with a certain lag time. SSAO protein was also highly detected in the cells by immunofluorescent 497‑P staining. Pre-treatment of UD-014, completely inhibited the induction of SSAO and the cellular damage. Interestingly, post-treatment of UD-014 WITHDRAWN before SSAO over-expression also inhibited the cellular damage and SSAO mRNA over-expression. To investigate whether these effects are due to metabolites of methylamine, H2O2 was added to GMVECs. H2O2 also caused increased SSAO expression and cytotoxicity. SSAO upregulation may be a key to the damage of vascular endothelial cells in the presence of its substrates and contribute to the microvascular injury in diabetic complication. SSAO may be an intriguing therapeutic target for diabetic nephropathy and chronic kidney diseases by directly protecting the kidney from oxidative stress.

500‑P Suppression of High Mobility Group Box1 Improves Kidney Injury in Streptozotocin-Induced Diabetic Rats HONG ZHANG, XIANGCHENG ZHANG, RU ZHANG, WEI LI, Huaian, China, Xuzhou, China 498‑P Aim: High mobility group box 1 (HMGB1) has been implicated in the patho- Expressions of Integrins Are Upregulated in in Early- genesis of autoimmunity diseases, atherosclerosis, and diabetes. However, Stage Diabetic Nephropathy and Cultured Podocytes Stimulated the mechanism of HMGB1 on diabetic nephropathy remains unclear. Here, with TGF-1 we investigated the potential role and mechanism of HMGB1 on renal injury KAICHIRO SAWADA, MASAO TOYODA, HITOMI MORIYA, MORITSUGU KIMURA, in diabetic rats. TAKEHIKO WADA, MASAFUMI FUKAGAWA, Isehara, Japan Methods: In streptozotocin (STZ)-induced rat model, we evaluated urine injury plays an important role in the onset and progression of albumin creatinine ratio (UAC), morphologic changes of renal tissues. The diabetic nephropathy (DN). Downregulation of α;3β;1-integrin expression in levels of pro-inflammatory factor, chemokines, and cell adhesion molecules, podocytes has been thought to be associated with podocyte detachment type IV collagen in kidney tissues or serum were measured by quantitative from the glomerular . However, in our previous study, RT-PCR or ELISA. Furthermore, the expressions of HMGB1 and its receptors, the analysis of the expression level of integrin in podocytes of early and mitogen-activated protein kinases (MAPKS) and nuclear transcription factor advanced DN patients showed an upregulation of α;3- and β;1-integrins Kappa B(NF-κB) in kidney were assessed. In addition, we studied the effect in early-stage. To elucidate the mechanism of podocyte detachment in the of the HMGB1 inhibitor glycyrrhizin (GA) on the kidney in diabetic rats. course of DN progression, we analyzed the expression levels of α;3-, α;5-, Results: Diabetic rats showed elevated levels of UAC, TGF-β1 and col- α;V- and β;1-integrins in isolated glomerular from kidneys of diabetic rats pro- lagen IV, and increased expressions of TNF-α, IL-6, IL-1β, MCP-1 and ICAM-1 duced with streptozotocin (STZ) injection. Quantitative RT-PCR showed that in kidney. Importantly, we found the expressions of HMGB1, receptor for the expression of these integrins progressively increased during 2 months advanced glycation end products (RAGE) and toll-like receptor 4 (TLR-4) in after STZ injection and gradually decreased thereafter, which seemed to the kidney tissue were significantly upregulation, and the levels of phos- reproduce the process in DN patients. Immunohistochemistry showed that phorylated ERK, JNK and p38MARK and NF-κB activation were significantly α;3-integrin was accumulated chiefly in podocytes, whileα ;5-, α;V- and increased. Oral administration of GA ameliorated UAC, renal injury, and β;1-integrins were dispersed in whole glomerular. We also detected upregu- decreased levels of TNF-α, 1L-6, IL-1β, MCP-1, ICAM-1, TGF-β1 and collagen lations of α;3-, α;5-, α;V- and β;1-integrins in in vitro culture of podocyte IV. Furthermore we found the levels of HMGBI, RAGE and TLR4 in kidney tis- (HSMP: heat-sensitive mouse podocyte) treated with transforming growth sue in diabetic rats were inhibited by GA treatment. After GA treatment, the

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A131 COMPLICATIONS—NEPHROPATHY—BASIC AND EXPERIMENTAL SCIENCE

ERK and p38MAPK phosphorylation and NF-κB activation were significantly VAP-1 activity in the plasma was measured on the last day in the STZ rats. reduced while phosphorylation of JNK was not affected. VAP-1 activity increased about 2-fold compared to that of the control rats. Conclusions: Our findings indicated that HMGB1 may induce renal injury UD-014 inhibited the elevated plasma VAP-1 activity dose-dependently, but and inflammatory responses in diabetic rats through RAGE/TLR4-NF-κB did not. We also observed that VAP-1 was expressed in the glom- activation. eruli in the kidney using immunofluorescence staining, and the site of VAP-1 Supported By: National Natural Science Foundation of China (81200595, expression co-localized with the up-regulated site of α-SMA, a marker of 81400807); Six High-Peak Talents Project of Jiangsu Province (WSN-101); Jiangsu transformed mesangial cells, in the STZ rats. Provincial Commission of Health and Family Planning (F201549/H201667) These results suggest that UD-014 has the potential to be a therapeutic POSTERS

Complications agent to ameliorate kidney functions in diabetic nephropathy. Acute and Chronic 501‑P 504‑P Role of Dietary Balance on Diet-Restriction-Mediated WITHDRAWN Renoprotection in Diabetic Nephropathy SHOHEI YOSHIDA, KOSUKE YAMAHARA, SHINJI KUME, NAOKO TAKEDA, MASAMI CHIN-KANASAKI, HISAZUMI ARAKI, DAISUKE KOYA, YUSUKE ADA- CHI, SHIN ICHI ARAKI, HIROSHI MAEGAWA, Otsu, Japan, Kahoku, Japan, Kawa- saki, Japan Diet restriction (DR) exerts renoprotective effects in diabetic nephropa- 502‑P thy, yet it is unclear what dietary nutrient is responsible for DR-mediated PBI-4050 Prevents Destruction of β-Cells and Islets, and Diabetic renoprotection. An interesting recent study reported that an imbalance Nephropathy in NOD Mice, a Model of Type 1 Diabetes between dietary essential and nonessential amino acids (EAA and NEAA) LYNE GAGNON, KATHY HINCE, FRANÇOIS SARRA-BOURNET, MIKAËL TREM- negated DR-mediated lifespan elongation in Drosophila. We hypothesized BLAY, MARIE-PIER CLOUTIER, SYLVIE LÉTOURNEAU, PIERRE LAURIN, BRIGITTE that the dietary amino acid imbalance affects DR-mediated renoprotection GROUIX, FRANÇOIS LEBLOND, Laval, QC, Canada in diabetic nephropathy. To this end, we examined the effects of recovering Introduction: PBI-4050, a novel first-in-class orally active compound which either all dietary amino acids, only NEAA, or only EAA on DR-mediated reno- is currently in a phase II clinical trial, significantly reduced glycated hemo- protection in diabetic db/db mice. Ten-week-old db/db mice were divided globin (HbA1c) after 12 and 24 weeks of treatment in patients with type 2 into five groups: ad libitum; simple DR; DR + all amino acids (AAA); DR + diabetes and metabolic syndrome with elevated HbA1c despite anti-hyper- NEAA; and DR + EAA. Dietary intake in the DR groups was restricted by 40% glycemic treatment. In the present study, we examined whether PBI-4050 of that of the ad libitum group. Groups were followed until 22 weeks old. DR may prevent diabetes and diabetic nephropathy in NOD mice, a model of completely prevented mesangial expansion with periodic acid-Schiff stain- type 1 diabetes. ing and partly, but significantly, ameliorated albuminuria and podocyte foot Methods: NOD/ShiLtJ female mice (8 weeks of age) received vehicle process effacement in db/db mice. Recovering dietary amino acids, regard- (water) or PBI-4050 (200 mg/kg/day) by daily gastric gavage for 23 weeks. less of group, neither altered whole-body glucose and lipid metabolism nor Results: Mice were housed in sterile conditions and did not develop dia- canceled DR-mediated renoprotective effects in db/db mice. Recovering betes after 7 weeks of treatment. Mice were then housed in a non-sterile dietary NEAA alone surprisingly and additionally ameliorated residual albu- environment and 4 weeks later started to develop diabetes. However, mice minuria, accompanied by longer podocyte foot processes in diet-restricted treated with PBI-4050 did not develop diabetes. OGTT was also normalized db/db mice; findings not observed in the DR + AAA and DR + EAA groups. in PBI-4050 treated mice. NOD mice treated with PBI-4050 maintained nor- In conclusion, dietary NEAA appeared to exert an additional renoprotec- moglycemia despite of a reduced number of islets. In fact, PBI-4050 reduced tive effect on this diet-restricted mouse model of diabetes although the by 30% the number of severely damaged islets. In kidney, PBI-4050 protects responsible molecular mechanism remains unclear. The balance in dietary against lesions by reducing glomerular volume and mesangial cross sec- amino acids can influence DR-mediated renoprotective effects in diabetic tional area. Furthermore, tubular lesions were absent in PBI-4050-treated nephropathy. These results shed new light on the development of effective mice. Tubular dystrophy as PAS granular accumulation was constantly pres- diet therapy in diabetic nephropathy. ent in proximal tubules of NOD control mice and treatment with PBI-4050 completely abrogated that dystrophy. 505‑P Conclusion: This data suggests that PBI-4050 prevents destruction Dipeptidyl Peptidase-4 Plays a Pathogenic Role in BSA-Induced of β-cell and islets in type 1 diabetes model, and also prevents diabetic Kidney Injury in Diabetic Mice nephropathy. YUTA TAKAGAKI, KEIZO KANASAKI, MAKOTO KATO, DAISUKE KOYA, Uchinada, Japan, Kahoku, Japan, Tokyo, Japan 503‑P induces tubular epithelial cells (TECs) damage and influ- UD-014, a Novel, Selective, Orally Available, Long-Acting Vascular ences the prognosis of diabetic kidney disease (DKD). Epithelial-mesen- Adhesion Protein-1 Inhibitor, Improves Albuminuria in a Strepto‑ chymal transition (EMT) program of TECs could be important player for zotocin (STZ)-Induced Diabetic Nephropathy Model in Rats the extracellular matrix (ECM) production in kidney cells. We have already MASARU SHINOHARA, KOUJI ITOH, TETSUO KAWAGUCHI, HIROYOSHI reported that the interaction between dipeptidyl peptidase-4 (DPP-4) and KAWADA, AKISHI NINOMIYA, KENICHI KOMORI, NORIAKI IWASE, SHIGERU integrin β1 (INTβ1) induced endothelial-mesenchymal transition in dia- USHIYAMA, Ube, Japan betic kidney. Caveolin-1 (CAV1), the protein known to interact with either Vascular adhesion protein-1 (VAP-1) that is also known as semicarbazide- DPP-4 or INTβ1, is the scaffolding protein within caveolar membranes and sensitive amine oxidase (SSAO) is recognized to increases in plasma of regulates cellular homeostasis. Here we found that DPP-4 inhibition sup- patients with inflammation-associated diseases, and is considered a poten- pressed the EMT program in proteinuria-damaged TECs of type 1 diabetic tial therapeutic target for various inflammatory diseases, including diabetic mice via disruption of the interaction between DPP-4, INTβ1 and CAV1. complications. UD-014 is a novel, potent and orally active VAP-1 inhibitor We utilized proteinuria-induced TECs damage model with intraperitoneal with an extended duration of action. The aim of this study is to investigate free fatty acid (FFA)-bound 30% bovine serum albumin (BSA) for 14 days efficacy of UD-014 on kidney function in a STZ-induced diabetic nephropathy in streptozotocin (STZ)-induced type 1 diabetic CD-1 mice (4 weeks after model in rats. STZ injection). DPP-4 inhibitor teneligliptin (TENE) was initiated with same periods. BSA injection induced TECs damage, characterized by tubular cell UD-014 inhibited recombinant human VAP-1 with an IC50 value of 3.2 nM. flattering and lumen dilation, tubular cell vacuolation and tubulointerstitial IC50 values of MAO-A and B (other classes of monoamine oxidase) inhibition were 6.1 μM and 12 μM, respectively. UD-014 was orally administrated at fibrosis, and TENE ameliorated such alterations. The expression of DPP-4, single doses (0.1-10 mg/kg) to normal SD rats. Plasma VAP-1 activity was INTβ1, CAV1, and EMT-relevant molecules (snail, slug and Zeb1 etc.) were inhibited dose-dependently up to 24 hrs. SD rats were subjected to type 1 increased with BSA injection and TENE restored all in diabetic mice. Immu- diabetes by an i.v. injection of streptozotocin (50 mg/mL/kg) and were orally nostaining showed BSA injection accelerated EMT program in proximal administered UD-014 (1 and 3 mg/kg) or losartan (30 mg/kg) once a day from tubular cells of diabetic mice; TENE ameliorated EMT program. EMT pro- the first day to 4 weeks after STZ treatment. Urinary albumin and urinary gram in damaged TECs can induce tubulointerstitial fibrosis via producing L-FABP, a marker of disorder and oxidative stress, were ECM. We also found TENE suppressed the interactions between DPP-4, reduced significantly by UD-014 treatment compared to the vehicle control. INTβ1 and CAV1 in HK-2 cell. These findings suggest that DPP-4 inhibition

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A132 COMPLICATIONS—NEPHROPATHY—BASIC AND EXPERIMENTAL SCIENCE can be relevant to combat DKD by targeting such EMT program induced by 508‑P the crosstalk between DPP-4, INTβ1 and CAV1. Ferroptosis Is Involved in Renal Tubular Cell Death in Diabetic Nephropathy 506‑P SEONGHUN KIM, BO YOUNG NAM, JIMIN PARK, MEIYAN WU, SUKYUNG SGLT2 Inhibitor Ameliorates Diabetic Nephropathy Independently KANG, JUNG TAK PARK, SEUNG HYEOK HAN, TAE HYUN YOO, SHIN WOOK of Its Glucose-Lowering Effect: A Novel Mechanism for Renopro‑ KANG, Seoul, Republic of Korea tective Effect Ferroptosis, an atypical iron-dependent cell death, is distinct from apop-

TOSHINOBU MAKI, SAYAKA MAENO, YASUTAKA MAEDA, NORIYUKI SONODA, tosis, necroptosis, and autophagy. Two molecules, glutathione peroxidase 4 POSTERS MASANORI WAKISAKA, YOSHIHIRO OGAWA, TOYOSHI INOGUCHI, Fukuoka, (GPX4) and glutamate/cystine antiporter (xCT), are known to be principally Complications Japan involved. Recently, ferroptosis has been reported to cause several kidney Acute and Chronic Several clinical studies have shown the beneficial effects of sodium- diseases. However, the impact of ferroptosis on tubular cell death under glucose cotransporter-2 (SGLT2) inhibitors on diabetic nephropathy. How- diabetic conditions has never been elucidated. In vitro, rat proximal tubu- ever, the molecular mechanism is poorly understood. Here we show that lar epithelial cells (NRK-52Es) were cultured in DMEM media containing 5.6 very low dose of (Cana), a SGLT2 inhibitor, ameliorates dia- mM glucose (normal glucose, NG) or NG + TGF-β1 (10 ng/ml) with or with- betic nephropathy independently of its glucose lowering effect. Cana was out ferroptosis inhibitors (Ferrostatin-1 and Liproxstatin-1) and iron chelator orally administrated to 12-week-old db/db mice for 2 or 8 weeks. Cana (3.0 (Deferoxamine). In vivo, 24 C57BL/6 mice were intraperitoneally injected mg/kg/day, conventional dose) significantly reduced plasma glucose and with saline (Control, C) (N=12) or STZ (50 mg/kg/d) for 5 consecutive days fructosamine levels, but very low dose of Cana (0.01 mg/kg/day) did not (diabetes, DM) (N=12), and 6 mice from each group were treated with Fer- affect body weight, blood pressure, plasma glucose, or fructosamine levels rostatin-1 (5 mg/kg/d) for 6 weeks. NRK-52Es and the mouse kidneys were in db/db mice. Of great interest, very low dose as well as conventional dose analyzed for GPX4, xCT, hypoxia-inducible factor (HIF)-1α, HIF-2α, heme of Cana significantly ameliorated albuminuria in db/db mice to control lev- oxygenase-1 (HO-1). Cell viability and lipid peroxidation (MDA) were also els. In 8 weeks treatment, renal mesangial expansion found in db/db mice evaluated in NRK-52Es. Compared to NG cells, the protein expressions of was also ameliorated by very low dose of Cana as similarly as conventional GPX4, xCT, HIF-1a, HIF-2a and HO-1 were significantly decreased in TGF-β1- dose. Next, renal mesangial cells (MCs) were isolated from kidneys of db/+ stimulated NRK-52Es. MDA levels were significantly increased along with mice. Western blotting analysis revealed the expression of SGLT2 in renal significantly decreased cell viability in TGF-β1-stimulated NRK-52Es. These mesangial cells. In addition, we examined the effect of Cana (0, 10, 100 nM) changes were significantly ameliorated by ferroptosis inhibition. Further- on high glucose level-induced reactive oxygen species (ROS) production in more, a significant decrease in xCT protein expression was observed in the cultured MCs. Exposure of MCs to a high glucose level for 72 hr significantly kidney of DM mice compared to C kidney, and these changes in DM kidney increased ROS production comparing to exposure to a normal glucose level. were significantly attenuated by Ferrostatin-1 treatment. These results sug- This increase was significantly inhibited by the treatment with 100nM Cana. gest that ferroptosis is involved in renal tubular cell death under diabetic In conclusion, very low dose of SGLT2 inhibitor may ameliorate diabetic conditions and that ferroptosis inhibition can be a promising therapeutic nephropathy independently of its glucose lowering effect. The inhibition of target in patients with diabetic nephropathy. SGLT2 in renal MCs may cause an inhibition of ROS production and this may be at least in part associated with the renoprotective effect of very low dose 509‑P of SGLT2 inhibitor. Very low dose of SGLT2 inhibitor may serve as an effec- , a Novel Peroxisome Proliferator-Activated Recep‑ tive therapeutic agent for diabetic nephropathy without various adverse tor Gamma Agonist, Attenuates Renal Fibrosis Caused by Unilateral effects caused by increased urinary glucose excretion. Ureteral Obstruction KWI HYUN BAE, GWON SOO JUNG, JUNG BEOM SEO, YUN A. JUNG, MI JIN 507‑P KIM, YEON KYUNG CHOI, HYE JIN HAM, KEUN GYU PARK, Daegu, Republic of Epigenetic Modifications in Diabetic Nephropathy Korea VIKRAM THAKUR, JOSUE ENRIQUEZ, POMPEYO QUESADA, ROBERTO SOLIS, Renal tubulointerstitial fibrosis is a common feature of the final stage of MAYRA GONZALEZ, MUNMUN CHATTOPADHYAY, El Paso, TX nearly all cause types of . Although classic peroxi- Nephropathy is the leading cause of end-stage renal disease in diabetic some proliferator-activated receptor-gamma (PPARγ) agonists have a pro- patients, unfortunately available treatments are inadequate. Although tective effect on diabetic nephropathy, much less is known about their direct the pathogenesis of diabetic nephropathy is multifactorial, only a limited effects in renal fibrosis. This study aimed to investigate possible beneficial number of studies have highlighted the importance of histone deacetylases effects of lobeglitazone, a novel PPARγ agonist, on renal fibrosis in mice (HDAC)s in renal dysfunction. HDACs-mediated epigenetic mechanisms play with unilateral ureteral obstruction (UUO). Through hematoxylin/eosin and critical roles in the homeostasis of histone acetylation and gene transcrip- Sirius red staining, we observed that lobeglitazone effectively attenuates tion. The distribution profiles of HDACs in the kidney and the subtypes of UUO-induced renal atrophy and fibrosis. Immunohistochemical analysis in HDACs that are associated with kidney dysfunction are still unclear. We conjunction with quantitative RT-PCR and Western blot analysis revealed measured HDAC activity using commercially available HDAC Activity/Inhi- that lobeglitazone treatment inhibited UUO-induced upregulation of renal bition Assay which shows an increase in total HDAC in kidney cortex of Smad-3 phosphorylation, alpha-smooth muscle actin, plasminogen activa- type 2 diabetic mice. This increase was inhibited by FK228 (Class I HDAC tor inhibitor-1, and type 1 collagen. In vitro experiments with rat mesangial inhibitor) but not by TSA (Trichostatin A; a pan HDAC inhibitor), suggesting cells and NRK-49F renal fibroblast cells suggested that the effects of lobe- that HDAC activity in diabetic kidney is more subtype specific. We investi- glitazone on UUO-induced renal fibrosis are mediated by inhibition of the gated the changes in specific HDAC expression in type 2 diabetic animals TGF-β/Smad signaling pathway. which also showed increases in HDAC2 and HDAC9 expression in kidney cor- In conclusion, the present study demonstrates that lobeglitazone has a tex along with increases in toll like receptor 4 (TLR4) and (IL)-1β. protective effect on UUO-induced renal fibrosis, suggesting that its clini- To further demonstrate the effect of specific HDACs on kidney inflamma- cal applications could extend to the treatment of nondiabetic origin renal tion in diabetes, we used NRK52E cell lines under hyperglycemic conditions disease. and inhibited the increase in HDAC2 with FK228 (strongly inhibits HDAC1 and HDAC2). The result demonstrates that treatment with FK228 decreases 510‑P HDAC2 as well as TLR4 expression under hyperglycemic condition which The Impact of Sirtuin 3 In Renal Tubular Cell Apoptosis Under Dia‑ suggests that HDAC2-TLR4 mediated inflammation may play a central role in betic Conditions the pathogenesis of diabetic nephropathy. Inhibition of HDAC2 also altered SEONGHUN KIM, JIMIN PARK, BO YOUNG NAM, MEIYAN WU, SUKYUNG KANG, the expression of the kidney damage marker, gelatinase-asso- JUNG TAK PARK, SEUNG HYEOK HAN, TAE HYUN YOO, SHIN WOOK KANG, Seoul, ciated lipocalin (NGAL) in hyperglycemic kidney cells. This study may lead Republic of Korea to the identification of specific HDAC associated inflammatory pathways in Reactive oxygen species (ROS) play important roles in diabetic nephropa- the progression of nephropathy in diabetic animals and may help develop a thy, via mediating renal tubular epithelial cell injury. Recently, the role of therapy to effectively treat this debilitating condition. mitochondrial sirtuins (SIRTs) has been implicated in numerous ROS-medi- Supported By: Texas Tech University Health Sciences Center ated diseases. SIRT3 located in mitochondria has been suggested to be involved in pathogenesis of various kidney diseases. However, the impact of SIRT3 on tubular cell apoptosis under diabetic conditions has never been elucidated. In vitro, rat proximal tubular epithelial cells (NRK-52Es) were

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A133 COMPLICATIONS—NEPHROPATHY—BASIC AND EXPERIMENTAL SCIENCE

cultured in DMEM media containing 5.6 mM glucose (normal glucose, NG) namics, proteinuria and metabolic parameters were assessed. All compari- or NG + TGF-β1 (10 ng/ml) with or without SIRT3 siRNA or plasmid SIRT3 sons are vs. the untreated control group. transfection. In vivo, 24 C57BL/6 mice were intraperitoneally injected with Results: After 11 weeks of treatment there were no significant differ- saline (Control, C) (N=6) or STZ (50 mg/kg/d) for 5 consecutive days (dia- ences in body weight, mean arterial pressure or heart rate among the treat- betes, DM) (N=6), and 6 mice from each group were treated with SIRT3- ment and control groups. Urinary protein excretion was lower in the 1, 3 expressing lentivirus (4 x 108 TU/w) (LV-SIRT3) for 6 weeks. NRK-52Es and and 10 mg/kg/day IW-1973 groups compared to controls (1.1±0.1, 1.1±0.1 and the mouse kidneys were analyzed for SIRT3 and apoptosis-related proteins 1.0±0.1 vs. 1.5±0.99 g/day; all p<0.05). Relative kidney (2.1±0.1 vs. 2.4±0.1 (Bax, Bcl-2 and cleaved-caspase 3). Immunohistochemical staining for SIRT3 g/g brain) and liver (31.8±1.2 vs. 34.8±1.2 g/g brain) weights were lower POSTERS

Complications was also performed with renal tissues. The protein expression of mitochon- in rats treated with 10 mg/kg/day IW-1973, while subcapsular brown adi- Acute and Chronic drial SIRT3 and Bcl2 was significantly decreased in TGF-β1-stimulated NRK- pose tissue weights were significantly higher in the 1, 3 and 10 mg/kg/day 52Es whereas, that of Bax and cleaved-caspase 3 protein was significantly IW-1973 groups (1.83±0.12, 1.76±0.11 and 1.59±0.05 vs. 1.26±0.08 g/g brain; increased. Similar changes in protein expression were also observed in NG all p<0.05). Fasted blood glucose was lower in the 10 mg/kg/day IW-1973 cells transfected with SIRT3 siRNA. In contrast, transfection with plasmid group (194±9 vs. 239±12 mg/dL; p<0.05), and oral glucose tolerance was SIRT3 significantly abrogated the changes in apoptosis-related protein improved in the 3 and 10 mg/kg/day IW-1973 groups. expression in TGF-β1-stimulated NRK-52Es. A significant decrease in SIRT3 Conclusion: IW-1973 reduced proteinuria and improved metabolic param- expression was demonstrated in the kidney of DM mice compared to C kid- eters without affecting hemodynamics in ZSF1 rats. This study suggests that ney. In addition, there were significant increases in TUNEL-positive tubular evaluation of IW-1973 for the treatment of diabetic nephropathy is warranted. epithelial in DM kidney, and these changes were significantly attenuated by LV-SIRT3 treatment. These results suggest that SIRT3 plays a protective role 513‑P in diabetic conditions and that SIRT3 can be a therapeutic target in patients Protects Podocytes against Apoptosis via Autophagy with diabetic nephropathy. Related to miR-383-5p in a Mouse Model of Diabetic Nephropathy SHAN SHAN HUANG, DA FA DING, SHENG CHEN, CHENG LONG DONG, XIAO 511‑P LONG YE, YA MIN FENG, QIANG YOU, JIA QING SHAO, YI BING LU, Nanjing, SGLT2 Inhibitor Therapy Improves Blood Glucose but Does Not China, Yancheng, China Prevent Albuminuria nor Glomerular Hypertrophy in Diabetic db/db Podocyte apoptosis coincides with albuminuria onset and precedes podo- Mice cytopenia in diabetic nephropathy (DN). However, there is a lack of effective YUMI TAKIYAMA, TOSHIHIRO SERA, MASANORI NAKAMURA, TSUYOSHI therapeutic drugs to protect podocytes from apoptosis. Resveratrol is a non- YANAGIMACHI, RYOICHI BESSHO, YUKIHIRO FUJITA, MASATO HOSHINO, KEN- flavonoid polyphenol with various pharmacological effects and it has attrac- TARO UESUGI, YASUAKI SAIJO, NAOTO YAGI, MASAKAZU HANEDA, Asahikawa, tive increased research attention in the field of DN for its potential value in Japan, Fukuoka, Japan, Saitama, Japan, Hyogo, Japan kidney protection, however, few studies have been conducted on the role of SGLT2 inhibitors represent a promising strategy in the treatment of diabe- resveratrol in podocytes and the underlying mechanism is not fully under- tes by targeting renal proximal tubules. To investigate the impact of SGLT2 stood. This study investigates whether resveratrol can relieve the apoptosis inhibitor on the number and volume of glomeruli in the whole kidneys, we of podocytes and reveals the mechanism of resveratrol action. We demon- explored CT imaging of diabetic mice using synchrotron radiation micro-CT strated that resveratrol relieved a series of indicators of DN and attenuated in SPring-8. We also analyzed the spatial distribution of the glomeruli in apoptosis of podocytes in db/db diabetic model mice. In addition, resveratrol the ten-segmented layer from the centroid to the surface of the kidney. We induced autophagy in both db/db mice and human podocytes. Furthermore, found a weak interaction between db/db and -treated db/db inhibition of autophagy by 3-methyladenine (3-MA) and autophagy gene 5 mice in the distribution of glomerular number (p=0.050, by two-way ANOVA), (Atg5) shRNA reversed the protective effects of resveratrol on podocytes. suggesting the remodeling of glomerular distribution driven by the treat- Besides, mice kidney samples were subjected to microarray analysis to iden- ment of SGLT2 inhibitor. Unexpectedly, luseogliflozin-treated db/db mice tify differentially expressed microRNAs. We found when db/db mice treated had larger glomeruli and less glomeruli compared with non-treated db/db with resveratrol, the expression of miR-383-5p caused the most obvious mice, suggesting compensatory glomerular hypertrophy for the glomerular decrease relative to db/db mice. Besides, the expression of miR-383-5p was loss. Luseogliflozin ameliorated hyperglycemia and renal hypertrophy, but also downregulated after treatment with resveratrol in podocytes. Overex- not albuminuria. Indeed, luseogliflozin exhibited the mitigation of glomeru- pression of miR-383-5p significantly blocked the increase in autophagy of lar damage, but augmented tubular injury as vacuolar change. Intriguingly, podocytes and attenuation of HG-induced apoptosis of podocytes induced luseogliflozin enhanced hypoxia marker, pimonidazole staining in the outer by resveratrol. While, the suppression of miR-383-5p further increased stripe of outer medulla, suggesting hypoxia caused by SGLT1-mediated com- autophagy and attenuated apoptosis in podocytes. Together, our results pensatory reabsorption. In addition, electron microscopic analysis revealed indicate that resveratrol effectively attenuates high glucose-induced apop- that luseogliflozin attenuated the thickening of gomerular basement mem- tosis via the activation of autophagy in db/db mice and podocytes, which brane, but decreased the number of in renal proximal tubules, involves miR-383-5p. Thus, this study reveals a new possible strategy to and increased the number of granular cells with large and confluent vesicles treat diabetic nephropathy. with low density in juxtaglomerullar apparatus, which indicates prolonged Supported By: National Natural Science Foundation of China (81100577 to strong stimulation of renin-producing cells and suggests the potential D.F.D.), (81270896 to Y.B.L.); Six Talent Peaks Project in Jiangsu Province (2015- involvement of renin angiotensin system (RAS) activation with SGLT2 inhi- WSN-016 to D.F.D.), (2013-WSN-049 to Y.B.L.) bition. Taken together, SGLT2 inhibitor therapy might require the combined inhibition of RAS and SGLT1 for renoprotection in diabetes. 514‑P Supported By: Japan Society for the Promotion of Science (15K09372); Japan Metabolomic and Transcriptomic Studies in Diabetic Kidney Dis‑ Diabetes Foundation; Asahikawa Medical University ease in Mice and Humans Identify Impaired Mitochondrial Function MANJULA DARSHI, SATOSHI MIYAMOTO, BENJAMIN VAN-ESPEN, YASMIN 512‑P AGHAJAN, RINTARIO SAITO, WENJUN JU, PIERRE-JEAN SAULNIER, MATTH- IW-1973, a Soluble Guanylate Cyclase Stimulator, Inhibits Progres‑ IAS KRETZLER, ROBERT NELSON, KUMAR SHARMA, La Jolla, CA, Ann Arbor, MI, sion of Diabetic Nephropathy in the ZSF1 Rat Model Phoenix, AZ ALBERT T. PROFY, COURTNEY SHEA, ELISABETH LONIE, GUANG LIU, G. TODD Background: Previously we identified 13 urinary metabolites as a metabolomic MILNE, MARK G. CURRIE, JAIME L. MASFERRER, Cambridge, MA signature of diabetic kidney disease (MSDKD) in humans. In our current study we Background: Diabetic nephropathy is associated with endothelial dys- sought to determine the metabolomic signature of a progressive (Alport) and a function and reduced nitric oxide (NO) signaling. IW-1973, a soluble guanyl- non-progressive kidney disease mouse model (db/db). Furthermore, we analyzed ate cyclase (sGC) stimulator in clinical development, has been shown to dis- correlation of mitochondrial gene expression in DKD patients with GFR. tribute extensively to tissues, enhance NO-cyclic guanosine monophosphate Methods: GC-MS/MS based targeted organic acid analysis was used (cGMP) signaling, and mediate hemodynamic, anti-inflammatory, and anti- to quantify urinary metabolites. from Alport (n=10) and db/db (n=8) along fibrotic effects in animal disease models. This study assessed the effects of with their respective controls. Kidney cortical tissues were analyzed for IW-1973 in the ZSF1 model of diabetic nephropathy. changes in mitochondrial protein levels by Western blot. Gene expression Methods: Obese, diabetic ZSF1 rats (N=9 per group) received 0, 1, 3 or 10 was performed in renal biopsy specimens with laser captured glomerular mg/kg/day IW-1973 in food for 11 weeks. Body and tissue weights, hemody- and tubulointerstitial samples from diabetic kidney disease patients.

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A134 COMPLICATIONS—NEPHROPATHY—CLINICAL AND TRANSLATIONAL RESEARCH Results: Both Alport and db/db mice showed significant changes in sev- & 516‑P eral mitochondrial metabolites reported previously in patients with progres- Serum Pigment Epithelium-Derived Factor Predicts Development of sive diabetic kidney disease (MSDKD). At day 56 the Alport mice showed Kidney Dysfunction in Type 2 Diabetes: The Veterans Affairs Diabe‑ significant reduction in 7 urinary metabolites. Similar to the Alport mice, the tes Trial (VADT) Study db/db mice showed significant reduction in several mitochondrial metabo- KELLY J. HUNT, ALICIA J. JENKINS, DANIELLE STEVENS, DONGXU FU, JIAN lites including succinic acid, citric acid and aconitic acid. Analysis of mito- XING MA, RICHARD L. KLEIN, MADONA AZAR, SARAH ZHANG, MARIA F. LOPES- chondrial proteins by immunoblotting revealed dramatic reduction in several VIRELLA, TIMOTHY J. LYONS, VADT INVESTIGATORS, Charleston, SC, Fitzroy, Aus- proteins associated with oxidative phosphorylation and structure in d56 tralia, Belfast, United Kingdom, Oklahoma City, OK POSTERS

Alport mice with no significant changes in db/db mice. Additionally, PGC1α Serum pigment epithelium-derived factor (PEDF) is an adipokine which has Complications expression levels were significantly reduced only in d56 Alport mice kidney been associated with insulin resistance, diabetes and diabetic vascular compli- Acute and Chronic tissue compared with WT suggesting severe mitochondrial damage along cations. In a subset of the VADT cohort, we evaluated serum PEDF as a predictor with the progression of the disease. Transcriptomic analysis of kidney biop- of macro-albuminuria (ACR ≥300 g/mg creatinine) and chronic kidney disease sies of DKD patients revealed a positive correlation with complex I genes (CKD) stages 3 (GFR <60 ml/min on two occasions) and stage 4 (GFR <30ml/ and PGC1α with GFR, suggesting decrease in mitochondrial biogenesis and min on one or two occasions). Serum PEDF was measured on 972 participants function in human diabetic nephropathy. (median 1.87 y after enrolment), and repeated in 741 (median 3.76 y after enrol- Supported By: National Institute of Diabetes and Digestive and Kidney Diseases ment). Participants were followed a median of 5.74 years after enrolment with (DP3-DK094352); JDRF; U.S. Department of Veterans Affairs a median follow-up of 2.01 y following each PEDF measurement. In multivariate event time models which used both measurements of serum PEDF, serum PEDF predicted events either prior to the next PEDF measurement (i.e., time between COMPLICATIONS—NEPHROPATHY—CLINICAL AND first and second measurement) or prior to study end (i.e., time between final TRANSLATIONAL RESEARCH measurement and study end). Increased serum PEDF was significantly associ- ated with the subsequent development of kidney dysfunction, as shown (Table). Over roughly a 2 year period, serum PEDF may serve as a predictive biomarker Moderated Poster Discussion: Markers and Mechanisms of Clinical for advanced nephropathy in people with type 2 diabetes. Diabetic Nephropathy (Posters: 515-P to 520-P), see page 21. Table. Cox Proportional Hazard Models for Renal Outcomes for a One Stan- dard Deviation Increase in PEDF. Uses PEDF Data Measured at Two Time & 515‑P Points. The Association of Glycosphingolipids with Chronic Kidney Dis‑ ease in Type 1 Diabetes MARIA F. LOPES-VIRELLA, NATHANIEL BAKER, KELLY J. HUNT, DANIELLE R. STE- VENS, SAMAR M. HAMMAD, RICHARD L. KEIN, Charleston, SC There is mounting evidence that altered glycosphingolipid (GSL) metabo- lism may contribute to complications in diabetes. This study proposes to assess whether plasma GSL (lactosyl and hexosylceramide) levels, collected at baseline and closeout of the DCCT study can predict both progression to macroalbuminuria (MA; AER≥300 mg/24 h) and chronic kidney disease (CKD; GFR <60 ml/min) in type 1 diabetes. The presence of MA and CKD was assessed up to EDIC year 18. A sub-cohort of 516 DCCT/EDIC patients was selected. This sub-cohort was representative of the entire DCCT/EDIC cohort. Logistic regression models were used to assess the effect of GSL levels on the odds of developing MA or CKD during EDIC. Our results show that low levels of total and very long chain lactosylceramides (L-Cer) mea- sured at both DCCT baseline and closeout were significantly associated Supported By: American Diabetes Association (7-12-CT-46 to T.J.L.) with progression to MA but not with CKD. In contrast, high levels of hexosyl (H-cer) both at DCCT baseline and closeout were significant pre- & 517‑P dictors of CKD (Table 1). Renal Functional Decline Is Associated with Decrease of Glomeru‑ In conclusion, our data show that low levels of very long chain L-Cer are lar Filtration Surface Reflecting Increase of Mesangial Expansion associated with the development of MA similarly to our previous findings in Normo- and Microalbuminuric Type 2 Diabetes with very long chain ceramides. CKD is however associated with high levels TATSUMI MORIYA, MADONNA MATSUBARA, KEI HAYAMA, YUKI YOSHIDA, of H-Cer. Further studies are needed to determine whether GSL may help dif- MOTOSHI OUCHI, Sagamihara, Japan, Shimotsuga, Japan ferentiate early kidney dysfunction with a predominant tubular component This study addresses whether renal structural changes at the early from more advanced glomerular dysfunction. stage of diabetic nephropathy (DN) in type 2 diabetic patients is related Table 1. Associations between Plasma Glycosphingolipid Measures at to renal functional development using serial renal biopsies. Ten type 2 dia- DCCT Closeout and Progression to Chronic Kidney Disease During EDIC. betic patients (7 men, 48±8 yrs, known duration 12±8 yrs, HbA1c 7.3±1.3%, Odds Ratios are for a One Standard Deviation Increase in Each Glycosphin- 121±16/74±10 mmHg, glomerular filtration rate [GFR] measured by iohexol golipid Measure. injection 122±23 ml/min/1.73 m2 [mean±SD]) showing normo- or microal- buminuria participated. One-hr biopsy materials from living related renal transplant donors served as normal controls. Light and electron microscopic morphometric analyses provided quantitative glomerular and interstitial structural changes. Mean glomerular volume (MGV), glomerular basement membrane (GBM) width, and mesangial volume fraction [Vv(Mes/glom)] were significantly larger than controls. Average follow-up was 6.1±2.4 (3.0- 11.9) yrs. Urinary albumin excretion (UAE) and GFR were determined every year. Post follow-up, a second renal biopsy was performed. UAE, GFR, blood pressure and frequency of diabetic retinopathy (DR) did not change between baseline and follow-up as a group. Eight of 10 cases showed individual decrease of GFR. MGV, GBM width, Vv(Mes/glom) and filtration surface per glomerulus [Sv(PGBM/glom)] did not change. The changes in morphometric data did not correlate with the change of UAE. However, annual decrease of Sv(PGBM/glom) positively correlated with GFR decrease during 6-year Supported By: National Institutes of Health/National Institute of Diabetes and follow-up with renal biopsy (r=0.71, p=0.02). Annual change of [Sv(PGBM/ Digestive and Kidney Diseases glom)] negatively correlated with that of Vv(Mes/glom) (r=0.68, p=0.03). In renal biopsy-proven normo- and microalbuminuric type 2 diabetic patients,

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A135 COMPLICATIONS—NEPHROPATHY—CLINICAL AND TRANSLATIONAL RESEARCH GFR decrease is associated with decreased glomerular filtration surface, & 520‑P reflecting mesangial expansion during 6-year observation. These could be Apps Ameliorates Diabetic Nephropathy Progression by Transcrip‑ initial changes in early DN possibly occurring without worsening of albu- tional Repression of TGFβ1 through Interaction with Nrf2 minuria and DR. PAN GAO, LILIANG LI, JUNLI LIU, Shanghai, China Transforming growth factor beta 1 (TGFβ1) is a well-distinguished media- & 518‑P tor of progressive renal fibrosis in diabetic nephropathy (DN). Based on our Association of Renal Effects of with Baseline Daily previous work, therapeutics against TGFβ1 signaling had potent treatment values. Herein, we further reported that Apps, in association with Nrf2, POSTERS Urinary Sodium Excretion in Type 2 Diabetes Mellitus Complications KIYOHIDE NUNOI, YUICHI SATO, KOHEI KAKU, HIDEAKI SUGANAMI, Kurume, exhibited potential renal protective roles by transcriptional repression Acute and Chronic Japan, Okayama, Japan, Tokyo, Japan of TGFβ1 signaling in DN. Apps was significantly upregulated in the early Sodium-glucose co-transporter 2 (SGLT2) facilitates reabsorption of glu- stage of DN and mesangial cells with short-time exposure of high glucose. cose and sodium and its inhibition improves glycemic status and glomerular Apps bound to the specific region of TGFβ1 promoter and transcriptionally hyperfiltration. SGLT2 inhibitors should improve renal function, but their repressed its expression, which was blunted by mutagenesis of the binding effects at different levels of sodium intake are unclear. This study evaluated sites. Overexpression of Apps using AAV-delivered plasmid system amelio- the renal effects of tofogliflozin (TOFO) in type 2 diabetes mellitus (T2DM) rated, while knockout of Apps with AAV-cre worsened the oxidative stress according to baseline daily urinary sodium excretion (U-Na). The Kawasaki and diabetic kidney injury in vivo. Moreover, Apps synergistically interacted formula was used to calculate U-Na in 775 T2DM patients receiving TOFO with Nrf2 in a time- and dose-dependent manner to suppress the expression in 2 phase 3 studies. To assess renal effects according to baseline sodium of TGFβ1 in DN. After activation by Nrf2, Apps was recruited into the nuclear intake, subjects were divided into 4 groups by baseline U-Na quartiles. of mesangial cells and inhibited the transcription of TGFβ1. Our study sug- Adjusted variables were evaluated using analysis of covariance to examine gested the protective role of Apps in the early stage of DN and might provide differences among groups, with the group as a fixed effect and baseline novel clues for the prevention and treatment of DN in clinic. values and estimated glomerular filtration rate (eGFR) as covariates. The Figure. 775 subjects were divided into Quartile 1 (Q1) (n=193) and Q2-4 (each with n=194). Baseline U-Na values (Q1: median 140, Q2: 186, Q3: 224, and Q4: 284 mmol/day) and eGFR (mean 79, 81, 86 and 90 ml/min/1.73 m2) differed significantly, while baseline HbA1c and blood pressure were similar. TOFO concurrently reduced HbA1c, weight, and blood pressure. Significant differ- ences were observed in eGFR at week 4 (Q1: mean -1.6, Q2: -3.1, Q3: -4.7, and Q4: -5.3, p<0.05), 24 (1.9, -0.8, -1.7, and -2.0, p<0.05), and 52 (4.1, 2.7, 1.4, and 1.6 ml/min/1.73 m2, p<0.05). Two weeks after cessation of TOFO, eGFR increased significantly in all groups from week 52. Additionally, in subjects with albuminuria (urine albumin:creatinine ratio [UACR] ≥30 mg/gCr), a simi- lar time course of eGFR values followed baseline U-Na levels, while a similar degree of reduction in UACR among groups was maintained. Improvement in glomerular hyperfiltration with TOFO may depend on baseline sodium intake. & 519‑P Attenuates Diabetic Nephropathy and the Activation of the Inflammasome in Mice with T2DM YUMEI YE, YOCHAI BIRNBAUM, HSIU CHIUNG YANG, MANDEEP BAJAJ, Galves- ton, TX, Houston, TX, Mölndal, Sweden SGLT2 inhibitors and DPP-4 inhibitors (DPP-4i) are used to treat T2DM. DPP-4i attenuate Nlrp3 inflammasome activation in the kidney. SGLT2 inhi- bition reduces inflammation and attenuates the progression of diabetic nephropathy (DN). The effects of Dapagliflozin (Dapa) on the activation of the Nlrp3 inflammasome and the combined effect of SGLT2 and DPP- 4i on T2DM-induced inflammasome activation and progression of DN has not been previously studied. We assessed whether Dapa attenuates the inflammasome activation and progression of DN in T2DM mice and whether this effects can be augmented by adding DPP-4i Saxagliptin (Saxa). Male BTBR ob/ob and wild type (WT) mice received vehicle, Dapa or Dapa+Saxa for 8 wks. Serum BUN in the WT mice was 16.9±0.8 mg/dl. It increased to 55.7±2.8 mg/dl in the BTBR mice. Dapa alone reduced BUN to 31.4±1.2 mg/dl. A greater effect was seen in the Dapa+Saxa combination (24.8±0.8 mg/dl). Serum creatinine was 0.16±0.02 and 1.01±0.04 mg/dl in the WT and BTBR mice, respectively. Dapa and Dapa+Saxa attenuated the increase to 0.65±0.02 and 0.40±0.03 mg/dl, respectively. Serum cystatin C increased in the BTBR mice (3.9±0.1 vs. 0.6±0.2 ng/ml). Dapa (2.4±0.1) and Dapa+Saxa (1.4±0.1) attenuated this increase. Kidney weight increased in the BTBR Supported By: National Natural Science Foundation of China mice. Dapa reduced kidney/body weight ratio in the BTBR mice. Dapa+Saxa tended to have greater effect, but the difference was not significant. mRNA 521‑P levels of NALP3, ASC, IL-1β, IL-6, Caspase-1, TNFα, collagen-1 and collagen-3 Arteriolar Hyalinosis Predicts Albuminuria Increase and GFR significantly increased in the kidneys of the BTBR compared to the WT mice. Decline in Normo-Microalbuminuric Dapa alone and to a greater extent, Dapa+Saxa attenuated the activation of TATSUMI MORIYA, MADOKA MATSUBARA, KEI HAYAMA, YUKI YOSHIDA, the inflammasome. Yet, the combination did not result in greater attenuation MOTOSHI OUCHI, Sagamihara, Japan, Shimotsuga, Japan of the collagen-1 and collagen-3 mRNA levels. Glomerular basement membrane (GBM) thickening and mesangial expan- Conclusions: Dapa attenuates T2D-induced activation of the inflamma- sion are risk factors for urinary albumin excretion (UAE) increase and GFR some and progression of DN. Adding Saxa to Dapa augmented attenuation decline in types 1 and 2 diabetic patients. Above morphometric analysis of the inflammasome, but had no significant effect on kidney weight or col- requires electron microscopic (EM) samples, and what light microscopic (LM) lagen-1 and -3 mRNA levels. histological factors, including arteriolar hyalinosis, affect UAE increase and Supported By: AstraZeneca GFR decline remain unclear. We examined 29 type 2 diabetic patients by per- cutaneous renal biopsy (22 men, 49±10 yrs, known duration 12±7 yrs, HbA1c 8.2±1.9%, 120±16/73±11 mmHg, GFR, measured by iohexol injection, 119±27

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A136 COMPLICATIONS—NEPHROPATHY—CLINICAL AND TRANSLATIONAL RESEARCH ml/min/1.73 m2 [mean±SD]; 15 normoalbuminuria; 14 microalbuminuria) to was calculated using the Modification of Diet for Renal Disease equation. clarify histological factors affecting GFR and UAE during 8.0±3.5 yrs follow- Chronic kidney disease was defined as an eGFR < 60 ml/min/1.73m2. Multi- up. EM morphometric analyses gave quantitative glomerular structural variate logistic regression analyses were performed to assess the associa- changes including mesangial fractional volume [Vv(Mes/glom)]. LM tissues tions between CKD and potential factors. provided glomerular and interstitial changes including index of arteriolar Results: The mean age was 54.5 years (range 21-92 years) and the median hyalinosis (IAH). Annual GFR and UAE were measured. 1. Vv(Mes/glom) neg- duration of diabetes was 5.0 years ranging from newly diagnosed to 31 years. atively correlated with baseline and follow-up GFR but not UAE both times; Over half of the patients; 213/370 (57.6%) and 232/370 (62.7%) had an average 2. IAH positively correlated with UAE and negatively correlated with GFR at blood pressure > 140/90mmHg and poor glycemic control (HbA1c >7%) respec- POSTERS follow-up, while IAH did not correlate with UAE or GFR at baseline; 3. There tively. Of the 370 patients enrolled, 31/370 (8.4%) had CKD. Only 18/370 (4.9%) Complications were no significant differences in clinical or morphometric data except IAH had diagnosis of CKD documented in their charts. Level of education (OR=3.60, Acute and Chronic scores among patients with IAH <2.0 and ≥2.0. GFR at follow-up in IAH≥2.0 95% CI 1.23-10.55; p=0.02) and duration of diabetes >10 years (OR=0.26, 95% (77.2±36.2 ml/min/1.73 m2) was decreased from baseline (122.6±28.1, CI 0.01-0.67; p=0.01) were independently associated with CKD. p=0.025) and less than IAH <2.0 (112.9±21.8, p=0.005) not decreased from Conclusion: The prevalence of CKD by estimated eGFR was low com- baseline (117.3±27.1); 4. Stepwise regression analysis showed that only IAH pared to most previous studies. Almost half of patients with CKD are not affected the GFR at the follow-up. Arteriolar hyalinosis, seen by LM, is an documented resulting to potential of prescription errors and drug toxicity. additional histological predictor for albuminuria increase and GFR decline in Substantial number of our patients had uncontrolled hypertension and poor normo- and microalbuminuric type 2 diabetic patients. glycemic control making them at increased risk CKD. This calls for more tight measures to achieve target goals. 522‑P Magnetic Resonance Detection of Kidney Iron Deposition by Hap‑ 524‑P toglobin (Hp) Genotype in Type 1 Diabetes (T1D): Results from the Elevated Kallistatin Levels in Patients with Diabetic Nephropathy Kirha Study and Diabetic Mice TINA COSTACOU, TREVOR J. ORCHARD, CHAN HONG MOON, KYONGTAE BAE, YANHUI YANG, XUEMIN HE, RUI CHENG, JIAN XING MA, Oklahoma City, OK LINDA FRIED, RHOBERT W. EVANS, Pittsburgh, PA Kallistatin is an endogenous tissue kallikrein inhibitor and has multiple Hp’s main role is to bind free hemoglobin (Hb), reducing its oxidative functions independent of its interactions with kallikrein, including anti- potential. The Hp-Hb complex formed is cleared by macrophage CD163. In angiogenesis, antioxidant, anti-inflammatory effects, and so forth. Previ- diabetes, impaired Hp 2-2 - Hb CD163 clearance and abnormal glomerular ously, we have reported a significant increase of circulation kallistatin in permeability allow the large Hp 2-2 - Hb complex to cross the barrier, where diabetic patients and a decrease of vitreous level in patients with diabetic its redox active iron leads to cellular toxicity. Hp genotype 2-2 was shown retinopathy. However, the implication of kallistatin in diabetic nephropathy to predict renal function decline in 2 large T1D cohorts though no human (DN) was unknown. In the present study, 36 diabetic patients with DN, 32 data exist on iron deposition by Hp. In this pilot study, we tested the thesis diabetic patients without DN, and 30 healthy controls were enrolled, and of greater kidney iron in T1D with Hp 2-2. We used renal Blood Oxygen Level the correlation between serum kallistatin levels and clinical parameters Dependent Magnetic Resonance Imaging (BOLD MRI) to estimate iron level were analyzed. Serum kallistatin was measured by ELISA. Renal kallistatin in the and medullar of adults with childhood onset T1D (15 Hp protein level was detected by Western blot analysis in difference diabetic 1-1 and 15 Hp 2-2 carriers of similar age (53 yrs), duration (45 yrs) and sex murine models including Akita, OVE26, db/db mice and the control mice. (50% men)). Total kidney iron was estimated as the sum of the cortex and Diabetic patients (with and without DN) had higher levels of circulation kal- medullar iron. Albuminuria was defined as urinary albumin to creatinine ratio listatin (mean±SD) than healthy controls (32.4±7.2ug/ml vs. 24.2±5.2ug/ml, >30 mg/g in 2 of 3 samples. Total kidney iron did not differ by sex or Hp but p<0.01). There was no significant differenece in kallistatin levels between was higher in those with albuminuria (p=0.05). This association appeared DM patients without DN and healthy controls (P>0.05). Kallistatin levels in confined to Hp 2-2 carriers as no difference in kidney iron deposition was diabetic patients with DN were significantly increased compared to those seen by albuminuria with Hp 1-1 (Table). Our pilot supports the hypothesis of in diabetic patients without DN (37.8±4.7ug/ml vs. 26.7±4.6ug/ml, p<0.001). increased kidney iron deposition with Hp 2-2 in T1D with albuminuria. This For the diabetic patients, there was a positive correlation between the thesis merits evaluation in larger studies. serum kallistatin level with albumin/creatinine ratio (ACR) (r=0.398, p<0.01) Table. Kidney Iron Deposition (sec-1) by Albuminuria Status and Hp Genotype. and serum LDL-cholesterol (r=0.291, p<0.05). However, there was no correla- tion between the serum kallistatin level with BMI, HbA1C, BUN or serum Hp 1-1 carriers Normoalbuminuria Albuminuria p-value creatinine levels. All of the three diabetic animal models showed elevated (n=9) (n=5) renal kallistatin expression levels compared to their age and genetic back- Total kidney iron 81.4 (9.7) 84.5 (5.0) 0.51 ground-matched nondiabetic controls. These observations suggested that Cortex iron 48.4 (5.6) 49.2 (3.5) 0.78 increased levels of kallistatin could become a biomarker for diagnosis or Medullar iron 33.0 (4.6) 35.4 (3.1) 0.33 prognosis of DN. Hp 2-2 carriers Normoalbuminuria Albuminuria p-value (n=11) (n=4) 525‑P Total kidney iron 80.1 (8.2) 90.6 (5.9) 0.04 Diabetic Nephropathy Clinical Trajectories among Groups of Patients with Type 2 Diabetes: A Latent Class Analysis Cortex iron 47.3 (4.5) 52.6 (3.1) 0.05 ZHOU ZHOU, HONGBO YANG, RUIXUAN JIANG, ERNEST LAW, MELVIN S. MUN- Medullar iron 32.8 (4.8) 38.0 (3.5) 0.07 SAKA, RAAFAT SEIFELDIN, Boston, MA, Chicago, IL, Deerfield, IL Background: Diabetic nephropathy (DN) is a prevalent complication of type 2 diabetes (T2DM), and increased mortality, morbidity, and costs have 523‑P been linked to worsening DN severity. Patients with T2DM are clinically het- Prevalence of Chronic Kidney Disease and Associated Factors erogeneous in their disease severity, treatment, and co-morbidities, which among Patients Attending a Tertiary Diabetes Clinic in Botswana may result in differential DN progression courses. GODFREY M. RWEGERERA, THATO MOSHOMO, MAREA GAENAMONG, Aims: To (1) identify patient groups with distinct clinical profiles of T2DM ADERONKE T. OYEWO, SIVASOMNATH GOLLAKOTA, ANTHONY MASAKA, Gabo- disease severity and (2) explore the association between T2DM disease rone, Botswana severity and DN progression or reversal. Introduction: Diabetes mellitus is one of the most common contributors Methods: U.S. Adults with ≥2 T2DM diagnoses and ≥2 urine albumin (UA) to chronic kidney disease (CKD) globally. The objective of this study was to tests (n=23235) within the Truven MarketScan database (2004-14) were estimate the prevalence of CKD among diabetes mellitus (DM) patients. We included. Latent class analysis (LCA) was used to group patients based on also aimed to determine factors associated with presence of CKD among T2DM-related complications, comorbidities, and diabetic therapy. DN sever- the study population. ity was categorized into normo-, micro-, and macroalbuminuria based on UA. Methods: A cross-sectional study was conducted at the block 6 diabetes The risks of DN progression (change to a more severe DN category) and clinic in Gaborone, capital city of Botswana. Demographic and clinical data reversal (change to a less severe DN category) were compared between were obtained from patients through interviews and from dockets using a all T2DM severity profiles using Kaplan-Meier analyses and log-rank tests. standard questionnaire. Data was collected over a period of two months Results: Four clinically distinct T2DM patient profiles were identified: from July to September 2015. Estimated glomerular filtration rate (eGFR) healthiest and low treatment (46.5% of sample); low comorbidity and high

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treatment (29.0%); moderate comorbidity and high insulin use (9.7%); and 528‑P high comorbidity and moderate treatment (14.8%). The 5-year progres- Associations between Hypertension and End-Stage Renal Disease sion rates were 11.8%, 18%, 16.5%, 27.7%, respectively. Compared to the (ESRD) in People With and Without Diabetes healthiest and low treatment group, all other groups were associated with ATSUSHI FURUYA, KAZUYA FUJIHARA, MASAHIRO ISHIZAWA, MASAHIKO an increased risk of progression (all p<0.001). The low and high comorbidity YAMAMOTO, YASUHIRO MATSUBAYASHI, SATOSHI MATSUNAGA, CHIE ITO, groups were associated with increased risk of reversal (both p<0.001) com- MIEKO KOISHI, NAUTA YAMANAKA, KIMINORI KATO, SATORU KODAMA, HIRO- pared to the healthiest patients. HITO SONE, Niigata, Japan, Tokyo, Japan Conclusions: This exploratory study used LCA to identify patient groups

POSTERS While hypertension is the strongest predictor of nephropathy except gly-

Complications with varying T2DM severity and evaluated their risks of DN progression and cemia among people with diabetes mellitus (DM), its role in the develop- Acute and Chronic reversal. Future prospective studies are needed to confirm the significance ment of ESRD has not been analyzed in detail. Also whether associations of these groups on health and cost outcomes. between hypertension and ESRD differ between those with and without DM have not been clarified. We analyzed a nation-wide claim-based database of 526‑P 146,139 participants in 2008-15. Participants were classified as having nor- Association between Serum Galectin-3 and Advanced Glycation mal glucose tolerance (NGT) or DM based on fasting plasma glucose, HbA1c, End Products in Type 2 Diabetes and prescriptions (130,575 with NGT; 15,564 with DM). During the study KATHRYN C.B. TAN, SAMMY W.M. SHIU, YING WONG, Hong Kong, China period, 59 and 21 participants in the NGT and DM categories, respectively, Galectin-3, a member of the multifunctional galectin family, acts a broad- started dialysis. In both categories, the baseline SBP was higher in those spectrum biological response modifier and is involved in tissue fibrosis, who started dialysis than in those who did not. Multivariate Cox regres- immunity, and inflammatory response. The role of galectin-3 in diabetic com- sion model showed that SBP was a significant and independent predictor of plications is unclear and animal studies have provided evidence for protec- the start of dialysis in both NGT and DM categories, with quite comparable tive and detrimental functions of galectin-3. Although galectin-3 is a media- hazard ratios (HRs). Participants with SBP ≥160 mmHg had a 16.7-fold risk tor of inflammation and fibrosis, it is also a scavenger receptor for advanced for dialysis in the NGT group and a 20.8-fold risk for dialysis in the DM group. glycation end products (AGEs) and stimulates their degradation. We have Using SBP ≤129 mmHg in the NGT category as the reference, participants investigated whether serum galectin-3 level is related to circulating AGEs with SBP ≥160 mmHg had a 21.7-fold risk for dialysis in the NGT group and and/or inflammation in type 2 diabetic patients with and without chronic a 17.7-fold risk for dialysis in the DM group (Table). These findings show that kidney disease (CKD). 270 type 2 diabetic patients (30% with CKD and eGFR hypertension affects the risk of ESRD to the same degree regardless of the <60 ml/min/1.73m2) and 230 controls were recruited. Serum galectin-3 and presence of diabetes. AGEs were assayed by ELISA and plasma high sensitivity C-reactive protein Table. Cox Regression Analysis of Variables for Incidence of Dialysis Accord- (CRP) was measured by immunoturbidimetric assay. Diabetic patients had ing to Glucose Tolerance Status. higher HbA1c, AGEs (4.46 ± 1.06 unit/ml (CKD+); 4.10 ± 0.98 (CKD-); 3.56 ± NGT DM 0.99 (control); ANOVA p<0.01) and CRP levels than controls. Serum galec- tin-3 was highest in those with CKD (9.07 ± 2.64 ng/ml (CKD+); 7.48 ± 2.36 HR (95% CI) P-value HR (95% CI) P-value (CKD-); 5.36 ± 1.65 (control); ANOVA p<0.01). Serum glaectin-3 correlated Male 3.94 (1.78-8.75) <0.01 1.69 (0.47-6.09) 0.43 with AGEs (r=0.34, p<0.01), eGFR (r=-0.35, p<0.01) and age (r=0.25, p<0.01) Age, 5-y increment 1.28 (1.10-1.50) <0.01 0.88 (0.66-1.18) 0.40 but there were no associations with duration of diabetes, HbA1c or CRP. The BMI, 1 kg/m2 increment 0.82 (0.68-0.98) 0.03 1.07 (0.86-1.34) 0.52 association with AGEs remained significant even after excluding subjects with CKD (r=0.30, p<0.01). On linear regression analysis, serum AGEs and WC, 10 cm increment 1.42 (0.75-2.69) 0.29 0.54 (0.22-1.33) 0.18 eGFR were independent determinants of serum galectin-3 after adjusting for HDL cholesterol, 10 mg/dL increment 0.69 (0.56-0.85) <0.01 0.62 (0.40-0.95) 0.03 age, gender, body mass index, and smoking, accounting for 10% and 12% of LDL cholesterol, 10 mg/dL increment 0.84 (0.76-0.92) <0.01 0.96 (0.85-1.09) 0.54 the variability respectively (p<0.01). TG, 10 mg/dL increment 0.99 (0.96-1.02) 0.51 1.02 (1.00-1.05) 0.09 In conclusion, serum galectin-3 was increased in type 2 diabetic patients with and without CKD, and serum level was associated with AGEs and renal HbA1c, 1% increment 0.16 (0.07-0.34) <0.01 1.31 (1.06-1.61) 0.01 function but not with inflammation. The role of galectin-3 in diabetic compli- Smoking 0.42 (0.22-0.79) 0.01 0.37 (0.13-1.04) 0.06 cations in humans remains to be determined. Vascular disease 8.64 (4.23-17.6) <0.01 5.64 (1.77-18.0) <0.01 ≤129 mmHg Reference <0.01 Reference <0.01 527‑P SBP 130-159 mmHg 6.37 (3.55-11.4) <0.01 4.72 (1.31-17.0) 0.02 Prognostic Significance of Home Pulse Pressure for Progression of 160 mmHg 16.7 (5.47-51.3) <0.01 20.8 (5.02-86.5) <0.01 Diabetic Nephropathy: Kamogawa-HBP Study ≥ NORIYUKI KITAGAWA, EMI USHIGOME, SHINOBU MATSUMOTO, CHIKAKO ≤129 mmHg Reference <0.01 0.80 (0.21-3.07) 0.75 OYABU, YUKIKO FUKUDA, SAORI MAJIMA, MAI ASANO, MUHEI TANAKA, SBP 130-159 mmHg 6.83 (3.86-12.1) <0.01 3.90 (1.50-10.2) 0.01 MASAHIRO YAMAZAKI, MICHIAKI FUKUI, Kyoto, Japan ≥160 mmHg 21.7 (7.23-65.2) <0.01 17.7 (5.60-56.2) <0.01 Pulse pressure (PP) has been noted as a potentially independent risk factor for micro- and macrovascular diabetic complications. We aimed to exam- NGT: normal glucose tolerance; DM: diabetes mellitus; HR: hazard ratio; CI: confidence interval; BMI: body mass index; WC: waist circumference; ine the prognostic value of home-measured PP (home PP) in patients with HDL/LDL: high-density/low-density lipoprotein; TG: triglyceride; HbA1c: type 2 diabetes. This study is a two-year prospective cohort study of 1,414 hemoglobin A1c; SBP: systolic blood pressure. patients with type 2 diabetes. Home blood pressure measurements were performed for 14 consecutive days. We defined the progression of diabetic 529‑P nephropathy when the diabetic nephropathy stage advanced to the higher Association between Retinal Vascular Structure and Diabetic stage during the two years. Using logistic regression analyses, we inves- Nephropathy tigated the relationship between home PP and home systolic blood pres- LEI KUANG, BENNY ZEE, JACK LEE, ERIC HUI, SHUK YUN LEUNG, SUM LEE, sure (SBP) in the morning and in the evening, and the progression of diabetic DAVID OWENS, REBECCA THOMAS, Hong Kong, China, Wales, United Kingdom nephropathy. Furthermore, we measured area under the receiver-operating Microvascular impairment is a major cause for diabetic nephropathy characteristic curve (AUC) to compare the predictive ability of the progres- (DN). Retinal vascular images allow us to observe microvascular change sion of diabetic nephropathy of home PP and home SBP. During the two year noninvasively thus it has been proposed as a new tool for DN detection. In study, the progression of diabetic nephropathy was observed in 94 patients. order to understand the relation between retinal vascular change and DN, a The adjusted odds ratio of home PP and home SBP to the progression of case control study was designed based on 709 diabetes patients recruited diabetic nephropathy was 1.23 (95% CI, 1.01 to 1.49, P = 0.032) and 1.14 (95% from the Wong Siu Ching Family Medicine Clinic of the Hong Kong Hospi- CI, 0.99 to 1.33, P = 0.077), respectively. AUC of home PP and home SBP to tal Authority New Territories East Cluster. Subjects were divided into three the progression of diabetic nephropathy in home PP was 0.624 (0.665-0.679; groups based on the eGFR level as group 1: >90 ml/min/1.73m2; group 2: P < 0.001). The optimal cut-off points, sensitivity and specificity for home 60-90 ml/min/1.73m2; and group 3: <60 ml/min/1.73m2 respectively. Clini- PP which are associated with progression of diabetic nephropathy were cal and retinal vascular characteristics were compared among DN groups 57.7 mmHg, 0.649 and 0.580, respectively. Our findings implicate, for the using one-way ANOVA. The study showed that in addition to traditional first time, home PP as an independent predictor of progression of diabetic risk factors for DN such as longer diabetes duration (p<0.0001), higher nephropathy in patients with type 2 diabetes. age (p<0.0001), BMI (p=0.038) and systolic blood pressure (p=0.019), more

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A138 COMPLICATIONS—NEPHROPATHY—CLINICAL AND TRANSLATIONAL RESEARCH severe DN were associated with increased retinal arterial curvature tortu- control showed that the areas under the ROC are 0.545, 0.822 and 0.732 osity (p=0.012) and branching coefficient (p=0.029). On the contrary, lower respectively. HDL-Cholesterol (p=0.001), decreased venous caliber (Central retinal venous Conclusions: We conclude that increased FEVDBP contributes to the equivalent, CRVE) (p=0.002), fractal dimension (p<0.0001) and branching mechanisms of VDD in T2DM. The strong associations of FEVDBP with gly- angle (p=0.001) for both arteriole and vennule were associated with greater cemic control and DN suggests that this index could play a wider role in the DN severity. Our study supported the possibility of using retinal vascular pathogenesis and/or detection of diabetic complications. characteristics in DN detection. Statistical modeling regarding DN evalu- Supported By: Kuwait Foundation for the Advancement of Science (2011-1302-01) ation will be further studied based on the results of this study and further POSTERS validation will be conducted using a separate population from the Diabetic 532‑P Complications Eyes Screening Wales (UK) program. Nonalbumin Proteinuria Predict the Progression of Type 2 Diabetic Acute and Chronic Nephropathy Better than Albuminuria 530‑P JONG HO KIM, SEO YOUNG OH, EUN HEUI KIM, MIN JIN LEE, YUN KYUNG JEON, A T-Cell Mediated Cytokine Milieu Might Be an Indicator of β-Cell SANG SOO KIM, BO HYUN KIM, JIN MI KIM, IN JOO KIM, Busan, Republic of Inflammation and Loss in New-Onset of Diabetes after Renal Trans‑ Korea plantation Objective: Albuminuria is generally accepted to be a sensitive marker of MOHAMED JAHROMI, PARUL AGARWAL, OSAMA GHAITH, NASHWA OTHMAN, diabetic nephropathy. The aim of this study was to compare whether nonal- NARAYANAN NAMPOORY, Kuwait City, Kuwait bumin proteinuria or albuminuria lead to better prediction of the progression New-onset of diabetes after transplantation (NODAT) is a serious meta- of type 2 diabetic nephropathy. bolic complication of diabetes. Although dysfunction is considered Methods: In this retrospective observational study, the urine albumin- the main contributing factor in the developments of NODAT, the precise to-creatinine ratio (ACR) and the nonalbumin protein-to-creatinine ratio pathogenesis has not been identified yet. While several cytokines have been (NAPCR) were measured in 325 patients with type 2 diabetes and estimated suggested to be involved in the inflammation of beta cells in diabetes, only a glomerular filtration rates (eGFR)≥ 30 mL/min/1.73m2. The patients were few studies have examined beta cell dysfunction in NODAT. In this study we divided into four groups according to cutoff points of urinary ACR (30 mg/g) examined the association between NODAT and functionally significant SNPs and NAPCR (119 mg/g). The renal outcomes were chronic kidney disease in interferon-gamma and IL-4 genes. This will enable us to examine TH1/ (CKD) progression and accelerated eGFR decline. TH2 cytokines milieu which might be related to inflammation due to beta Results: During the 48 months follow-up period, 25 (7.7%) patients showed cell dysfunction. A total of 126 renal transplant recipients were included CKD progression and 69 (21.2%) patients showed accelerated eGFR decline. without . The association between NODAT development After adjusting for ten clinical parameters including ACR, the group with was analyzed using the key immunosignatures cytokines such as interferon- NAPCR levels above 119mg/g exhibited a higher cumulative incidence of gamma and IL-4 genes. SNPs in these genes were amplified using ARMA- CKD progression (hazard ratio, 5.43; P = 0.006) and accelerated eGFR decline PCR technology compared to kidney transplant recipient without diabetes (hazard ratio, 1.90; P = 0.024). In patients with normoalbuminuria, the group after a year, non-NODAT. Results were analyzed using Fisher’s exact test. with NAPCR levels above 119mg/g also exhibited a higher cumulative inci- There was a strong association among TT compared to AA and AT geno- dence of CKD progression (hazard ratio, 21.73; P = 0.005) and accelerated types of interferon-gamma in NODAT vs. non-NODAT, p=0.009. However, eGFR decline (hazard ratio, 1.87; P = 0.085). there was no statistically significant association between IL-4 CC, CT and TT Conclusions: Nonalbumin proteinuria is a better predictor of the progres- genotypes in cases compared to control group. The data was adjusted for sion of CKD than albuminuria in patients with type 2 diabetes. age at onset and gender, however, no substantial difference was noted. This is the first report suggesting NODAT as TH1 mediated diabetic complication. 533‑P The data suggest that inflammation of beta cells might play a crucial role in Progression of Diabetic Nephropathy in Association with Platelet the pathogenesis of NODAT post . It is highly recom- Activating Factor Receptor Gene Expression in Peripheral Blood mended to further this study and also to be repeated by other centers using Mononuclear Cells different ethnicities. SAHAR GHAVIDEL DARESTANI, MIHOKO KURANO, KIYOHISA YAMAMOTO, Supported By: Dasman Diabetes Institute; Kuwait Foundation for the Advance- ATUSHI SHINNAKASU, TAKAHISA DEGUCHI, YOSHIHIKO NISHIO, AIKO ment of Sciences ARIMURA, HIROSHI HASHIGUCHI, Kagoshima, Japan In our previous study, we have shown that the level of platelet activat- 531‑P ing factor receptor (PAFR) gene expression in peripheral blood mono-nuclear Fractional Excretion of Vitamin D Binding Protein as a Novel Marker cells (PBMCs) is closely associated with the urinary albumin to creatinine of Vitamin D Status in Subjects with Type 2 Diabetes ratio (ACR) and flow mediated dilatation of forearm (FMD). However, the role NABILLA ABDELLA, OLUSEGUN A. MOJIMINIYI, Safat, Kuwait, Kuwait City, of PAFR expression in PBMCs in joining endothelial dysfunction and diabetic Kuwait nephropathy remains overlooked. Neither the factors affecting PAFR gene Introduction: Vitamin D deficiency (VDD) plays significant roles in the expression in PBMCs nor the mechanisms linking diabetic nephropathy and pathogenesis and complications of type 2 diabetes mellitus (T2DM). Vitamin arterial dysfunction are clearly understood. To elucidate the findings from D-binding protein (VDBP), a 58-kDa glycoprotein, is a significant determinant our previous study and to clarify this mechanism, we followed the partici- of biologically active 25(OH) Vitamin D (25(OH)D). Studies showed increased pants for one year. Ninety-five patients were recruited from hospital out- urine excretion of VDBP in patients with diabetic nephropathy (DN) resulting patient clinic and 88 of them (normoalbuminuric=41, microalbuminuric=24, from postulated mechanisms linked to renal tubular damage. We evaluate macroalbuminuric=23) were followed for one year. Blood samples for analy- the utility of the Fractional Excretion of VDBP (FEVDBP) as an index of VDD sis of mRNA expression in PBMCs and urine samples for evaluation of ACR and DN. were collected at the beginning and at the end of one year. Expression of Methods: 25(OH)D, HbA1c, serum and urine concentrations of VDBP, cre- PAFR mRNA in PBMCs was significantly higher in macroalbuminuric patients atinine were measured in 405 (129M, 276F) T2DM patients. Ratio of urine compared to normoalbuminurics (p=0.000). Pearson correlation revealed a microalbumin to creatinine was determined to categorize subjects as nor- strong positive association between changes in ACR and changes in PAFR moalbuminuric (NAO); microalbuminuric (MIA) and macroalbiminuric (MAA). only in microalbuminuric patients (r=0.628, p=0.001). Furthermore, changes FEVDBP was calculated as 100 x (UrineVDBP x SerumCreat)/(SerumVDBP in PAFR level were negatively correlated with changes in eGFR in macroalbu- x UrineCreat). Univariate and multivariate analyses were used to compare minuric patients (r= -0.47, p=0.03). Thus changes in PAFR gene expression in study subjects grouped by Vitamin D status, glycemic control and degree of PBMCs are associated with progression of nephropathy either in microalbu- microalbuminuria. minuric or macroalbuminuric patients. Considering previous findings demon- Results: VDD (<50nmol/L; n = 237) or insufficiency (VI) (50-75 nmol/L; strating interactions between platelet activating factor (PAF) on endothelial n=84) was prevalent. Mean FEVDBP in subjects with normal 25(OH)D, VI cells and PAFR on PBMCs and the association of endothelial dysfunction and VDD were 5.5, 5.4 and 8.5 respectively; mean FEVDBP in NAO, MIA with elevated expression of PAFR in PBMCs, our findings also contribute to and MAA were 3.7, 23.3 and 55.9 respectively. Significant correlations better understanding the development of macrovascular complications in of FEVDBP were with age (r=0.38), glucose (r=0.42), HbA1c (r=0.46), urine patients with nephropathy. microalbumin:creatinine ratio (r=0.56) and significant negative correlation Supported By: Japan Ministry of Education, Culture, Sports, Science and Tech- with serum albumin (r = -0.30). Receiver operating Curve (ROC) analyses of nology the use of FEVDBP for detection of VDD, microalbuminuria and poor glycemic

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534‑P show the associations with the annual decline rate in eGFR and the progres- Renal Failure and Mortality in Patients with Type 2 Diabetes, a sion of CKD stage (r=0.322, P=0.0015). Real-Life Study In conclusion, urine clusterin reflects tubular damage in the early-stage MANUEL J. ROMERO JIMÉNEZ, EVA N. GUTIÉRREZ CORTIZO, FRANCISCO J. of DKD. The increase of urine clusterin along with albuminuria could be an CABALLERO GRANADO, ELENA SÁNCHEZ RUIZ-GRANADOS, FRANCISCO HER- independent predictive marker for the progression of DKD in type 2 diabetes. RERO MACHANCOSES, ANTONIO LEÓN JUSTEL, PILAR RODRÍGUEZ ORTEGA, Supported By: American Diabetes Association (7-12-BS-094 to Y.B.K.); Pusan ISABEL GONZÁLEZ CARMELO, Huelva, Spain National University Hospital (2010-3)

POSTERS The prevalence of renal failure (RF) among hospitalized patients diag- Complications nosed with type 2 diabetes (T2D) is unknown. Determining this prevalence 536‑P Acute and Chronic and its associated mortality will improve the therapeutic approach offered Hypertriglyceridemia Is a Residual Risk Factor Associated with to our patients. New-Onset DKD in Type 2 Diabetic Patients without Hypertension An analytical-descriptive and retrospective study of 3148 type 2 diabetic LEE TAO-CHUN, Pingtung, Taiwan patients admitted to the Huelva University Hospital Complex during 2013 Background: Under standard care, risk factors to new-onset diabetes kid- with or without RF (groups 1 and 2) was conducted. The glomerular filtration ney disease (DKD) were not clear, particularly in patients with type 2 diabe- rate was calculated using the CKD-EPI formula. tes mellitus (T2DM) without hypertension. The aim of this study is to reveal A total of 57.3% of patients had RF. Based on the classification of stages residual risk factors associated with new-onset DKD in T2DM patients with- of renal function 14.20% of patients were classified as stage 1 and 28.60% out hypertension. as stage 2; 17.5% of patients were on stage 3a, 17.30% were on stage 3b, Material and Methods: Non-albuminuric normotensive T2DM patients 14% were on stage 4, and 8.4% of patients were on stage 5. with estimated GFR (eGFR) ≥ 60 ml/min/1.73m2 who had followed in the Tai- The average hospital stay of group 1 was 11.34 days (SD = 11) com- wan Diabetes Shared Care Program were included. The development of DKD pared with 9.16 days (SD = 9) for group 2, which was significantly different was defined as the new-onset of albuminuria or eGFR < 60 ml/min/1.73m2. (t = -5.905, p < .01). Group 1 had more readmissions (OR = 1.2976, 95% confi- Multivariate cox-regression analysis was used to identify the risk factor dence interval [CI] = 1.0951-1.5375, p < .01). Moreover, 17.7% of group 1 died associated with new-onset DKD. during the first year after admission, while in group 2 was 6.2% (OR = 2.6453, Results: 563 T2DM patients with mean age of 58.4 were included. Dur- 95% CI = 2.2051-3.1734, p < .01). ing the 4.8-year follow-up period, there were 67 patients (11.9%) recog- The presence of RF among hospitalized patients with T2D was high and nized as the new-onset DKD group. There were no significant differences was associated with longer hospital stays, increased hospital readmissions in age, gender, smoking status, diabetes disease duration, body mass index, and mortality. RF limits the treatment of T2D, which is essential for develop- mean HbA1c, mean total cholesterol, mean LDL-C and mean diastolic pres- ing therapeutic approach during admission and at discharge. sure between the two groups. However, the new-onset DKD group had a Table. Prevalence and Mortality in T2D Hospitalized. significant higher mean value of systolic pressure, triglycerides and lower mean HDL-C levels. After multivariate cox-regression analyses, the results GFR Stage 1 Stage 2 Stage 3a Stage 3b Stage 4 Stage 5 normal (GFR >90) (GFR 60-90) (GFR 60-45) (GFR 45-50) (GFR 30-15) (GFR <15) indicated triglycerides ≥200 mg/dl is the only residual metabolic risk fac- tor associated with new-onset DKD with OR of 3.52 (95% C.I.: 1.48~8.34; Age 66.34 57.77 70.59 73.41 75.45 75.83 74.31 P=0.004). Sex (Female) 49.3% 50.4% 48.7% 42.8% 44.9% 43.5% 42.9% Conclusion: In T2DM patients without hypertension under standard diabe- Mean stay 9.16 9.76 8.86 10.11 10.45 12.26 14.18 tes management, hypertriglyceridemia is an independent risk factor associ- Hospital 20.6% 20.6% 20.6% 21.4% 21.7% 29.3% 33.5% ated with new-onset DKD. readmission Death at 2.1% 2.2% 2% 4.2% 6.1% 10% 9.8% 537‑P 1 month Urinary Trefoil Factor 3 Levels in Association with Albuminuria in Death at 11.4% 11.9% 11.1% 15.6% 18.9% 32.4% 44.4% Patients with Early Stages of Diabetic Nephropathy 6 months NAOTO TERAMI, DAISUKE OGAWA, TAKASHI HATANAKA, HIROMI TACHIBANA, HITOSHI SUGIYAMA, JUN WADA, Okayama, Japan, Fukuyama, Japan, Tsuyama, Death at 14.4% 14.6% 14.3% 21% 25.6% 36.7% 52.3% 1 year Japan Trefoil factor (TFF) are coexpressed with mucins in the gastro- intestinal tract including stomach and colon, and they are also expressed in kidney tubules. Serum levels of TFF3 have been reported a possible bio- 535‑P marker of gastric cancer. Previous study reported that urinary TFF3 (uTFF3) Urine Clusterin Is a Tubular Damage Marker that Predicts the Pro‑ levels were reduced, and urinary albumin levels increased in response to gression of Diabetic Kidney Diseases renal tubular injury in mice. In this study, we determined whether uTFF3 is SANGSOO KIM, SANG HEON SONG, JONG HO KIM, YUN KYUNG JEON, BO an efficient biomarker in patients with early stages of diabetic nephropathy. HYUN KIM, MIN CHEOL KANG, SUNG WAN CHUN, MICHELLE CHUNG, YONG Spot urine samples were obtained from 79 male and 64 female type 2 dia- KI KIM, IN JOO KIM, YOUNG BUM KIM, Boston, MA, Busan, Republic of Korea betic patients (n=143) at Okayama University Hospital. The levels of uTFF1, Knowledge of urine clusterin’s action in the development of chronic kidney uTFF2, and uTFF3 were measured quantitatively by specific ELISAs to analyze disease (CKD) including diabetes kidney disease (DKD) remains incomplete. the correlation between uTFF1, uTFF2, uTFF3 and various clinical parameters. The objective of this study was to evaluate the clinical implication of urine The level of uTFF3 significantly increased in diabetic patients with micro- clusterin on the development and/or progression of DKD in type 2 diabetes. albuminuria compared to those with normoalbuminuria (p=0.0139). In con- 159 type 2 diabetic patients and 20 nondiabetic subjects with estimated trast to the level of uTFF3, the level of uTFF1 or uTFF2 did not significantly 2 glomerular filtration rate (eGFR)≥ 60 mL/min/1.73 m were enrolled. The elevate in diabetic patients with microalbuminuria compared to those with baseline values of urine clusterin and tubular damage markers (kidney injury normoalbuminuria. We investigated a multiple logistic regression analysis molecule-1 [KIM-1], Neutrophil gelatinase-associated lipocalin [NGAL], liver- of predictor of albuminuria for five parameter; HbA1c > 7.0%, triglyceride type fatty acid binding protein [LFABP] and non-albumin protein-to-creat- > median, uTFF1 > median, uTFF2 > median, uTFF3 > median. A parameter inine ratio [NAPCR]) were measured. The primary outcome was an annual with the primary large Odds ratio (OR) for early diabetic nephropathy was decline rate in eGFR and secondary outcomes were the development of CKD uTFF3 (OR: 2.34, 95% CI: 1.00 to 5.69, P = 0.0488). In addition, uTFF3 level was stage 3 or greater and a persistence/progression of albuminuria. The median significantly correlated with tubulointerstitial injury marker; urinary L-FABP follow-up duration of enrolled patients was 3.0 (1.0-5.9) years. Baseline (r = 0.5855, p < 0.0001), α1MG (r = 0.7559, p < 0.0001), β2MG (r = 0.6552, clusterin levels in urine were significantly increased in type 2 diabetic sub- p < 0.0001) and NAG (r = 0.5105, p < 0.0001). These data indicate that uTFF3 jects compared with those of nondiabetic subjects (P=0.002). The levels of is a predictor of microalbuminuria and tubulointerstitial injury in patients urine clusterin had a strong correlation with other tubular damage markers with diabetes mellitus. Further studies are necessary to elucidate whether (all P<0.001). A positive correlation between the annual rate of decline in uTFF3 can predict the progression of diabetic nephropathy. eGFR and urine clusterin after adjusting for clinical confounding factors was detected (r=0.258, P=0.007). Multivariate analysis further indicated that urine clusterin correlated with the development of CKD stage 3 or greater and persistence/progression of albuminuria, respectively (both P=0.032). In type 2 diabetic subjects with albuminuria, urine clusterin still remained to

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538‑P 540‑P Nonalbuminuric Diabetic Kidney Disease in Type 1 Diabetes: Asso‑ Renin Angiotensin System (RAS) Activation and Nephropathy ciation with Major Vascular Outcomes Risk and All-Cause Mortality Resistor Status in Long-Standing Type 1 Diabetes: The Canadian MONIA GAROFOLO, ELEONORA RUSSO, ROBERTO MICCOLI, DANIELA LUC- Study of Longevity in Diabetes CHESI, LAURA GIUSTI, GIUSEPPE DANIELE, GIUSEPPE PENNO, STEFANO DEL JULIE A. LOVSHIN, GENIEVE BOULET, YULIYA LYTVYN, MOHAMMED FAROOQI, PRATO, Pisa, Italy LEIF E. LOVBLOM, VESTA LAI, LESLIE CHAM, JOSEPHINE TSE, ALANA WEISMAN, Nonalbuminuric Diabetic Kidney Disease (Alb-DKD) was described in HILARY KEENAN, NARINDER PAUL, VERA BRIL, BRUCE A. PERKINS, DAVID Z.I.

T1D in DCCT/EDIC, in the 2nd Joslin Kidney Study and in FinnDiane. Rate CHERNEY, Toronto, ON, Canada, Quebec City, QC, Canada, Hamilton, ON, Canada, POSTERS of Alb-DKD and association with the EURODIAB score for major vascular Boston, MA Complications outcomes (based on age, HbA1c, WHR, A/C and HDL) and all-cause mortality Aims: Physiological mechanisms that protect against the development of Acute and Chronic were evaluated in a cohort of 774 T1D (M/F 53/47%, age 40.2±11.7 yrs, DD nephropathy in some patients (“resistors”) and not in others (“non-resistors”) 19.3±12.2 yrs, HbA1c 7.8±1.2%). Out of 774 T1D, 692 (89.4%) had no DKD, 53 are poorly defined in those with longstanding T1D. We aimed to determine (6.8%) DKD stages 1-2, 12 (1.6%) Alb+DKD stages ≥3 and 17 (2.2%) Alb-DKD whether RAS activation differed between non-resistors, resistors and stages ≥3. Alb-DKD account for 20.7% of all DKD and for 58.6% of DKD healthy controls (HC). stages ≥3. Out of 774 T1D, 466 (60.2%) had low EURODIAB score (<16), 205 Methods: Patients with prolonged durations of T1D (n=75, duration >50 (26.5%) intermediate score (IS, 16-20) and 103 (13.3%) high score (HS, ≥21). years) and age/gender-matched HC (n=75) were studied. The T1D cohort was Rate of HS was: no DKD, 9.1%; DKD stages 1-2, 34.0%; Alb-DKD, 64.7%; stratified into 49 resistors (eGFR >60mL/min/1.73m2 and <30 mg/day urine Alb+DKD, 91.7% (p<0.0001). In a follow-up of 8.25±2.34 yrs (median 7.58, albumin) and 26 non-resistors. Renal hemodynamic function (GFRINULIN, renal IQR 6.47-9.75), 40 (5.2%) T1D died with a rate of 6.26 x1000 pts/yr. Mortal- plasma flow [RPF], renal blood flow [RBF], renal vascular resistance [RVR]) ity increased with DKD phenotypes: no DKD, 3.0%; DKD stages 1-2, 15.1%, was measured at baseline and after angiotensin II (AngII) infusion. The pre- HR 4.504 (1.992-10.186); Alb-DKD, 29.4%, HR 8.573 (3.222-22.815); Alb+DKD, specified primary endpoint for RAS activation was defined by RVR change in 50.0%, HR 20.683 (8.292-51.587) (p<0.0001). Accounting for age (HR 1.070; response to AngII. 1.044-1.098, p<0.0001) and gender (M: HR 1.922; 0.969-3.813, p=0.061), HRs Results: Non-resistors had lower GFR (93±15 vs. 108±16 [resistors, for all-cause mortality vs. no DKD were: DKD stages 1-2, 3.841 (1.686-8.750, p<0.001], 106±19 [HC, p=0.005] ml/min/1.73m2), lower RPF and RBF and p=0.001); Alb-DKD, 2.970 (1.018-8.662, p=0.046); Alb+DKD, 7.441 (2.781- higher RVR and plasma renin at baseline, with a higher ratio of afferent (RA) 19.911, p<0.0001). In a second model, accounting for gender and the score to efferent (RE) arteriolar resistance compared to resistors and HC. Non- (IS: HR 3.346, 1.201-9.321, p=0.021; HS: HR 11.736, 4.437-31.044, p<0.0001), resistors had attenuated RVR responses to AngII indicating the highest HRs for mortality were: DKD stages 1-2, 2.571 (1.113-5.939, p=0.027); level of endogenous RAS activation (19±9% vs. 29±14% [resistors, p=0.008], Alb-DKD, 2.772 (0.968-7.943, p=0.058); Alb+DKD, 4.584 (1.691-12.421, 30±18% [HC, p=0.009]). As hypothesized, non-resistors and resistors had p=0.003). In T1D, Alb-DKD accounts for 20% of all DKD and is associated similar responses on RE compared to HC to AngII, however unexpectedly, with an increased risk of major outcomes and an high all-cause mortality, the response to AngII on RA differed significantly in non-resistors compared to the same extent as stages 1-2 DKD. The highest risk and mortality were to resistors and HC. observed in Alb+DKD. Intensive cardiovascular prevention strategies should Conclusions: Though RAS activation clearly underlies development of be applied to these subjects. nephropathy, these results identify the key role of renal afferent arteriolar resistance in nephropathy pathogenesis. This implies that RAS inhibition 539‑P (which targets renal efferent arteriolar tone) may be insufficient for nephro- Hemorheological Approach for Screening Diabetic Nephropathy in protection and that additional strategies targeting renal afferent arteriolar Type 2 Diabetes tone require further study in T1D. SANG BAE LEE, JUNG HYE KIM, KAHUI PARK, YU SIK KIM, DA WOON HAN, Supported By: JDRF BOMIN KOH, SU JEONG PARK, HO SEON PARK, JI HONG YOU, JI SUN NAM, JONG SUK PARK, CHUL WOO AHN, KYUNG RAE KIM, Seoul, Republic of Korea 541‑P Background: Hemorheologic alterations or changes in blood viscosity have Is There a Causal Relationship between Hypertriglyceridemia, Cor‑ been suggested to play a role in the pathogenesis of diabetic microvascular onary Heart Disease, and Diabetic Nephropathy in Type 1 Diabetes? complications. We measured various hemorheologic parameters in type 2 LARS STECHEMESSER, CAROL FORSBLOM, NICOLAE M. PANDURU, IIRO TOP- diabetes patients and assessed their possible role as a diagnostic tool for PILA, PER-HENRIK GROOP, FINNDIANE STUDY GROUP, Salzburg, Austria, Helsinki, diabetic nephropathy. Finland, Bucharest, Romania Methods: Four hundred-seventy patients with type 2 diabetes were Background and Aims: Dyslipidemia is a major risk factor for coronary included in this study. Hemorheologic parameters, including erythrocyte heart disease (CHD) and diabetic nephropathy, the most common causes deformability, elongation index (EI), critical shear stress (CSS), and aggrega- of premature death in type 1 diabetes (T1D). Data whether hypertriglyc- tion index (AI) were measured using microfluidic hemorheometer. Various eridemia is causally related to these diabetic complications are scarce. We metabolic parameters were assessed from fasting blood samples and uri- therefore investigated if hypertriglyceridemia is causally related to CHD and nary albumin to creatinine ratio was used to assess diabetic nephropathy. diabetic nephropathy in T1D. Results: There were significant differences in Elongation index at 3 Pas- Methods: This cross-sectional analysis was part of the prospective, ongo- cal (EI at 3Pa), Fibrinogen/EI, and shear stress among patients in different ing Finnish Diabetic Nephropathy Study (FinnDiane). Between 1994 and 2015 stages of chronic kidney disease (all p<0.05), EI at 3 Pa, Fibrinogen/EI, and data were obtained from 4481 T1D patients in more than 80 hospitals or shear stress significantly differed among the groups. Fibrinogen/EI differed health centers across Finland. The potentially causal effect of triglycerides between normal or CKD 1 and CKD 2 patients. In multiple regression analy- on CHD and stages of nephropathy was explored using Mendelian Random- sis, Fibrinogen/EI at 3Pa was an independent predictor of albumin to creati- ization (MR) and a genetic risk score based on 96 SNPs associated with nine ratio independent of age, ESR, hematocrit, HbA1c, and body mass index triglycerides in the . MR analysis was performed using the GMM (β=0.101, p<0.05). Also, critical time, critical stress, fibrinogen/EI at 3Pa, estimation of structural mean models for binary and the two-stage least CSS/EI et 3Pa, ad fibrinogen/CSS at 3Pa were significantly different among squares method for continuous outcome. patients at different stages of diabetic nephropathy (all p< 0.05). Among the Results: CHD was present in 8.3% of the population (n=4481, females variables, Fibrinogen/EI at 3Pa showed area under the ROC curve of 0.721, 47.8%, age 41±12.7 yrs, diabetes duration 25.1±13.1 yrs, HbA1c 8.4±1.4%). 18% suggesting 860 mg/dL% as a cut off point for diabetic nephropathy with the had CKD stage 3-5, 12.5% microalbuminuria, 13.8% macroalbuminuria and sensitivity of 74% and specificity of 62%. 9.7% had end stage renal disease. Triglycerides (p<0.001) but not the genetic Conclusions: Fibrinogen/EI is a sensitive parameter measured via point- risk score (p=0.464) correlated with eGFR. However, instrumental variables of-care testing for screening diabetic nephropathy in patients with type 2 regression showed no causal effect of triglycerides on eGFR (p=0.470). Tri- diabetes. glycerides (p<0.001) and genetic risk score (p=0.002) correlated with CHD. Notably, MR showed a causal effect of genetically elevated triglycerides on CHD with a 2.5-fold elevated risk for each SD-elevation of log Triglycerides. Conclusions: In addition to confirming that triglycerides are highly corre- lated with CHD, our data also suggest that there is a causal link between triglycerides and CHD in T1D. In contrast, we found no causal relationship between triglycerides and diabetic nephropathy in this population.

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542‑P known whether eGFR is associated with future changes in insulin sensitivity Statin Use and Renal Function Decline in Patients with Diabetes or insulin secretion. Therefore, we aimed to examine the relation of baseline EUGENE HAN, YONGIN CHO, MINYOUNG LEE, KWANG JOON KIM, YONG HO eGFR to longitudinal changes in insulin sensitivity index (SI) and acute insulin LEE, BYUNG WAN LEE, BONG SOO CHA, CHUL HOON KIM, EUN SEOK KANG, response (AIR) in 739 IRAS participants without diabetes at baseline (age, Seoul, Republic of Korea 40-69 years; median follow-up, 5.2 years [range, 4.6-6.6 years]; and 115 inci- Although the beneficial effects of statin treatment in dyslipidemia and ath- dent cases of type 2 diabetes). We calculated eGFR by the Modification of erosclerosis have been well studied, there is limited information regarding Diet in Renal Disease equation at the baseline visit, and measured SI and AIR by the frequently sampled intravenous glucose tolerance test at the base-

POSTERS the renal effects of statins in diabetic nephropathy. We aimed to investigate

Complications whether and which statin could help maintain renal function in Asian patients line and follow-up visits. Individuals with stage 3 CKD (eGFR 30-60 ml/min Acute and Chronic with diabetes. We enrolled 484 patients with diabetes who received statin per 1.73 m2) had a smaller longitudinal increase in log-transformed AIR than treatment for more than one year. Individuals treated with moderate inten- those with normal CKD (≥90 ml/min per 1.73 m2), but a similar longitudinal sity statin dose either atorvastatin (10-20 mg/day) or rosuvastatin (5-10 mg/ decline in log-transformed SI (Table). day) were included. The primary outcome was changes in glomerular filtration In conclusion, since insulin sensitivity declines with age, subjects with rate (GFR), during statin treatment. Rapid renal decline was defined >3% GFR stage 3 CKD have a limited ability to augment their first phase insulin secre- reduction in a year. Both statin treatment improved serum lipid levels. GFR was tion and thus be at higher risk of developing diabetes. slightly, but significantly decreased in both statin groups (from 80.3 ml/min/1.73 Table. Baseline and Longitudinal Changes in SI and AIR by CKD Categories. m2 to 78.8 ml/min/1.73 m2; P=0.012 for atorvastatin, from 79.1 ml/min/1.73 m2 2 eGFR 41.8 - 59.9 eGFR 60 - 89.9 eGFR 90.0 - 239.1 to 76.1 ml/min/1.73 m ; P=0.001 for rosuvastatin). More rapid GFR decline was ml/min × 1.73 m2 ml/min × 1.73 m2 ml/min × 1.73 m2 observed in rosuvastatin group compared to those with atorvastatin (48.7% vs. 38.6%, P=0.029). Multiple logistic regression analyses demonstrated more n 55 496 188 rapid renal function loss in rosuvastatin group (OR=1.60, 95% CI=1.06-2.42) Baseline log-SI 1.09 ± 0.07, 1.04 ± 0.02, 0.96 ± 0.04 (Ref.) compared to atorvastatin group after adjustment for other confounding factors. (× 10-4 min-1 . μU-1 . ml-1) p = 0.100 p = 0.080 In conclusion, moderate intensity dose of rosuvastatin treatment was Δlog-SI adjusted for - 0.30 ± 0.05, - 0.28 ± 0.02, - 0.26 ± 0.03 (Ref.) associated with rapid GFR loss compared to atorvastatin. Our findings sug- baseline log-SI p = 0.547 p = 0.539 gest that atorvastatin treatment in patients with diabetes might be more Baseline log-AIR 3.98 ± 0.10, 3.92 ± 0.03, 3.93 ± 0.05 (Ref.) beneficial to preserve GFR than rosuvastatin. However, further prospective (μU/ml) adjusted for p = 0.710 p = 0.789 and randomized trials are needed to demonstrate this effect compared to baseline log-SI placebo and other statins. Δlog-AIR adjusted 0.05 ± 0.07, 0.23 ± 0.02, 0.23 ± 0.04 (Ref.) for baseline log-SI and p = 0.035 p = 0.956 log-AIR and log-S 543‑P Δ I Plasma Uric Acid and Renal Function in Adolescents Compared Δlog-AIR adjusted 0.03 ± 0.07, 0.23 ± 0.02, 0.24 ± 0.04 (Ref.) with Adults with T1D for More than 50 Years for baseline fasting and p = 0.011 p = 0.816 YULIYA LYTVYN, MOHAMMED A. FAROOQI, JULIE A. LOVSHIN, GENEVIEVE 2h glucose, BMI, log-SI, and log-AIR, and log-S BOULET, VESTA LAI, JOSEPHINE TSE, LESLIE CHAM, ERIK LOVBLOM, ALANNA Δ I WEISMANN, HILLARY KEENAN, MICHAEL BRENT, NARINDER PAUL, VERA BRIL, Δlog-SI and Δlog-AIR indicate longitudinal change in log-transformed SI and ETIENNE SOCHETT, BRUCE A. PERKINS, DAVID CHERNEY, Toronto, ON, Canada, AIR, respectively. Boston, MA Supported By: National Heart, Lung, and Blood Institute (HL-47889, HL-47890) Higher plasma uric acid (PUA) levels, even within the normal range, are associated with lower GFR and lower renal plasma flow (RPF) in young adults 545‑P with T1D. To determine the timeline for these relationships, we examined the Urinary Type IV Collagen as a Predictor for a 30% Decline in eGFR in association between PUA levels and renal hemodynamic function in adoles- Young Type 1 Diabetic Patients with Normoalbuminuria cents vs. adults with ≥50 years of T1D from the Canadian Study of Longev- MIWA MORITA, KO HANAI, YASUKO UCHIGATA, Shimane, Japan, Tokyo, Japan ity. PUA, GFR (inulin) and RPF (para-aminohippurate) were measured during a Morphological changes such as glomerular basement membrane thickening euglycemic clamp in 37 adolescent patients with 9.6±2.6 years of T1D com- and mesangial matrix expansion have been demonstrated to appear before the pared to 54.2±5.6 years in 63 older T1D adults. PUA levels were compared incidence of microalbuminuria in diabetes. Type IV collagen (T4C) is the main to a historical cohort of uncomplicated young T1D adults 18 to 40 years old. component of these extracellular matrix, and increased levels of urinary T4C are Gomez’s equations were used to estimate afferent (RA) and efferent (RE) arte- considered to be associated with increased shedding from glomeruli, because riolar resistances and glomerular hydrostatic pressure (PGLO). The relationships of its high molecular weight. Here, we aimed to examine the hypothesis that between PUA and renal hemodynamic parameters were evaluated by univari- urinary T4C levels are a predictor for a 30% decline in eGFR, accepted as a surro- able linear regression correlation coefficients. PUA levels were higher with gate endpoint of end-stage renal disease, in young type 1 diabetic patients with longer T1D duration: 197±53 in adolescents vs. 234±62 in uncomplicated young normoalbuminuria. A historical cohort study was conducted, which included adults vs. 281±83µmol/L in patients with ≥50 years of T1D (p<0.001). While normoalbuminuric patients diagnosed with type 1 diabetes before the age PUA levels did not associate with renal hemodynamic parameters in adoles- of 30 years, who were <40 years old at baseline. Patients with < 18 years old cents, in patients with ≥50 years of T1D higher PUA correlated with lower GFR and a diabetes duration of <5 years were excluded. Overall, 170 patients were (r=-0.42, p=0.001), lower RPF (r=-0.26, p=0.040), higher renal vascular resis- enrolled. The endpoint was ≥30% decline in eGFR from baseline. To examine the tance (r=0.25, p=0.050) and lower renal blood flow (r=-0.29, p=0.020). In association of urinary T4C levels with the endpoint, we used the multivariate Longevity Study participants, higher PUA tended to correlate with higher Cox proportional hazard model analysis. The covariates were the followings: RA (r=0.35, p=0.070) but did not correlate with RE or PGLO. The relationship age, sex, BMI, systolic and diastolic blood pressure, logarithmically urinary albu- between higher PUA with lower GFR and RPF observed in young T1D adults min-to-creatinine ratio, HbA1c, eGFR, total cholesterol, logarithmically TG, use is absent in adolescents with shorter T1D duration, but is evident after long- of renin angiotensin system blocker. During the mean follow-up period of 12.2 standing T1D. Thus, T1D may modify the association between PUA and renal ± 5.1 years, 49 patients reached the endpoint. Higher levels of logarithmically hemodynamic function, suggestive of renal vasoconstriction and ischemia. urinary type IV collagen-to-creatinine ratio were associated with the endpoint Further work is required to determine whether pharmacological PUA lowering (hazard ratio: 4.121, 95% CI 1.301-13.059, p= 0.016). Next, when patients were prevents renal vasoconstriction, thereby improving long-term renal outcomes. classified into 2 groups by the median of urinary type IV collagen-to-creatinine Supported By: JDRF (17-2013-312 to Y.L.); Canadian Diabetes Association (to Y.L.) ratio levels, the hazard ratio of patients with high T4C levels (vs. those with low T4C levels) for the endpoint was 2.536 (1.384-4.649, p= 0.003). 544‑P In conclusion, urinary T4C levels may be a predictor for a 30% decline in Subjects with Chronic Kidney Disease (CKD) Have a Diminished eGFR in young type 1 diabetic patients with normoalbuminuria. Acute Insulin Response over a Five-Year Period: The Insulin Resis‑ tance Atherosclerosis Study (IRAS) CARLOS LORENZO, ANTHONY J. HANLEY, MARIAN REWERS, STEVEN M. HAFF- NER, San Antonio, TX, Toronto, ON, Canada, Aurora, CO We have previously reported that changes in estimated glomerular filtra- tion rate (eGFR) may precede the onset of type 2 diabetes. However, it is not

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546‑P 548‑P Hyperglycemia-Induced NETs Promotes Renal Fibrosis Perfluoroalkyl Substances and Kidney Function in Chronic Kidney HU JINBO, SR., LI XUAN, SR., CHUAN PENG, SR., RUFEI GAO, QIFU LI, Chongqing, Disease, Anemia, and Diabetes China BAQIYYAH N. CONWAY, KIM INNES, TINA COSTACOU, JOHN ARTHUR, Morgan- Diabetic nephropathy (DN) is characterized by fibrosis. Neutrophil extra- town, WV, Pittsburgh, PA, Little Rock, AR cellular traps (NETs), which composed of decondensed chromatin and anti- Anemia often complicates chronic kidney disease (CKD), leading to insuf- microbials, was released from after hyperglycemia stimulation. ficient tissue oxygenation and ultimately hypoxic injury, a factor thought to

Whether NETs promotes diabetic renal fibrosis were examined here. underlie progression from CKD to renal failure. Perfluorocarbons are potent POSTERS

In vivo, the levels of serum creatinine and urinary albumin to creatinine oxygen transporters that have been used in synthetic blood development; Complications ratio (UACR) of STZ-induced diabetic mice (16 weeks) were increased com- however, data are scarce on their relationship with kidney function and CKD, Acute and Chronic pared to controls significantly. Renal tissues from donors of normoglycemic especially in diabetes where anemia and hypoxia are more common. We subjects, patients with diabetes and diabetic kidney disease (DKD) were investigated the relationship of serum perfluoroakyl acids (PFAS) with kid- collected. Periodic Acid-Schiff stain (PAS) and Sirius-Red stain further con- ney function and variation by diabetes and anemia status. Data on 53,650 firmed renal fibrosis in diabetic mice or patients with DKD. In diabetic mice, adults (5,210 with diabetes) were obtained from the C8 Health Project. CKD NETs-related markers MPO and Cit-histone3 immunostaining inside renal was defined as an eGFR <60 mL/min/1.73 m2. Four PFAS were investigated: tubules were increased accompanied with a higher expression of α-SMA perfluorohexane sulfonate (PFHxS), perfluoroctanoic acid (PFOA), perfluoroc- and a lower expression of E-Cadherin. In patients with diabetic kidney dis- tane sulfonate (PFOS), and perfluorononaoic acid (PFNA). Linear regression ease, fibrosis of renal tubules was also accompanied by NETs formation. analysis was used to estimate the relationship between each of the PFAS In Vitro, it was further explored whether hyperglycemia-induced NETs with eGFR. Multinomial logistic regression was used to assess this relation- promotes fibrosis of HK-2 (human proximal tubule epithelial) cells. Neutro- ship with stages of kidney function, with eGFR stratified into the following phils isolated from peripheral blood of health donors were treated with categories: ≥90, 60-89, 45-59, 30-44, <30. Multivariable models included normal glucose (NG, 5 mmol/L), high glucose (HG, 25 mmol/L), phorbol age, sex, diabetes duration (in diabetes specific analyses), BMI, lipids, WBC, 12-myristate 13-acetate (PMA, 25 nmol/L), high glucose with chloroquine and CRP. Each PFAS positively was associated with eGFR among those with (CQ, 5 ummol/L), respectively in 4 hours. Sytox Green and dsDNA quantifica- CKD or anemia; this was strongest among those with both CKD and ane- tion showed NETs could be induced by HG through autophagy. In HK-2 cells, mia, followed by those with CKD uncomplicated by anemia. These relation- exposure from hyperglycemia-related NETs (100ng/mL, 24 h) resulted in a ships were more pronounced among those with diabetes (all p<0.01). In the decreased level of E-Cadherin (epithelial cell marker), and increased levels absence of CKD or anemia, PFAS inversely associated with eGFR. Among of Snail/pSmad3/Smad3, which determined by Western Blot. These results those with both anemia and diabetes, when further stratified by CKD stage, indicated that HK-2 cells underwent a partial epithelial-mesenchymal transi- compared to an eGFR <30, ORs (95% CI) for an eGFR ≥90, 60-89, 45-59, and tion (EMT) after exposure from hyperglycemia-related NETs. 30-45, respectively, were 3.4 (2.1-5.2), 2.7 (1.9-3.8), 3.1 (2.2-4. 6), and 2.0 In conclusion, hyperglycema-induced NETs might promote diabetic kidney (1.4-3.0) for each ng/dL increase in PFHxS. Similar results were observed for fibrosis. each PFAS. PFAS may have beneficial effects on kidney function in CKD and Supported By: National Key Clinical Specialties Construction Program of China; diabetes, especially when anemia is present. National Natural Science Foundation of China (81670785, 81370954); Fundamental Supported By: National Institutes of Health (U54GM1049) Science & Advanced Technology Research of Chongqing (CSTC2015JCYJBX0096) 549‑P 547‑P Kidney Tubular Biomarkers in Type 1 Diabetes Patients with Kidney The Long-Term Effects of Intensive Lifestyle Intervention on Renal Disease in the DCCT/EDIC Study Function in Patients with Diabetes in a Real-World Clinical Prac‑ IAN H. DE BOER, XIAOYU GAO, IONUT BEBU, ANDREW HOOFNAGLE, JOHN M. tice: A Five-Year Longitudinal Study LACHIN, ANDREW D. PATERSON, BRUCE A. PERKINS, AMY SAENGER, MICHAEL AHMAD AL MARADNI, SYLVIA ROSAS, ADHAM MOTTALIB, DAVID M. POBER, W. STEFFES, BERNARD ZINMAN, MARK E. MOLITCH, DCCT/EDIC RESEARCH CATALINA MORALES, OSAMA HAMDY, Boston, MA GROUP, Seattle, WA, Washington, DC, Rockville, MD, Toronto, ON, Canada, Diabetes (DM) and obesity are known risk factors for chronic kidney dis- Minneapolis, MN, Chicago, IL ease. We investigated the long-term effects of weight loss on renal function Background: Biomarkers of tubulointerstitial function may provide insights in obese patients with DM in real-world clinical practice. into kidney damage and prognosis that are complimentary to estimated glo- We evaluated 104 patients with obesity and type 1 (13.4%) and type 2 merular filtration rate (eGFR) and albuminuria. DM (86.6%) with a mean age of 53.8±10.0 yrs and mean diabetes duration Methods: We performed a cross-sectional case-control study of 86 DCCT/ of 10.7±9.8 yrs who enrolled in the Weight Achievement and Intensive Treat- EDIC participants with type 1 diabetes. Cases were defined by incident sus- ment (Why WAIT) program, a 12-week intensive lifestyle intervention (ILI) at tained eGFR <60 ml/min/1.73m2 with urinary albumin excretion rate (AER) the Joslin Diabetes Center. At one year, we stratified participants accord- >300mg/24 h. Controls were defined by persistent eGFR> 90 ml/min/1.73m2 ing to their % weight loss into: group A, who maintained ≥7% weight loss and AER <30 mg/24 h and. We measured biomarkers reflecting tubular (49.1%) and group B, who maintained <7% weight loss (50.9%) and prospec- injury, inflammation, fibrosis, secretion, and synthetic function using stored tively followed them for 5 years to study the impact of weight maintenance biospecimens collected at the time of incident eGFR <60 ml/min/1.73m2 (for vs. weight regain on renal function. cases) or at the same study year (for controls). At 5 years, group A lost 18.2±25.4lb (7.5±10.1%, p<0.01) and group B Results: Mean age was 50.5 years and mean duration of diabetes was lost 9.3±21.9lb (3.4±8.4%, p<0.01). A1c dropped in group A (-0.27±0.95%, 30.1 years. Mean eGFR was 38.7 vs. 103.4 ml/min/1.73m2 for cases vs. con- p<0.001) and increased in group B (+0.93±1.78%, p<0.001). Systolic BP trols, respectively, and mean AER was 1980 vs. 10 mg/d. In addition, the improved in both groups but was only statistically significant in group B (from distributions of each measured renal tubular biomarker differed significantly 128.6±14.3 mmHg to 121.4±15.4 mmHg, p<0.05). Serum creatinine decreased (Table). in group A (from 0.92±0.19mg/dL to 0.85±0.23mg/dL, p<0.01) and in group B Conclusions: In type 1 diabetes, marked abnormalities in kidney tubular (from 0.92±0.21mg/dL to 0.84±0.26mg/dL, p<0.01). eGFR increased in group biomarkers accompany reduced eGFR and albuminuria. Longitudinal studies A (from 86.0±28.4mL/min/1.73 m2 to 97.7±37.7mL/min/1.73 m2, p<0.001) and are needed to determine the time course over which these biomarkers change in group B (from 84.3±24.9mL/min/1.73 m2 to 97.2±37.4 mL/min/1.73 m2, relative to eGFR and albuminuria and to determine whether these biomarkers p<0.01). Urinary microalbumin/creatinine ratio did not change from baseline are associated with kidney disease progression and complications. in both groups. All improvements in renal function remained significant after adjustment for A1c and systolic BP. Maintenance of weight loss at one year was associated with a significant improvement in A1c at 5 years. Improvements in eGFR and serum creati- nine continue long-term; irrespective of weight loss maintenance or weight regain after ILI.

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Table. Cell Contents are Mean (SD). Results: TGF-beta1 increased mRNA and protein of MCP-1 in NRK-52E cells in association with reduction in BAMBI mRNA. By contrast, metfor- Controls Cases p-value for (N=43) (N=43) difference min decreased mRNA and protein of MCP-1 without affecting cell viability. Knockdown of BAMBI increased, while overexpression of BAMBI reduced Urine 21.2 (8.7) 5.3 (2.8) <0.0001 basal level of MCP-1 and TGF-beta1-induced MCP-1. Metformin and TGF- (mcg/g Cr) beta1 synergistically enhanced the phosphorylation of AMPK. Pretreat- Urine monocyte chemoattractant 122.6 (99.7) 595.6 (859.5) <0.0001 ment with metformin blocked TGF-beta1-induced upregulation of MCP-1 protein-1 (ng/g Cr) and downregulation of BAMBI, in association with reduced basal and TGF- POSTERS

Complications Urine galectin-3 (mcg/g Cr) 52.3 (35.3) 168.0 (145.0) <0.0001 beta1-induced Phosphorylation of ERK1/2. Phosphorylation of MEK/ERK1/2 Acute and Chronic Plasma soluble tumor necrosis 1021.7 (256.4) 3694.9 (1288.9) <0.0001 was reduced in NRK-52E cells overexpressing BAMBI. Furthermore, U0126, factor receptor-1 (pg/mL) a specific inhibitor of MEK1/2, completely blocked TGF-beta1-induced MCP-1 Plasma galectin-3 (ng/mL) 11.0 (5.3) 5.6 (1.9) <0.0001 expression. Conclusion: In rat renal tubular epithelial cells, metformin inhibits basal Plasma arginine-citrulline ratio 7.7 (2.8) 5.6 (1.9) 0.0004 and TGF-beta1-induced MCP-1 expression, probably via BAMBI-mediated (mcg/mcg) suppression of MEK/ERK1/2 signaling. Urinary cinnamoylglycine 317.4 (149.5) 76.7 (70.7) <0.0001 Supported By: National Natural Science Foundation of China (81370847); Merck clearance (mL/min) Serono Diabetes Research Foundation Supported By: National Institute of Diabetes and Digestive and Kidney Diseases COMPLICATIONS—NEUROPATHY 550‑P Urinary Afamin Levels Predict Decline of Renal Function in Patients with Type 2 Diabetes Moderated Poster Discussion: Mechanisms and Potential Treatment ERI TAKAHASHI, HIROYUKI UNOKI-KUBOTA, AKINORI OKUMURA, RITSUKO for Diabetic Neuropathy and Cognitive Impairment (Posters: 552-P to 557-P), YAMAMOTO-HONDA, TOMOKO SHIGA, HIROSHI KAJIO, SHIGEO YAMASHITA, see page 21. MITSUHIKO NODA, YASUSHI KABURAGI, Tokyo, Japan, Saitama, Japan The proportion of patients with diabetic nephropathy (DN), which is the most common cause of end-stage renal disease, has progressively increased & 552‑P during the last few decades. In this study, we applied label-free quantita- Diabetic Peripheral Neuropathy as an Independent Predictor of Cardio­ tive proteomic analysis to search for novel urinary proteins predicting DN. vascular and Renal Outcomes: Post-hoc Analysis of the ALTITUDE Two-Dimensional Image-Converted Analysis of Liquid Chromatography and Trial Mass Spectrometry, 2DICAL, was used to profile urinary proteins which JELENA P. SEFEROVIC, NISH CHATURVEDI, BRIAN CLAGGETT, AKSHAY DESAI, were differentially excreted in normoalbuminuric and macroalbuminuric STEVEN HAFFNER, JOHN J.V. MCMURRAY, HANS-HENRIK PARVING, SCOTT D. patients with type 2 diabetes mellitus (T2DM) (n=6/each). A total of 2602 SOLOMON, MARC A. PFEFFER, Boston, MA, London, United Kingdom, San Antonio, peptide sequence peaks were detected, and 1198 peaks displayed signifi- TX, Glasgow, United Kingdom, Copenhagen, Denmark cant alterations in excretion between them, resulting in the identification We assessed whether the Michigan Neuropathy Screening Instrument (MNSI) is predictive of adverse outcomes in a cohort of 8606 type 2 diabetes of 96 distinct proteins. Among them, urinary levels of 9 proteins (α-1-acid mellitus (T2DM) patients and chronic kidney disease (CKD), cardiovascular glycoprotein 1, α-1-antichymotrypsin, α-1-antitrypsin, α-1B-glycoprotein, (CV) disease, or both enrolled in the Aliskiren Trial in Type 2 Diabetes Using afamin, CD44 antigen (CD44), Ig α-1 chain C region, Lysosome-associated membrane glycoprotein 2 (LAMP2), and serotransferrin) were validated to Cardiorenal Endpoints (NCT00549757). Diabetic peripheral neuropathy be differentially excreted by multiple reaction monitoring analysis using the (DPN) was classified using points from the screening MNSI questionnaire (Q) urine from normoalbuminuric (n=19), microalbuminuric (n=19), and macroal- and exam (E) in four categories: none (Q=0; E=0); mild (Q≥1 and ≤3, E<2.5); buminuric (n=15) patients with T2DM, and healthy control subjects (n=24) moderate (Q≥4, E<2.5) and severe (E≥2.5). Proportional hazards regression in the independent sample set. Moreover, urinary levels of three proteins models were used to assess the association between MNSI category and (afamin, CD44, and LAMP2) were significantly associated both with urine CV and renal outcomes (variables adjusted for shown in footnote to Table 1). albumin to creatinine ratio (ACR) and eGFR, which have not been previously At screening, 7616 (88.5%) of patients had DPN (none 11.5%, mild 45.1%, implicated in DN. We measured afamin in the urine of 204 patients with moderate 21.5%, severe 21.9%). Patients who had any category of DPN had T2DM who did not yet have significant kidney disease (ACR < 300 mg/g or longer duration of T2DM, higher BMI, HbA1c, and urine albumin to creatinine ratio, lower eGFR, more frequently used insulin and had more self-reported eGFR change rate ≤ 3.3%/year). Urinary afamin to creatinine ratio was sig- nificantly elevated in the patients who progressed to more severe DN stages retinopathy and nephropathy compared to those without DPN. All catego- ries of DPN were associated with higher risk of CV and renal outcomes, even or eGFR change rate ≥ 3.3%/year than in the patients who did not progress. Thus, the afamin to creatinine ratio may be useful to predict patients with when adjusted for multiple demographic and CV risk factors (Table 1). MNSI T2DM at risk of nephropathy before the development of microalbuminuria or is a strong predictor of adverse outcomes in patients with T2DM and CKD, reduced kidney function. CV disease, or both, seemingly independent of other known risk factors. Supported By: Japan Society for the Promotion of Science Table 1. Multivariable Modeling.

551‑P Metformin Inhibits TGF-beta1-Induced MCP-1 Expression via BAMBI-MEK/ERK1/2 Pathway in Renal Epithelial Cells SHANYING LIU, YAN LI, YING LIANG, DIEFEI LIANG, ZIJIAO SONG, Guangzhou, China Aim: Metformin has been reported of renal protective effects by inhibiting TGF-beta-induced profibrotic responses, the mechanisms remains unclari- fied. MCP-1 has been recognized as a key mediator in the progression of kidney diseases and can be induced by TGF-beta1 in renal parenchyma cells. This study aimed to investigate the effects of metformin in TGF-beta1- induced MCP-1 expression and the underlying mechanisms in a rat renal Supported By: Novartis tubular epithelial cell line NRK-52E. Methods: Cultured NRK-52E cells were challenged with TGF-beta1 and metformin alone or in combination. The mRNA of MCP-1 and BAMBI was evaluated by qPCR. The protein level of MCP-1 was measured by ELISA. BAMBI, total and phosphorylated MEK1/2, ERK1/2, and AMPK were evalu- ated by Western blot. Down- and upregulation of BAMBI were achieved by RNA interference and lentiviral-mediated overexpression of BAMB, respec- tively. Cell viability was analyzed with cck-8 reagents.

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A144 COMPLICATIONS—NEUROPATHY & 553‑P (10mg/kg/day by gavage) from onset of diabetes for 8 weeks. Nerve function Docosahexaenoic Acid Promotes Neurite Outgrowth through PI3K and structure was assessed at regular intervals throughout the study. At and JNK-Mediated Signaling Pathways in Neuro2a Cells study end, insulin had normalized blood glucose and HbA1c levels in diabetic YASUAKI TATSUMI, AYAKO KATO, NAOKO KONDO, TATSUHITO HIMENO, mice and prevented motor nerve conduction slowing, tactile allodynia, paw MASAKI KONDO, YOSHIRO KATO, HIDEKI KAMIYA, JIRO NAKAMURA, KOICHI thermal hypoalgesia, depletion of epidermal and dermal fibers, depletion of KATO, Nagoya, Japan, Nagakute, Japan corneal stromal and sub-basal nerve plexus fibers and impaired Barnes maze We previously demonstrated that the docosahexaenoic acid (DHA) performance. NSI-189 was without discernable effect on general physiol- induced the anti-oxidative enzyme, heme oxygenase-1 (HO-1) through the ogy or nerve function in control mice and did not alter the severity of sys- POSTERS nuclear factor erythroid 2-related factor 2 (Nrf2) and protected immortal- temic diabetes. NSI-189 given from onset of diabetes significantly (P<0.05) Complications ized adult mouse Schwann (IMS32) cells from oxidative stress. It has improved all indices of peripheral nerve function and structure compared to Acute and Chronic been reported that DHA has cerebral neuroprotective effects and it may vehicle-treated diabetic mice. It also significantly (P<0.05) prevented loss of contribute to the prevention and treatment of dementia. In this study, we long-term memory in the Barnes maze test. Quantification of neuronal vol- examined the effect of DHA on neurite outgrowth of murine neuroblastoma ume in the CA1, CA3 and dentate gyrus regions of the hippocampus revealed Neuro2a cells which were derived from sympathetic cells in peripheral ner- a significant (P<0.05) loss of volume in diabetic mice compared to control vous system. To evaluate the effect of DHA on neurite outgrowth, Neuro2a mice that was prevented or attenuated by insulin. NSI-189 also significantly cells were cultured with or without DHA for 72 hours and were analyzed by (P<0.05) attenuated loss of volume in the CA1 and dentate gyrus. As NSI-189 immunofluorescence staining. In the results, 2.5 μM DHA significantly pro- is currently in clinical trial for major depressive disorder, the rapid transla- moted neurite outgrowth by 1.5-fold compared with control. Furthermore, tion of this therapy for clinical investigation against diabetic neuropathy and 2.5 μM 4-hydroxy hexenal (4-HHE), an end-product of DHA peroxidation, encephalopathy is possible. significantly promoted neurite outgrowth by 2.1-fold compared with control. To examine whether DHA induces neurite outgrowth through the phos- & 556‑P phoinositide-3-kinase (PI3K) or c-Jun N-terminal kinase (JNK) pathways, we Role of Caveolin-1 in the Formation of Tau Hyperphosphoryla‑ evaluated neurite outgrowth in the presence or absence of PI3K inhibitor tion Mediated by mTOR/S6k Signalling in Hyperglycemia: A Novel LY294002 or JNK inhibitor SP600125. Neurite outgrowth promoted by DHA Molecular Mechanism of Diabetes-Induced Cognitive Dysfunction was significantly inhibited either by LY294002 or by SP600125. These results JING WU, SHANLEI ZHOU, LINHUA PI, XIAJIE SHI, LINGRAN MA, MINLI QU, XIN suggest that DHA not only protects Schwann cells from oxidative stress, but LI, DUANFANG LIAO, SHENGDAN NIE, JINJING PEI, SHAN WANG, Changsha, also promotes neurite outgrowth in neural cells via PI3K and JNK pathway China, Stockholm, Sweden and that DHA might be beneficial in the treatment of diabetic neuropathy. The abnormally hyperphosphorylated tau is thought to be implicated in diabetes-associated cognitive deficits. Caveolin-1 (Cav-1), the essential & 554‑P structure protein of caveolae, has been considered as a critical protein DLC2 Deficiency Improves Post-Injury Sciatic Nerve Regeneration in promoting growth and survival of neurons. In this study we found that in Type 1 Diabetic Mice a chronic exposure to high glucose (HG) directly decreases Cav-1 expres- RUI FENG, SOOKJA K. CHUNG, Hong Kong, China sion, increases tau phosphorylation and over-activates mTOR signalling Diabetic neuropathy is one of the main complications of diabetes and in the hippocampal neurons of the brains in diabetic rats. HG-induced tau causes significant morbidity in diabetic individuals. Previously, we have hyperphosphorylation was abolished by over-expression of Cav-1 in primary reported that mice with the deletion of a RhoGAP protein, Deleted in Liver hippocampal neurons, in contrast it is aggravated via mTOR/S6K signalling Cancer 2 (DLC2), possessed higher RhoA activity, and exhibited more severe pathwayby silencing Cav-1. Cav-1 expression was found to be independent hyperalgesia to formalin-induced noxious thermal and inflammatory pain. of mTOR signalling. Our results suggest that tau hyperphosphorylation and Furthermore, these painful responses were shown to be more exaggerated the sustained over-activated mTOR signalling under hyperglycemia maybe by the diabetic condition in DLC2-deficient (DLC2-/-) mice compared to those due to the suppression of Cav-1. Overall, the present study establishes a pro- of wild type (DLC2+/+) mice. Here, we hypothesized that the tumor suppres- tective role for Cav-1 against HG-associated tauopathies. Therefore, Cav-1 sor, DLC2, regulates axonal degenerative and regenerative ability under is a potential therapeutic target for diabetes-induced cognitive dysfunction. diabetic condition. Diabetic DLC2-/- mice showed lower blood glucose level Supported By: National Natural Science Foundation of China (81170754, with earlier onset of painful response after multiple low-dose streptozotocin 81402915, 81573480, 81470046); Key Research and Development Program of (MLDS) induced diabetes. Diabetic DLC2-/- mice showed accelerated nerve Hunan Province (2016JC2071) regeneration and functional recovery after transection of the sciatic nerve +/+ compared to the diabetic DLC2 . Moreover, significantly increased nerve & 557‑P conduction velocity was detected at 10 and 15 weeks after sciatic nerve Metformin Association with in Type 2 Diabetes transection, with higher myelinated fiber densities found at 15 and 20 weeks (T2D) Patients: Dose-Response and Diet Assessment Study -/- +/+ after transection in diabetic DLC2 mice compared to diabetic DLC2 TURKI J. ALHARBI, SR., AYLA TOURKMANI, NOUF ALHARBI, ABDULAZIZ mice. This may be the result of more effective inflammatory response in BIN RASHEED, HESHAM ALKHASHAN, OSAMA ABDELHAY, NAJEEBUDDIN -/- -/- DLC2 mice. Diabetic DLC2 mice displayed higher numbers of invading MOHAMMED, ABDULRAHMAN AL-ASMARI, ABDULAZIZ ALQAHTANI, ABDUL- macrophages in the distal post-transection nerve segments compared to RAHMAN ALRASHEED, Riyadh, Saudi Arabia those of wild type animals. Taken together, these results indicate that DLC2 Metformin is widely used in patients with T2D and can contribute to deficiency may be beneficial to nerve regeneration following transection in Vitamin B12 (B12) deficiency. This cross-sectional observational study was MLDS treated diabetic mice via an increase in the number of invading mac- conducted among 412 subjects with T2D grouped by their metformin use (93 rophages, resulting in a more efficient inflammatory response. non-metformin and 319 metformin users) at a primary care center in Saudi Supported By: Research Grants Council of Hong Kong Arabia. The objectives were to estimate the prevalence and risk of B12 defi- ciency in both group of interest. Baseline variables included age, gender, PPI & 555‑P intake, comorbid diseases, BMI, duration of T2D, metformin intake and dose, Therapeutic Efficacy of NSI-189 against Diabetic Neuropathy and and the average dietary B12 intake in each group using a Encephalopathy in Mice validated survey. Peripheral neuropathy was assessed using Toronto Clini- CORINNE G. JOLIVALT, ALEX MARQUEZ, CARLOS ANAYA, BETELHEM KIFLE, cal Neuropathy scoring System. B12 deficiency was defined < 156 pmol/l, NABEEL MUTTALIB, MICHAEL HEFFERAN, KARL JOHE, NIGEL A. CALCUTT, Homocysteine reference (4-15 umol/l). The differences between groups in La Jolla, CA, Rockville, MD the covariates were assessed using a two independent t-test, Fisher’s exact NSI-189 is a novel orally active neurogenic small molecule that penetrates test and the Chi-squared test of association. Mean age 57.8±0.6. The overall the CNS and stimulates neurogenesis, synaptogenesis and increased hip- prevalence of B12 deficiency was 7.77% (32 patients, 2 non-metformin users pocampal volume in mice. NSI-189 is currently in a Phase 2 clinical trial for and 30 metformin users). Homocysteine levels were higher in B12 deficiency major depressive disorder. The neurotrophic and neuroprotective properties group vs. the non-deficient group (93.8% vs. 32.6% respectively, P<0.01) of NSI-189 promoted us to determine efficacy of NSI-189 against indices of with mean level of 31.2 ±2.1. The overall prevalence of mild peripheral neu- neuropathy and encephalopathy in a mouse model of type 1 diabetes. Swiss ropathy was 20.7%. Adjusted multivariate logistic regression showed that Webster mice with streptozotocin-induced diabetes (90 mg/kg ip, 2 con- the duration of metformin use exceeding 4 years and a metformin dose more secutive days) were treated with insulin (sub-cutaneous pellet) or NSI-189 than 2 g increased the risk of B12 deficiency by approximately 6 and 22 folds respectively (OR = 6.35, P <0.01), (OR = 21.67, P <0.01). The best-fitted Logis-

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A145 COMPLICATIONS—NEUROPATHY

tic model showed high predictivity with AUC = 0.91 (0.86-0.96). In addition the hypothesis that sRAGE, by limiting the interaction of advanced glycation higher dietary B12 intake was positively associated with higher serum B12 end-product (AGE) with RAGE, can provide protection against levels, although the level of dietary B12 intake did not protect from B12 defi- AGE toxicity. ciency associated with metformin usage. In conclusion, our study suggests a role for a targeted screening for B12 560‑P deficiency in patients taking metformin dose more than 2 g over a period Optimal Quantitative Vibration Perception Threshold (VPT) for that exceed 4 years. Diagnosis of Diabetic Polyneuropathy (DPN)

POSTERS AHMED I.M. HANAFY, GHADA EL-KANISHY, OMAYMA SALEH, ADEL SHABANA, Complications 558‑P HALA ABD EL-HAFEZ, Mansoura, Egypt Acute and Chronic Anatomical Differences in the Association of Sudomotor Function Background: Quantitative measurement of VPT using Neurothesiometer and Cardiovascular Autonomic Function in Type 2 Diabetes Mellitus is a well-established tool to predict diabetic foot ulceration. VPT ≥ 25 volts is RAELENE E. MASER, M. JAMES LENHARD, RYAN T. POHLIG, P. BABU BALAGO- associated with 20% cumulative incidence of foot ulceration compared PAL, Newark, DE, Jacksonville, FL to 3% with VPT <15 volts. Many researchers use cut-off value of 25 volts to Autonomic nervous system (ANS) dysfunction is a frequent complication diagnose DPN. of type 2 diabetes mellitus (T2DM). Measures of ANS include heart rate Aim of the Study: To detect best cut-off value of VPT that can be used to variability (HRV) and sudomotor function. Reduced HRV is associated with diagnose DPN. increased risk of mortality. Sudomotor dysfunction, assessed by sweat Subjects and Methods: Presence of DPN was assessed in 100 adult type 2 gland function, is associated with the development of foot ulceration. Sweat diabetic patients using both nerve conduction study (NCS) and VPT. Abnor- glands are innervated by thin unmyelinated sympathetic C fibers and may be mality of one or more nerve conduction parameters in 2 or more nerves affected early in the pathological process. We evaluated 50 persons with (including sural nerve) is required to diagnose DPN. Receiver operating T2DM (age=59±11 years, 27 females). HRV was examined by RR-variation characteristic (ROC) curve for the performance of VPT in predicting NCS- during deep breathing (i.e., mean circular resultant (MCR), expiration/inspi- confirmed DPN was used to calculate sensitivity and specificity of different ration (E/I) ratio). The MCR and E/I ratio assess parasympathetic function. VPT values. Sudomotor function was measured by electrochemical skin conductance Table. Analysis of ROC Curve to Determine the Diagnostic Efficacy of VPT for (ESC) of hands and feet. Analyses of the data were performed using a step- Predicting the Diagnosis of NCS-Confirmed DPN. wise selection procedure regressing ESC hands and feet on a number of parameters (e.g., gender, race, reduced HRV, glycemic control, body mass Criterion AUC 95% CI of AUC P value Sensitivity Specificity PPV NPP (VPT equal or greater than) index). Thirteen persons had an abnormal MCR and/or E/I ratio. ESC of hands was lower for those with reduced HRV (61±20 vs. 50±19, p<0.05) after 25 volts 0.874 0.793- 0.955 <0.0001 41% 100% 100% 28% adjusting for race and gender. HbA1c was higher for those with reduced 10 volts 0.874 0.793- 0.955 <0.0001 79% 84% 96% 48% HRV but was not significant (p=0.59). Gender (p<0.05), race (p<0.001), and reduced HRV (p<0.05) were associated with reduced sudomotor function of Conclusion: VPT ≥10 volts is the best cut-off value to diagnose DPN. the hands (model R2=0.475, p<0.001). The ESC for the feet was not signifi- Although the specificity of VPT≥ 25 volts is 100%, its sensitivity is low and cantly lower for those with reduced HRV and only race was independently using this criteria can lead to underdiagnosis of DPN. associated in regression modeling. Reduced HRV appears to track with sudomotor dysfunction in the hands but not in the feet. The progression of 561‑P neuropathy is thought to occur as a distal to proximal disorder but this theory Fasting Plasma Glucose Is Associated with Development of is under challenge. Our results suggest that in T2DM dysfunction of small Alzheimer’s Disease nerve fibers occurs in the upper extremities before the lower. Reexamination OLOV ROLANDSSON, ANNA BACKESTRÖM, STURE ERIKSSON, Umeå, Sweden of the natural history of the progression of diabetic peripheral neuropathy Background: Type 2 diabetes (T2D) is associated with development of is warranted. Alzheimer’s disease (AD). We have previously found an association between fasting plasma glucose (fPG) and episodic memory, first memory to be 559‑P affected in the AD process. The aim of this nested case-control study was to Type 1 Diabetes and Hearing Loss: Audiometric Assessment assess if there was an association between fPG and AD in a population free and Measurement of Circulating Levels of Soluble Receptor for from AD and T2D at baseline. Advanced Glycation End Products Methods: We identified 187 persons (women 57%), who had developed AD MOHAMMED AL-SOFIANI, SARA MACLEOD, HUSAM GHANIM, NANCY in a longitudinal population based cohort. Controls (n=694) were matched for STECKER, HOWARD LIPPES, Baltimore, MD, Buffalo, NY sex, age, and date of sampling. At baseline, mean age was 60 (±0.4) years; Background: The impact of chronic hyperglycemia on highly vascular all participants performed an OGTT after overnight fasting. Blood pressure, organs such as kidney and retina results in DM being the leading cause of height, and weight were measured, and BMI calculated. Information on edu- and blindness among American adults. The impact of DM on cation, smoking, and physical activity was collected. Total cholesterol was a highly vascular organ such as the cochlea remains a topic of interest that analyzed on fasting blood samples. We used conditional logistic regression lacks rigorous investigation. to analyze the association between fPG and development of AD. Objectives: To study the audiometric pure-tone findings in type 1 diabetic Results: Diastolic blood pressure was lower among controls at baseline; (T1D) subjects and assess whether a significant correlation exists between no other differences were found in background variables. Cases developed hearing impairment, duration of DM, HbA1c level, and levels of serum and AD after mean time of 10.7 (±4.6) years. The association between fPG and urinary biomarkers. AD is displayed in Table 1. Methods: Participants with T1D aged 20 to 60 years old (n=30, mean age Conclusion: Fasting plasma glucose was associated with development 43 ±11.4 years) and 11 nondiabetic subjects aged 40 to 60 years old com- Alzheimer’s disease after adjustment for previously known confounders. Our pleted audiometric assessment. Hearing impairment was defined as a pure finding suggests that good metabolic control in T2D may reduce the develop- tone threshold greater than 25 dB at 0.5 -8 KHz. Levels of serum CRP, VEGF, ment of AD. and sRAGE and urinary isoprostane were measured in the diabetic subjects. Table 1. Association between Fasting Plasma Glucose and Development of Results: High frequency hearing loss was revealed in 19 diabetic subjects Alzheimer’s Disease. (63.3%). Fifteen out of the nineteen diabetics with high frequency hearing Crude odds ratio Odds ratio adjusted Odds ratio adjusted for education, BMI, loss (78.95%) had only mild hearing loss (i.e., pure tone threshold between (95% CI) for education diastolic blood pressure, and total cholesterol 25 -40 dB). Compared with diabetics with normal hearing, diabetics with (95% CI) (95% CI) high frequency hearing loss were older (48.16 vs. 36.2 years old, p=0.004) Fasting plasma glucose (mmol/l) 1.16 (1.02 - 1.32) 1.16 (1.02 - 1.33) 1.21 (1.05 - 1.40) and had a significantly longer duration of DM (30.7 vs. 21.2 years, p= 0.02), more prevalence of peripheral neuropathy (57.9 vs. 9.1%, p= 0.02), and sig- Supported By: Västerbotten County Council, Umeå, Sweden nificantly lower levels of sRAGE (1054.27 vs. 1306.83 pg/ml, p=0.03). Conclusion: Our results suggest that type 1 diabetics 60 years old and younger may show early high frequency hearing loss similar to early presby- cusis. High frequency hearing loss is positively associated with age, dura- tion of DM, and presence of peripheral neuropathy. Our results may support

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562‑P 564‑P Evidence for Abnormal Cardiac Repolarisation in Early Autonomic Diabetic Peripheral Neuropathy Is Associated with Coronary Artery Neuropathy: Time for Action? Calcification in Adults with Type 1 Diabetes: CACTI (Coronary MISBAH OLEOLO, DINESH SELVARAJAH, RAJIV GANDHI, GANESH RAO, IAIN Artery Calcification in Type 1 Diabetes) Study WILKINSON, JEFFERSON MARQUES, SOLOMON TESFAYE, Sheffield, United King- MAMTA JAISWAL, RACHEL SIPPL, RODICA POP-BUSUI, EVA FELDMAN, JANET dom, Florianópolis, Brazil SNELL-BERGEON, Ann Arbor, MI, Aurora, CO Aims: Diabetic cardiac autonomic neuropathy (CAN) is associated with an Coronary artery disease (CAD) mortality in adults with diabetic peripheral

increased risk of adverse cardiac events and sudden cardiac death. QT vari- neuropathy (DPN) exceeds the risk in those without DPN, perhaps due to POSTERS ability index (QTVI) is a measure of cardiac repolarization that is associated common risk factors and pathological processes. We explored the associa- Complications with myocardial electrical instability and arrhythmias. We therefore aimed tion between subclinical CAD as assessed by CAC (Coronary artery calcium) Acute and Chronic to study the relationships between QTVI and CAN. and DPN as assessed by Michigan Neuropathy Screening Instrument (MNSI) Methods: Using O’Brien’s tests and baroreceptor sensitivity (BRS) testing, in adults with type 1 diabetes (T1D) enrolled in the longitudinal CACTI study. 62 diabetes subjects (37 with T2DM, 38 males) were classified into three 243 subjects with T1D (45% males, 97% white) underwent a CAC scan groups: 22 with no CAN (No-CAN, 47±15 yrs), 28 subclinical CAN (53±12 yrs) using a Siemens 128-slice computed tomography scanner. DPN was defined and 12 established-CAN (57±14 yrs). We then analysed QTVI and indices of as MNSI exam score of ≥ 2. As shown in the Table, subjects with DPN heart rate variability (HF-parasympathetic and LF-sympathetic) in all sub- [n=91(37%)] were older, had a longer duration of diabetes, and had higher jects. HbA1c, systolic blood pressure and CAC scores and lower eGFR compared to Results: QTVI was significantly lower in subjects with No-CAN (-0.76±0.62) those without DPN. In multivariate logistic regression, DPN was associated compared to Subclinical-CAN (-0.11±0.56) and Established-CAN (0.28±0.86; with natural log CAC score independent of age, CAD risk factors, eGFR and ANOVA p<0.003). Parasympathetic and sympathetic activity were sig- glycemic control [OR= 1.18(1.03-1.35=) P=0.012]. An increase in prevalence of nificantly higher for No-CAN vs. Subclinical-CAN and Established-CAN, LF DPN was observed with higher CAC scores (21% prevalence among people (2.17±0.58 vs. 1.11±0.65 and 0.94±0.52 ms2, respectively; ANOVA p<0.001) with zero CAC, 35% among people with CAC score 1-100, 54% among people and HF (2.03±0.59 vs. 0.92±0.60 and 0.98±0.53 ms2; ANOVA p<0.001). with CAC score 100-400 and 68% among people with CAC score >400). There was a strong negative correlation between QTVI and sympathetic In conclusion, DPN was associated with subclinical CAD independent of (ρ=-0.844) and parasympathetic activity (ρ=-0.713; p<0.001). Moreover, CAD risk factors, glycemic control and renal function. BRS significantly (p<0.001) correlated with QTVI (-0.753), LF (0.758) and HF Table. Characteristics of Subjects with Type 1 Diabetes Stratified by Their (0.718). DPN Status. Conclusion: These results demonstrate a strong association between CAN Variable No DPN= 152 DPN= 91 P value and cardiac repolarisation abnormalities, which are recognized to increase the susceptibility to cardiac events. Alarmingly there is a clear demonstra- Age (years) 49 ±9 56 ±8 <0.0001 tion of significant abnormalities in early subclinical CAN that appears to be Duration (years) 36 ±8 40 ±9 0.0004 missed currently. The results of this study support the current ADA recom- SBP (mm Hg) 122 ±9 125 ±11 0.0077 mendation of annual CAN testing from diagnosis in T2DM and after 5 years Ln CAC score (Agatston units ) 1.87 ±2 4±3 <0.0001 in T1DM, which unfortunately is not widely implemented. Supported By: National Institutes of Health eGFR ( mL/min/1.73 m2) 87 ±18 77 ±22 <0.0001 HbA1c (%) 7.6 ±1.0 8.0 ±1.1 0.029 563‑P Serum Creatinine (mg/dl) 0.91 ±0.25 1.1 ± 0.86 0.0069 Differential Association of Cardiac Autonomic Function with Insu‑ Supported By: National Heart, Lung, and Blood Institute (R01HL61753, lin Sensitivity and Insulin Secretion in Recently Diagnosed Type 1 R01HL79611, R01HL113029) and Type 2 Diabetes DAN ZIEGLER, ALEXANDER STROM, GIDON BÖNHOF, SONJA PÜTTGEN, KÁLMÁN BÓDIS, CHRISTIAN HERDER, KARSTEN MÜSSIG, JULIA SZENDROEDI, 565‑P MICHAEL RODEN, GERMAN DIABETES STUDY GROUP, Düsseldorf, Germany It is unclear to which extent reduced insulin sensitivity and/or secre- WITHDRAWN tion contribute to the development of diabetic cardiovascular autonomic neuropathy (CAN) characterized by diminished heart rate variability (HRV). We sought to determine whether HRV indices are associated with measures of insulin sensitivity and secretion in 275 individuals with type 1 diabetes (T1D), 450 persons with type 2 diabetes (T2D) (diabe- tes duration ≤1 year), and 81 glucose-tolerant controls from the baseline cohort of the German Diabetes Study (GDS): (T1D/T2D/controls [mean±SD]: age: 36.0±11.8/52.7±10.5/47.4±13.8 years, male: 62/64/68%, HbA1c: 6.6±1.2/6.4±0.9/5.3±0.3%). Four time domain and frequency domain HRV measures each, reflecting vagal and/or sympathetic modulation over 3 h were determined during a hyperinsulinemic-euglycemic clamp. Insulin sensitivity was calculated by the M value, while fasting and glucagon- stimulated C-peptide and their ratio served as measures of insulin secre- tion. After adjustment for sex, age, BMI, and smoking, M value remained significantly lower in the groups with T1D and T2D compared to controls (8.6±3.2 and 6.1±2.6 vs. 10.8±3.9 mg/kg*min; both P<0.05). Multiple linear regression analyses showed that in the T1D group 5 out of 8 and in T2D par- ticipants 7 out of 8 HRV measures were positively associated with M value (e.g., SDNN T1D: β=0.193; T2D: β=0.218, both P<0.005; very low frequency (VLF) power: T1D: β=0.219; T2D: β=0.219, both P<0.001). The low-frequency/ high-frequency (LF/HF) power spectrum as an indicator of sympathovagal balance was positively associated with the C-peptide ratio in T1D partici- pants (β=0.221, P=0.001) and with stimulated C-peptide in those with T2D (β=0.119, P=0.016). In conclusion, in both well controlled recent-onset T1D and T2D, lower cardiac vagal activity is linked to insulin resistance, while predominant sym- pathetic tone is associated with higher insulin secretion. Supported By: German Ministry of Innovation, Science and Research of the State of North Rhine-Westphalia; German Federal Ministry of Health; German Federal Ministry of Education and Research; German Center for Diabetes Research

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566‑P This data does not support an association between large and small fiber Concurrent Diagnostic Validity of In Vivo Corneal Confocal Micros‑ impairment and the presence of abnormal BMD in longstanding T1D, but copy for the Diagnosis of Diabetic Polyneuropathy (DPN): A Pooled the relationship of such impairments with fall-risk as well as bone quality Multinational Analysis requires further study. BRUCE A. PERKINS, LEIF E. LOVBLOM, VERA BRIL, KATIE EDWARDS, NICOLA Supported By: JDRF (17-2013-312) PRITCHARD, ANTHONY RUSSELL, DANIÈLE PACAUD, KENNETH ROMANCHUK, JEAN MAH, ANDREW J.M. BOULTON, MARIA JEZIORSKA, ANDREW MAR- 568‑P

POSTERS SHALL, RONI M. SHTEIN, RODICA POP-BUSUI, EVA L. FELDMAN, STEPHEN I. Ethnic Differences in Structural and Functional Measures of Small- Complications LENTZ, MITRA TAVAKOLI, NATHAN EFRON, RAYAZ A. MALIK, Toronto, ON, Can- Fiber Function in Subjects with Type 2 Diabetes Acute and Chronic ada, Brisbane, Australia, Calgary, AB, Canada, Manchester, United Kingdom, Ann CAROLINA M. CASELLINI, HENRI PARSON, BRITTANY BINION, JOSHUA F. Arbor, MI, Exeter, United Kingdom EDWARDS, JESSICA R. WEAVER, MICHAEL BAILEY, CAROLINE LIU, LINDSEY Several small studies have suggested that corneal nerve fibre length PURCELL, IOANNIS PETROPOULOUS, RAYAZ A. MALIK, AARON I. VINIK, Norfolk, (CNFL) assessed using the non-invasive technique of corneal confocal VA, Doha, Qatar, Manchester, United Kingdom microscopy may serve as a valuable proxy measure of DPN. An international Corneal Confocal Microscopy (CCM) is a validated test for detecting pooled multicenter analysis involving 5 sites was used to establish the con- morphological alterations of small nerve fibers in the cornea. Ethnic dif- current validity and diagnostic thresholds of CNFL for DPN. ferences have been observed in functional measures of small fiber func- 1000 participants-516 with type 1 diabetes (T1D) and 484 with type 2 dia- tion (SFF), however, little is known about ethnic differences in CCM. The betes (T2D)-underwent CNFL quantification concurrently with determination aim of this study was to evaluate differences in structural and functional of DPN, based on electrophysiology-based consensus criteria. Automated measures of SFF in African American (AA) and Caucasian (C) subjects with image analysis protocols were used to quantify CNFL. Concurrent validity type 2 diabetes (T2DM). 50 subjects with T2DM were evaluated using: bilat- and diagnostic thresholds were determined from receiver operating char- eral eye CCM examination of the corneal nerve plexus using the Rostock acteristic curves and area under the curve (AUC) and were derived and cornea module of the Heidelberg Tomograph III; sudomotor function using validated in 50% random samples, stratified by diabetes type. Sensitivity Sudoscan™ that measures electrochemical skin conductance (ESC); time analyses were performed to address use of an imperfect reference standard and frequency domains of heart rate variability (HRV); skin blood flow (SBF) for DPN-definition. using continuous laser Doppler on the lower limb (LL) (32 and 44°C areas T1D and T2D sub-cohorts had a mean age of 42±19 and 62±10 years, diabe- under the curve (AUC)); and nerve conduction studies (NCS) We included 27 tes duration 21±15 and 12±9 years, and HbA1c 8.3±1.6 and 7.6±1.5%, respec- C and 23 AA subjects. Baseline group characteristics were similar barring tively. DPN prevalence was 31% in T1D and 53% in T2D. AUC for CNFL was significantly higher BMI and HDL in AAs and triglyceride levels in C. Cor- 0.77 in T1D and 0.68 in T2D (p<0.001). AUC was approximately reproduced neal nerve fiber density, branching and length were similar between groups in each validation set. In the derivation sets the optimal threshold was 12.5 (CNFD: 18.64±1.73 vs. 17.24±0.94 p=0.51; CNBD: 43.87±6.11 vs. 33.26±4.01 mm/mm2 for T1D and 12.3 mm/mm2 for T2D. In the merged cohort (N=1000), p=0.14; CNFL: 17.31±1.62 vs. 14.38±1.02 p=0.12). Conversely, AAs had worse the optimal threshold of 12.3 mm/mm2 identified cases with 67% sensitiv- sudomotor function (Feet ESC: 54.64±4.39 vs. 66.30±3.72 p=0.01), and worse ity and 66% specificity. The sensitivity analysis showed greater concurrent LL skin blood flow (32 AUC: 1887.2±247.43 vs. 3908.79±404.36 p=0.0004; validity. 44 AUC:8951.1±1931.3 vs. 14633.2±1836.5 p=0.02). HRV measures and NCS Despite use of an imperfect reference standard (based on large fiber mea- were similar between the groups. Sudomotor function and SBF showed dif- sures), we confirmed the concurrent diagnostic validity of CNFL (a small fiber ferences amongst ethnicities, contrasting with the absence of differences in test) in DPN, and validated the diagnostic thresholds, regardless of diabetes structural SFF in the cornea. This suggests functional as opposed to struc- type. These results support CNFL as a diagnostic end point in DPN and rein- tural differences may have significant consequences in the pathogenesis of force the need for longitudinal studies to establish its predictive validity. microvascular and neuropathic complications in AA. Our findings emphasize Supported By: National Institutes of Health (1DP3DK104386-01) the need for functional measures as markers of early dysfunction of micro- vascular complications. 567‑P Supported By: VeroScience Diabetic Neuropathy (DN) and Bone Mineral Density (BMD) in Patients with Long-Standing Type 1 Diabetes 569‑P OMAR N. ALHUZAIM, LEIF E. LOVBLOM, MOHAMMED A. FAROOQI, GENEVIÈVE Cardiovascular Autonomic Neuropathy and Renal Function in BOULET, ALANNA WEISMAN, JULIE A. LOVSHIN, YULIYA LYTVYN, HILLARY A. Type 1 Diabetes Participants in the PERL (Preventing Early Renal KEENAN, MICHAEL H. BRENT, NARINDER PAUL, VERA BRIL, DAVID Z.I. CHER- Loss in Type 1 Diabetes) Trial NEY, BRUCE A. PERKINS, Riyadh, Saudi Arabia, Toronto, ON, Canada, Boston, MA MAMTA JAISWAL, ERIC HENRICKS, ALESSANDRO DORIA, MICHAEL MAUER, Although inconsistent, studies have implied an association between RODICA POP-BUSUI, Ann Arbor, MI, Boston, MA, Minneapolis, MN diabetic neuropathy and low BMD in T1D, possibly related to the health of Cardiovascular autonomic neuropathy (CAN) may predict progression of afferent sensory fibers that directly innervate cortical and trabecular bone. diabetic nephropathy (DN) and more serious renal and cardiovascular out- We aimed to characterize the relationship between BMD and DN in patients comes. We evaluated the cross-sectional association between CAN and with longstanding T1D for ≥50 years using large and small-fiber measures DN in subjects with type 1 diabetes (T1D) and mild to moderate chronic of DN. kidney disease enrolled in the ongoing PERL trial which is testing the uric BMD was assessed in 75 participants with T1D using dual-energy X-ray acid lowering effect of allopurinol on kidney function decline. Measures absorptiometry; t-scores and the presence of normal BMD, osteopenia, and of DN included estimated glomerular filtration rate (eGFR), iohexol plasma osteoporosis were determined for the lumbar spine (LS) and femoral neck disappearance GFR (iGFR), and albumin excretion rates (AER). Measures of (FN). Neuropathy was assessed using clinical exam scales, functional tests CAN included resting heart rate, standard deviation of normal RR interval of large and small fibers including nerve conduction studies and heart rate (SDNN), and QTi (Bazzett formula) derived from resting ECG recordings (RR variability (HRV), and structural tests including corneal nerve fiber length interval extracted from digital ECG images by ECGSCAN software). Baseline (CNFL). The presence of neuropathy was also defined according to electro- analyses are presented for 154 T1D PERL participants with valid CAN and -based consensus criteria. Associations between BMD and neu- DN data (age 53 ± 11 years, 73% male, 82% non-Hispanic white, duration ropathy variables were assessed using linear regression. 36 ± 13 years, HbA1c 8±1%). Subjects in the lowest tertile of iGFR were older Participants had mean age 65±8 years, mean HbA1c 7.4±0.8% and (53±10 vs. 47±12 year, P = 0.0007), had longer duration of diabetes (39±13 41(55%) were female. DN by consensus criteria was present in the majority vs. 29±10 year, P = 0.001), lower diastolic blood pressure (69±11 vs. 74±7 of participants (89%). For the LS, 46(61%) participants had normal t-scores, mm Hg, P=0.047), elevated serum uric acid level (6.7±1.7 vs. 5.5±0.8 mg/dl, 23(31%) had osteopenia, and 5(7%) had osteoporosis. For the FN, 20(27%) P=0.0004) and lower log SDNN (2.5±0.7 vs. 3.0±0.8, P = 0.0054) as compared participants had normal t-scores, 47(63%) had osteopenia, and 8(11%) had to those in the upper tertile of iGFR. No differences in the heart rate and QTi osteoporosis. HBA1c was not different between LS groups, but those with were observed between the 3 groups. Lower SDNN correlated with lower osteoporosis in the FN had higher HbA1c (p<0.05). Sural nerve amplitude did iGFR (r=0.33, P <0.0001), lower eGFR (r=0.27, P = 0.0009) and higher AER not differ between groups according to both LS and FN definitions. CNFL was (r= -0.26, P=0.001). In multiple linear regression analysis there was a positive higher in those with osteoporosis in the LS, but no difference in FN groups association between log SDNN (independent variable) and iGFR (dependent was observed. A trend of abnormal HRV and presence of osteoporosis was variable) (β = 5.9, SE=1.4, R2=0.31, P=0.0039) independent of age, gender, observed (p=0.05). glycemic control, blood pressure, body mass index, and serum uric acid. T1D

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PERL participants with worse renal function presented with advanced mea- Figure. sures of CAN. Supported By: American Diabetes Association (1-16-PDF-051 to M.J.)

570‑P Relationship between Glycemic Variability and Baroreflex Sensi‑ tivity in Type 2 Diabetic Patients

DAISUKE MATSUTANI, HIROYUKI IUCHI, HIROFUMI SUZUKI, MASAYA SAKA- POSTERS MOTO, KAZUNORI UTSUNOMIYA, Tokyo, Japan Complications Many studies have shown that not only reducing the HbA1c but also Gly- Acute and Chronic cemic Variability especially accompanied with hypoglycemia is important to avoid cardiovascular disease (CVD) in type 2 diabetic patients. Recently, it is demonstrated that Baroreflex Sensitivity (BRS), which is a sensitive measure of cardiovascular autonomic function, is tended to be impaired in diabetic patients. However, the relationship between Glycemic Variabil- ity and BRS has not been elucidated yet. Here we show for the first time that there is inverse relationship between Glycemic Variability and BRS in type 2 diabetic patients. Glycemic Variability and BRS were measured in 60 hospitalized patients with type 2 diabetes patients. The BRS was measured as the slope of spontaneous changes in systolic blood pres- sure (BP) and pulse interval from 15 minutes BP and ECG recordings after 15 min supine rest on the hospitalized day. The Glycemic Variability was measured by attaching continuous glucose monitoring. Standard deviation (SD), coefficient of variation (CV) and mean amplitude of glycemic excur- sions (MAGE) were calculated using the data in 24 hours from 0:00 of the hospitalized day. Characteristics of the subjects are age 61±13 years (mean±SD), HbA1c 8.9±1.7%, body mass index (BMI) 25.5±4 kg/m2, SBP 111±12mmHg, duration diabetes 13±12 years, complicated hypertension 72% and eGFR 76.3±18ml/min/1.73m2. BRS were negatively correlated with SD, CV and MAGE (r=-0.511, p=0.003; r=-0.522, p=0.003; r=-0.582, p=0.004, respectively). Stepwise multiple regression model revealed that 572‑P Glycemic Variability (CV) were related to BRS (R-squared 0.54,β=-0.456, Oxidative Stress-Induced Neuritin Depression and Sciatic Nerve p=0.013) independently of age, SBP, BMI, CAVI. Interestingly, the number Dysfunction in Diabetic Rats of BRS was low in patients with hypoglycemia (n=7, P<0.01) also indepen- JIANBO LI, YINGDUAN ZHANG, CHUNHONG XI, YUCHENG WANG, Nanjing, dent from factors above. These findings suggest that BRS might be useful China predictive factor for CVD in type 2 diabetic patients. It’s necessary to pay Neuritin is a recently discovered neurotrophin in support of peripheral attention to BRS especially in case of increased Glycemic Variability. nerves. In this study, for the first time, we linked oxidative stress to neural neuritin reduction in association with peripheral nerve dysfunction in dia- 571‑P betic rats. Hyperglycemia was induced with streptozotocin in adult Sprague- Association and Predictive Model for Diabetic Neuropathy Using Dawley rats. A single dose of lipoic acid was intraperitoneally administrated. Retinal Image Characteristics Sciatic nerve malonaldehyde, sciatic nerve neuritin mRNA and peptide, and LEI KUANG, BENNY ZEE, JACK LEE, SHUK YUN LEUNG, SUM LEE, ERIC HUI, S/MNCVs (sensory/motor nerve conduction velocities) were measured, DAVID OWENS, REBECCA THOMAS, Hong Kong, China, Wales, United Kingdom respectively. We found that, at week 2 of the experiment, malonaldehyde Diabetic peripheral neuropathy (DPN) is one of the most common forms increased, neuritin mRNA and peptide contents decreased in sciatic nerves of diabetic microvascular complications. Though severe consequences are of saline treated-diabetic rats in contrast to normal rats (P < 0.05). How- caused by DPN, early identification is limited. Retinal vasculature analysis ever, malonaldehyde increase, along with neuritin decrease, was blocked has been proposed as a promising tool for diabetes complication assess- with lipoic acid treatment, compared to saline treatment: malonaldehyde ment. The aim of this study is to determine if retinal vascular characteristics (259.43 ± 12.91 vs. 328.75 ± 13.21 (nmol/g, P<0.01)), neuiritin mRNA (0.952 ± associate with DPN and if predictive model can be developed. A case-con- 0.023 vs. 0.774 ± 0.01 (arbitrary units, P < 0.05)) and neuiritin protein (0.541 ± trol study was designed based on 199 DPN cases and 510 controls recruited 0.012 vs. 0.46 ± 0.008 (arbitrary units, P < 0.05)), respectively. NCVs were from WSC Family Medicine Clinic of the Hong Kong Hospital Authority NT shown to be slow 2 weeks after nerve neuritin down-regulation in saline-, East Cluster. Clinical information as well as retinal vascular characteristics not lipoic acid-, treated rats in contrast to normal rats (P < 0.05). At week 6, such as fractal dimension, caliber and tortuosity were measured. Statisti- in contrast to saline-treated diabetic rats, these parameter changes were cal models to calculate the probability of DPN were built using both clinical delayed (although not completely prevented compared to normal controls) and/or retinal information. We found that previous stroke history (p=0.04), with lipoic acid treatment, including malonaldehyde (285.24 ± 13.02 vs. increased serum creatinine (p=0.008), retinal vessel tortuosity (p=0.0002) 372.11 ± 13.47 (nmol/g, P < 0.005)), neuiritin mRNA (0.892 ± 0.012 vs. 0.623± and decreased venous vessel caliber (p=0.0001) were associated with 0.010 (arbitrary units, P < 0.01)), neuiritin protein (0.485 ± 0.007 vs. 0.33 ± higher DPN probability. Model with only retinal vascular information have 0.005 (arbitrary units, P < 0.01)), SNCV (25.61 ± 4.01 vs. 20.7 ± 2.79 (m/s, a comparable performance (AUC=0.689) in predicting DPN as model using P < 0.05)) and MNCV (21.47 ± 2.68 vs. 16.26 ± 3.47 (m/s. P < 0.05)), respec- only clinical information (AUC=0.672). Adding retinal information into clinical tively. During the experiment, blood glucose levels were stable. model enhanced the model predictive ability (AUC=0.736). Further validation In conclusion, oxidative stress-induced down-regulation of neuritin will be conducted using data from the Diabetic Eyes Screening Wales (UK) expression was associated with peripheral nerve dysfunctions in diabetic program. rats. 573‑P The Mechanisms Underlying the Involvement of Spinal P2X7 Recep‑ tor in Painful Diabetic Neuropathy XU LAN, SUN HEPING, XU XIANG, CAO HONG, Wuxi, China, Shanghai, China Objective: We used the mice induced by streptozotocin (STZ), as an animal model of type 1 diabetes, to investigate the role of P2X7 receptor (P2X7R) in the occurrence and development of painful diabetic neuropathy. Methods: Male C57 mice and P2X7 receptor knockout mice (P2X7R KO), weighing 20-23g, overnight fasting for 12 h, were prepared for diabetic model by intraperitoneal injection STZ (100mg/kg). 7 days after STZ injec-

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tion, fasting blood glucose was measured and which >11.1mmol/L was 575‑P included into the diabetic group. We measured the behavioral responses to Thermal Imaging for Detection of Preclinical Diabetic Peripheral mechanical stimuli from 1 to 10 weeks after STZ injection. Western Blot was Neuropathy used to measure P2X7R expressions in lumbar spinal cord, and immunohis- MARK R. BURGE, PETER SOLIZ, ZYDEN JARRY, CHRISOPHER S. CALDER, NIRAN- tochemistry was applied to locate the P2X7R in spinal cords. At the point of CHANA MANIVANNAN, Albuquerque, NM 3 weeks after administration, it was measured the behavioral responses to This study was designed to assess the clinical value of thermal imaging mechanical stimuli after intrathecal injection of P2X7R selective antagonist for detection of diabetic peripheral neuropathy (DPN). Thermal imaging has A740003. demonstrated changes in microvascular function that correlates with nerve POSTERS

Complications Results: In both C57 mice and P2X7R KO mice, there were significantly function. We aim to show that this non-invasive method coupled with our

Acute and Chronic higher blood glucose in diabetic mice than in control mice. We found that novel thermal model is useful for detecting DPN. diabetic mice developed mechanical allodynia at early stage of diabetes. Foot pathology accounts for 25% of all hospital stays among DPN patients, During the period of mechanical allodynia, we detected increased expres- and the cost of its management in the United States is estimated at billions sions of P2X7R at the dorsal horn. The mechanical threshold increased sig- of dollars annually. The standards for identifying DPN include clinical exami- nificantly after intrathecal injection of P2X7R selective antagonist A740003. nation (monofilament and tuning fork tests), nerve conduction studies (NCS), P2X7R KO diabetic mice could still form mechanical allodynia, but the degree and quantitative sensory testing. While useful in the latter stages, these was lighter than C57 diabetic mice and the hyperalgesia lasted shorter. sensory tests lack sufficient sensitivity to detect changes during the early Conclusion: This study demonstrated that spinal cord P2X7R is involved stages of DPN. in the occurrence and development of painful diabetic neuropathy. Intra- Our thermal imaging system was tested on 137 subjects (46 nondiabetic thecal injection of P2X7R selective antagonist A740003 could significantly controls, 49 diabetes patients without known DPN, and 32 with known DPN decrease mechanical allodynia. Without of P2X7R, the degree of mechani- in age-matched groups). All subjects received comprehensive neurological cal allodynia was lighter and lasted shorter. We speculate that inhibition of examinations. Thermal videos of the feet taken before and after cooling P2X7R may become an effective therapy of PDN. using an infrared camera were subsequently analyzed using a novel combi- Supported By: WuXi AppTec (WX0302-B020507-160036-PB) nation of spatial-temporal features (Principal Component, angiosomes, and thermoregulation). Our thermal model identified known DPN patients with 574‑P a sensitivity and specificity of 97% and 95%, 92% and 63% among female Cardiac Autonomic Neuropathy Is Efficiently Diagnosed by Valsalva and male subjects respectively. Additionally, NCS were performed on 37 Maneuver Alone subjects, and these correlated well with our model (Spearman’s rho = 0.62; ERNESTO MADDALONI, ROSSELLA DEL TORO, ELVIRA FIORITI, GAIA TABACCO, 0.66 for females and 0.58 for males). Analysis of the neurological examina- GIUSEPPE DEFEUDIS, CLAUDIA CAGGIANO, SILVIA MANFRINI, PAOLO POZZILLI, tion showed that all female diabetic patients who have not been diagnosed Rome, Italy with DPN and classified as DPN by our thermal model also had examination Cardiac autonomic neuropathy (CAN) is a life-threatening diabetes com- findings consistent with DPN. plication. It often goes ignored as the recommended Ewing’s battery for CAN We conclude that detection of DPN using thermal imaging is feasible and diagnosis is time-consuming and difficult to perform. To evaluate if the num- may prove to be a clinically useful tool for use in primary care clinics, where ber of required tests could be reduced, we compared each test included in early detection of DPN is critically important and when intervention is most the Ewing’s battery and their combinations in 115 diabetic subjects with a cost effective. median age of 58 [interquartile range: 48-67] years. According to the Toronto Supported By: National Institutes of Health criteria, CAN was diagnosed in 53 subjects. After multivariable adjustments, CAN was associated with higher HbA1c and triglycerides levels, lower HDL 576‑P cholesterol and higher retinopathy rates (all p-values<0.05). No subject had Cardiac Vagal Tone, a Noninvasive Index of Parasympathetic Tone, positive orthostatic hypotension test. The age-adjusted area under the Is a Clinically Relevant Tool to Monitor Autonomic Neuropathy in receiver operating characteristic curve (ROC-AUC) of Valsalva maneuver Type 1 Diabetes Mellitus was significantly higher than deep breathing and lying to standing tests CHRISTINA BROCK, NIELS JESSEN, BIRGITTE BROCK, POUL ERIK JAKOBSEN, alone (Figure) and in combination (p<0.01). Diagnostic effectiveness did not TROELS KRARUP HANSEN, JESPER MOSGAARD RANTANEN, SAM RIAHI, differ between the full Ewing’s battery and Valsalva alone (p=0.48 for dif- YOANNA DIMITROVA, ANNA DONS JENSEN, QASIM AZIZ, ASBJØRN MOHR ference in ROC-AUC), Youden’s J statistic showing the best performance for DREWES, ADAM DONALD FARMER, Aalborg, Denmark, Aarhus, Denmark, London, CAN diagnosis at 80.5% sensitivity and 86.3% specificity with age-adjusted United Kingdom Valsalva results vs. 85.0% and 85.7% with the full battery. Objective: A novel noninvasive index of cardiac vagal tone (CVT) was In conclusion, Valsalva maneuver could be used alone to efficiently diag- compared against established parameters, hypothesizing that it would be nose CAN, allowing a fast and easy screening for a frequently unrecognized reduced in patients with type 1 diabetes. deadly diabetes complication. Research Design and Methods: Analysis included 130 resting 5-minute recordings of CVT (42 patients with and 23 without peripheral neuropathy, Figure. and 65 healthy participants). Patients with neuropathy had 24-hour analysis of heart rate variability (HRV) measured. Results: CVT was lower in those with neuropathy in comparison to those without neuropathy and healthy participants (p<0.0001). CVT correlated with the time and frequency domain parameters of HRV, which are estab- lished measures of parasympathetic function (r=0.79, p<0.0001 and r=0.72, p<0.0001 respectively). CVT was negatively associated with age (p=0.003), disease duration (p<0.0001), HbA1c (p=0.02) and cardiovascular risk score (p=0.02). Conclusions: This technique represents a novel, near patient and conve- nient method for evaluating parasympathetic function, potentially facilitat- ing earlier identification of patients who should undergo formal autonomic function testing.

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Figure. POSTERS Complications Acute and Chronic

Supported By: Novo Nordisk A/S; Empowering Industry and Research, Northern Jutland; Innovation Fund Denmark; Individuals, Disease and Society, Denmark (10-092786); Aalborg University (to C.B.); Danish Diabetes Academy (to A.D.F.); University Hospitals of North Midlands

577‑P Application of an Easy Quantitative Tactile Examination Method for Early Detection of Diabetic Neuropathy JUNICHI DANJO, SONOKO DANJO, HIDEYUKI SAWADA, YU NAKAMURA, Miki, Japan, Takamatsu, Japan Background: Diabetic neuropathy (DPN) is asymptomatic in the early phases, but can cause serious complications, such as amputation as it pro- gresses. Most DPN tests are cumbersome and qualitative. We aimed to develop an easy and quantitative method to measure the decrease in tactile sensation in DPN. 579‑P Brain Microglia Activation Is Associated with Diabetic Induced Methods: Using a novel micro-vibration actuator with shape-memory Neural Pain—Evidence from Pet Imaging Study alloy wires, we measured the tactile sensations in 66 type 2 diabetic out- SIYING LIU, HANGPING ZHENG, NA YI, LIJIN JI, SHUO ZHANG, XIUHONG LU, patients and 88 healthy controls in this cross-sectional study. Patients were CHUANTAO ZUO, YIMING LI, BIN LU, Shanghai, China evaluated for DPN using American Diabetes Association (ADA) criteria, Objective: Painful diabetic peripheral neuropathy (PDPN) is popular among Michigan Neuropathy Screening Instrument (MNSI), and Tactile Sensation diabetics. Microglia was reported as a key cell of PDPN. However, we are Threshold (TST) scores of the index and middle fingers determined using the still lack of direct evidence of the microglia activation in diabetic brain in new device. Patients were classified into probable DPN (n = 30) or non-DPN vivo. We aim to observe microglial activation in DPN rats’ brains by PET (n = 36) groups according to ADA criteria and into symptomatic (n = 25) or imaging and to compare the difference in painful and painless brains. asymptomatic (n = 41) groups using the MNSI. TST scores (range 1-30) were Methods: STZ induced diabetic SD rats were used to quantify thermal based on the micro-vibration stimulation intensity, which was controlled by and machinery hypersensitivity. [18F]DPA714 was used as a marker of central adjusting the pulse width, period, and voltage amplitude. We used propen- microglial activation in PET study. Animals were tested nerve conduction sity score weighting to compare the scores in the control group with those velocity (NCV) and then executed for immunohistochemistry (IHC) study. in each patient group. Age-matched animals were used as control. Results: The mean time for determining TST scores (~3 minutes/patient) Results: Significant decrease was observed at 4-6 week after STZ injec- did not differ significantly between groups. The TST score in each patient tion in thermal and mechanical threshold. In PET imaging (Figure 1A), microg- group was higher than that in the control group (P < 0.01). The scores for lia’s activity increased in diabetic especially in PDPN group (35.1% in cortex, right fingers were 20.0 ± 4.9, 11.6 ± 5.1, 19.4 ± 4.5, 15.6 ± 6.9, and 6.5 ± 78.9% in thalamus and 50.3% in whole brain). Iba-1 immunolabelled brain 5.7 for the DPN, non-DPN, symptomatic, asymptomatic, and control groups, sections showed the profound microglial proliferation and morphological respectively. The odds ratio; 95% confidence interval, and P value were transformation in different regions (Figure 1B). 2525; 131–48500, 0.00 in the DPN, 3.67; 0.51–26.6, 0.198 in the non-DPN, Conclusion: Microglia activation increased in PDPN rats’ brains compared 1513; 16.4–139443, 0.002 in the symptomatic, and 5.75; 1.24–26.5, 0.025 in to painless and control group. We are the first to give the direct evidence of the asymptomatic group, respectively. the existence of low-grade inflammation in the whole brain of diabetic rats Conclusions: Our novel device can be used for easy quantitative evalua- and observe microglia activation in PDPN. tion of tactile sensation in diabetic patients to enable the early detection of asymptomatic DPN. Figure 1. Supported By: Japan Society for the Promotion of Science; Merck Sharp & Dohme; Takeda Pharmaceutical Company Limited

578‑P

WITHDRAWN

Supported By: National Natural Science Foundation of China (81370884 to B.L.); Shanghai New Excellent Program (XYQ2013120 to B.L.); Fudan Zhuoxue Project (to B.L.); Pudong Municipal Commission of Health and Family Planning (PW2014D-2 to B.L.); Shanghai Science and Technology Committee Program (14411962200 to Y.L.)

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580‑P 582‑P Efficacy of Administration of ACE-Inhibition for Two Years on Dia‑ Insulin Resistance and Cardiovascular Autonomic Tone in Early betic Autonomic and Peripheral Neuropathy in Patients with Dia‑ Stages of Glucose Intolerance betes Mellitus RUMYANA DIMOVA, TSVETALINA TANKOVA, GEORGI KIRILOV, NEVENA CHA- TRIANTAFYLLOS DIDANGELOS, KONSTANTINOS TZIOMALOS, CHARALAMPOS KAROVA, GRETA GROZEVA, LILIA DAKOVSKA, Sofia, Bulgaria MARGARITIDIS, ZISIS KONTONINAS, IOANNIS STERGIOU, STEFANOS TSOTOU- Aim: The aim of the study was to assess the relation between indirect LIDIS, APOSTOLOS HATZITOLIOS, Thessaloniki, Greece markers of insulin sensitivity and insulin secretion and cardiovascular auto-

POSTERS Aim: The present open, parallel-group, placebo-controlled study was under- nomic dysfunction (CAD) in subjects at early stages of .

Complications taken to evaluate the effect of the angiotensin converting enzyme inhibitor Material and Methods: A total of 265 subjects (104 males, 161 females), Acute and Chronic 20 mg/day on definite diabetic autonomic neuropathy (DAN) and of mean age 44.7±12.4 years and mean BMI 30.5±6.2 kg/m2, at early stages peripheral neuropathy (DPN) in patients with diabetes mellitus (DM). of glucose intolerance were recruited. They were divided according to Patients and Methods: Sixty-three consecutive patients with DM (46% HOMA-IR - with HOMA-IR>2.5 and with HOMA-IR<2.5; and according to the males, 27 DMT1, mean age 52 years (range, 22 to 65 years) were studied. presence of CAD - with CAD and without CAD. Glucose tolerance was stud- Patients had definite DAN [abnormal results in 2 cardiovascular reflex tests ied during OGTT. Fasting immunoreactive insulin (IRI) was assessed using (CRTs)] and DPN. Patients with hypertension or coronary heart disease ECLIA method and HOMA-IR=(plasma glucose (mmol/l)×IRI(mIU/l))/22.5 and (based on scintigraphy) were excluded. Patients were randomized to receive HOMA-β=(20×IRI (mIU/l))/(plasma glucose (mmol/l)−3.5)% were calculated. quinapril (Group A, n = 31) or placebo (Group B, n = 32) for 2 years. The follow- CAD was assessed by ANX-3.0 (ANSAR, USA) using frequency-domain anal- ing methods for detecting DPN and DAN were used: Michigan Neuropathy ysis during standard clinical tests - deep breathing, Valsalva and standing. Screening Instrument Questionnaire and Examination (MNSIQ and MNSIE), Results: The prevalence of CAD was 15.2% in HOMA-IR>2.5 group in measurement of vibration perception threshold with biothesiometer (BIO) comparison to 9.2% in HOMA-IR<2.5 group (OR 1.67 (95% CI:0.76-3.64), and CRTs [R-R variation during deep breathing [assessed by expiration/inspi- p=0.036) and the number of abnormal tests per person was higher in sub- ration ratio (E/I), mean circular resultant (MCR) and standard deviation (SD)], jects with insulin resistance. No statistically significant difference in fasting Valsalva maneuver (Vals), 30:15 ratio and blood pressure response to stand- immunoreactive insulin levels and HOMA-β was established between the ing (orthostatic hypertension (OH))]. groups according to the presence of CAD. HOMA-IR was significantly higher Results: The following indices increased significantly in group A (base- in subjects with CAD (4.78 vs. 3.31, p=0.015). There was a trend towards line vs. final): E/I 1.11±0.06 vs. 1.23±0.12, MCR 18.1±6.2 vs. 38.7±20.5, SD elevated HOMA-IR value with the increase in the number of abnormal tests 31.1±11.9 vs. 56.6±23.0 (p for all comparisons < 0.05). We did not observe (p=0.001), and HOMA-IR was significantly higher in subjects with abnormal a significant change in all other indices: Vals 1.48±0.28 vs. 1.56±0.33, 30:15 deep breathing and standing test (p=0.010, p=0.008, respectively). 1.15±0.12 vs. 1.18±0.12, OH 16.0±11.8 vs. 10.4±6.1, MNSIQ, MNSIE and BIO Conclusion: Our results demonstrate increased prevalence of CAD in the 23±2 vs. 20±1.9. In group B, all CRT indices deteriorated significantly, except presence of insulin resistance in subjects at early stages of glucose dys- Vals, which did not change. MNSIQ, MNSIE and BIO did not change. metabolism. Probably HOMA-IR could be used as a predictive marker for the Conclusions: In the present study, DAN (mainly parasympathetic dysfunc- presence of CAD in this population. tion) improved after 2 years of treatment with quinapril. Improved autonomic balance may be of clinical importance in the long-term outcome of patients 583‑P with DM. Association of Sudomotor Function with Glucose Tolerance Status in Subjects with Normal Glucose Metabolism, Impaired Glucose 581‑P Metabolism, and Diabetes Mellitus Huperzine A Ameliorates the Impairment of Diabetes-Related Cog‑ TAE JUNG OH, SOO LIM, SEO YOUNG LEE, YE SEUL YANG, MYOUNG JIN JI, SE nitive Performance Involving Neuronal Insulin Signaling Pathway HEE MIN, JAE HYUN BAE, KYOUNG MIN KIM, JAE HOON MOON, SUNG HEE in Mice CHOI, HAK CHUL JANG, Seongnam, Republic of Korea, Seoul, Republic of Korea XIAOQIU XIAO, HONGYING WANG, XIAOPING WEI, YI HOU, XUEMEI CAO, XIAO- Sudomotor function represents small-fiber nerve function which can be QIN SHI, JIBIN LI, HAIYAN ZHANG, XICAN TANG, Chongqing, China, Shanghai, observed in the early course of diabetes mellitus. SUDOSCAN is a device to China assess the sudomotor function by measuring electrochemical skin conduc- Objective: Huperzine A (Hup A), a lycopodium alkaloid extracted from tance (ESC). In this study, we measured feet ESC in subjects with normal glu- the Chinese herb Huperzia serrata, is a potent and reversible inhibitor of cose metabolism (NGM; n = 101), impaired glucose metabolism (IGM; n = 38), acetylcholinesterase (AChE), which is one of the main drugs clinically for and diabetes mellitus (DM; n = 159). Michigan neuropathy screening instru- Alzheimer’s disease (AD). Type 2 diabetes (T2D) and AD share several com- ment (MNSI) and 10-g monofilament test were performed in the subjects mon pathophysiological features such as impairment in brain insulin signal- with DM. Sudomotor dysfunction was defined when feet ESC levels were ing, chronic inflammation, and cognitive dysfunction. Here, we explored less than 60 μS. Mean ESC levels were 71.4±11.6, 69.9±11.7, and 65.0±21.6 whether Hup A treatment can improve metabolic and cognitive dysfunctions μS in NGM, IGM, and DM subjects, respectively. The sudomotor dysfunction in high fat diet (HFD) induced obese mice or genetic ob/ob mice. was observed in 9.9% in NGM, 18.4% in IGM, and 32.1% in DM subjects Methods: Male C57 BL/6 mice were randomly divided into HFD and nor- (P = 0.008). Among the DM subjects, MNSI score was significantly higher mal diet (LFD). HFD and ob/ob mice were administrated with Hup A at doses in subjects with sudomotor dysfunction than those with normal sudomo- of 0.1 and 0.3 mg/kg/day, respectively, for three months. Body weight was tor function (3.3±2.6 vs. 0.3±0.5, P <0.001). In addition, the subjects with monitored and glucose homeostasis was determined by glucose tolerance sudomotor dysfunction had lower pressure sensation assessed by the 10-g test (GTT). After 10 weeks of Hup A treatment, mice were submitted to the monofilament test than those with normal function. Age, duration of DM, novel object recognition test to assess short- and long-term memory, and and fasting plasma glucose levels were negatively correlated with feet then Morris water maze to assess the spatial memory. Insulin and Aβ42 lev- ESC levels. In contrast, eGFR showed positive correlation with feet ESC els in brain were measured using ELISA kits. Akt, p-Akt and β-site amyloid levels. In multiple linear regression analysis, old age, low BMI, impaired precursor protein cleaving enzyme 1 (BACE1) in cortex were tested via West- renal function, and higher fasting plasma glucose levels were independently ern blotting. associated with sudomotor dysfunction. The present study demonstrates Results: Treatment of Hup A had no significant effect on body weight that sudomotor function gradually deteriorates from NGM to IGM and DM. and peripheral glucose metabolism in obese mice. Low dose of Hup A sig- Moreover, the sudomotor function was profoundly decreased in the poorly nificantly improved spatial learning, recognition and memory of obese mice. controlled, elderly DM subjects with impaired renal function. Further pro- Insulin levels and the expression of p-Akt in the cortex were significantly spective study is needed to evaluate whether sudomotor dysfunction can be increased after Hup A treatment. In addition, high dose of Hup A could sig- restored by intervention such as controlling hyperglycemia. nificantly decrease the expression of BACE1, but Hup A has almost no effect on the change of Aβ42 in cortex. Conclusion: While Hup A had no significant effects on body weight and peripheral glucose metabolism, it could enhance the activity of neuronal insulin signaling pathway, which may possibly contribute to its improvement on cognitive dysfunctions associated with T2DM. Supported By: National Natural Science Foundation of China (81270947, 81570763)

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584‑P 586‑P Cardiovascular Autonomic Dysfunction Predicts Future Hemoglo‑ Vibrotactile Sense in Children with Type 1 Diabetes bin A1c Variability in Subjects with Type 2 Diabetes ERIK ISING, LARS B. DAHLIN, HELENA ELDING LARSSON, Malmö, Sweden YEOREE YANG, EUN YOUNG LEE, JAE HYOUNG CHO, KUN HO YOON, BONG YUN Introduction: Diabetic peripheral neuropathy (DPN) is a devastating com- CHA, SEUNG HWAN LEE, KYUNG MIN KO, Seoul, Republic of Korea plication to DM, potentially leading to diabetic foot ulcers. Studies using Objective: We examined the association between cardiovascular auto- electrophysiology shows high occurrence of subclinical DPN among children, nomic neuropathy (CAN) and future hemoglobin A1c (HbA1c) variability in but it is unclear when and how to start screen children with T1D for DPN.

subjects with type 2 diabetes. Aim: To investigate whether evaluation of the vibrotactile sense, using POSTERS

Methods: Eligible subjects were the type 2 diabetes patients who con- a multi-frequency method, in the right hand and foot can detect underly- Complications ducted the cardiovascular autonomic function test (AFT) between October ing sensory neuropathy in children with T1D, and whether the DPN has any Acute and Chronic 2008 and September 2011. HbA1c measurements after the date of AFT were correlation to, for example, gender, age, duration of disease and metabolic collected. Finally, subjects with at least 4-year follow-up after AFT and had control measured as HbA1c values. ≥ 6 HbA1c recordings were selected in this analysis. Subjects were catego- Methods: Vibration perception thresholds (VPTs), resulting in curves rized by the baseline AFT score; normal, early, and severe CAN group. HbA1c and numeric values, were evaluated using a VibroSense Meter. VPTs were variability was measured as the standard deviation (SD) of serial HbA1c related to normative data obtained from healthy children, and evaluated in measurements (HbA1c-SD), the coefficient of variation of HbA1c (HbA1c-CV), relation to different characteristics. and the adjusted SD (adj-HbA1c-SD). Subjects were 8-18 years old T1D patients. Subjects that failed to produce Results: A total of 681 subjects were included (normal 294, early 314, at least one visibly evaluable curve were excluded. Z-values of >2.0 were and severe CAN 69). Compared to the normal and early group, the severe considered pathological. Subjects needed at least 3 pathological frequen- CAN group showed older age, longer duration of diabetes, higher mean of cies at the same site in order to claim the examined site as pathological. HbA1c, lower GFR, more diabetic retinopathy, and more use of insulin, and Results: 73 children (boys = 39) with mean age 13.2 [8.39-17.96] years antiplatelet agent. Also, all the HbA1c variability indices increased as the and duration of T1D of 5.9 [0.54-14.58] years met the inclusion and exclu- severity of CAN group was higher. Multivariable logistic regression analysis sion criteria. On index and little fingers, 5/73 (6.8%) and 4/73 (5.5%) children showed that severe CAN group was significantly associated with the higher respectively had pathological values. On metatarsal heads one and five, Hba1c variability after the adjusting for the multiple covariates included age, 9/73 (12.3%) children had pathological values on each site. In total 15/73 sex, BMI, diabetes duration, mean HbA1c, heart rate, GFR, and insulin use (21.0%) children had at least one pathological site and 3 (4.1%) had patho- (OR [95% CI]; higher HbA1c-SD, 2.776 [1.434-5.376]; higher HbA1c-CV, 2.066 logical values on all sites examined. Presence of pathological values on all [1.114-3.831]; higher adj-HbA1c-SD, 2.513 [1.302-4.848]). With the subgroup sites correlated to the height of the subject (p = 0.011) but not to gender, analysis, this association was more prominent with subjects who had longer duration of disease or HbA1c. duration of diabetes (>10 years), and low mean value of HbA1c (< 7%). Conclusion: Our findings suggests that DPN, reflected by impaired vibro- Conclusions: CAN is associated with a future Hba1c variability in subjects tactile sense, is present among children with T1D. Since up to 21% of the with type 2 diabetes. Cardiovascular autonomic dysfunction could have bad children showed signs of impaired vibrotactile sense, it may be important to influence on the future glycemic status interactively, making the vicious screen children with T1D for early detection of DPN. cycle. Supported By: Swedish Childhood Diabetes Foundation

585‑P 587‑P Impaired Pulmonary Function in Patients with Recent-Onset Type 2 Effects of Acute Glycemic Stress on Measures of Cardiovascular Diabetes Autonomic Neuropathy in Type 1 Diabetes MARTIN RÖHLING, DOMINIK PESTA, KALMAN BODIS, VOLKER BURKART, LYNN ANG, MAMTA JAISWAL, KARA MIZOKAMI-STOUT, CYNTHIA PLUNKETT, KLAUS STRASSBURGER, JULIA SZENDROEDI, KARSTEN MÜSSIG, MICHAEL JACOB REISS, RODICA POP-BUSUI, Ann Arbor, MI RODEN, Düsseldorf, Germany We aimed to assess the effects of acute glycemic stress on measures Reduced lung function has been associated with deterioration of glyce- of cardiovascular autonomic neuropathy (CAN), and subsequent arrhythmo- mic control and diabetes-related oxidative stress in long-standing type 2 genesis risk in type 1 diabetes (T1D) subjects with various stages of disease. diabetes. We hypothesized that patients with recent-onset type 2 diabe- Subjects with T1D with mild (N=9) and without complications (N=6), mean tes already exhibit abnormalities in pulmonary function when compared to age 39±16 years, diabetes duration 25±14 years, HbA1c 8.2±1.2%), and age- glucose-tolerant humans and that it associates with hemoglobin A1c (HbA1c) matched healthy controls (HC)(N=6) underwent sequentially euglycemic and maximum oxidative consumption (VO2max). Patients with type 2 diabetes (100±20mg/dl), hyperglycemic (300±20mg/dl) and hypoglycemic (45±10mg/ and a known diabetes duration <1 year (n=34, mean±SD; body-mass index dl) clamp studies to mimic real life scenarios. All clamps were done in the -2 -1 (BMI): 30.8±5.6 kg*m , HbA1c: 46.0±7.7 mmol*mol ) had comparable age, same day after an overnight fast in the clinical research center while holding sex, BMI and height as glucose-tolerant humans (n=26, BMI: 29.6±4.3 kg*m-2, any other medications except insulin. CAN was assessed by time [standard -1 HbA1c: 36.0±3.3 mmol*mol ). Lung function was assessed by spirometry deviation of normal RR interval (SDNN), root mean square of successive comprising forced vital capacity (FVC), forced expiratory volume in one sec- differences (RMSSD)] -and frequency-domain [low frequency (LF), high fre- ond (FEV1) and the ratio of FEV1 to FVC (FEV1/FVC). VO2max, HbA1c and smoking quency (HF) and their ratio (LF/HF)] indices of heart rate variability (HRV) status were determined. Analyses were adjusted for sex, age, BMI, height derived from continuous ECG recordings during this sequence using the and smoking status. Patients with type 2 diabetes had a lower pulmonary ANX 3.1 (ANSAR Inc. Philadelphia, PA). Indices of HRV were similar between function as assessed by FEV1 (2.58±0.72 vs. healthy humans 3.29±0.81 l) T1D with no complications and HC during all clamp conditions. However and FEV1/FVC (0.67±0.12 vs. 0.76±0.08) (both P<0.001) as well as a lower several indices of HRV were significantly different during euglycemic (LF -1 -1 2 2 VO2max (20.3±5.4 vs. 24.0±5.9 ml*min *kg , P<0.05). Among the patients, 1.9±1.9 vs.6.4 ±1.3 ms ; HF 0.7±1.2 vs.3.3±1.2ms ; SDNN 39±19 vs.79±12ms; FEV1 correlated positively with VO2max (r=0.40, P<0.05) and FEV1/FVC cor- RMSSD 16±13 vs.42±17ms, p<0.05 for all), hyperglycemic (LF 1.3±1.5 vs.5.7 2 2 related negatively with HbA1c (r=-0.49, P<0.01). Healthy humans showed no ±2.5 ms ; HF 0.5±0.7 vs.3.3±1.3ms ; SDNN 48±33 vs.92±33ms; RMSSD 14±9 2 correlations of any parameter of lung function with HbA1c or VO2max. Fur- vs.49± 31ms, p<0.05 for all), and hypoglycemic (LF 2.2±1.5 vs.6.7 ±0.1 ms ; 2 2 thermore, regression analyses using FEV1/FVC as a dependent and HbA1c as HF 0.4±0.2 vs.3.9±0.9 ms ; LF/HF 5.2±4 vs.14± 5 ms , p<0.05) clamp stud- independent variable yielded that group differences in FEV1/FVC could be ies in T1D with mild complications compared with those without complica- explained by different HbA1c levels. tions respectively. This preliminary data suggest that acute glycemic stress In conclusion, patients with recent-onset type 2 diabetes exhibit reduc- affects cardiovagal balance in T1D patients especially in those with compli- tions in some features of pulmonary function, which are at least in part due cations, which may potentially explain the link between CAN and cardiac to glucotoxicity. arrhythmias. Supported By: National Heart, Lung, and Blood Institute; National Institute of Diabetes and Digestive and Kidney Diseases

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588‑P 590‑P Vitamin D and Cerebral Metabolites in Painful Diabetic Neuropathy Diabetic Neuropathy Is Associated with Oxidative Stress and Endo‑ IAIN D. WILKINSON, PALLAI RAPPAI SHILLO, DINESH SELVARAJAH, RICHARD thelial Activation in Patients with Type 1 Diabetes A.E. EDDEN, SOLOMON TESFAYE, Sheffield, United Kingdom, Baltimore, MD JAN ŠKRHA, JR., TOMÁŠ PELCL, JAN ŠOUPAL, MARTA KALOUSOVÁ, MARTIN Objective: Lower vitamin D levels have been demonstrated in painful PRÁZNÝ, JAN ŠKRHA, Prague, Czech Republic compared to painless diabetic peripheral neuropathy (DPN). The mechanis- Aim: Oxidative stress (OxS) is believed to be an important factor in the tic basis for this remains unclear. Pre-clinical models report an association development of vascular damage in diabetes. The aim of this study was to

POSTERS between vitamin D deficiency and prominent changes in mouse behavior and evaluate the relationship between the severity of diabetic neuropathy and

Complications brain neurochemistry. The current study examined potential relationships markers of OxS and endothelial activation. Acute and Chronic between human vitamin D levels, neuropathic pain status and thalamic neu- Methods: Our study involved 53 patients with T1 diabetes (aged 37 ± 15 rometabolites on Magnetic Resonance Spectroscopy (MRS). yrs). Neuropathy was evaluated by vibration perception threshold (VPT) Methods: Forty-four patients with type 2 diabetes (T2DM) (14 Painful- measured repeatedly by Biothesiometer on both feet, and mean results >15 DPN, 15 Painless-DPN and 15 No-DPN) and 15 nondiabetic healthy volun- V were considered as impaired VPT. OxS was evaluated by malondialdehyde teers (HV) underwent detailed clinical and neurophysiological assessment (MDA) and antioxidative ascorbic acid (AA) plasma concentration. Mark- to determine their neuropathy composite score [NIS(LL)+7 and Douleur Neu- ers of endothelial activation (ICAM-1, VCAM-1, vWF, soluble receptor for ropathique 4 score (DN4)]. 25(OH)-Vitamin D was measured between May- advanced glycation endproducts /sRAGE/), routine biochemical parameters September and all subjects had seasonal sunlight exposure and daily activity and anthropometric data were measured in all patients. recorded. Spectral-edited, single-voxel, proton-MRS was used at 3T to yield Results: Impaired VPT (iVPT) was present in 10 patients (VPT 28.3 ± 11.1 thalamic glutamate/glutamine (Glx) metabolite resonance information rela- V). Significantly higher ICAM-1 and VCAM-1 were observed in iVPT group tive to that of water in each subject. as compared with normal VPT patients (ICAM-1: 266 ± 75 vs. 214 ± 48 pg/l, Results: The mean ages of the study groups were not significantly differ- p<0.01; VCAM-1: 858 ± 200 vs. 686 ± 186 pg/l, p<0.02). Similarly, significantly ent (painless-DPN=59±10; painful-DPN=60±7; no-DPN=57±7 and HV=55±10 higher MDA was observed in iVPT group (0.83 ± 0.41 vs. 0.63 ± 0.16 µmol/l, yrs; ANOVA p>0.05). Subjects with painful-DPN had the lowest vitamin D p<0.05), whereas AA was strongly reduced in these patients in comparison levels (painful DPN=35.8±17.1; painless-DPN=56.1±28.7; No-DPN=46.8±19.2 to normal VPT patients (46 ± 22 vs. 98 ± 37 µmol/l, p<0.0005). Both sRAGE and HV=58.5±27.8 nmol/l ANCOVA p<0.01). Glx levels followed a similar and vWF did not differ between groups (sRAGE: 1115 ± 398 vs. 1074 ± 388 trend (1.27±0.23, 1.37±0.14, 1.34±0.21, 1.45±0.36; ANOVA p>0.05). There ng/l, NS; vWF: 151 ± 114 vs. 138 ± 109%, NS). Significant inverse relationship was a significant correlation between vitamin D and Glx in patients with was found between VPT and AA (r=-0.48, p<0.0001), while VPT was posi- painful-DPN only (r=0.75, p<0.01). tively associated with MDA (r=0.37, p<0.01). Inverse relationship between Conclusion: This study is the first demonstration of an association MDA and sRAGE was observed (r=-0.35, p<0.01). between vitamin D deficiency and neurochemical changes in the brains of Conclusion: Higher markers of endothelial activation and OxS and lower patients with T2DM who have developed painful DPN. A larger study is antioxidative capacity in patients with impaired VPT supports the idea of needed to confirm these findings and to assess the impact of vitamin D sup- their involvement in the pathogenesis of diabetes complications. For future plementation on thalamic Glx in conjunction with neuropathy status in DPN. treatment, it should be therefore useful to target antioxidative systems to Supported By: Wellcome Trust UK delay the onset of diabetic neuropathy. Supported By: Agency for Healthcare Research of the Czech Republic (P25/ 589‑P LF1/2, 15-26705A) Corneal Confocal Microscopy Detects a Small-Fibre Neuropathy in Patients with LADA 591‑P UAZMAN ALAM, IOANNIS N. PETROPOULOS, HASSAN FADAVI, OMAR ASGHAR, The Influence of Glycemic Variability on the Peripheral Nerve Dam‑ ANDREW MARSHALL, MARIA JEZIORSKA, SHAZLI AZMI, NICOLA PRITCHARD, age in the Experimental Diabetic Rats GEORGIOS PONIRAKIS, KATIE EDWARDS, CIROUS DEHGHANI, SANGEETHA KYUNG AE LEE, YU JI KIM, HEUNG YONG JIN, HONG SUN BAEK, TAE SUN PARK, SRINIVASAN, MITRA TAVAKOLI, ANDREW J.M. BOULTON, NATHAN EFRON, Jeonju, Republic of Korea RAYAZ MALIK, Manchester, United Kingdom, Brisbane, Australia, Doha, Qatar, Chronic hyperglycemia is well known to be harmful to peripheral nerve Exeter, United Kingdom fibers. However, there is little supporting evidence to determine whether Latent Autoimmune Diabetes in Adults (LADA) is often misdiagnosed, and glycemic variability (GV) is a risk factor of diabetic peripheral neuropathy a prolonged period of poor glycemic control may increase the risk of micro- (DPN) and control of GV is beneficial for DPN. Therefore, in this study, we vascular complications. We have undertaken a detailed assessment of neu- performed the comparison of peripheral nerve damage according to differ- ropathy in control subjects (C) (n=33), subjects with type 2 DM (T2DM) (n=33) ent insulin regimen in experimental diabetes. Animals were divided into 4 and LADA (n=33) by quantifying: neuropathy disability score (NDS), pero- groups (n=6-8) according to the intervention as follows: Normal (Group 1), neal and sural nerve conduction velocity and amplitude (PMNCV, PMNAmp, diabetes (DM, Group 2), DM + once daily insulin glargine (Group 3), DM + SSNCV, SSNAmp), cold threshold (CT), warm threshold (WT), intraepidermal once daily insulin glargine with twice daily (Group 4). DM nerve fibre density (IENFD), corneal nerve fibre density (CNFD), branch den- was induced by intraperitoneal streptozotocin injection (60mg/kg). After 12 sity (CNBD) and fibre length (CNFL). weeks, diverse sensory thresholds were assessed and biochemical param- There was no significant difference in age (51.0±14.8 vs. 57.3±6.2 vs. eters and immunohistochemistry comparison were performed at 24 weeks. 52.7±12.2years, P=NS), duration of diabetes (T2DM: 15.8±8.4 vs. LADA: In the results, response threshold for von frey monofilament was lower in 11.9±9.8years, P=NS) or blood pressure. HbA1c (38.2±3.5 vs. 55.6±10.3 vs. DM group as compared with normal group and insulin treatment blunted 75.4±24.2mmol/mol, P=0.003) was higher, total cholesterol (5.1±1.0 vs. 3.8±1.0 this decrease in group 4 with statistical significance (P<0.05) that was not vs. 4.6±1.1mmol/l, P=0.001), BMI (28.8±5.7 vs. 31.0±4.7 vs. 27.1±4.0kg/m2, observed in group 3. Tail flick response was showed reversed pattern and P=0.003) and triglycerides (1.7±0.8 vs. 2.1±1.4 vs. 1.3±0.4mmol/l, P=0.001) therapeutic responses were not observed in both group 3 and group 4. Ran- were lower in LADA compared to T2DM. There were no significant dif- dall Selitto response showed improved threshold after insulin treatment in ferences in NDS (1.0±1.7 vs. 2.3±2.0 vs. 2.8±3.1, P=NS), PMNCV (46.2±6.6 both group 3 and group 4. Quantitative comparisons of peripheral nerve by vs. 43.2±8.2 vs. 41.8±6.6m/s, P=NS), PMNAmp (4.3±2.0 vs. 5.1±7.3 vs. intraepidermal nerve fiber density (IENFD) were decreased in DM group com- 3.6±2.5mV, P=NS), SSNCV (49.9±4.5 vs. 44.9±9.6 vs. 43.1±6.9m/s, P=NS), pared to normal (11.47±1.2 vs. 15.9 ±0.89, p<0.05) and IENFD was improved SSNAmp (17.7±9.6 vs. 12.2±9.9 vs. 12.1±7.2µV, P=NS), WT (37.4±3.5 vs. after insulin treatment in group 3 and group 4. However, this increased trend 40.3±8.3 vs. 41.7±4.5°C, P=NS), CT (28.0±1.7 vs. 26.8±3.2 vs. 24.0±6.4°C, did not reach to the significant difference (13.77±0.80, 14.31±0.89, p>0.05) P=NS) or IENFD (10.5±3.9 vs. 6.8±3.9 vs. 4.8±3.4no/mm, P=NS) between and did not different in between group 3 and group 4. The present results LADA and T2DM. However, CNFD (26.0±7.8 vs. 23.8±9.5 vs. 17.5±7.4no/mm2, demonstrated different insulin regimen was showed similar effects on the P=0.03), and CNBD (31.8±15.1 vs. 38.3±23.7 vs. 22.3±17.3no/mm2, P=0.003) DPN except von Frey filament test. Further studies are needed to elucidate were significantly lower with no difference in CNFL (15.3±3.8 vs. 15.0±4.8 vs. the potential role of GV on the peripheral neuropathy. 12.9±4.0mm/mm2, P=NS) between LADA and T2DM. Despite, comparable age, duration of diabetes and cardiovascular risk fac- tors, poorer glycemic control in subjects with LADA was associated with a significant small fibre neuropathy, which was detected using CCM. Supported By: JDRF (8-2008-362)

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592‑P However, the relationship between retinal neurodegeneration and diabetic neuropathy has never been investigated, so far. The aim of the present study was, therefore, to test the hypothesis that retinal neurodegeneration could WITHDRAWN be viewed as a neuropathy, affecting the retinal parenchyma, similar to peripheral diabetic neuropathy. Methods: 19 T1DM patients with no symptoms/signs of peripheral poly- neuropathy, without DR or with mild non-proliferative DR, and without peripheral neuropathy, and 14 healthy controls (C) matched for age and gen- POSTERS

der were enrolled. All subjects underwent the following electrophysiologi- Complications

cal tests: standard nerve conduction studies (NCS), incremental motor unit Acute and Chronic number estimation (MUNE) from abductor hallucis (AH), with assessment of AH average SMUP size (aSMUP), and multifocal electroretinogram measur- ing amplitude density (Amp) and implicit time (IT) of 4 macular quadrants. Results: AH MUNE was significantly decreased in T1DM (p <0.0001), and aSMUP significantly increased (p 0.007), vs. C. A positive correlation between Amp in nasal and inferior quadrant and AH MUNE (Pearson cor- relations: r 0.537, p 0.03; 0.606, p 0.01, respectively) was observed in T1DM patients. Conclusion: The finding of an association between neuroretinal dysfunc- tion and early peripheral motor unit decline, before the onset of peripheral neuropathy, suggests the hypothesis that neuroretina can be viewed as a new target for the early identification of diabetic neuropathy.

595‑P Inflammatory Markers and Progression to Kidney Dysfunction: Dif‑ ferent Assessment Windows in Type 1 Diabetic Subjects NATHANIEL L. BAKER, KELLY J. HUNT, MARIA F. LOPES-VIRELLA, DANIELLE R. STEVENS, RICHARD L. KLEIN, GABRIEL VIRELLA, THE DCCT/EDIC RESEARCH GROUP, Charleston, SC The objective of this study was to determine if biomarkers of inflammation and endothelial dysfunction predicted the development of kidney dysfunc- tion and their predictive time-frame. Biomarkers were measured at 4 time points over 28 years of DCCT/EDIC treatment and follow-up in 1396 type 1 593‑P diabetes patients. In addition to traditional inflammation biomarkers (CRP, The Comparison of Neuroprotective Effect between R-Alpha-Lipoic fibrinogen) we measured Interleukin 6 and soluble Acid and Racemic Mixture of Alpha-Lipoic Acid in the Experimental receptors-1 and 2 (sTNFR-1/2), markers of endothelial dysfunction (sICAM-1, Diabetes sVCAM-1, sE-selectin), and fibrinolysis (total and active PAI-1). Subjects had HEUNG YONG JIN, KYUNG AE LEE, YU JI KIM, HONG SUN BAEK, TAE SUN PARK, GFR > 60 and AER < 200 mg/24 h at entry and kidney dysfunction was defined Jeonju, Republic of Korea as progression to chronic kidney disease (CKD, stage 3 or worse) and mac- Alpha-Lipoic Acid (ALA) seems to delay or reverse peripheral diabetic neu- roalbuminuria (AER >e 300 mg/24 h). Models were developed to determine ropathy through its multiple antioxidant properties. Most widely available the utility of each biomarker to predict events at 3 and 7 year intervals. Mul- commercial form of ALA consists of a 50/50 mixture of the R- and S- enan- tivariate event time models indicate that increased levels of inflammation tiomers that are mirror images of each other and is called a ‘racemic’ mixture markers, endothelial dysfunction and of fibrinolysis were associated with (RS-ALA). Previous studies have been reported R-alpha-lipoic acid (R-ALA) increased risk of subsequent development of CKD stage 3 or worse (Table shows superior antioxidant effects to its racemate. Therefore, present study 1). While some biomarkers (TNFR, sE-selectin, fibrinogen) were significantly is aim to compare neuroprotective effect between R-ALA and RS-ALA. Ani- predictive of CKD over both 3 and 7 year intervals, l, PAI-1 was only predic- mals were divided into 6 groups (n=6-8) according to the intervention as fol- tive for the 3 year interval. This illustrates the need to select appropriate lows: Normal, Normal+R-ALA, Normal+RS-ALA, diabetes (DM), DM+R-ALA, biomarkers when designing clinical trials. DM+RS-ALA. DM was induced by intraperitoneal streptozotocin injection Table 1. Associations between Biomarker Measures and Time to Progres- (60mg/kg) and both R-ALA and RS-ALA were administered orally (100mg/kg). sion to Chronic Kidney Disease during Long and Short Follow-Up Window After treatment, diverse sensory thresholds were assessed and immu- Lengths. Data are Reported as Hazard Ratios and Associated 95% Confi- nohistochemistry comparison was performed at 24 weeks. In the results, dence Intervals. response thresholds for von frey monofilament and Randall-Selitto test were lower in DM group compared to normal group. Both of R-ALA and RS-ALA blunted these decrease, but there was no significant difference between R-ALA and RS-ALA. Hot plate response did not show improved threshold after R-ALA or RS-ALA treatment. Quantitative comparisons of peripheral nerve by intraepidermal nerve fiber density (IENFD) were decreased in DM group compared to control group and IENFD was improved after R-ALA or RS-ALA treatment in DM group significantly (P<0.05). However, there was no significant difference between R-ALA and RS-ALA. In conclusion, both R-ALA and RS-ALA treatment showed neuroprotec- tive effect in experimental diabetes, but the difference is not significant between the groups in this study.

594‑P Neuroretinal Dysfunction Is Associated with Early Peripheral Motor Unit Loss in Type 1 Diabetes Mellitus FABIANA PICCONI, GIORGIA MATALUNI, LUCIA ZICCARDI, ANTONIO DI RENZO, Supported By: National Institutes of Health (R01DK081352) MARIACRISTINA PARRAVANO, SARA COLUZZI, DORINA YLLI, PATRIZIO PASQUALETTI, LAURA CHIOMA, ILARIA MALANDRUCCO, DAVIDE LAURO, GIROLAMA ALESSANDRA MARFIA, SIMONA FRONTONI, Rome, Italy Aim: The predictive role of early retinal neurodegeneration in the develop- ment of clinically detectable microvascular damage is now widely accepted.

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596‑P 598‑P Cerebral Microvascular Smooth Muscle Cells Trigger Neuronal Effect of Niclosamide Ethanolamine Mild Uncoupling Treatment Dysregulation in Chronic Diabetes Mellitus by Upregulating Vascu‑ on the Development of Diabetes and Peripheral Neuropathy in the lar Adhesion Protein-1 and Methylglyoxal Production BKS-db/db Mouse KESHORE R. BIDASEE, FADHEL M. ALOMAR, XIA JIANXUN, POONAM JOSHI, RUI LUCY M. HINDER, JOHN HAYES, CAREY BACKUS, SUBRAMANIAM PENNATHUR, FANG, HUANGUI H. XIONG, JESSE L. COX, JAIPAUL SINGH, Omaha, NE, Dammam, FRANK C. BROSIUS, III, EVA L. FELDMAN, Ann Arbor, MI Saudi Arabia, New York, NY, Shandong, China, Preston, United Kingdom Aim: Hyperglycemia and hyperlipidemia contribute to diabetic peripheral

POSTERS Impairments in neuronal functions are increasingly being recognized in neuropathy (DPN) in type 2 diabetes (T2D). Niclosamide ethanolamine (NEN)

Complications individuals with diabetes mellitus (DM). These deficits negatively impact daily attenuates this diabetic phenotype in db/db mice (1). The current study Acute and Chronic living activities by compromising mental flexibility, information processing investigated the effect of NEN treatment on the development of T2D, DPN, and memory. Supra-physiologic production of the reactive dicarbonyl species and dorsal root ganglia (DRG) neuron bioenergetic dysfunction in db/db mice. methylglyoxal (MG) has emerged as a leading candidate since it promotes Methods: Male BKS-db/db and db/+ controls were fed control or NEN- hypoxia, oxidative stress, inflammation and vascular leakage/microbleeds; containing chow (AIN-93M ± 1500 ppm NEN) from 6-12 wk, and 6-24 wk. At conditions that impair neuronal functions. Why/how MG increases in brain 12/24 wk T2D was assessed via body weight, fasting blood glucose, and gly- during DM remains undefined. Here autopsied brains from chronic type 1 cated hemoglobin. At 12/24 wk DPN was assessed via sensory/motor nerve and type 2 DM patients and type 1 DM rats were used to assess expres- conduction velocities (NCVs), thermal hindpaw withdrawal latency, and sion/activities of the MG synthesizing and degradation enzymes, vascular intraepidermal nerve fiber density (IENFD). At 12 wk mitochondrial function adhesion protein-1 (VAP-1) and glyoxalase-I (Glo-I), respectively. Glo-I was was determined in primary DRG neurons using the Seahorse XF Analyzer. also increased in cerebral microvascular smooth muscle cells (cSMCs) and Results: Unexpectedly, NEN treatment did not lower body weight or neurons of DM rats to determine if it would ameliorate impairment in evoked hyperglycemia in db/db mice. Treatment did not prevent NCV deficits or excitatory postsynaptic potentials (eEPSPs) in CA1 of hippocampal slices. In IENFD loss at 12/24 wk, and induced thermal hyperalgesia in db/db mice brains from patients and rats with DM, VAP-1 expression and activity were at 12 wk. Mitochondrial oxidative metabolism was downregulated in 12 wk upregulated in cSMCs, Glo-I was reduced in cSMCs and neurons and MG db/db DRG neurons, with decreased resting ATP production (maintained cou- was increased. Targeted increase of Glo-I in cSMCs and neurons of using a pling), and decreased maximal and spare respiratory capacities. Treatment custom-designed adeno-associated virus, blunted impairments in initial slope uncoupled mitochondria in db/+ and db/db neurons, and potentiated the and paired pulse ratio of eEPSPs induced by DM. Glo-I restoration in DM rats db/db metabolic reprogramming. also attenuated VAP-1 upregulation in cSMCs, lowered brain MG, prevented Conclusions: NEN treatment did not improve T2D or DPN phenotypes in cerebral hypoxia, leakage/microbleds, and reduced oxidative stress/inflam- db/db mice, suggesting earlier intervention may be required (1). Whether mation. It also blunted impairments in attention, decision-making, motor metabolic reprogramming in db/db neurons is a compensatory mechanism to function, and orofacial and forelimb movements. From these new data we limit reactive oxygen species generation during increased substrate avail- conclude that cSMCs are triggering neuronal dysregulation by upregulating ability, or whether compromised ATP production contributes to nerve dys- VAP-1 and MG. They also suggest that restoring Glo-I in cSMCs and neurons function remain to be determined. However, the data suggest that directly could be a viable strategy to blunt neuronal dysregulation in DM. targeting neuronal mitochondria with uncoupling drugs may worsen DPN. Supported By: National Institutes of Health (P30GM103335); Gene Therapy Supported By: National Institutes of Health (DP3DK094292, R24082841); JDRF; Resource Program (1053); University of Nebraska Medical Center A. Alfred Taubman Medical Research Institute

597‑P 599‑P Development of Peripheral Neuropathy in Female C57BL6/J Mice Therapeutic Effects of Conditioned Media from Cultured Dental Fed a High-Fat Diet Pulp Stem Cells in Diabetic Polyneuropathy PHILLIPE D. O’BRIEN, JOHN M. HAYES, CAREY BACKUS, EVA L. FELDMAN, Ann KEIKO NARUSE, ERIKO MAKINO, MASAKI HATA, MAIKO OMI, NOBUHISA NAKA- Arbor, MI MURA, MEGUMI MIYABE, TOMOKAZU SAIKI, KAZUNORI SANGO, TATSUHITO Peripheral neuropathy (PN) is a common complication in patients with HIMENO, HIDEKI KAMIYA, JIRO NAKAMURA, KEN MIYAZAWA, SHIGEMI GOTO, impaired glucose tolerance and type 2 diabetes (T2D). As female mice fed a TATSUAKI MATSUBARA, Nagoya, Japan, Tokyo, Japan, Nagakute, Japan high fat diet (HFD) display a degree of protection against HFD-induced meta- We previously demonstrated that the transplantation of progenitor/stem bolic changes, we hypothesized that HFD-fed female mice would also exhibit cells improved diabetic polyneuropathy. Especially, dental pulp stem cells resistance to developing PN. Male and female C57BL6/J mice were fed (DPSCs), a kind of mesenchymal stem cells, are a favorable stem cell source, either a standard diet (10% kcal fat; Ctrl) or high fat diet (60% kcal fat; HFD) because they can be isolated from extracted teeth by orthodontic reasons from 5 wk. At 16, 24, and 36 wk, neuropathy phenotyping was performed in youth and cryopreserved until the time of need. However, despite the effi- and complemented with metabolic measures, including insulin tolerance cacy of cell therapy there are also potential risks which may limit clinical use. testing (ITT). Neuropathy phenotyping consisted of hindpaw latency to heat To conquer these limitations, we evaluated the therapeutic effects of con- stimulus, motor and sensory nerve conduction velocity (NCV), and terminal ditioned media from cultured DPSCs (DPSC-CM) in diabetic polyneuropathy. intraepidermal nerve fiber (IENF) counts. Assessment of insulin resistance DPSCs were isolated and cultured from six week-old male Sprague-Dawley through ITT demonstrated that during early HFD feeding, female HFD mice rats. DPSCs secreted abundant factors, such as angiogenic and neurotrophic exhibited relatively normal insulin responsiveness, while male HFD mice factors. DPSC-CM promoted the neurite outgrowth of dorsal root ganglion exhibited insulin resistance. Despite this finding, 16 wk female HFD mice dis- neurons and increased the viability and myelin-related protein expression of played a similar pattern of PN to that of their male counterparts, with similar Schwann cells. Next, we evaluated the effects of DPSC-CM administration fold-changes in hindpaw latency and sensory and motor NCV (Table). There- in diabetic polyneuropathy. Eight weeks after the STZ injection to SD rats, fore, although female HFD mice exhibit resistance to HFD-induced metabolic DPSC-CM were injected into the unilateral hindlimb skeletal muscles. Four changes, they display a robust PN comparable to male HFD mice suggesting weeks after DPSC-CM injection, neurophysiological measurements were that systemic insulin resistance alone does not mediate PN. assessed. Diabetic rats showed significant delay of sciatic motor/sensory Table. Phenotyping of C57BL6/J Mice at 16 Weeks. nerve conduction velocity (MNCV/SNCV) and decrease in sciatic nerve blood flow (SNBF) compared with the normal rats. DPSC-CM significantly amelio- Male Female rated MNCV/SNCV and SNBF compared with non-treatment diabetic rats. * = p < 0.05 Ctrl HFD Ctrl HFD In conclusion, DPSC-CM could be a promising tool for treatment of dia- Body weight (g) 29.18 36.95* 21.19 28.09* betic polyneuropathy. Fasting glucose (mg/dl) 238.8 304.6* 169.9 249.8* Supported By: Japan Ministry of Education, Culture, Sports, Science and Technology 16,497 28,883* 13,927 16,887 (area under curve analysis) Hindpaw latency (s) 3.43 3.46 2.56 2.66 Motor NCV (m/s) 61.70 52.16* 55.37 49.56* Sensory NCV (m/s) 22.94 18.49* 22.48 18.13* Supported By: 1DP3DK094292, 1R24082841NNF, 14SA0006

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A156 COMPLICATIONS—RETINOPATHY COMPLICATIONS—RETINOPATHY & 602‑P Microvascular Complications in Young Adult Onset Type 2 Diabetes: Independent Associations with Increased Retinopathy Moderated Poster Discussion: Diabetic Retinopathy—New Insights TIMOTHY L. MIDDLETON, MARIA I. CONSTANTINO, LYNDA MOLYNEAUX, and Directions (Posters: 600-P to 605-P), see page 21. STEPHEN M. TWIGG, TED WU, JENCIA WONG, Sydney, Australia It is now appreciated that type 2 diabetes (T2D) with onset in young adult- 600‑P hood is an aggressive condition. However, data on long term complication & outcomes in comparison to later onset T2D are few. To compare outcomes Visual Function Tests Reveal Neuroretinal Dysfunction in Early Dia‑ POSTERS by age of T2D onset we abstracted data relating to 4457 clinical encoun- Complications

betic Retinopathy Acute and Chronic KATHERINE A. JOLTIKOV, VINCIUS M. DE CASTRO, JOSE R. DAVILA, ROHIT ters from the RPAH Diabetes Centre Database (1990-2016). To account for ANAND, SAMI KHAN, NEIL FARBMAN, GREGORY R. JACKSON, CHRIS A. JOHN- increasing risk of complications by T2D duration, data were stratified into SON, THOMAS W. GARDNER, Ann Arbor, MI, Hummelstown, PA, Iowa City, IA duration bands: 10-15, 15-20 and 20-25 years. Binary logistic regression Diabetic retinopathy (DR) disrupts the entire neurovascular unit, so we analysis was performed to compare microvascular complication rates (reti- hypothesize that quantitative visual function tests may serve as effective nopathy [Ret], microalbuminuria [MA], peripheral neuropathy [PN]) in age of tools to diagnose and monitor early diabetic neuroretinal disease. Fifty-one diagnosis bands (15-29, 30-39, 40-49, 50-59, 60-69 years). Adjusted odds subjects with diabetes (24 without diabetic retinopathy [no DR], 15 with ratios (OR) after accounting for current age, gender, smoking, BMI, updated mild nonproliferative diabetic retinopathy [mild NPDR], 12 with moderate- HbA1c, systolic BP and cholesterol are presented (Figures). The odds of Ret to-severe [moderate NPDR]) and 18 healthy controls underwent refraction, after 10-15 years of T2D were greatest in those with T2D onset 15-30 years e-ETDRS visual acuity, fundus photography, and the quick contrast sensitiv- (OR 6.0, p=0.02). Similar results were seen in 15-20 and 20-25 year duration ity function (qCSF) method (Adaptive Sensory Technology). Three perimetric bands, a pattern not seen in models of MA or PN. Early onset T2D appears tests were performed; SITA standard and SITA-SWAP (Carl Zeiss Meditec) to be a more aggressive disease especially with respect to the microvas- using the 24-2 algorithm, Rarebit Perimetry (RBP) using the fovea testing cular complication of Ret even accounting for duration and traditional risk strategy, and frequency doubling perimetry (FDP) (Carl Zeiss Meditec) using factors, including HbA1c exposure. Whether factors such as greater growth the 10-2 and 24-2 algorithms. Student’s t-test and ANOVA were used for sta- hormone bioactivity in youth may account for this relationship remains to tistical analysis. All diabetic groups had reduced e-ETDRS scores and areas be determined. under the log of CSF compared to controls (p = 0.0081 and p < 0.0001, respec- Figures. tively), but there was no difference between the no DR and NPDR groups. SITA and SITA-SWAP mean deviation (MD) and foveal threshold (FT) values were reduced in subjects with moderate NPDR compared to mild NPDR, no DR, and controls (SITA-MD; p < 0.0001, FT; p = 0.0028) (SITA-SWAP-MD; p < 0.0001, FT; p = 0.0004). RBP foveal testing revealed that both NPDR groups had reduced mean hit ratios (MHR) compared to controls and no DR (p = 0.0004). FDP 24-2 showed reductions in MD and FT in mild NPDR compared to no DR (MD; p = 0.03, FT; p = 0.04), but no difference between mild and moderate NPDR. FDP 10-2 also showed reduced MD and FT (10° around the fovea) in mild NPDR compared to no DR, and additionally distinguished between mild and moderate NPDR (p < 0.0001 for both). This multimodal testing analysis reveals new insights into the progressive disruption of the neurovascular unit in dia- betes, and may lead to improved means to test novel therapies. Supported By: Research to Prevent Blindness; A. Alfred Taubman Medical Research Institute; National Eye Institute (EY20852) & 601‑P Blocking Diabetes-Induced LOX Overexpression Prevents Retinal Vascular Lesions BRIAN SONG, DONGJOON KIM, DAYEUN LEE, SAYON ROY, Boston, MA Upregulation of lysyl oxidase (LOX), a cross-linking enzyme, is linked to vas- cular basement membrane (BM) thickening and compromised BM ultrastruc- ture. However, whether it contributes to cell apoptosis and vascular leakage in diabetic retinopathy (DR) is unclear. Here we examined whether blocking diabetes-induced LOX overexpression prevents retinal vascular lesions. Four groups of Sprague-Dawley rats were used in the study: nondiabetic, diabetic, diabetic intravitreally (IV) injected with LOX siRNA, and diabetic IV injected with scrambled (scram) siRNA, as control. After 1 month of diabetes, the lat- ter 2 groups received 3 injections of LOX siRNA or scram siRNA, respectively at monthly intervals. At the end of study, eyes were enucleated, retinas iso- lated and subjected to trypsin digestion to isolate retinal capillary network. Retinal capillaries were stained with PAS and hemotoxylin, and random areas of the network imaged to assess acellular capillaries (AC) and pericyte ghosts (PG). To assess vascular leakage, extravasation of FITC-dextran was evalu- ated in retinal capillaries. Western blot (WB) analysis was performed to con- firm LOX reduction via LOX siRNA injection. WB analysis indicated IV injected LOX siRNA significantly reduced retinal LOX expression in the diabetic rats compared to that of untreated diabetic rats by 33%; p<0.05; scram siRNA had no effect. In diabetic retinas, AC and PG were significantly increased 2.3-fold and 2.6-fold, respectively compared to those of control. Importantly, diabetic rats treated with LOX siRNA exhibited significant decrease in AC and PG counts compared to those of untreated diabetic rats by 27% and 55%; p<0.05, respectively; scram siRNA had no effect. Furthermore, diabetic rats treated with LOX siRNA showed significant decrease in FITC extravasation compared to that of untreated diabetic rats (38%; p<0.05). Findings from this study suggest that diabetes-induced LOX overexpression contributes to apoptosis and vascular leakage associated with DR. Supported By: National Eye Institute (EY025528)

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& 603‑P & 605‑P Closing the Care Gap Using an End-to-End Diabetic Retinal Exam Ten-Year Follow-Up of Background Diabetic Retinopathy within a Solution in Primary Care Community-Based National Screening Programme in Wales MARK COSTLEY, SUNIL GUPTA, St. Louis, MO, Pensacola, FL REBECCA L. THOMAS, RACHEL V. NORTH, DAVID R. OWENS, Swansea, United CoxHealth serves over 24,000 patients with diabetes. In early 2016, their Kingdom, Cardiff, United Kingdom HEDIS compliance rate for diabetic retinal eye exams (DRE) was only 32%. The aim of this study was to examine the progression of diabetic retinopa- To address this care gap, CoxHealth required a solution that would be easy thy (DR) in persons with non-sight-threatening DR (NSTDR e.g., background POSTERS

Complications for physician use with standardized workflows and an automated process. DR [BDR]) over a 10 year period.

Acute and Chronic In June 2016, CoxHealth System partnered with IRIS to create an end-to- Data from persons with diabetes attending Diabetic Eye Screening Wales end solution for diabetic eye exams in primary care. The solution needed to (DESW) between 2003 and 2013 were evaluated. Survival analysis (Kaplan not only perform the highest quality images but create a workflow process Meier and Cox regression) were used to explore the progression from and systems integration from patient identification to reimbursement. In ten NSTDR to sight-threatening DR requiring referral to hospital eye services. weeks, IRIS implemented a solution in fourteen clinic locations, including A total of 1,538 people with type 1 diabetes and 17,407 with type 2 diabe- primary care and endocrinology. In the first 5 months of the program, 2,443 tes with NSTDR were included in the analysis. Of those with type 1 diabetes patients received the DRE. 400 of those patients (16.3%) were diagnosed 712 had minimum BDR and 826 had Moderate BDR and of those with type 2 with diabetic retinopathy that would otherwise have gone undiagnosed. Of diabetes 10,553 had minimal BDR and 6,854 had moderate BDR. In type 1 those patients, 26 were diagnosed with vision-threatening disease (6.5%). diabetes there was a 2 fold increased risk of progression to Sight-threaten- A total of 800 eye pathologies were identified and these patients will be ing DR in those with moderate compared to minimal BDR when adjusted for referred to an ophthalmologist for treatment they otherwise would not have age, gender and duration of diabetes (Table). Similarly there and a 2.9 fold received. By creating smart workflow processes and integrating the solu- increased risk of progression to referable DR in type 2 diabetes. tion with their electronic medical record (EMR), discrete data is integrated Extending the screening interval from 1 to 2 years in persons with type 2 directly back to the patients’ charts, allowing PCPs to see end organ dam- diabetes with minimal BDR appears justified (progression rate of 0.1% in the age and create more aggressive care plans. With early detection of disease, first 12 months). However, for persons with type 2 diabetes with moderate CoxHealth has improved patient outcomes and reduced cost-of-care. They BDR and type 1 diabetes with any severity of BDR annual screening should have impacted RAF scores, HEDIS measures and payer metrics, resulting be retained. in improved STAR ratings. Based on initial data, implementing a DRE solu- Table. Cox Regression Analysis for the Progression to Sight-Threatening DR. tion not only closed a care gap for CoxHealth patients with diabetes, but has also been found to identify end organ damage in patients with HA1c multivariate multivariate levels that were considered in-control. These early findings further support Age 0.99 (0.97, 0.996) Age 0.98 (0.98, 0.99) the importance of the DRE in allowing physicians to better manage their Gender Gender diabetic population. Male NS Male NS Female Female & 604‑P Duration of DM Duration of DM Renin-Angiotensin System (RAS) Regulates Vascular Repair in Dia‑ /=27 0.62 (0.42, 0.91) >/=9 2.15 (1.91, 2.42) YAQIAN DUAN, ELENI BELI, SERGIO LI CALZI, JUDITH QUIGLEY, TATIANA SALA- ZAR, THAO LE PHUONG TRINH, GAVIN OUDIT, MARIA B. GRANT, Indianapolis, IN, Baseline DR status Baseline DR status Edmonton, AB, Canada Min BDR 1.00 Min BDR 1.00 Mod BDR 2.08 (1.67, 2.59) Mod BDR 2.85 (2.61, 3.11) Angiotensin-converting enzyme 2 (ACE2) is the primary enzyme of the vasoprotective axis of the renin angiotensin system (RAS). Most of the Treatment of DM RAS components are present in the bone marrow, including ACE2 and its Diet 1.00 downstream peptides, Ang-(1-7) and alamandine. We tested the hypothesis OHA 2.10 (1.71, 2.56) Insulin 3.59 (2.88, 4.47) that the protective arm of the RAS axis acts to maintain homeostasis of the Other 2.84 (2.28, 3.54) diabetic bone marrow stem cell compartment and to regulate the reparative function of the bone marrow cells. Both processes could influence the devel- Supported By: Wellcome Trust UK opment of diabetic retinopathy. Diabetic ACE2 knockout (KO)/C57BL/6-Ins2 (Akita) mice were examined at 3, 6 and 9 months of diabetes and compared 606‑P to age-matched controls. Both ACE2KO-Akita and Akita cohorts showed Retinal and Choroidal Vascular Features in Patients with Diabetes reduced retinal thickness by optical coherence tomography at 9 months Imaged by OCT-Based Microangiography of diabetes (p<0.0001). Absence of ACE2 in 9-month diabetic mice led to KASRA REZAEI, QINQIN ZHANG, CHIEH LI CHEN, RUIKING WANG, Seattle, WA an accelerated increase in acellular capillaries (p=0.0185). Electroretino- Purpose: Diabetic retinopathy is the leading cause of blindness in patients gram also showed persistent deterioration of the neural retina in 9-month aged 20-64 years. Early detection of diabetic retinopathy may help to bet- ACE2KO-Akita mice (p=0.0089), with no compensatory improvement as ter control of diabetes and prevent progression of retinopathy. This study observed in Akita alone. In the ACE2KO-Akita bone marrow, increased describes the OCT based microangiography (OMAG) imaging of different numbers of short term repopulating hematopoietic stem cells (HSCs) were stage of diabetic retinopathy. observed compared to the Akita group at 3-month of diabetes, while at Methods: Patients at different stages of diabetic retinopathy, diagnosed 9-month of diabetes, the absence of ACE2 also caused a reduction of long by fundus examination and fluorescein angiography (FA), underwent imaging term repopulating HSCs in the diabetic bone marrow. Reparative function with Cirrus-5000 SD-OCT-based angiography prototype (Carl Zeiss, Med- studies showed that ACE2KO exacerbated diabetes-induced impairment of itec, USA). To visualize the retinal vascular plexus in different depth layers, lineage-c-kit+ HSC migration (p=0.0019) and proliferation (p=0.0071) as early a semi-automated retinal layer segmentation algorithm was developed to as 3-month of diabetes. HSCs from both early and late stage diabetic mice, segment the retina and choroid into different depth layers. pretreated with Ang-(1-7) or alamandine showed restored migration and pro- Results: OMAG images showed clearer microvascular maps of the ret- liferation. These data suggested that development of diabetic retinopathy ina and choroid compare with fundus images and FA. In addition, OMAG was associated with loss of the protective arm of RAS in HSCs supporting provides depth-resolved retinal microvascular networks organized within that loss of ACE2 exacerbates vascular dysfunction in diabetic mice. three physiological layers. The results also demonstrated the ability of OCT- Supported By: R01HL110170 angiography prototype system to distinguish different stages of diabetic retinopathy. Conclusions: OMAG is a non-invasive imaging modality that demonstrated capability to generate detailed retinal microvascular maps that can be valu- able in the early diagnosis and therapeutic invention of DR.

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Figure. have checked the enhanced images. Diagnosis becomes more obvious and the grading more comfortable. Another limitation for the diagnosis is that images of the same patient acquired for different examinations cannot be directly super- imposed. Indeed, the eye of the patient is never in the exact same position, the image is a projection of a 3D scene into the plane of the sensor, the optics of the camera creates a radial deformation and the colour of the image may have changed. We have developed an automatic method to superimpose eye fundus images acquired in the same position (nasal or macular). It is based on contrast POSTERS

standardisation, matching of salient points and a deformation model taking into Complications

account two radial distortions. We have performed tests for 69 patients with Acute and Chronic pairs of retinal examinations acquired in good conditions at an interval of one year with and without the same camera. A similar test has been performed on 5 patients with 20 pairs acquired in harsher conditions. A minimum of 96% of pairs were correctly superimposed. This is an important step towards the longitudinal analysis of large public health databases. 609‑P Diabetic Retinopathy among Insured Adults with Diabetes, U.S., 2010-2014 STEPHEN R. BENOIT, Atlanta, GA Diabetic retinopathy (DR) is the leading cause of blindness among work- Supported By: National Eye Institute (R01EY024158); Carl Zeiss Meditec, Inc.; ing age adults in the U.S. Despite the disability caused by DR, prevalence Research to Prevent Blindness and severity data are scarce. Using Truven Health MarketScan databases, we identified a cohort of adults 20-64 years old with diabetes (DM) (defined 607‑P as ≥2 outpatient visits or ≥1 inpatient visit with a DM code, or ≥1 DM drug Smoking, Diabetes Status, and Incident Vision Loss among Middle- prescription) who had continuous insurance coverage from 2010-2014. We Aged and Older U.S. Adults used previously defined claims definitions for DR, DM type, and eye exam SHILPA J. REGISTER, Birmingham, AL codes. Proliferative DR (PDR) was used as a marker for severity. We calcu- Smoking and diabetes are independently associated with a high risk lated the 5-year period prevalence of DR overall and by number of annual of visual impairment. However, the joint effect of these factors on visual eye exams, stratified by DM type. Among the cohort of 239,358 persons impairment is unclear, as prior studies reported mixed results. This study with DM, 5.0% had type 1. The overall prevalence of DR for type 1 and evaluates the effect of smoking and glycemic status on incident vision loss type 2 DM was 41.4% and 14.1%, respectively. Of those with DR, 42.1% with among adults from the national Reasons for Geographic and Racial Differ- type 1 and 22.9% with type 2 had PDR. The prevalence of DR and PDR was ences in Stroke study. higher among patients with more consistent annual eye exams (Figure). The Black and white adults (N=26,156), age 45 years and older completed an proportion of people with no eye exams during the study period was 19.5% in-home examination with blood sample collection and baseline question- for type 1 and 30.6% for type 2 DM. Overall, DR prevalence for type 2 DM naires (2003-07). Glycemic status was divided as diabetes (FBG>=126mg/dL; was lower than previously reported in population-based studies. However, our BG>=200mg/dL; or use of insulin or oral medications), prediabetes (FBG=100- data only included diagnosed DR for an insured cohort with continuous care. 125mg/dl; BG=140-199mg/dL), or normal glucose tolerance (FBG<100mg/dl; Patients with DR were more likely to have annual eye exams. These findings BG<140mg/dL). Participants were followed every 6 months through 2016 to highlight the utility of claims data for surveillance and the continued gap in DM assess self-reported vision loss using a validated questionnaire. Cox propor- preventive eye care. tional hazards regression was used to assess the association of smoking status with incident vision loss, stratified by glycemic status. During the follow-up period, 2,757 participants reported incident vision loss. In models adjusted for demographics and cardiovascular risk factors, the association of smoking with incident vision loss was modified by age and glycemic status. Among those age 45-59 years, non-smokers with diabetes had an increased risk of vision loss (HR=1.79, 95% CI=1.39-2.31) compared with non-smokers with normal glucose tolerance, while there was no asso- ciation for smokers with diabetes (HR=1.13, 95% CI=0.69-1.85). For those age 60-74 years, there was an increased risk for smokers with diabetes (HR=1.48, 95% CI=1.08-2.03) and non-smokers with diabetes (HR=1.45, 95% CI=1.26-1.68). In contrast, for those age 75 years and older, there was an increased risk for those with prediabetes who smoke (HR=1.61). The association of smoking with incident vision loss varying by age and glycemic status suggests that the nature of aging and the mere presence of diabetes may play an important role. Supported By: 2P60MD000502-11 608‑P Enhancing Eye Fundus Images for Diabetic Retinopathy Screening GUILLAUME NOYEL, MICHEL JOURLIN, MICHEL SMANS, REBECCA L. THOMAS, SIMON ILES, GAVIN BHAKTA, ANDREW CROWDER, DAVID R. OWENS, PETER BOYLE, Lyon, France, Saint Etienne, France, Swansea, United Kingdom, Pontypridd, United Kingdom, Cardiff, United Kingdom Many eye fundus images present strong variations of contrast which can be a limitation to the diagnosis of the retinopathy. Either some lesions are not taken into account or only a limited part of the domain of the image can be read. Graders have to manually adjust the contrast, which is tedious and not easily reproducible. We have developed an automatic system, which standardises the colour contrast across the whole domain of the image. The method is consis- tent with the physical principles or image formation and ensures that the colour aspect of lesions such as micro-aneurysms or anatomical structures such as veins are similar. It is more powerful than the existing grey level methods. We have tested our approach on several thousand images acquired in good or in harsher conditions. Some were bright while others were dark. Expert graders

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610‑P 612‑P A1C, Glycated Albumin, and Fructosamine Do Not Detect Retinal Gan‑ The Relationship between Serum Vitamin D and Diabetic Retinopa‑ glion Cell Layer Thinning in Africans: The Africans in America Study thy in Chinese Community Type 2 Diabetic Patients AKSHAR B. ABBOTT, DAVID J. VALENT, MADIA RICKS, JEAN N. UTUMATWISHIMA, JUN LI, DONGLI XU, HONG FANG, YUJIE YAN, BAODONG YAO, PU LIU, WEIJIE DAVID B. SACKS, EMILY Y. CHEW, ANNE E. SUMNER, Bethesda, MD WANG, Shanghai, China Early detection of diabetic eye disease would be beneficial. With diabetes, Background: Vitamin D has a possible pathogenetic role in diabetic com- retinal ganglion cell (RGC) layer thickness determined by Spectral Domain plications, given the role of vitamin D in retinal health, the purpose of this

POSTERS Optical Coherence Tomography (SD-OCT) is thinner than normal. It is unknown study was to evaluate the relationship between serum vitamin D and dia-

Complications whether nonfasting markers of glycemia such as A1C, glycated albumin (GA) betic retinopathy in Chinese community type 2 diabetic patients. Acute and Chronic or fructosamine are able to detect early changes in RGC layer thickness and Methods: A total of 310 type 2 diabetic patients both enrolled in the thereby yield benefit from early intervention. Data on diabetic eye disease Minhang District Bio-bank and the Minhang District Management Program in Africans is scant yet diabetes in Africa is reaching epidemic proportions. during Feb 2012 to Dec 2014 were included in this study. Of these patients, This pilot study was designed to determine whether A1C, GA or fructosamine 105 were diagnosed with diabetic retinopathy according to their fundus are associated with decreased RGC layer thickness in Africans. Therefore 85 oculi results, 205 were patients without diabetic retinopathy, considered as Africans (58% [49/85] male, mean age 42±9, [mean±SD], range 25-62y; BMI cases and controls, respectively. The patients’ general data and biochemical 28.1±4.6, range 19.9-42.4 kg/m2) living in America but born in sub-Saharan parameters were collected, detected and analyzed. Africa had OGTT with A1C determined by HPLC and GA and fructosamine by Results: The patients with diabetic retinopathy had significantly lower serum enzymatic assays. Minimum value RGC layer thickness was calculated by the 25-hydroxyvitamin D (25-OHD) levels (t = 3.465, P < 0.001) and higher prevalence Zeiss Cirrus SD-OCT segmentation algorithm. Elevated values of A1C, GA and of 25-OHD deficiency (< 20 ng/mL) (χ2 = 5.313, P = 0.021) than the controls. In fructosamine were defined as:≥ 5.7%, ≥13.94%, 235 µmol/L, resp. Based on multivariate logistic regression analysis, lower 25-OHD levels remained inde- the OGTT, the prevalence of diabetes (DM), prediabetes and normal glucose pendently associated with diabetic retinopathy (OR = 0.957, 95% CI: 0.931 tolerance was 11% (10/85), 26% (22/85) and 63% (53/85) resp. Average mini- ~ 0.984, P = 0.002), after adjustment for sex, age, age at onset of diabetes, mum RGC layer thickness was greatest in the group with prediabetes (81.52 duration of diabetes, body mass index, waist-hip ratio, systolic blood pressure, µM), less in the group with normal glucose (80.86 µM) and least in the group diastolic blood pressure, fasting plasma glucose, total cholesterol, triglyceride, with DM (78.75µM), but these differences were not significant (P =0.52). With high-density lipoprotein cholesterol and low-density lipoprotein cholesterol. The an A1C cutoff of ≥5.7%, average minimum RGC layer thickness did not differ serum 25-OHD level was weakly negatively associated with the T2DM duration above or below the cutoff (P =0.27). At GA value ≥13.94%, average minimum (r = -0.116, P = 0.042), and positively related with the LDL-C (r = 0.114, P = 0.045). RGC layer thickness did not differ above or below the cutoff (P =0.16). Similarly, Conclusions: Low serum 25-OHD level was independently associated with at a cutoff of 235 µmol/L, fructosamine did not detect a difference in average diabetic retinopathy in Chinese community type 2 diabetic patients. minimum RGC layer thickness above or below the cutoff (P=0.21). This pilot Supported By: Minhang District Scientific and Technical Committee, Shanghai study suggests that as single tests, A1C, GA and fructosamine are not associ- ated with RGC layer thinning in Africans. 613‑P Supported By: National Institutes of Health The Impact of Marital and Socioeconomic Status on the Risk of Dia‑ betic Retinopathy 611‑P YUYA TAKANO, YURIA NAMIKI, HIROTO SASAKI, HIKARU TAKAMINE, KOJI INA- Metabolic Stress and O-GlcNAc Modification Regulate the Selec‑ ZUMI, YUKO MUROHASHI, URU NEZU OSADA, Yokohama, Japan tive Sensitivity of Retinal Vascular Cells to Hyperglycemia Family support in the home and socioeconomic welfare are important for ZAFER GUREL, NADER SHEIBANI, CHRISTINE M. SORENSON, Madison, WI long-term management of diabetes. We conducted a cross-sectional study Diabetic retinopathy is a major cause of blindness in middle-aged people. It is in diabetic patients admitted to our hospital from April 2015 to October 2016. closely linked to the chronic hyperglycemia exposure during diabetes. Hypergly- We investigated patients were 1) currently single, and 2) receiving the public cemic conditions have adverse effects on many tissues and cell types, includ- welfare assistance and tested these impact on the clinical outcomes such as ing retinal vascular cells. Specifically, the early loss of retinal pericytes (PC) is HbA1c (NGSP) and diabetic retinopathy at the timing of their admission. As the induced by hyperglycemia. However, the underlying mechanisms responsible result, effective responses were obtained from 377 out of 419 patients. Among for selective sensitivity of retinal PC to hyperglycemia are undefined. Hypergly- these, 225 (60%) were male, mean age was 64.1±14.5 years, diabetes dura- cemia activates several metabolic pathways, including the hexosamine biosyn- tion was 10.5±10.0 years, BMI was 25.2±5.0 kg/m^2, and HbA1c was 9.7±2.1%. thetic pathway (HBP). The product of HBP serves as substrate for O-linked β-N- As for marital status, 65 were single (49 males, 16 females) and the remaining acetylglucosamine (O-GlcNAc) modification. O-GlcNAc modification affects a 312 were married (176 males, 136 females). Twenty three patients (16 males, wide range of proteins by altering their stability, activity and protein interac- 7 females) were receiving public welfare assistance. There were 10 single tions. We recently reported that retinal PC exhibit an increase in O-GlcNAc patients (7 males, 3 females) with receiving the public welfare assistance. modification compared with retinal endothelial cells (EC) and astrocytes under These patients were younger (age 45.4±8.0 vs. 64.6±14.3 years old, p<0.01) high glucose conditions. Furthermore, we detected a decrease in migration and their HbA1c was higher (HbA1c 11.2±2.7 vs. 9.7±2.1%, p=0.03) than the oth- and an increase in the apoptotic rate of retinal PC, not only under high glu- ers. Furthermore, the prevalence of diabetic retinopathy was higher in single cose conditions but also under treatment with O-GlcNAc modification inducing patients receiving welfare assistance (70.0% vs. 15.0%, p<0.01). In reference to agents in normal glucose conditions. Our results indicated that p53 stabilization Married not receiving welfare assistance group, the odds ratio on diabetic reti- by O-GlcNAc modification may be responsible for the selective early loss of nopathy was 1.03 (95% CI:0.53-1.97) in Single not receiving welfare assistance, PC during diabetes. Recently, we have detected differences in glucose utiliza- 3.11 (95% CI:1.01-9.58) in Married receiving welfare assistance and 6.23 (95% tion of retinal PC and EC, including higher glucose uptake, increased oxidative CI:1.57-24.7) in Single receiving welfare assistance, respectively. Multivariate metabolism, and lower citrate levels in PC. These observed effects of hypergly- analysis adjusted by sex, duration, BMI, CPR index also indicated that Single cemia on specific retinal vascular cell types has enhanced our understanding of receiving welfare assistance were independent risk factors for diabetic reti- the role glucose metabolism and O-GlcNAc modification play during diabetes. nopathy (β 1.01, SE 0.39, odds ratio 7.61 (95% CI: 1.72 -40.5)). Gaining a better understanding of these mechanisms will allow us to determine In conclusion, lack of family support and socioeconomic indigence might both cell specific targets and stages for potential intervention to prevent and/or additively increase the risk on the poor glycemic control and diabetic com- slow down the progression of diabetic retinopathy. plications. Supported By: Research to Prevent Blindness; Retina Research Foundation; National Institutes of Health (R24EY022883, P30EY016665); Environmental Protec- 614‑P tion Agency (83573701)

WITHDRAWN

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615‑P 617‑P The Association Study between Serum Total Bilirubin and Diabetic Risk Score for Diabetic Retinopathy Microvascular Complications STINE BYBERG, MARIT EIKA JØRGENSEN, MICHAEL LARSEN, HENRIK LUND- LIU YAN, PANG MENGDUAN, ZHANG RUI, GONG SIQIAN, MA YUMIN, HAN ANDERSEN, DORTE VISTISEN, Copenhagen, Denmark, Gentofte, Denmark XUEYAO, JI LINONG, Peking, China Screening for diabetic retinopathy (DR) is recommended to ensure timely Low serum total bilirubin concentration in patients with type 2 diabetes detection of sight-threatening retinal lesions. Several risk factors for DR are mellitus Complicated by Microvascular Diseases. We conducted this study known, but updated risk scores are needed.

to investigate the association between serum total bilirubin level (TBIL) We included all type 1 and type 2 diabetes patients treated at an outpa- POSTERS and microvascular complications in patients with type 2 diabetes mellitus tient clinic from 1/1/2001 to 1/1/2016. Patients with DR at baseline were Complications (T2DM). 894 hospitalized patients with T2DM were investigated. TBIL was excluded. Outcome was incident DR, graded using an adaptation of the Early Acute and Chronic compared between patients with and without microvascular complications Treatment Diabetic Retinopathy Scale (ETDRS). We included baseline values (Micro) (diabetic retinopathy (DR) and diabetic nephropathy (DN). Subjects of; HbA1c, age, eGFR, BMI, diabetes duration, diabetes type, systolic and dia- were assigned to quartiles based on serum TBIL (Quartile (Q) 1 <8.9μmol/L; stolic blood pressure, sex, total cholesterol, LDL, HDL, VLDL, smoking, exer- Q2 8.9-11.5μmol/L; Q3 11.5-13.9μmol/L; Q4 >13.9μmol/L). Logistic regression cise, alcohol consumption and systemic interventions (ACE-inhibitors, other analyses were performed to examine the effects of various factors on Micro. blood-pressure lowering drugs, lipid-lowering drugs). Using random forest The age, duration of diabetes, BMI, HbA1c, TBIL of the patients were 56.7 ± and survival tree analysis, we identified potential risk factors and interac- 12.7 years, 10 (5, 16) years, 25.5±3.6 kg/m2, 9.1±2.1%, and 11.6±3.8 μmol/l, tions to include in the final prediction model. We then fitted a Poisson model respectively. Of patients with 894 T2DM, 207 and 185 were complicated with all significant risk factors for DR using backwards selection. by DR and DN, respectively. TBIL was significantly lower in patients with We included 5712 patients, were 1649 (29%) developed DR during follow- DR (10.5±3.6 vs. 12.0±3.9 μmol/l, p<0.001, and DN (10.0±3.7 vs. 12.1±3.8 up. Mean follow-up was 3.7 years (SD 3.6 years). The survival tree indicated μmol/l, p < 0.001) than in those without complications. The prevalence of several interactions. The final Poisson model is shown in Table. Area under DR and DN was significantly lower in Q4 compared with the other three the curve was 0.67 (95% CI: 0.65-0.69). quartiles (Ptrend < 0.001). The results of the logistic regression analyses Using novel statistical methods, we developed a model of DR develop- were as follows: TBIL (OR = 0.536, 95% CI 0.331-0.869, p=0.011), HbA1c, ment in an exploratory study population using recent data. The causal rela- duration of diabetes, triglyceride and systolic blood pressure were shown to tion between blood pressure and DR should be explored further. be independent determinants of Micro. Serum total bilirubin might function Table. Rate Ratios with 95% CI for Risk Factors for Incident DR in People protectively against microvascular complications in T2DM patients. with Diabetes. Supported By: National High-Technology Research and Development Pro- gram of China (2012AA02A509); Beijing Science and Technology Committee (Z141100007414002, D131100005313008) Predictor variable RR (95% CI) p-value Antihypertensive medication used at 1.19 (1.06-1.36) 0.005 616‑P first screening visit Prevalence and Characterization of Retinopathy in Children with Type 2 DM (vs. type 1) 0.88 (0.79-0.98) 0.01 Type 1 Diabetes (T1D) Using a Non-mydriatic Fundus Camera Diabetes duration (added risk for each 1.21 (1.16-1.26) <0.001 CHELSEA ZIMMERMAN, BRITTANY BRUGGEMAN, LAURA M. JACOBSEN, additional 10 years) AMANDA LAPORTE, JANET SILVERSTEIN, Gainesville, FL Smoking status <0.001 Diabetic retinopathy (DR) is the leading cause of blindness among young Smoker at baseline (vs. non-smoker) 1.28 (1.15-1.43) adults. More than 90% of individuals with T1D will develop DR within 20 years after diagnosis, but most cases are not diagnosed until more than 5 years Ex-smoker at baseline (vs. non-smoker) 1.00 (0.73-1.38) after onset. Major risk factors for DR include time since or age at diagnosis HbA1c (10 mmol/mol) 1.07 (1.04-1.09) <0.001 and severity of hyperglycemia. Previous studies in pediatric patients with T1D Age at first visit (added risk for each 1.04 (1.00-1.07) 0.02 have reported conflicting results, with some studies showing minimal risk in additional 10 years) patients less than 15 years of age while others report DR within 3 years of diagnosis. Therefore, while the incidence of DR is high after 20 years of T1D, incidence in the pediatric T1D population is unclear. Additionally, adherence to DR screening guidelines in children remains poor. Our study utilized the Digi- 618‑P tal Retinography System (DRS), a non-mydriatic fundus camera, to assess the prevalence of DR in children with T1D, evaluate risk factors, and test the DRS WITHDRAWN as a screening tool with hopes of improving compliance, as it takes 1 minute to perform in the office. Of 317 T1D subjects, age 9-26 years (average 14.8 years, SD±3.7), 10 cases of DR were identified (2 background, 7 mild and 1 moderate non-proliferative DR). Our cohort, which includes subjects from clinic, diabetes camp and Children with Diabetes conference, had a DR prevalence of 3.2%, at an average age of 20.2 years, the youngest 17.1 years old. In subjects with DR, the average duration of T1D was 12 years (SD±4.6 years, range 6.2-20.0), and 3-year average HbA1c (where available) was 9.3% (range 6.7-14.0%) com- pared to 8.7% in the non-DR group. DKA at diagnosis was reported in 67% of subjects with DR and 47% without. Lastly, ours and other studies support fundal photography as an effective screening tool, and in-clinic imaging has the potential to decrease barriers to screening. In summary, our large cohort of young people shows older adolescents and young adults (> 17 years old) with longer T1D duration (>6 years) are at risk for DR development in contrast to earlier guidelines.

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619‑P 621‑P Prevalence of Diabetes Eye Disease and Its Correlation with Renal Findings from a Pilot Diabetic Retinopathy Screening Service in the Function in a Chain of Community Clinics in India South West Regional Health Authority in Trinidad and Tobago: A KALPANA DASH, SUNIL KUMAR CH, AFTAB AHMED, SAMBIT DAS, BALAJI Basis for a National Programme JAGANMOHAN, SUREKHA TIPPISETTY, VAMSI KRISHNA KOLUKULA, KRISHNA NALISHA MONROE, REBECCA L. THOMAS, VISHWAMITRA RAMDATH, RONNIE G. SESHADRI, APOLLO SUGAR RESEARCH GROUP, Raipur, India, Hyderabad, India, BHOLA, DAVID R. OWENS, San Fernando, Trinidad and Tobago, Swansea, United Bhubaneswar, India, Bangalore, India, Chennai, India Kingdom

POSTERS Background: Diabetic Eye Disease (DED), a complication of type 2 diabetes A pilot diabetic retinopathy (DR) screening project was initiated as part of

Complications Mellitus (T2D) frequently coexists with renal disease. The purpose was to the Johns Hopkins International (JHI) Trinidad and Tobago Health Sciences Acute and Chronic determine the prevalence of DED and its association with glomerular filtra- Initiative project, a 5-year plan which included a co-ordinated diabetes care tion rate (GFR) in a community clinic population across sugar clinics in India. programme, to reduce the increasing burden of the disease and its related Methods: Apollo Sugar Clinics has a total of 45 clinics with over 100 complications. Approximately ~180,000 people have diabetes in Trinidad health care providers, and 36500 diabetes patients with 21000 EMR pen- (14.5%). etrations in 6 months, and over 6000 fundus photographs. Total 1547 (935 A register of people with diabetes in the SWRHA area has been estab- males and 612 females) T2D patient’s EMR were retrospectively analyzed. lished since 2015 which included approximately 25,000 people with diabe- Patients were grouped into GFR ≤60 and GFR >60 mg/ml/1.73m2. Statistical tes. Between 2013 and 2016 a total of 7 sites were used for screening with a analysis was done using SPSS version 20, with significance p≤ 0.05. team of 4 screeners conducting screening using Canon CR2 cameras. Visual Results: In 1547 T2D patients, 204 (13.1%) had DED with mean (SD) age acuity was measured using Snellen charts and 2X45 degree digital retinal 56.7 (10.0) years; 122 (59.8%) males and 82 (40.2%) females. Among 1547 images were captured per eye following dilation. Images were transferred patients we could calculate GFR in 443 patients. The mean GFR was 91.2 to a central grading office for primary grading. Secondary and tertiary grad- mg/ml/1.73 m2, GFR >60 and ≤60 was observed in 388 (87.5%) and 55 ing was performed by ophthalmologists. (12.5%), respectively. DED was observed in 79 (17.8%) patients, mean (SD) Over a 3 year period a total of 6,143 people were offered appointments age 55.5 (13.8) years; among them 49 (62%) males and 30 (38%) females. GFR with 5,591 attending screening (91% attendance rate). The majority had was significantly lower in patients with DED compared to patients without type 2 diabetes (97.3%) with 58.8% having diabetes for less than 10 years. DED (79.4 vs. 93.8 mg/ml/1.73m2; p=0.000). Prevalence of DED in patients 29.3% of those screened required referral to ophthalmology with 9.5% hav- with GFR ≤60 was higher compared to patients with GFR >60 (38% vs. 15%; ing sight-threatening eye diseases. DR accounted for 47.1% of hospital refer- p=0.000). However, the overall analysis didn’t show any gender variation. rals, 3.3% due to DR +/- other lesions and 49.6% due to other lesions only Conclusion: This is one of the few studies in India that reported the preva- (cataract and glaucoma were the main causes). lence of DED from a community based setting based on fundus photographs Currently the pilot programme has screened ~20% of diabetic subjects and correlating them with GFR, thus diabetes kidney disease (DKD). DED has in SWRHA over a 3 year period. Based on a 30% referral rate extrapolation clinical significance while evaluating the DKD in a community setup. The to the remaining population in this region means that an additional ~6000 correlation of DED with low GFR (OR of 3.5) can suggest presence of DKD people could require referral to ophthalmology with ~10% deemed urgent. in a community, thus, reaffirming the significance of DED screening in com- Further extrapolation to the anticipated population with diabetes in Trinidad munity diabetes practices. Implementing these findings into clinical decision and Tobago then a National DR screening programme could expect to detect pathways will improve quality of health care delivery. 18,000 with sight-threatening eye diseases and an equal number with non- DR lesions requiring treatment. 620‑P Elevated Heart Rate Predicts Incident Retinopathy in Adolescents 622‑P with Type 1 Diabetes Undiagnosed Musculoskeletal Complication in Type 2 Diabetes: VALLIMAYIL VELAYUTHAM, MARIA CRAIG, PAUL BENITEZ-AGUIRRE, YOON HI Diabetic Cheiroarthropathy and Its Association with Diabetic Reti‑ CHO, GERALD LIEW, KIM C. DONAGHUE, Westmead, Australia, Sydney, Australia nopathy Objective: Cardiac autonomic dysfunction is a frequent complication of PRAVAT THATOI, MANOJ PARIDA, BIDYUT DAS, Cuttack, India diabetes mellitus, which may occur before or along with other microvas- Introduction: Diabetic cheiroarthropathy is one of the musculoskeletal cular complications. We hypothesise that cardiac autonomic dysfunction complications of diabetes mellitus manifested in hands and also associated predates retinopathy in adolescents with type 1 diabetes mellitus. with other diabetic complications e.g., diabetic retinopathy. In diabetic chei- Methods: This is a longitudinal study of patients with T1 diabetes (n=1083) roarthropathy there is thick, tight, waxy skin reminiscent of scleroderma. seen in diabetes complication assessment service at The Childrens’ Hospital Limitation of movement of small of hands e.g., metacarpophalangeal, at Westmead from 2009-2016 (median follow-up of 2.5 years). Heart rate proximal and distal interphalangeal joints and sclerosis of sheaths (HR) and Heart rate variability (HRV) parameters were derived from a 10 causing flexion contractures of the fingers. The prayer sign, that is the minute ECG recording using Labchart Pro: Standard deviation of mean NN patient’s inability to press their palms together completely without a gap intervals (SDNN) where NN = adjacent QRS complexes; root mean squared indicates presence of cheiroarthropathy. difference of successive NN intervals (RMSSD)-estimates of overall HRV; Aim: To know the association of cheiroarthropathy with type 2 diabetes low/high frequency (LF:HF) ratio; geometric measure-Triangular index (TI). mellitus and with diabetic retinopathy. Early retinopathy is defined as the presence of at least one microaneurysm Methods: After obtaining consent from the patients of cheiroarthropa- or haemorrhage from seven standard fields. Patients were classified as hav- thy, for a period of one year (1st October 2015-30th September 2016) who ing normal or abnormal HR and HRV based on their age and sex matched were presented to OPD of SCB medical college and hospital controls. Statistical analysis was performed using generalised estimating Cuttack were screened for diabetes mellitus through Fasting and two hours equations with covariates HbA1c and duration for the outcome of incident post prandial , HbA1c. Those patients, who were detected with retinopathy. diabetes mellitus undergone fundoscopic examination for retinopathy. Results: Adolescents with diabetes had a mean age of 16 ± 6 years, Result: Basing on clinical examination and prayer sign of hands 22 patients median duration of 7 years (IQR 4.2-9.8). Incident retinopathy developed in were detected with cheiroarthropathy, who had never been through the 17%. Reduced HRV measures were significant predictors of retinopathy only screening of diabetes mellitus. But after screening for diabetes mellitus 20 in univariable analysis. In multi variable analysis, elevated mean heart rate patients were detected to be diabetes mellitus. Amongst the 20 diabetic was a significant predictor of incident retinopathy [OR 1.57 95% CI (1.15, patients 4 were having diabetic retinopathy. 2.15)], HbA1c [1.26(1.15, 1.35)], duration [1.18(1.13, 1.23)]. Conclusion: The association of cheiroarthropathy in diabetes mellitus is Conclusion: Elevated heart rate but not reduced heart rate variation significant. Its prevalence depends upon duration of diabetes mellitus along predicts incident retinopathy in adolescents with type 1 diabetes, indepen- with higher level of average fasting blood sugar and HbA1c, contradict- dently of duration and metabolic control. Autonomic vascular tone may con- ing the previous studies. Diabetic retinopathy often present with diabetic tribute to development of diabetic retinopathy. cheiro­.

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623‑P inconsistent. The purpose of this study was to correlate anthropometric Reproducibility of Tablet-Based Bayesian Adaptive Estimation of parameters to the level of DR. the Contrast Sensitivity Function (BCSF) with Tools for Distance Materials and Methods: A sample of type 2 diabetes mellitus (T2D) Compliance in Diabetic Patients patients and nondiabetic healthy controls was prospectively and consecu- JAE W. RHEE, LUIS LESMES, LLOYD PAUL AIELLO, JENNIFER K. SUN, Boston, MA tively recruited. Participants underwent a comprehensive examination, BCSF is a promising method for evaluating visual function changes due to including ophthalmoscopy of the posterior segment after pharmacological diabetic eye disease independent of age and visual acuity. Utilization of this mydriasis, anthropometric measurements and bioelectrical impedance. They were divided into six groups according to the severity of the disease: non- technique for home monitoring necessitates a portable device with tools to POSTERS enforce viewing standards to provide robust results. We evaluated a novel, diabetics, T2D patients without DR, with mild non-proliferative DR (NPDR), Complications tablet-based BCSF test at multiple viewing distances and with and without moderate NPDR, severe NPDR and proliferative DR (PDR). After testing for Acute and Chronic camera-based distance estimation and software prompts to enable viewer a normal distribution of variables, ANOVA test was applied between all compliance. Diabetic (DM) and non-DM participants with good vision (Snel- groups and post-hoc tests among the control group and the other ones. len equivalent > 20/25) were tested on a Sentio Home Vision Tester. Area Results: 163 subjects were included. Basal characteristics of the whole under the letter/contrast size function (AULCSF) and Contrast Sensitivity group, expressed as mean and SD, were: age 67.24 ± 8.07 years, BMI 30.69 ± Function (CSF) Acuity were estimated with 25 trials over spatial frequencies 6.57 kg/m2 and HbA1c 7.10 ± 1.45%. Anthropometric data are shown in the from 1.5-18 cpd. 15 patients (7 non-DM and 8 DM), tested with a chinrest at Table. a 14” standard distance, had mean±SD test-retest differences of 0.03±0.15 Conclusion: Patients with a more advanced stage of the disease had a for AULCSF and 0.002±0.135 for CSF Acuity. Average test duration per eye greater BMI and abdominal circumference (except for the PDR group), neck was 3.3min±22s. Without chinrest, participants (N=7) maintained viewing circumference and visceral fat rate. distance within +/- 2” of the prescribed standard on 60% of trials without Table. Anthropometric Parameters (Expressed as Mean ± SD). prompting. With distance estimation and prompting, +/- 2” was maintained BMI Neck Abdominal Waist hip Trunk fat Visceral on 80% of trials, and +/-3” for 92%. To evaluate impact of distance vari- (kg/m2) circumference circumference ratio (%) fat rate ability on BCSF, 8 additional DM patients were tested at 12,” 14” and 16”. (cm) (cm) Differences in AULCSF at 12” and 16” as compared to 14” were 0.05±0.08 Nondiabetic subjects 28.04 ± 4.53 35.68 ± 3.64 96.08 ± 10.52 0.95 ± 0.06 38.47 ± 8.69 10.91 ± 4.47 and -0.02±0.19, respectively, corresponding to mean 4.6% and -1.9% differ- ences in CS threshold across the entire spatial frequency range. These data T2D without DR 29.89 ± 4.28 40.17 ± 4.62 (*) 105.14 ± 10.26 (*) 1.02 ± 0.68 (*) 39.67 ± 6.73 15.80 ± 5.72 (*) suggest that tablet BCSF with distance compliance tools is readily utilized T2D with mild NPDR 32.15 ± 10.12 (*) 42.22 ± 4.11 (*) 107.87 ± 13.13 (*) 1.01 ± 0.48 (*) 38.5 ± 8.27 16.33 ± 6.23 (*) with excellent reproducibility. The distance prompt enables good compli- T2D with moderate 31.85 ± 6.32 (*) 40.43 ± 4.03 (*) 108.0 ± 12.26 (*) 1.01 ± 0.48 (*) 37.15 ± 9.73 15.72 ± 4.85 (*) ance with a standard 14” testing distance, and BCSF results are minimally NPDR affected over a 4” range of movement. Future studies will directly evaluate T2D with severe 34.62 ± 6.6 (*) 41.84 ± 3.38 (*) 114 ± 13.05 (*) 1.01 ± 0.05 (*) 42.55 ± 10.11 17.10 ± 6.28 (*) the use of BCSF for home monitoring and determination of treatment need NPDR for diabetic ocular complications. T2D with PDR 30.96 ± 3.42 41.96 ± 2.81 (*) 105.54 ± 10.57 1.02 ± 0.05 (*) 37.52 ± 9.89 18.12 ± 7.42 (*) Supported By: National Eye Institute (R01EY024702); JDRF (3-SRA-2014-264-M- R); Beatson Childcare Ambassador Program Foundation; Massachusetts Lions Eye (*). P-value < 0.05. Research Fund; Adaptive Sensory Technology 626‑P 624‑P Relationship between the Stage of Diabetic Retinopathy and Serum The Specific Signature of Diabetes and Diabetic Retinopathy in the Levels of Inflammatory and Angiogenic Markers Human Retinal Transcriptome GUAYENTE VERDES, FRANCISCO BARTOL-PUYAL, BEATRIZ ABADIA, NAIARA PATRICE E. FORT, JEROME E. ROGER, YANG SHAN, Ann Arbor, MI, Orsay, France ROMERO-SANCHEZ, FRANCISCO-JAVIER ACHA-PEREZ, BEATRIZ LARDIES, Diabetic retinopathy, the major ocular complication associated with dia- ANTONIO FERRERAS, Teruel, Spain, Zaragoza, Spain betes, remains the primary cause of vision loss in the working age popula- Background and Aims: Some studies have suggested a link between tion. Using non-targeted transcriptome analysis approaches, several groups serum levels of inflammatory and angiogenic mediators and the presence have identified specific regulatory pathways affected in diabetic rodents. and severity of diabetic retinopathy (DR). The purpose of this study was to Use of those animal models has yielded critical discoveries relative to the correlate plasma levels of biomarkers to the level of DR. general changes affecting the retinal transcriptome, but those are limited Materials and Methods: A sample of type 2 diabetes mellitus (T2D) by the absence of a macula and the limited difference of peripheral vs. cen- patients and nondiabetic age-matched controls was recruited. Patients tral retina in rodents. These limitations are critical as diabetic retinopathy were prospectively and consecutively enrolled from the department of oph- is a regional disease that affects the retina non-homogeneously, as dem- thalmology, whereas controls were recruited from hospital staff, relatives onstrated by macular , peripheral non-perfusion and regional loss of and healthy volunteers. They all underwent a comprehensive medical and receptor fields in diabetic patients. Using non-fixed, freshly isolated retinal ophthalmological history and examination, including an ophthalmoscopy tissues from human donors, with and without diabetes and with or without of the posterior segment after pharmacological mydriasis and laboratory retinopathy, we used RNA deep sequencing to assess the transcriptional testing. Participants were divided into six groups according to the sever- changes affecting the retina. In this study, we independently analyzed the ity of the disease: nondiabetics, and T2D patients without DR, with mild transcriptome of the macular, perimacular and peripheral regions of the non-proliferative DR (NPDR), with moderate NPDR, with severe NPDR and retina (n=6) in order to identify the regional impact of diabetes. Over 500 with proliferative DR. After testing for a normal distribution of variables, genes were statistically significantly affected (p>0.05) with region specific Pearson’s correlations were calculated. patterns as a function of the disease state. Principal component analysis Results: 163 subjects were included in the statistical analysis. Basal char- confirmed the clustering of the samples while pathway analysis using the acteristics of the whole group, expressed as mean and SD, were: age 67.24 ± GeneGo/MetaCore integrated software identified specific inflammatory, 8.07 years, BMI 30.69 ± 6.57 kg/m2 and HbA1c 7.10 ± 1.45%. Correlations are metabolic and neuroglial regulatory pathway, some of which were validated expressed through Pearson coefficient r: homocysteine 0.146 (p 0.011), apoB by qRT-PCR on additional samples (n≥7 per group). This study offers the first -0.225 (p<0.001), C-reactive protein (CRP) 0.130 (p 0.025), N-terminal pro- regional analysis of the pathophysiological mechanisms of diabetic reti- brain natriuretic peptide (NT-proBNP) 0.176 (p 0.002), prostaglandin E2 0.029 nopathy with a high potential of identification of novel therapeutic targets. (non-significant -NS-), erythrocyte sedimentation rate (ERS) 0.252 (p<0.001), Supported By: Diabetic Complications Consortium; Eversight somatostatin 0.280 (p<0’001), IGF-1 -0.004 (NS). Conclusion: The severity of diabetic retinopathy correlated positively with 625‑P homocysteine, CRP, NT-proBNP, ERS and somatostatin. An increasing stage Relationship between the Stage of Diabetic Retinopathy and of the disease was associated with fewer levels of apoB. No association Anthropometric Measurements was found with prostaglandin E2 neither with IGF-1. GUAYENTE VERDES, MARIA JIMENEZ-SANMARTIN, PILAR CALVO, BEATRIZ ABADIA, FRANCISCO BARTOL-PUYAL, ALEJANDRO SANZ-PARIS, ANTONIO FER- RERAS, Teruel, Spain, Zaragoza, Spain Background and Aims: Some studies have reported a relationship between diabetic retinopathy (DR) and obesity, although these results are

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DIABETIC DYSLIPIDEMIA 629‑P (HDL-C/apoA-I): A Multi-Vessel Risk Marker in Women with Type 2 627‑P Diabetes Effect of Rosuvastatin on Apolipoprotein-Defined Lipoprotein Sub‑ MICHEL P. HERMANS, SYLVIE A. AHN, MICHEL F. ROUSSEAU, Brussels, Belgium classes (ADLS) in Patients with Type 2 Diabetes with Possible Sex‑ The role of HDL-C as CV risk factor remains controversial, despite small- ual Dimorphism in Responses dense/dysfunctional HDLs being linked to metabolic syndrome (MetS) and MISTI J. LEYVA, ALBINA GOSMANOVA, PETAR ALAUPOVIC, CHRISTOPHER E. T2DM. To clarify the epidemiological risk of HDLs of different sizes/densi-

POSTERS ASTON, TIMOTHY J. LYONS, Oklahoma City, OK, Belfast, United Kingdom ties, without direct measurement, we adjusted HDL-C to its main apolipopro- Complications Background: Macro- and microvascular complications cause a majority of tein (apoA-I), yielding a (HDL-C/apoA-I) ratio. The latter not only estimates Acute and Chronic diabetes-related morbidity and mortality, and dyslipidemia mediates risk. In type the average size of HDLs, but also provides indices as to their number, cho- 2 diabetes mellitus (T2DM) ‘statin’ treatment is standard of care, but its effects lesterol load, and density. To assess the usefulness of this ratio, we strati- on detailed lipoprotein subclasses are poorly defined. Apolipoprotein-based fied 280 Caucasian T2DM women according to (HDL-C/apoA-I) quartiles (Q), lipoprotein subclasses (ADLS) define lipoproteins by qualitative apolipoprotein to determine how it segregates cardiometabolic risk, β-cell function, glyce- content, reflecting function and atherogenicity. ApoA-I-containing particles mic control, and vascular complications. Five parameters were derived from comprise lipoprotein A-I (LpA-I) and lipoprotein A-I:A-II (LpA-I:A-II), and apoB- the combined determination of HDL-C and apoA-I, namely HDL size; HDL containing particles, lipoprotein B (LpB), lipoprotein B:C (LpB:C), lipoprotein B:E number; cholesterol load per particle (pP); apoA-IpP; and HDL density. An (LpB:E), lipoprotein B:C:E (LpB:C:E), and lipoprotein A-II:B:C:D:E (LpA-II:B:C:D:E). adverse cardiometabolic profile characterized Q-I and Q-II patients, whose Effects of statins on plasma concentrations of apolipoproteins also merit study. HDLs were denser (0.80 and 0.77) and apoA-I-depleted (78 and 88 mg/µmol). Purpose: To determine the levels of apoA-I- and apoB-containing ADLS, and Q-III patients had HDLs whose characteristics were close to controls (den- plasma apolipoproteins A, B, E and CIII before and 6 weeks after rosuvastatin sity: 0.74; apoA-IpP: 97 mg/µmol). Compared to Q-I, Q-IV pts had HDLs of treatment in men and women with T2DM. supernormal size/composition (density: 0.70; apoA-IpP: 113 mg/µmol) and Methods: ADLS and apolipoprotein concentrations were measured in a more favorable phenotype in terms of fat distribution; insulin sensitivity 40 fasting T2DM adults (19 men, 21 women) before and after rosuvastatin, (64% vs. 41%), MetS score/prevalence (3.3/5 and 79% vs. 4.7/5 and 100%), 10 mg/day, for 6 weeks. β-cell function (32% vs. 23%); exogenous insulin (44 vs. 89 U/day), and glyce- Results: Rosuvastatin reduced all five apoB-containing subclasses in both mic control (HbA1c: 56 vs. 61 mmol/mol), associated with lower prevalence men and women, but had no effect on LpA-I or LpA-I:A-II. Proatherogenic of micro-/macrovascular complications: all-cause 47% vs. ApoC-III levels were reduced in men, but not in women. Reductions in LpA- 61%; retinopathy 22% vs. 34%; all-cause macroangiopathy 19% vs. 31%; and II:B:C:D:E and ApoB were significantly greater in men than women. There was CAD: 6% vs. 24% (all p<0.05). The (HDL-C/apoA-I) can easily stratify T2DM a greater reduction of apoC-III bound to apoB-containing lipoproteins (CIII-HP) women according to metabolic phenotype, macrovascular and coronary than apoC-III bound to apoA-containing lipoproteins (CIII-HS). In both genders damage, β-cell function, microangiopathic risk, and retinopathy frequency. rosuvastatin lowered apoE, while apoAII was reduced only in women. This ratio is a versatile and readily available marker of cardiometabolic sta- Conclusion: Rosuvastatin was effective in reducing all atherogenic apo- tus and vascular complications in T2DM. B-containing ADLS in men and women with T2DM, but had no effect on anti- atherogenic apo-A-containing ADLS. A reduction in apo-CIII was observed 630‑P in men only. Dyslipidemia and Associated Demographic and Clinical Factors in Supported By: AstraZeneca Young Adults with Type 1 Diabetes ELIE ABED, BRENTON M. LABARBERA, JUSTIN D. DVORAK, JAMES T. LANE, 628‑P JONI K. BECK, YING ZHANG, MITALI TALSANIA, Oklahoma City, OK One-Year Real-Time Clinical Safety and Effectiveness of Saroglita‑ Young adults (YA) with type 1 diabetes (T1DM) have high cardiovascular zar in Patients with Diabetic Dyslipidemia mortality. We studied prevalence of dyslipidemia (DLD) in YA with T1DM and AMITESH KUMAR CHATTERJEE, MINAL MOHIT, ASHOK JAISWAL, Jalpaiguri, factors affecting DLD as it is a modifiable risk factor for cardiovascular dis- India, Jaipur, India, Ahmedabad, India ease (CVD). A cohort of YA with T1DM is being recruited to study associated , a dual PPAR α/γ agonist, is approved in India for the treat- factors of glycemic control during the transition from pediatric to adult care. ment of hypertriglyceridemia in type 2 diabetes not controlled with statins. 62 YA (45% females, 88% Caucasian with a mean age of 18) have baseline This is an observational study conducted to evaluate the real time clinical data collection of lipid profiles. In cross-sectional analysis, we compared safety and effectiveness of Saroglitazar in patients with diabetic dyslipid- demographics (age, sex, race) and clinical characteristics [body mass index emia (DD). Total 851 patients with DD and baseline triglycerides (TG) >150 (BMI), hemoglobin A1c (A1c), TSH, serum creatinine, duration of T1DM, use mg/dL, were enrolled in this study of which 764 completed 1 year follow- of lipid lowering agents and physical activity (PA)] between YA with and up. All these patients were prescribed Saroglitazar 4mg once daily over and without DLD. DLD was considered present if a YA had 1 or more lipid abnor- above the on-going antidiabetic and statin therapy. Safety and effective- malities. ADA cutoffs for DLD were followed. DLD was present in 44 YA ness were evaluated every 3 months till one year follow-up. At baseline, the (71%); 22 had 1 lipid disorder, 12 had 2, 7 had 3, and 3 had 4 lipid disorders. mean age of study population was 53 years, average duration of diabetes High LDL-C was the most prevalent lipid disorder (27 YA), followed by low was 11.23 years, and mean body weight was 66.77 kg. At 1 year follow-up, HDL-C, high triglyceride, and high total cholesterol levels. YA with vs. with- there was significant reduction (p<0.0001) observed in TG, and non HDL-C out DLD had significantly higher BMI (27.93 ± 10.49 vs. 20.20 ± 2.13, P<0.01) level (Table). Other lipid parameters such as LDL-C and total cholesterol (TC) and worse HbA1c (9.88 ± 2.30 vs. 8.31 ± 1.49, P<0.05). Age, sex, race, dura- were also significantly reduced (p<0.0001). Saroglitazar, in combination with tion of T1DM, TSH, and serum creatinine were similar between both groups. other antidiabetic medications, significantly reduced HbA1c from 11.33% at Grouped by their number of lipid disorders, YA with 2 or more lipid disorders baseline to 9.07% at 1 year (p<0.0001). Saroglitazar treatment was found to vs. YA with no or 1 lipid disorder reported less PA (27.7% stated they did 30 be weight neutral. No major adverse event was reported during follow-up. minutes of vigorous or moderate PA per day vs. 69.9%, p<0.05), and were Saroglitazar is a potential add on therapeutic option for the treatment of more likely to be on a statin (18.2% vs. 0, p=0.01). Among YA with T1DM, the hypertriglyceridemia in type 2 diabetes not controlled with statins. prevalence of DLD is high. Higher BMI and A1c were associated with DLD. Table. Change in Lipid and Glycemic Parameters at 1-Year Follow-Up. Values YA with multiple lipid disorders had less PA and were more likely to receive are in Mean ± SD. statin. This study provides evidence that lifestyle factors, glycemic control, and DLD were closely related among YA with T1DM. Longitudinal data will Laboratory parameters Baseline 1-year Absolute change % change P value follow-up at 1-year at 1-year assist in future interventions to improve glycemic control and lipid manage- ment for the prevention of CVD. TG (mg/dL) 295.1±101.87 162.24±55.67 -132.85 -45.86% <0.0001 LDL-C (mg/dL) 140.05±31.38 111.85±24.87 -28.2 -19.56% <0.0001 TC (mg/dL) 234.63±41.9 182.98±29.63 -51.65 -20.53% <0.0001 Non HDL-C (mg/dL) 199.34±42.45 144.53±30.5 -54.8 -27.71% <0.0001 HbA1c (%) 11.33±2.19 9.07±1.75 -2.27 — <0.0001 Fasting plasma glucose (mg/dL) 187.99±35.79 131.64±25.03 -56.35 -28.69% <0.0001 Postprandial plasma glucose (mg/dL) 351.55±69.99 224.99±44.79 -126.56 -34.85% <0.0001

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631‑P icosapent ethyl (IPE; Vascepa) in women and none have reported results spe- Concentration and Progression of Retinopathy cifically in women with HTG and DM2. IPE is high-purity prescription eicosa- with Type 2 Diabetes pentaenoic acid (EPA) ethyl ester approved at 4 g/d as an adjunct to diet for SHO OSONOI, HIROSHI MURAKAMI, KOKI MATSUMURA, KOTA MATSUKI, ERI TG ≥500 mg/dL. The 12-week ANCHOR study randomized 702 statin-treated SATO, JUTARO TANABE, YUKI MATSUHASHI, MIYUKI YANAGIMACHI, MAKOTO patients (39% women; 73% with DM) at increased CVD risk with TG 200-499 DAIMON, Hirosaki, Japan mg/dL despite LDL-C control (40-99 mg/dL) to IPE 2 g/d, 4 g/d, or placebo. Objective: Serum fatty acids (FAs) have been reported as involved in vari- This post-hoc analysis includes 146 women with DM2 (97% white, mean age 62 y) randomized to IPE 4 g/d (n=74) or placebo (n=72). IPE reduced TG with- ous metabolic disorders such as type 2 diabetes (DM) with unsaturated FAs POSTERS having positive, and saturated and trans-FAs having negative effects on out increasing LDL-C, and improved other potentially atherogenic lipid, lipo- Complications DM. However, association of FAs with diabetic complications has been little protein, and inflammatory parameters vs. placebo (Figure). IPE increased EPA Acute and Chronic examined. levels in plasma (+639%; P<0.0001; n=49) and RBC (+599%; P<0.0001; n=47) Research Design and Methods: Serum fatty acid (FA) composition was vs. placebo. Safety and tolerability for IPE were generally similar to placebo. analyzed for 107 type 2 diabetic patients (age: 54 ± 15 years) using liquid In conclusion, in women with DM2 at high CVD risk with persistent HTG chromatography-tandem mass spectrometry. Association between each FA despite statin use, IPE 4 g/d reduced potentially atherogenic parameters with concentration and diabetic complications (both micro- and macro angiopa- safety generally similar to placebo. Potential CVD benefits of IPE are being thies) was evaluated. tested in approximately 8000 men and women at high CVD risk in REDUCE-IT. Results: Among 24 serum FAs detected, palmitoleic acid concentration Figure. was found to be associated with a diabetic complication, or diabetic reti- nopathy (DR): It was lower in proliferative-DR (PDR) and pre-PDR than in simple-DR (SDR) and non-DR (NDR) (p<0.01), and was decreasing along with the progression of DR (81.3 ± 39.2, 66.1 ± 30.3, and 62.3 ± 28.4 for NDR, non- PDR, PDR, respectively; p for trend=0.02). Conclusions: Although palmitoleic acid have been reported to influence pancreatic β-cell survival, insulin secretion, and insulin response, and, thus, seems to be beneficial for progression of DM, associa- tion between increase in palmitoleic acid concentration and progression of DR was first to be reported here. Furthermore, since reduction of oxidative stress of rodent retinal pigment epithelium by intravitreal injection of palmi- toleic acid has been reported, our result further warrants studies examin- ing the mechanisms involved in the beneficial effects of palmitoleic acid on diabetic retinopathy.

632‑P An Intervention to Serum Triglyceride Improves the Quality of Lipo‑ protein and Endothelial Function in Patients with Type 2 Diabetes ATSUSHI SHINNAKASU, SAHAR G. DARESTANI, KIYOAKI YAMAMOTO, MIHOKO KURANO, HIROSHI ARIMURA, AIKO ARIMURA, AKIRA KIKUTI, HIROSHI HASHIGUTI, TAKAHISA DEGUTI, YOSHIHIKO NISHIO, Kagoshima, Japan 634‑P The effects of an intervention to serum triglyceride (TG) levels on the lipo- Angiopoietin-Like Protein-4 Is an HDL Component for HDL Metabo‑ protein profile and endothelial function were evaluated. Fifty patients with lism and Function in Nondiabetic Participants and Type 2 Diabetic type 2 diabetes treated with statins were recuited and divided into the group Patients treated with a combination of ezetimibe and fenofibrate (E/F group) or the YINGMEI FENG, LONGYAN YANG, CAIGUO YU, XUHONG WANG, SHASHA YUAN, group treated with statins (Statin group). Before and at 12 weeks after each LIJIE ZHANG, JIANAN LANG, DONG ZHAO, Beijing, China treatment, a meal test to assess postprandial dyslipidemia was performed. Rationale: Angiopoietin-like protein-4 (angptl-4) is a potent lipoprotein The lipoprotein profile was analyzed with gel permeation high-performance lipase inhibitor and present in HDLs in mice and human. However, it is not liquid chromatography, and the endothelial function was assessed using flow- known whether angptl-4 in HDLs could participate HDL metabolism and mediated dilation (FMD) at the forearm. In the E/F group, the level of low-den- affect its function in T2DM. sity lipoprotein (LDL) cholesterol was similar to that of Statin group (91.6±15.4 Methods: Nondiabetic (n=201) and T2DM patients (n=185) were analyzed vs. 90.7±21.6 mg/dL, P = 0.87) while that of total TG decreased (100.2±49.0 in the study. Angptl-4 levels and HDL components were measured. HDLs vs. 145.4±64.0 mg/dL, P < 0.01). In addition, the E/F group showed an increase were isolated from all subjects to assess cholesterol efflux to HDLs or sub- in the number of high-density lipoprotein (HDL) particle in very small fraction jected to endothelial lipase (EL)-expressing HEL293 cells to study the kinetic (5.7±0.7 vs. 4.8±0.9 µM, P < 0.05) and a decrease in the number of LDL in very of EL hydrolysis in vitro. Multivariate-adjusted analysis was performed to small fraction (0.16±0.04 vs. 0.20±0.06 µM, P < 0.05). The combination but not study the association between angptl-4 in HDLs and HDL components and the statin treatment significantly improved the FMD from 5.5±2.4 to 6.5±2.2% function in nondiabetic participants or diabetic patients, respectively. (P < 0.05). A stepwise multiple linear regression analysis showed that changes Results: The levels of angptl-4 in the plasma and HDLs were higher in in the cholesterol level in the very small fraction of HDL was a significant and T2DM patients than nondiabetic controls (p<0.0001 for both). When ang- independent predictor for determining the improvement of FMD (β=0.438, ptl-4-containing HDLs were subjected onto HEK 293 cells expressing EL, P < 0.01). Thus, the treatment with a combination of ezetimibe and fenofi- phospholipid and triglyceride were dramatically hydrolyzed in HDLs with brate reduced the serum TG effectively and improved endothelial function in medium- and high-levels of angptl-4 when compared with those having patients with type 2 diabetes. Compared with the treatment with statin, the lowest angptl-4 levels in HDLs in T2DM patients (p<0.05 for phospholipid; treatment with the combination improved the quality of lipoprotein such as a p<0.01 for triglyceride), but it did not occur in HDLs isolated from healthy decrease in the number of small fraction of LDL and an increase in the number controls (p>0.62 for both). Cholesterol hydrolysis was comparable and inde- of small fraction of HDL, suggesting the clinical importance of the intervention pendent of angptl-4 levels in HDLs in both groups. Multivariate-adjusted to the serum TG in patients with type 2 diabetes. analysis demonstrated that the change of apoA-I, SAA and the percentage of cholesterol efflux per one doubling increase of angptl-4 in circulation or 633‑P HDLs in nondiabetic participants and T2DM patients (nondiabetic: p<0.05 for Icosapent Ethyl in Statin-Treated Women with Persistent High Tri‑ all; T2DM p>0.10 for all). glycerides and Diabetes Mellitus: ANCHOR Study Subanalysis Conclusions: Angptl-4 in HDLs protected HDLs from hydrolysis and main- ELIOT A. BRINTON, JOHN R. GUYTON, CHRISTIE M. BALLANTYNE, SEPHY PHILIP, tained HDL function in nondiabetic controls. Resulting from increased circu- RALPH T. DOYLE, JR., REBECCA A. JULIANO, LORI MOSCA, Salt Lake City, UT, lating angptl-4 levels in T2DM, angptl-4 levels in HDLs were elevated that Durham, NC, Houston, TX, Bedminster, NJ, New York, NY compromised its inhibitory effect on EL, leading to increased HDL hydrolysis Hypertriglyceridemia (HTG) and diabetes mellitus type 2 (DM2) are both and dysfunction. stronger predictors of CVD in women than men, but few randomized, well- Supported By: National Natural Science Foundation of China (81470566, controlled clinical lipid-lowering studies have tested the TG lowering of 81670765)

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A165 DIABETIC DYSLIPIDEMIA

635‑P 637‑P Glucagon-Like Peptide-1 Receptor Agonists Reduce Serum LDL Metabolomic Changes during the 18-Month Copenhagen Insulin and Cholesterol Levels via LDL Receptor in Mice and Type 2 Diabetes Metformin Therapy (CIMT) Trial Patient TOMMI SUVITAIVAL, NARGES SAFAI, ASHFAQ ALI, PETER SPÉGEL, MAHMOUD YUKIKO HASEGAWA, MIKA HORI, TOMOKO NAKAGAMI, MARIKO HARADA- AL-MAJDOUB, BENDIX CARSTENSEN, HENRIK VESTERGAARD, MARTIN RID- SHIBA, YASUKO UCHIGATA, Tokyo, Japan, Osaka, Japan DERSTRÅLE, THE CIMT TRIAL GROUP, Gentofte, Denmark, Lund, Sweden, Malmö, Background: Glucagon-like peptide-1 receptor agonists (GLP-1RAs) were Sweden, Copenhagen, Denmark, Bagsværd, Denmark

POSTERS reported to change lipid profiles in type 2 diabetes (T2DM) patients, but their Effects of 18-month metformin vs. placebo treatment on the blood metabo-

Complications effects remains not to be established. Here we examined whether GLP-1RAs lome were studied in 372 T2DM patients participating in the randomized, con- Acute and Chronic affect cholesterol metabolism in mice and T2DM patients. trolled CIMT trial. Plasma samples were analyzed with ultra-high performance Method: 1) C57BL/6J mice at 10 weeks of age were randomly divided liquid chromatography coupled to a quadrupole time-of-flight mass spectrom- into two groups (n=8/group), and one group was injected intraperitoneally eter, uncovering relative levels of amino acids, acyl-carnitines, lipids and other with GLP-1RA, 10 nmol/kg of exendin-4 for 5 days. The other group received metabolites. saline as vehicle controls. Serum lipid profiles, hepatic cholesterol contents, Standardized data consisting of the metabolites, clinical variables and labora- hepatic genes and proteins related to cholesterol metabolism were mea- tory measurements were analyzed with a two-way repeated measures ANOVA sured. 2) We retrospectively assessed serum lipid levels, HbA1c and BMI in model inferred with a linear mixed effects model in R, taking into account the 103 T2DM patients at baseline and follow-up after initiating GLP-1RA. We treatment group, time point and their interaction, and adjusting for insulin treat- selected 214 T2DM patients without GLP-1RA treatment as controls. ment. Variables with a multiple testing-adjusted p<0.1 effect were reported. Result: 1) Exendin-4 treatment significantly decreased serum VLDL- and The metformin group had a larger improvement in plasma glucose and LDL-cholesterol levels in C57BL/6J mice and upregulated hepatic SREBP2- HbA1c, a larger increase in uric acid, and a larger decrease in glycerophos- related genes/proteins including LDLR proteins. 2) The serum LDL-C level phocholine between baseline and follow-up when compared to placebo decreased by 4.4% (p=0.008) in the GLP-1RA patients and by 0.3% (p=0.42) (Figure). The placebo group had a broad increase in amino acids and their in the controls. The GLP-1RA patients’ reduction rate of LDL-C was signifi- derivatives as well as in acyl-carnitines, lipids, other metabolites, LDL and cantly greater than that of the controls (p=0.045). HbA1c and BMI were sig- BMI, and clear changes in branched-chain amino acids. Nearly all of these nificantly decreased in the GLP-1RA group. There were no significant differ- increases were smaller or not seen in the metformin group. The results indi- ences in HDL-C or triglycerides between the GLP-1RA and control groups. cate that metformin helped maintain a better glucose control, which was In GLP-1RA group, patients treated with statins (which upregulate LDLR), broadly reflected in the levels of amino acids and lipids. LDL-C significantly decreased by 6.8%; it decreased by 2.9% in those with- Figure. out statin (p=0.296). Conclusion: Our data demonstrate that GLP-1RAs upregulate LDLR, and then decrease the serum LDL-C level in T2DM patients and the reduction of serum LDL-C by GLP-1RAs is amplified by the use of statin. Combinations of statin/GLP-1RA could ameliorate hypercholesterolemia with T2DM.

636‑P Early Dynamic Changes in the Serum Lipidome of Type 2 Diabetes Subjects in Response to RYGB HAN WU, VIOLETA RAVERDY, HAI H. BUI, PAUL MILLIGAN, AJIT REGMI, MARK FARMEN, MING SHANG KUO, FRANÇOIS PATTOU, MELISSA K. THOMAS, India- napolis, IN, Lille, France Mechanisms by which induces remission of type 2 dia- betes (T2D) are incompletely understood, but post-RYGB improvements in insulin sensitivity precede maximal weight loss. To identify molecular phe- notypes associated with early, dynamic metabolic improvement, we ana- lyzed fasting blood samples from 48 T2D subjects at 0, 1 and 3 months after RYGB. We measured RYGB-dependent lipid biomarker changes with multiple assay platforms. In a mass spectrometry-based targeted sphingolipid panel, 33 dihydroceramides, ceramides, and complexed ceramides were detected. Multiple sphingolipid levels changed from pre-operative baselines. Dihydro- ceramide C24:0, Ceramide 23:0, and Ceramide 24:0 significantly decreased by 1.4-1.7 fold at 1 and 3 months, and HexosylCeramide (C18:0) levels signifi- cantly increased by 1.5 fold (adj p<0.05). Many RYGB-dependent lipid bio- markers were associated with baseline metabolic phenotypes. For example, pre-RYGB Ceramide 24:0 levels correlated with baseline fasting glucose levels. In shotgun lipidomics assays we detected 380 triglyceride (TG) and 224 non-TG related lipids and daughter ions. Significant decreases in large numbers of lipid biomarker levels were observed at 1 and 3 months follow- ing bariatric surgery across a range of phosphatidylcholine, lysophospha- tidylcholine, phosphatidylethanolamine, phosphatidylinositide, cholesterol ester, triglyceride, and ceramide lipid classes. A smaller proportion of lipid biomarkers detected in shotgun lipidomics assays increased post RYGB, including 24:2 and Hexosylceramide 24:1 levels that were significantly increased at 1 and 3 months (1.17-1.38 fold; adj p<0.05). Our find- ings indicate that substantial remodeling of and metabolism occurs post-RYGB before considerable weight loss is achieved. We suggest Supported By: Novo Nordisk Foundation that multiple lipid signaling pathways contribute to adaptation of metabolic phenotypes in T2D subjects following bariatric surgery. 638‑P Supported By: Innovative Initiative Joint Undertaking (115317) Serum PCSK9 Concentration Responses to Acute Oral or Intrave‑ nous Glucose Challenge in Subjects with Metabolic Disorders YINGYUN GONG, YIZHE MA, DANDAN HU, WEN GUO, ZHENZHEN FU, PANPAN YANG, YING ZHOU, AIJIE HUANG, JINXIANG FU, JINGYU GU, TAO YANG, HON- GWEN ZHOU, Nanjing, China The role of proprotein convertase subtilisin/kexin 9 (PCSK9) in the meta- bolic process beyond cholesterol metabolism is far from clear. This study

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A166 FOOT CARE—LOWER EXTREMITIES aimed to investigate how serum PCSK9 concentration changes after acute The clinical features, complications of diabetes and comorbidities were col- glucose challenge. Firstly, totally 32 middle aged patients with diagnosed lected from the first ulceration event register and then followed for the next diabetes were assigned to undergo a 75-g oral glucose tolerance test 5 years evaluating DFU recurrence and amputation within this period. Autho- (OGTT). Blood samples were collected at 0, 30, 60 and 120 min for mea- rization was obtained from the Hospital’s ethics committee. Predictive anal- suring glucose, insulin as well as PCSK9 levels. Secondly, 19 lean and 31 ysis was performed with Kaplan Meyer curves, long rank test and risk using overweight or obese participants were enrolled in an intravenous glucose Hazard Ratio and Cox regression. P value <0.05 and 95% confidence interval. tolerance test (IVGTT) lasting for 60 min. Serum PCSK9 concentration were Results: A total of 626 clinical records were reviewed and 111 were detected by commercial ELISA kits. Serum PCSK9 levels were plotted across included. DFU recurrence was 36.6%, amputation 21.4%, general mortality POSTERS respective time points for each participant, and the variation patterns were 26.7%, and -associated mortality 5,35%. Only 14.4% had A1c <7%, and Complications classified into “rise,” “fall” or other according the curve shape. Under oral 62.2% continued to use oral antidiabetics. Survival analysis showed that at Acute and Chronic glucose challenge, 10 and 18 manifested with rise or fall pattern respec- 24.3 months of follow-up, 50% of the population presented DFU recurrence. tively in diabetes, and the “fall” group showed a significantly higher fasting At 8.6 months, 50% need some type of amputation. The long-rank test was serum PCSK9 concentration and higher serum insulin levels at all the time significant for previous heart attack (HA) (DFU recurrence p = 0.00, amputa- points. During IVGTT, subjects with higher basal PCSK9 level were more tion p = 0.02), pharmacological treatment (DFU recurrence p = 0.01, amputa- likely presented with fall vs. rise pattern after glucose challenge. Besides, tion p = 0.03), and peripheral arterial disease (PAD) (p = 0.00) and stroke subjects in fall group had increased BMI (35.71±8.82 vs. 27.15±8.29 kg/m2, (p=0.05) only in amputation cases. Cox regression analysis confirmed asso- P=0.003), total cholesterol (5.01±0.88 vs. 4.35±0.73 mmol/L, P=0.01), LDL-c ciation for previous HA and pharmacological treatment in DFU recurrence. (3.29±0.78 vs. 2.54±0.67 mmol/L, P=0.001) and worse glucose tolerance than Conclusions: The cardiovascular history (previous HA, PAD and stroke) and those in rise group. Pearson correlation analysis further showed that PCSK9 pharmacological treatment are prognostic factors for DFU recurrence and concentration correlates with serum insulin level (P<0.05 at 0, 60; P<0.001 at amputation. 4 min; P=0.002 at 10 min), but not with glucose levels. Serum PCSK9 charac- terized a robust change during acute oral or intravenous glucose challenge, & 641‑P and the variate pattern differs between people with different metabolic (HD) Is a Stronger Indicator of Patient Survival fol‑ status. Serum PCSK9 concentration has a positive correlation with serum lowing Lower Limb Amputation (LLA) than the Presence of Diabetes insulin level in both lean, overweight and obese subjects. Mellitus (DM) Supported By: National Natural Science Foundation of China SATOSHI FUNAKOSHI, YUTAKA MORI, KAZUNORI UTSUNOMIYA, JYUNICHIRO HASHIGUCHI, TAKASHI HARADA, TOMOYA NISHINO, Nagasaki, Japan, Komae, 639‑P Japan, Tokyo, Japan Atorvastatin Preserves eNOS Function in Rat Glomerular Endothe‑ Objective: To evaluate hemodialysis (HD) patients for life prognosis fol- lial Cells under Conditions of Hyperglycemia during LDL Oxida‑ lowing lower limb amputation (LLA) according to the presence of diabetes tion Ex Vivo mellitus (DM) or the duration of HD in these patients. R. PRESTON MASON, SAMUEL C.R. SHERRATT, HAZEM DAWOUD, ROBERT F. Methods: A total of 51 HD patients underwent LLA between 1996 and JACOB, HAIDAR ALHUMAID, FARINA J. MAHMUD, TADEUSZ MALINSKI, Boston, 2016 for post-LLA survivals were analyzed. MA, Beverly, MA, Athens, OH Results: Mean age, 54 ± 22.5 years; men/women, 37/14; and DM/non-DM, Atorvastatin has been shown to reduce cardiovascular events and 36/15. As shown in the Figure, a comparison of post-LLA survival between preserve renal function in diabetic patients in a manner not predicted by the DM and non-DM patients showed that the DM patients had significantly changes in lipid levels. To better understand the basis for potential cho- longer survival at 3.2 years than the non-DM (1.8 years). Again, an analy- lesterol-independent actions, we tested the effects of atorvastatin active sis of survival by HD duration showed that while those with HD durations metabolite (ATM) on glomerular endothelial cells (ECs) isolated from WKY ≤ 5 years included more DM patients (n = 26) than non-DM (n = 2) and had rats and exposed ex vivo to hyperglycemic conditions (300 mg/dL) with LDL a mean survival of 3.15 years, those with HD durations ≥ 10 years included (11 mg/dL) undergoing oxidation. ECs were assayed for nitric oxide (NO) and more non-DM patients (n = 10) than DM patients (n = 1) and had a signifi- peroxynitrite (ONOO-) release following LDL pretreatment with equimo- cantly shorter mean survival of 0.83 years, the shortest among the 3 groups lar ATM (10.0 µM) or rosuvastatin vs. vehicle. Changes in NO and ONOO− compared. release were measured using porphyrinic nanosensors following stimulation Conclusions: HD patients with DM had gradually shorter survival follow- with calcium. The effects of the statins on oxidation of LDL were determined ing LLA, while all HD patients without DM died within 3 years of LLA, sug- by measuring the formation of malondialdehyde (MDA). The results showed gesting that LLA may represent only an event of interest occurring at some that ATM inhibited LDL oxidation by >90% (p<0.001) compared to rosuvas- point in the natural history of atherosclerosis among HD patients with DM tatin or vehicle. As a result, LDL pretreated with ATM did not cause any but the final stage of the disease among non-DM HD patients, i.e., those significant loss in EC function or NO bioavailability under oxidation condi- with long-standing vascular calcification. tions. By contrast, exposure of ECs with LDL pretreated with rosuvastatin Figure. produced a 39% reduction in NO release (195 ± 16.3 to 119 ± 9.37 nM, p<0.001) and decreased the NO/ONOO− ratio, an indicator of eNOS coupling efficiency, by 44% (0.465 ± 0.0623 to 0.258 ± 0.0287, p<0.001) compared to control ECs. These data suggest that atorvastatin preserves glomerular endothelial benefits under disease-like conditions of hyperglycemia and LDL oxidation which may be of particular interest to patients with diabetes.

FOOT CARE—LOWER EXTREMITIES

Moderated Poster Discussion: Stepping Into Discussion on the Diabetic Foot (Posters: 640-P to 645-P), see page 21. & 642‑P & 640‑P Nrf2/Keap1 Redox Pathway Regulates Multipotency and Regenera‑ Prognostic Factor for Diabetic Foot Ulcers Recurrence and Amputa‑ tive Capacity of Stromal Cells tion: Follow-Up of Five Years PIUL S. RABBANI, MARC SOARES, SOPHIA HAMEEDI, ROHINI KADLE, CAMILLE ISABEL A. PINEDO-TORRES, ALFONSO A. RAMIREZ-SABA, SR., WILFOR AGU- KIM, RITA A. SARTOR, BUKHTAWAR WAQAS, ADNAN MUBASHER, YEE C. LOW, IRRE-QUISPE, Lima, Peru MARIA H. KOWZUN, DANIEL CERADINI, New York, NY Objective: To determine the prognostic factors for diabetic foot ulcers The mechanisms underlying management of reactive oxygen species (DFU) recurrence and amputation in a National Hospital of Lima, Perú. (ROS) are emerging as crucial factors in cell integrity. The role of redox and Material and Methods: Historical cohort. We included all clinical records metabolic cues in regulation of bone marrow-derived multipotent stromal from January 2005 to June 2016 of patients with type 2 diabetes and foot cells (BMSCs) is relatively unexplored. Through comparison of BMSCs from ulcers. Patients with immunodeficiencies and varicose ulcers were excluded. wild type mice and diabetic mice, the latter with a known redox and meta-

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A167 FOOT CARE—LOWER EXTREMITIES

bolic disorder, we found that the cytoprotective Nrf2/Keap1 pathway is dys- codes (CPT 97597-97598; 11042-11047), and incision and drainage codes (CPT regulated in diabetic BMSCs. Nrf2 is basally active in BMSCs, but chronic 10060-10061; 10140; 10120) were also analyzed for trends when a diabetic ROS exposure in diabetes leads to irregular inhibition of Nrf2 by Keap1, foot complication was identified. Linear interpolation and extrapolation alters metabolism and limits BMSCs to premature adipogenic differentia- were performed. The results demonstrate an approximate 191% increase tion. Upon knockdown of the repressor Keap1, upregulation of Nrf2-directed in number of outpatient encounters for patients with diabetic foot compli- transcription restores redox and subsequently, metabolic homeostasis. cations after PS was established compared to prior, with an approximate Gene expressions of antioxidants downstream of Nrf2, NQO1, HO1, and, 128% increase in the amount of emergency services provided for this cohort. GSR, are significantly upregulated in response to Nrf2 activation to explain Procedure codes for diabetic foot rose substantially (139 vs. POSTERS

Complications the recaptured homeostasis. Normalized Nrf2/Keap1 signaling restores mul- 726 patients). In contrast, inpatient encounters for higher level amputations

Acute and Chronic tipotent cell properties in diabetic BMSCs to allow tri-lineage differentia- decreased significantly (1271 year vs. 88/year; p <0.05) after PS and fre- tion along osteogenic, chondrogenic and adipogenic paths. Following exci- quency of minor amputations remained relatively unchanged. These findings sional wounding in diabetic mice skin, we seeded BMSCs into wound beds support that inclusion of podiatric services in the management of diabetic and immediate periphery. Diabetic BMSCs with activated Nrf2 resulted in foot, is associated with a significant reductions in the rates of major lower closing by 20.75±2.36 days, significantly reduced compared to the extremity amputation and a shift towards wound care and limb salvage. 31.25±1.5 days necessary for untreated diabetic wounds (p<0.05) and in the range of the 22±2.4 days with wild type BMSC-seeded diabetic wounds. This 645‑P 60.9% decrease in pathologic time to closure of diabetic wounds is reflected in larger areas of in diabetic wounds seeded with Nrf2- WITHDRAWN induced diabetic BMSCs, as well as increased CD31+ neovascularization, compared to untreated wounds. Our data strongly suggest that metabolic and redox integrity, regulated by the Nrf2/Keap1 pathway, are essential for facilitating the function of BMSCs in promoting closure of diabetic wounds. Supported By: American Diabetes Association (1-16-ACE-08 to D.C.) & 643‑P Depression, Physical Activity, and Healing LORETTA VILEIKYTE, BIING JIUN SHEN, STEVEN BROWN, ANDREW J.M. BOUL- TON, ROBERT KIRSNER, NEIL REEVES, RYAN T. CREWS, Manchester, United King- dom, Singapore, Singapore, Miami, FL, North Chicago, IL An international study of 79 (46 UK/33 U.S.) persons with type 2 diabetes (66 male; age 57±10 yrs) and plantar diabetic foot ulcers (Texas Classification 1A:57; 1B:9; 2A:12; 2B:1) examined the role of depression and physical activ- ity in diabetic foot ulcer (DFU) healing over a 6-week period (35±10 days). Demographic, disease and psychological determinants of physical activity were also explored. Depression was assessed with a seven-item Hospital Anxiety and Depression Scale (HADS-D). Offloading adherence (OA) was assessed objectively by activity monitors. Daily step count (3335±1987) was recorded by a waist worn uniaxial accelerometer. DFUs were offloaded with a removable cast walker (worn during 59±22% of subjects’ activity). Controlling for baseline DFU size, daily step count and OA were each signifi- cantly associated with smaller DFU at 6wks (β=-.16; p<.05 and β=-.19; p<.05 respectively). However, in multivariate analyses controlling for OA, the asso- ciation of daily step count with DFU healing was no longer significant, though larger baseline wound size (β=.74; p<.001) and U.S. site (β=.22; p<.01) were independently predictive of larger DFU size at 6wks, while better OA (β=-.15; p<.05) remained significantly independently predictive of smaller DFU size at 6wks. At the bivariate level, age and HADS-D were significantly associ- 646‑P ated with lower step count (r=-.23 and r=-.28 respectively, p<.05). In multi- Effect of Short- and Long-Term Diabetes Control on In-Hospital and variate analyses only HADS-D was significantly independently associated One-Year Mortality Rates in Hospitalized Patients with Diabetic with lower step count (β=-.23; p<.01). These data indicate that offloading- Foot adherent weight-bearing physical activity has no independent detrimental MARINA SHARGORODSKY, Holon, Israel effect on healing. Depression, whilst having no direct effect on DFU healing, Introduction: The present study was designed to investigate the effect is an important determinant of diminished physical activity and should be of short- and long-term diabetes control on hospital outcomes including: in- targeted as part of comprehensive DFU management, especially in light of hospital and one year mortality rates, length of hospital stay and rate of findings linking depression to neuropathic unsteadiness - a key predictor to repeated admission in hospitalized patients with diabetic foot. offloading non-adherence and, in turn, impaired foot ulcer healing. Methods: The study group consisted of 341 type 2 diabetic patients hospi- Supported By: National Institutes of Health talized from January 2008 through December 2012 in Wolfson Medical Cen- ter due to the diagnosis of diabetic foot. All glucose measurement performed & 644‑P during hospitalization are stored in the patients’ health records. Adequate Role of Podiatry in Preventing Severe Diabetic Foot Complications short-term glycemic control was defined as average glucose levels during BRIAN M. SCHMIDT, CRYSTAL M. HOLMES, RODICA POP-BUSUI, Ann Arbor, MI admission in the range 110-180 mg/dL. HbA1c values below 7% was defined The objective was to evaluate the effects of in situ podiatry services (PS) as adequate long-term glycemic control. and the risk of major diabetic foot complications and amputations at a ter- Results: The average glucose levels during hospitalization were 179±45 tiary care center. We analyzed patient encounters with diabetes mellitus mg/dL and 39.6%±21.2% of the measurements were between 110-180 mg/ and a concomitant diabetic foot complication from two different 5-year peri- dL. Mean admission HBA1c levels were 8.43%±2.26%, and 31% of the val- ods, preceding and following the establishment of podiatry service based ues were below 7%. The mean length of hospital stay was 24.3±22.6 days, upon diagnosis and procedural codes appointed during each encounter and 15.0% of the patients needed surgical intervention during admission, the organized by encounter type: emergency, inpatient, or outpatient. These in-hospital mortality rate was 10.3%, and the rate of 1-year readmission codes were processed through an internal coding database, which included was 25.1%. In regression models, adequate diabetes control during hospi- over 100 million patient encounters. The codes for diabetes mellitus (ICD-9 talization was marginally associated with reduced in-hospital mortality (OR 250.xx) and a code of at least one diabetic foot complication including: 0.454, 95% confidence interval 0.186-1.103, p=0.081) and significantly with of foot or ankle (ICD-9 730.xx), ulcer foot (ICD-9 707.xx), cel- one year mortality (OR 0.269, 95% confidence interval 0.707-0.101, p=0.009). lulitis, abscess, and , were included. Procedural codes for higher- However, adequate diabetes control during hospitalization did not affect the level amputations, minor lower extremity amputation, wound debridement

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A168 FOOT CARE—LOWER EXTREMITIES length of hospital stay or rate of repeated admission. HbA1c levels were not associated with any of the prognostic factors. Conclusions: Improved glucose control during admission (levels between 110-180 mg/dL) is associated with reduction of one year mortality. HbA1c levels had no impact on these parameters in hospitalized patients with dia- betic foot.

647‑P POSTERS Complications

WITHDRAWN Acute and Chronic

649‑P Foot Care Practice and Knowledge Interventions TIMETHIA J. BONNER, Atlanta, GA Purpose: Lower extremity amputations are some of the most debilitat- ing complications of type 2 diabetes (T2DM). T2DM-related lower extremity amputations have severe consequences in terms of overall management, physical restrictions, psychosocial concerns, and financial hardships. Com- promised , nerve damage, and ulceration are thought to be the root causes of lower extremity amputations. Diabetes self-management education (DSME) and lifestyle behavior change (LBC) play a critical role in how individuals with T2DM manage foot care. For those with at-risk limbs, LBC may reduce the progression of end-stage disease, which is responsible for the bulk of health care costs, morbidity, mortality, and decreased quality 648‑P of life. We reviewed empirical studies to examine the landscape of T2DM- Hydrogen Sulfide Improves Diabetic Wound Healing in ob/ob Mice related interventions in the U.S. between 1989 and 2012. HUICHEN ZHAO, YUANTAO LIU, Qingdao, China Results: 31 studies were identified: case series, qualitative studies, cross- Aim: To investigate the role of hydrogen sulfide (H2S) in impaired wound sectional studies, cohort studies, randomized controlled trials, and surveys healing in ob/ob mice and to explore the possible mechanisms. examined foot care knowledge interventions in the U.S. Learning outcomes Methods: Full-thickness skin dorsal wounds were created on ob/ob mice measured were self-efficacy scores, self-care knowledge scores, and self- and C57BL/6 control mice. Cystathionine-γ-lyase (CSE) expression and H2S care scores. Behavioral outcomes measured were foot self-care exams and production were determined in granulation tissues of the wounds. Effects of daily foot inspection. Clinical outcomes measured fluctuated across the NaHS, a donor of H2S, on wound healing was evaluated. Inflammation and studies, but the most commonly measured outcomes included the risk of angiogenesis in granulation tissues of the wounds were examined. amputation, presence of ulceration, or presence of a foot lesion. Results: CSE expression in both mRNA (Figure A) and protein (Figure B) Conclusion: There are very few interventions addressing foot care knowl- levels were significantly reduced in granulation tissues of wounds in ob/ edge and self-care skills for people with T2DM, and there is no standardized ob mice compared with control mice. H2S content was also decreased in instrument to measure knowledge and skills. This study attempts to high- granulation tissues of ob/ob mice (Figure C). NaHS treatment significantly light the scarce number of interventions surrounding foot care knowledge improved wound healing in ob/ob mice (Figure D), which was associated and skills, as well as the inconsistency in the kinds of intervention that mea- with reduced neutrophil and macrophage infiltration (Figure E), decreased sured learning, behavioral, and clinical outcomes. production of tumor necrosis factor (TNF-α), interleukin (IL)-6 (Figure F). NaHS treatment decreased MMP-9 (Figure G), whereas increased vascular- 650‑P like structures in granulation tissues of wounds in ob/ob mice (Figure H). Prospective Cohort Study of Diabetic Foot Osteomyelitis: Patient Conclusion: CSE down-regulation may play a role in the pathogenesis of Characteristics Association with Clean Margin and Outcome diabetic impaired wound healing. Exogenous H S could be a potential agent 2 BRIAN M. SCHMIDT, JAMES WROBEL, Ann Arbor, MI to improve diabetic impaired wound healing by attenuating inflammation The objective was to evaluate patient characteristics and outcomes asso- and increasing angiogenesis. ciated with proximal histopathologic margins. Data was prospectively col- Figures. lected on 33 diabetics having ablative forefoot surgery over fourteen months. Data collected was pooled for analysis. A stratified analysis of key elements was performed. Participants had assessment of diabetes features includ- ing HbA1c, monofilament testing, previous foot ulcer and amputation record, diabetic foot classification, blood counts, inflammatory markers, imaging studies, non-invasive vascular studies, use, creatinine, and serum albumin values. A standardized method to retrieve intraoperative bone margins was implemented. Negative outcomes included major lower extremity amputation, dehiscence, re-ulceration, re-amputation, or death. Our results demonstrate when histopathologic viable proximal bone margins are attained, there is 86.2% less likelihood of readmission. Patient char- acteristics including absolute toe pressures, wound surface area, platelet counts, albumin levels, creatinine, and RBC/MCV/MCH were significantly different in the viable margin vs. dirty margin group. We conclude that sev- eral patient characteristics are significant to predict attainment of clean intraoperative bone margins and once viable margins are attained, rates of readmission are reduced.

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A169 FOOT CARE—LOWER EXTREMITIES

Table. Results: the sample had 116 subjects (65.8 ± 9.0 y; 75% girls; 28.4 ± 5.6 BMI). According to race, they were white, black, brown, yellow and indig- Variable Name Proximal Margin Clean Proximal Margin Dirty p-value enous 8.6%, 37.9%, 50.9%, 1.7% e 0.9% respectively. The median of DM Age 56.44 ± 14.08 59.25 ± 15.81 0.14 diagnosis was 8.7 ± 6.6 y. Smokers were 26.7%, 87.0% hypertensives and Sex, M/F 23/4 6/0 0.42 28.1% had history of other cardiovascular disease. The screening for risk of Toe Pressure, mmHg 153 91 0.03 diabetic foot with CAM and DMG was positive in 53.4% and 56% respec- tively. The agreement between both methods was 51.7% (k= 0.739). Surface Area, cm3 1.72 8.30 0.009 Conclusion: Similar results were obtained when comparing tests with POSTERS

Complications Readmission (%) 0.14 0.75 0.02 CAM and DMF that shows DMF as a real alternative to CAM and can prob-

Acute and Chronic Creatinine, mg/dL 1.20 ± 0.50 2.54 ± 0.311 0.03 ably be consolidated as a new tool in diagnosing foot at risk in diabetic Albumin 3.7 ± 0.29 3.0 ± 0.28 0.01 population. 653‑P 651‑P Ultrasound Evaluation of Sural Nerve in Diabetic Cutaneous Neu‑ The Possible Role of Trekking Poles to Improve Posture, Balance, ropathy: A Centre-Based Preliminary Study from Nepal and Plantar Pressures Distribution in Diabetic Patients SHYAM S. PARAJULY, TIRTHA L. UPADHYAYA, ANANDA B. SHRESTHA, SARVESH PIERGIORGIO FRANCIA, ROBERTO ANICHINI, MASSIMO GULISANO, LEONARDO GYAWALI, ROSHAN PANGENI, Pokhara, Nepal, Bhaktapur, Nepal BOCCHI, GIUSEPPE SEGHIERI, FILADELFIO PUGLISI, ANNA TEDESCHI, ALESSAN- Nerve examination by ultrasound in the country like Nepal is a very new DRA DE BELLIS, Florence, Italy, Pistoia, Italy factor. People still believe that ultrasound is done for only obstetric and for It is known that patients with diabetes can show postural and functional gall/renal stone evaluation. Almost all doctors are busy with their clinical impairments increasing the risk of falls and foot ulcers. Aim of this study practice rather than research. That’s the reason why our country is left far was to evaluate the effect of trekking poles on balance and forces distribu- behind in research and article publications. With advanced and emerging tion on the foot plantar surface of diabetic patients in standing position. tools and their applications in health care, high frequency probe of ultra- In 15 subjects with diabetes (males/females: 9/6; type 1/2: 7/8), mean sound can be used for morphological study of nerves. This study shows the age 54.95±15.49 yrs, duration of diabetes 21.64±13.10 yrs, mean HbA1c study of sural nerves (SN) and its morphological changes in the diabetic 7.69±1.04%, body mass index (BMI) 25.65±4.26 kg/m2, were evaluated: mus- patients. cle strength (Jamar hand grip), joint mobility of the ankle in plantar and dorsal Methods: This was a centre based prospective study. Fifty two, type 2 flexion (inclinometer), orthostatic plantar pressure distribution and postural diabetic patients and fifty two healthy volunteers were recruited for the control with and without trekking poles (P-walk baropodometric and stabilo- study. Diabetic patients which are considered to be having peripheral cuta- metric analysis, BTS, Italy), posture on the sagittal plane (images of patients neous neuropathy were further evaluated by high frequency (12.5 MHz) lin- in quiet standing). A paired Student t-test was used to determine the dif- ear probe ultrasound. Thickness and width of SN was measured. ference in the means calculated with or without the use of trekking poles. Results: Of the total 52 diabetic patients, with mean age of 56.03 years The patients showed a reduction of joint mobility greater than the group of (ranging from 36 to 79 years of old) with the history of diabetes and under 30 healthy controls matched for age (58.42±5.97 yrs) and BMI (26.26±3.43 medication for 2 months to 30 years were enrolled for the study. The thick- kg/m2): 133.72±18.43° vs. 103.34±31.77°; p <0.01. In standing, patients with ness of SN in diabetic patients ranges from 1.2 mm to 2.8 mm with the mean diabetes showed a greater load on the right foot (dx 44.49±3,91%) while thickness of 1.9 mm whereas the width of the SN varied from 1.8 mm to the load on the forefoot was about 52.45±6,61%. The use of trekking poles 4.7 mm with the mean width of 2.8mm. Significant difference of thickness resulted in an improvement of the weight distribution between the two and width of SN was found in healthy volunteer group (n=52), mean age of limbs (dx 46.40±4.57%) p <0.01, in addition to a mean velocity reduction of 23 years with the age ranging from 15-38 years of old. SN mean thickness the center of pressure (2.53 vs. 1.97 mm/sec), p <0.05. The use of trekking was 1.6 mm with the nerve thickness ranging from 1.4mm to 1.9mm and the poles resulted in a not significant variation of plantar loading at the forefoot mean width of 2.0 mm, width ranging from 1.8 mm to 2.3 mm. level: 51.90±7.85%. The preliminary results of this pilot study showed that Conclusion: Our centre based preliminary results suggested that there the use of trekking poles can contribute in improving the distribution of pres- were significant changes in nerve morphology in diabetic patients. In the sure between the two limbs with possible positive effects on the energy country like Nepal, with the limited technology and research platform this expenditure required to maintain the orthostatic posture without inducing kind of study can make real difference and aid in diagnosis of diabetic cuta- the loading shift forward on foot’s plantar surface. neous neuropathy.

652‑P 654‑P Domestic Monofilament: Is This a Real Alternative to Evaluate Risk Profile and Follow-Up Details of Diabetic Foot Ulcer Patients Admit‑ of Diabetic Foot? ted at a Time in a Diabetic Foot Care Center ANA LUISA OLIVEIRA, ALINE S. SANTOS, GABRIELA H.M. OLIVEIRA, MARIA SREEHARI G.K., ASWATHI V., VIJAYAKUMAR G., REVATHI V., Pandalam, India, SARA S.C. CARNEIRO, ATILA OLIVEIRA, NARA BERNARDES, VITORIA A.M. Kulanada, India SILVA, KELLY A. OLIVEIRA, ANTONIO CESAR OLIVEIRA, ANA MAYRA OLIVEIRA, All the patients with diabetic foot ulcer admitted at a time in a diabetic Campinas, Brazil, Feira de Santana, Brazil, Salvador, Brazil foot care centre were included in the study. 109 patients were enrolled. 1 Diabetic foot is a complication of diabetes mellitus (DM) and one tech- dropped out. 63.9% were males and 36.1% females. Mean age of the partici- nique used to clinically evaluate foot at risk is the Semmes-Weistein 10g pants was 62.17 + 11.018 years and mean duration of diabetes was 15 + 7.98 monofilament that is not widely available. Therefore options should be years and the mean age of onset was 46.74 + 11.79 years. Past history of foot tested such as domestic monofilament (DMF) which is constructed using ulcer was present in 47.2% of patients and 12.1% had history of amputation fishing line once it has similar properties and is less expensive. at various sites before the present ulcer. 94.4% claimed use of foot wear but Objective: Evaluate the efficacy of DMF to detect foot at risk and validate only 30% used protective foot wear. Most common site of ulcer was big toe it as an alternative test to 10g monofilament. (36.8%) with plantar aspect more commonly involved (23%). Majority were Methods: Cross-sectional study applied in DM subjects presenting more Wagner grade 3 ulcers (48.6%) and 55% had osteomyelitis. all patients had than 10 years of diagnosis and no history of vascular surgery. Tests were peripheral neuropathy and 57% had either PVD or arterial stiffening. Ceftri- made using commercially available 10 g monofilament (CAM) and DMF. First, axone tazobactam was the most commonly used antibiotic (31.8%) followed both were applied perpendicularly to the surface of the patient’s skin (hand, by ampicillin cloxacillin (20.6%) and meropenam (16.8%). Ciprofloxacin was elbow or forehead) with enough force to bend it to present the technique given to all patients and clindamycin capsules were used in 50% of patients. to the subjects. The filaments were then applied to the projections of the Majority of the grade 3 ulcers (42.3%) took 1-2 months for complete healing. ventral portions of hallux, 3rd finger and projection of the 1st, 3rd and 5th 30.8% of grade 3 ulcers and 62.55 of grade 4 ulcers took 2-6 months for metatarsal heads. The sensitivity was positive when the subject answered healing. 8 are still nonhealing. 33.3% of grade 4 ulcers and 18.2% of grade correctly to all applications and negative when at least one was incorrect. 5 ulcers needed some form of amputation in the hospital while only 5.8% of Foot at risk was defined when the sensitivity was abnormal. To validate the grade 3 ulcers needed amputation. Out of the 58 patients with osteomyelitis, DMF, it was necessary total agreement between the answers regarding the only 8 needed amputation. perception of the stimulus performed.

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A170 FOOT CARE—LOWER EXTREMITIES

Table. number of inflammatory cells, degranulated mast cells and elevated numbers of both M0 non-polarized and alternatively activated M2 macrophages, but Frequency Percent not classically activated M1 macrophages. These findings were corroborated Past History of Foot Ulcer 51 46.8% with microarray gene expression analysis. We thus show that augmented Past History of Amputation 13 11.9% inflammation and SP levels correspond to improved DFU healing. Use of Protective Foot Wear 31 28.4% Presence of Osteomyelitis 59 54.1% 657‑P

Delivering Mast Cell Stabilizers: A Novel Therapeutic Approach for POSTERS

Amputation in the Hospital for the Present Ulcer 10 9.2% Complications Diabetic Wound Healing Received Foot Care Education 90 82.6% SEEMA DANGWAL, ANA TELLECHEA, GEORGIOS THEOCHARIDIS, SUN LIJUN, Acute and Chronic Presence of Neuropathy 108 100% DAVID MOONEY, THEOHARIS THEOHARIDES, MANOJ BHASIN, ARISTIDIS VEVES, Boston, MA Diabetic foot ulcerations (DFU) represent one of the most debilitating and 655‑P costly complications of diabetes mellitus (DM), which lacks effective treat- Negative Pressure Wound Therapy Effect in Diabetic Foot Ulcer ment. Our previous studies show that in humans and mice, DM is associated May Be Mediated through Differential Gene Expression with increased skin mast cell (MC) degranulation, and that MC stabilization SEBASTIAN BORYS, AGNIESZKA LUDWIG-GALEZOWSKA, PRZEMYSLAW with systemic (i.p.) disodium cromoglycate (DSCG) improves wound healing in KAPUSTA, MICHAL T. SEWERYN, JULITA MACHLOWSKA, TERESA KOBLIK, DM mice. Here, we tested a new formulation delivering novel MC stabilizers JERZY HOHENDORFF, BEATA KIEC-WILK, PAWEL WOLKOW, MACIEJ T. MALECKI, (MCS) topically to the skin or wounds of DM mice. To that end, we devel- Kraków, Poland oped 12 indole-derived compounds and tested them in vitro. MCS-01 was the Diabetic foot syndrome (DFS) frequently leads to amputations. Negative most effective in inhibiting MC degranulation, and reducing pro-inflammatory pressure wound therapy (NPWT) has been successfully used for ulcerations mediator TNF-α and IL-8 release from human MC. Next, we developed algi- in DFS. However, its molecular mechanisms of action are not understood. nate dressings for topical sustained delivery of MCS-01 and applied them We assessed the effect of NPWT on gene expression. We included 21 to the shaved dorsum of DM mice for 10 days either before (pre-) or after type 2 diabetes (T2DM) patients with foot ulcers treated with NPWT and (post-) wounds were introduced. We evaluated the effect of topical MCS-01 8 T2DM patients treated by conventional debridement. The groups were pre- or post-wound on MC degranulation and wound healing in DM mice, and similar in terms of age: 69.0 ± 8.3 vs. 67.5 ± 4.3 years, sex: 80.9% vs. 75.0% compared it to i.p. DSCG pre-wound, as performed before. Topical MCS-01 male, T2DM duration: 14.7 ± 7.1 vs. 14.9 ± 6.0 years (p=NS for all compari- pre-wound reduced MC degranulation to the same degree as i.p. DSCG when sons), and other basic clinical characteristics. Tissue samples were obtained compared to placebo-treated mice. In agreement with our previous findings, from the bottom of the ulceration at two time points: before the therapy i.p. DSCG accelerated wound closure and increased re-epithelialization in DM was started and after 8 days. Total RNA was extracted and gene expression mice when compared to placebo, while similar results were observed with profiling was performed by means of Illumina human gene expression arrays. pre- and post-wound topical MCS-01. Systemic DSCG and topical MCS-01 Differential expression analysis of mRNAs was performed using the stan- polarized macrophage activation towards the regenerative M2 phenotype in dard R Bioconductor pipeline based on the ‘limma’ package. We identified the peri-wound area of DM mice. RNA sequencing data revealed upregulation 6 genes with differential expression (p<0.05) in the NPWT group, but not in of NFKB and STAT3 in MCS-01-treated groups when compared to placebo. Of the controls, between the two time points (after Benjamini-Hochberg cor- interest, NFKB correlated with wound closure. Taken together, our results rection for multiple testing). Expression of only one of them - RRP7A, which strongly suggest that topical treatment with the novel MC stabilizer MCS-01, is involved in rRNA processing - increased over 2-fold after treatment (logFC either pre- or post-wound, can ameliorate DM wound healing and this may be = 0.322, p=0.03), while the remaining 5 genes were downregulated. Two a promising therapeutic strategy for DFU. of the differentially regulated genes, CYP27A1 (logFC=-0.57, p=0.02) and Supported By: National Institutes of Health CLYBL (logFC=-0.08, p=0.03), associate with mitochondrial function. Two other genes, SRGAP3 (logFC = -0.14, p=0.01) and TRAPPC6A (logFC = -0.12, p=0.03), are associated with and Golgi apparatus, 658‑P Multidrug Resistant Bacteria: An Increasing Complication of Dia‑ respectively. Finally, the KIAA1683 gene (logFC = -0.1, p=0.03) encodes a betic Foot protein interacting with Calmodulin (CaM) messenger protein. ELISABETTA IACOPI, ALBERTO COPPELLI, CHIARA GORETTI, ENRICO TAGLIA- In summary, we found initial evidence that NPWT effect in diabetic foot FERRI, CARLO TASCINI, FRANCESCO MENICHETTI, ALBERTO PIAGGESI, Pisa, Italy ulcer may be mediated through differential gene expression. This finding Multigrug resistant (MDR) complicate diabetic foot ulcers (DFU) requires further confirmation. severely worsening their prognosis. The aim of this study was to assess Supported By: National Science Center of Poland (2013/11/B/NZ5/03298) the prevalence of MDR phenotypes of different bacterial strains in diabetic patients followed by our diabetic foot clinic from January 2001 until Decem- 656‑P ber 2014. We retrospectively analysed 7.826 culture results from deep The Role of Inflammation and in Diabetic Foot Ulcer wound specimens in diabetic patients (M/F: 6.065/1.761. Age: 63.2±11.7 Healing yrs) followed by our outpatient Clinic for DFU. From all bacterial strains we GEORGIOS THEOCHARIDIS, SEEMA DANGWAL, DIMITRIOS BALTZIS, ARISTIDIS selected those more prevalent and we analysed antimicrobial sensitivity VEVES, Boston, MA, Hannover, Germany pattern in relation to the more widespread antibiotic resistance pheno- Diabetic foot ulcers (DFU) constitute a serious health burden for dia- types. In particular, we evaluated the prevalence of aureus betic patients, significantly undermining their quality of life and leading to (SA), (PA) and Enterobacteriae (EB), sorting out extended hospitalization and to more than 80,000 lower extremity amputa- Methicillin-Resistant SA (MRSA), PA resistant to Ciprofloxacin (CiproRPA) tions per annum in the U.S. alone. There is evidence to suggest that the skin and Carbapenem (CRPA), EB resistant to Ciprofloxacin (CiproRE) or Extended of diabetic patients contains decreased levels of substance P (SP), a neuro- Spectrum Beta Lactamase producers (ESBL). To test if the MDR pattern peptide that functions as a neuromodulator and neurotransmitter. Recently changed overtime, we divided the obtained results in two groups: the first we demonstrated that SP expression is reduced in diabetic mouse skin and (Group A) included those from 2001 until 2007 while the second (Group B) treatment with SP enhanced wound healing in diabetic animals. We have from 2008 until 2014. SA was detected in 2.483 specimens in Group A and also previously shown in diabetic patients with DFU that increased mark- in 2.131 in Group B (NS), the presence of MRSA was 58.7% in Group A and ers of inflammation are associated with impaired wound healing. To further 51.2% in Group B (NS). PA was observed in 1.428 specimens in Group A and delineate the role of neuropeptides and inflammation in impaired wound heal- 1.783 in Group B (p<0.03): in particular, CiproRPA was detected in 45.1% of ing we completed a cross-sectional study including 23 healthy nondiabetic cultures in Group A and 64.1% in Group B (p< 0.04) while CRPA in 32.7% in subjects, 39 diabetic patients with DFU and 50 diabetic patients without Group A and 34.2% in Group B (ns). The presence of EB was detected in 1.516 DFU. The DFU patients were additionally divided into two groups: those who specimens in Group A and 2.032 in Group B (p<0.001); CiproRE prevalence completely healed the ulcer (healers) and those who did not (non-healers). was 28.0% in Group A and 47.7% in Group B (p< 0.02) while ESBL prevalence Increased levels of SP at baseline were associated with a quicker time for was 23.0% in Group A and 39.7% in Group B (p< 0.05). completion of healing. The concentrations of VEGF and IFNγ also correlated In conclusion, our data confirmed the high prevalence of MDR bacteria with improved healing. At the exit visit, the levels of TNFα and IL-8 in healers infections in DFU and their increasing overtime, stressing the importance of were increased. In addition forearm biopsies of healers exhibited increased a close monitoring of antimicrobial drugs susceptibility.

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A171 FOOT CARE—LOWER EXTREMITIES

659‑P Methods: We determined the diagnostic performance of nociception measurement in a prospective manner. Cases were patients either with dia- WITHDRAWN betes or prediabetes with neuropathy by the 1988 San Antonio Conference on Diabetic Neuropathy Consensus Statement. Controls were patients with lower back pain without neuropathy. All tests were made in a blinded man- ner. Sensibility and nociception were evaluated by measurement of needle pain threshold (NPT). A 23-gauge beveled needle was coupled to a 3 mL syringe barrel and weight was applied adding 2 grams to the other end at POSTERS

Complications each time. The lower weight that caused pain was registered. The differ-

Acute and Chronic ences in the minimum weights required to cause pain between the groups were compared and a ROC curve was constructed to determine the cutoff point from which the nociception measurement differentiates patients with and without polyneuropathy, and diagnostic accuracy tests were made. Results: 147 subjects were screened, from which 72 patients met inclu- sion criteria for neuropathy (age 59.56±12.46 years). Sixteen were paired controls (49.87±10.21 years). NPT was determined in 5.4 grams for each foot. Right foot 5.4 grams (AUC 0.927, 95% CI 0.826 to 0.995), left foot 5.4 grams (AUC 0.869, 95% CI 0.843 to 0.995). At this threshold point: sensibility 74.4% (CI 95% 0.636-0.832), specificity 97.1% (CI 95% 0.771-0.997), PPV 99.1% (CI 95% 0.919-0.999), NPV 47.1% (CI 95% 0.317-0.631). Conclusions: An objective assessment of NPT for nociception is a low- cost useful screening method for diabetic neuropathy. If these findings can be replicated in a larger population, it could be clinically useful as a screen- ing method for asymptomatic diabetic neuropathy.

662‑P WITHDRAWN

660‑P A Predictive Model to Identify Individuals with Diabetes at High Risk for Developing Foot Wounds Using Administrative Data and Medical Records MOLU SHI, DAVID STEENHARD, YANTING DONG, SHANNON HORSLEY, TODD PREWITT, STEPHANIE WEIDENBORNER, VIPIN GOPAL, Louisville, KY Foot wounds are one of the most severe complications among people with diabetes. Approximately 25% of individuals with diabetes are at risk of developing a foot wound over their lifetime, adversely affecting their health and well-being. A leading cause of hospitalizations of individuals with diabe- tes, foot wounds precede 84% of lower extremity amputations and can lead to lower survival rates, with post-diagnosis 3-year mortality rate as high as 28%. In this study, a predictive model was developed to determine the risk of developing a foot wound in the next year for individuals with diabetes enrolled in a Medicare Advantage plan. A broad set of data sources were used, including administrative medical and pharmacy claim information, lab data, demographics, and health risk survey data. In addition, text analyt- ics were applied to extract relevant information, such as vital signs, clinical conditions, family history, medications, etc., from medical records and nurse notes. More than 3,000 risk factors from over 1 million individuals were extracted, such as clinical conditions, healthcare resource utilization (HRU), demographics, and lifestyle. Various feature selection and machine learning algorithms were used. Area under curve (AUC) was used to measure model performance and select the final model. The selected model achieved an AUC of 0.81 with a lift of 6.1 for foot wound incidence among the top 5% of individuals by their predicted risk compared to random selection. Top predic- tors included past HRU, in particular diabetes-related HRU, previous wound, peripheral vascular disease, neuropathy, podiatrist visits, heart failure, and others. Work is currently underway to apply this model to identify high risk 663‑P individuals for disease management programs to enable early intervention. Neutralization of miR-615 Accelerates Angiogenesis and Diabetic Wound Healing 661‑P BASAK ICLI, DENIZHAN OZDEMIR, XIN LIU, GIORGIO GIATSIDIS, KEVIN CROCE, Diagnostic Accuracy of Nociception Evaluation for Diabetic DENNIS ORGILL, MARK FEINBERG, Boston, MA Neuropathy Development of new blood vessels Microvascular dysfunction and inef- DIEGO ESPINOZA-PERALTA, JULIO MARTINEZ-SALAZAR, CESAR ALEJANDRO fective angiogenesis associated with diabetes may lead to a range of com- ARCE-SALINAS, JOSEFINA AGUAYO-ARMENDARIZ, JIMENA HERNANDEZ- plications including peripheral artery disease or impaired dermal wound ALARCON, JORGE DAVID ROGEL-MANZANARES, Hermosillo, Mexico, Mexico healing. is critical for tissue repair in response to injury in ischemic vascular City, Mexico diseases such as peripheral artery disease (PAD), or diabetic wound heal- Background: At the time of diagnosis, up to 20% of patients with diabetes ing. MicroRNAs (miRNAs) are small, single-stranded, non-coding RNAs that present distal symmetric polineuropathy. Metabolic changes that generate regulate the expression of target genes in a variety of pathophysiological nerve damage could be present at a threshold below 126 mg/dl. processes including cardiovascular disease. Using a microarray profiling Objective: We built a tool to assess nociception​​ in patients at risk of poly- approach, we identified that miR-615 (miR-615) expression was significantly neuropathy, diagnostic performance was evaluated. increased in plasma of patients with acute coronary syndromes (ACS) isch-

For author disclosure information, see page A751. & Moderated Poster Discussion ADA-Supported Research A172 DIABETESCATEGORY EDUCATION emic cardiovascular disease compared to healthy controls, and decreased knowledge and barriers to addressing inpatient hyperglycemia, 2) institu- in response to proangiogenic stimuli (e.g., VEGF, bFGF) in endothelial cells tional protocol and guideline development for inpatient glycemic manage- in vitro. In response to high glucose in HUVECs, in diabetic wounds of the ment, 3) real-time severe hyperglycemia alert per the Pharmacy Expert db/db mice and in skin biopsies from diabetic patients, miR-615 expression System for daily analysis, 4) with “Just in Time Teaching” to prescribers in was significantly increased. Functionally, overexpression of miR-615 mark- response to severe hyperglycemia alerts, 5) hospital correction order set edly inhibited endothelial cell growth, migration, NO release, and network- change from upper limit 350mg/dl to 299mg/dl. Glycemic data was collected tube formation, whereas miR-615 inhibition had the opposite effects. from the electronic health record (EHR) through a system-wide database. Mechanistically, in the presence of VEGF, miR-615 overexpression in ECs Baseline data was obtained between January and September of 2015. The decreased Akt- and eNOS-phosphorylation, whereas miR-615 inhibition had rate of severe hyperglycemia events was 37.69 per 1000 at risk patient days. the opposite effects. In-vivo inhibition of miR-615 using LNA-anti-miR-615 Outcome data was collected between October 2015 and November 2016. significantly increased wound healing in diabetic db/db mice. Furthermore, The QI interventions reduced the rate of severe hyperglycemia to 30.27 per using human skin as in a model of dermal wound healing model 1000 at risk patient days, a 20% reduction. of human skin , inhibition of miR-615 significantly increased CD31 Figure. staining reflecting enhanced angiogenesis. Elucidation of the role of miR-615 in angiogenesis may provide a novel therapeutic approach to treat impaired angiogenesis in the setting of diabetes. Supported By: American Diabetes Association (1-16-JDF-046 to B.I.) POSTERS

DIABETES EDUCATION Behavioral Medicine, Clinical Nutrition, Education, and Exercise Moderated Poster Discussion: Community Strategies to Improve Care Delivery (Posters: 664-P to 669-P), see page 16. & 664‑P Real-World Clinical Inertia amongst T2DM Patients: Clinical, Physi‑ cian, and Patient Viewpoints VICTORIA HIGGINS, ANDREA LEITH, MELISSA KELLY, AMY ELLAMS, Maccles- field, United Kingdom A recent survey exploring real-world treatment patterns between 2000- 2015 found HbA1c control has not improved post 2008, one reason being attributable to clinical inertia. This analysis examines potential factors asso- ciated with clinical inertia in a real-world setting. Data was drawn from 2016 U.S./EU T2DM Adelphi Disease Specific Pro- gramme (DSP). Diabetes specialists and primary care physicians (PCPs) com- Supported By: Foundation for Barnes-Jewish Hospital pleted physician-reported forms for the next 10 consulting patients. Patients completed their own form. Current therapy duration vs. current HbA1c was & 666‑P applied to classify 3 groups: 1) dynamic: <6 month treatment change; any Ocho Pasos a la Buena Salud (Eight Steps to Better Health): A Suc‑ HbA1c; 2) non-dynamic/controlled: >6 months; <7.5%; 3) non-dynamic/ cessful Community Group Education Program for Latino Adults With uncontrolled: >6 months; >7.5%, i.e., clinical inertia. Adherence was mea- or At Risk for Type 2 Diabetes sured via the validated, patient-reported Morisky Medication Adherence MARY CONNEELY, SANDY ANDREWS, CEARA AXELROD, WENDY BEVIER, Scale (MMAS-8). ELOISA CHAVEZ, JENIFER SWARTZENTRUBER, DAVID KERR, Santa Barbara, CA A total of 352 specialists, 501 PCPs and 7487 patients were included. Of Type 2 diabetes (T2DM) is more prevalent in Latino adults compared to these, 32% were dynamic, 52% non-dynamic/controlled and 16% clinically non-Hispanic whites. Unfortunately, Latinos face serious health care dis- inert. Clinically inert patients, compared to dynamic, were more likely to be parities resulting in challenges for them in achieving optimal control of their consulting a PCP (55% vs. 45%), >65 years old (42% vs. 31%), retired (44% vs. diabetes. Ocho Pasos (Eight Steps) is an education program designed for 33%), have a caregiver (10% vs. 7%), have consulted a healthcare practitio- low-income, low-literacy Spanish-only speaking adults with or at risk for ner more often per annum (4.6 vs. 3.7), present with longest diabetes dura- T2DM. Classes are taught over 9 weeks in Spanish in a community setting. tion (8.5 vs. 5.0 [years]), be more likely to experience hypoglycemic episode The core activity focuses on transitioning to diabetes-friendly habits based on current regimen (21% vs. 10%), have lowest adherence (20% vs. 16%) on dietary modifications compatible with culturally appropriate food tradi- and highest physician-perceived CV-risk (29% vs. 24%) coupled with high- tions. Other aspects cover T2DM causes, self-monitoring, stress manage- est number of comorbidities (3.4 vs. 2.8). No observed differences between ment, and physical activity using images, food models, and other visual and gender, BMI, smoking status. tactile teaching aids. Pasaportes de Salud (Wellness Passports) monitor Clinically inert patients represent a group of older diabetics presenting key health measures and progress through the class. Between July 2015 with lower adherence and a higher degree of risk factors (CV and hypogly- and 2016, 273 Latino adults were enrolled [78% women), mean (±SD) age cemic events). Despite frequent consultation yet no therapy intervention 48.7 (±11.1) years]. For 60 individuals, there were pre-class and post-class in past 6 months, identification and understanding of these patients could values for HbA1c, BMI, and waist circumference (WC). For 16 individuals, further improve personalization of treatment pathways to achieve optimal HbA1Cc was ≥6.5%, between 5.7-6.4% for 20, and <5.7% for 24. Overall, diabetes control. HbA1c decreased by 4% (6.3±1.4% to 6.0±1.0%). For those with HbA1c in the diabetes range, it decreased by 10% (8.1±1.7% to 7.2±1.3%). For those & 665‑P with a prediabetes HbA1c, it decreased by 4% (6.0±0.2% to 5.7±0.3%). All A System Approach to Glycemic Management and Prevention of changes were significant P<.01. Overall, BMI did not change significantly, Severe Hyperglycemia in the Non-Critically Ill Hospitalized Patients 31.5±6.3 kg/m2 to 31.7±6.1 kg/m2. However, WC did decease significantly MARY C. BLACKBURN, CYNTHIA HERRICK, PAULINA CRUZ BRAVO, MICHAEL (P<.01), 103.9±14.5cm to 98.0±12.2cm. For those with HbA1c ≥6.5%, WC ELLIOTT, KEVIN HEARD, ELIZABETH O’CONNOR, KARIN STERL, GARRY S. TOBIN, decreased from 113.0±15.7cm to 102.6±6.6 cm. Ocho Pasos is effective for St. Louis, MO delivering community-based diabetes education targeting low income, low Severe hyperglycemia (>299mg/dl) in inpatients with or without diabetes literacy Latinos with an established diabetes diagnosis or at risk for develop- is associated with poor hospital outcomes, including infections, prolonged ing T2DM. length of stays, and poorer outcomes after the hospital stay.1-3 We designed a quality improvement (QI) project at Barnes-Jewish Hospital (BJH), an 1100 bed tertiary care-teaching hospital to reduce severe hyperglycemia and improve glycemic management. The QI team planned and coordinated the following interventions: 1) provider survey to understand practitioners’

ADA-Supported Research & Moderated Poster Discussion For author disclosure information, see page A751. A173