bluuebunny/crossref_metadata_2025 huggingface dataset:
1940-P: Disruption of the Exocyst Trafficking Complex Results in Impaired Glucose Uptake in Skeletal Muscle
Authors: FUJIMOTO BRENT , CARTER LAMAR T. , WONG ADRIAN M. , PITTS MATTHEW W. , VILLIGER ROSS K. , YOUNG MADISON , FOGELGREN BEN , POLGAR NOEMI
Published: June 2019

Objectives: Skeletal muscle cells are responsible for 80-90% of the insulin-induced glucose uptake in the body. Insulin signaling in skeletal muscle results in the targeted trafficking of glucose transporter type 4 (GLUT4) onto the cell membrane, enabling glucose uptake. Insulin resistant cells show defects in insulin-induced GLUT4 exocytosis. The eight-protein exocyst complex has an essential role in the insulin-induced exocytosis of GLUT4 vesicles in cultured adipocytes but it is not known if the exocyst-mediated molecular mechanism is conserved in other, insulin-responsive tissues, such as the skeletal muscle. We hypothesized that the exocyst complex is essential for the insulin-induced exocytosis of GLUT4-containing vesicles in skeletal muscle as well and that the exocyst is a master regulator of glucose homeostasis in insulin-responsive tissues.

Methods/Results: We have generated a tamoxifen-inducible skeletal muscle-specific knockout mouse strain of exocyst central subunit Exoc5 (Exoc5-SMKO) to assess the exocyst’s role in glucose homeostasis in vivo. Exoc5 knockout does not affect grip strength, motor coordination or locomotor activity levels in these animals. Both male and female Exoc5-SMKO mice present with elevated fasting glucose levels, as compared to control littermates. Glucose tolerance testing revealed an impaired glucose clearance in Exoc5-SMKO mice, while insulin tolerance, fasting insulin levels, and A1C levels were similar between knockouts and controls.

Conclusion: Our findings suggest that Exoc5 and the exocyst are necessary for insulin-stimulated glucose uptake in skeletal muscle. Ongoing work will further investigate the molecular mechanism of exocyst-mediated GLUT4 trafficking in skeletal muscle.

Disclosure

B. Fujimoto: None. L.T. Carter: None. A.M. Wong: None. M.W. Pitts: None. R.K. Villiger: None. M. Young: None. B. Fogelgren: None. N. Polgar: None.

Funding

National Institute of General Medical Sciences (5P20GM113134)
1946-P: Insulin-Like Growth Factor-1 Protects against the Detrimental Effects of Advanced Glycation End Products and High Glucose in Myoblastic C2C12 Cells
Authors: KANAZAWA IPPEI , ADACHI NAOKO , TANAKA KEN-ICHIRO , TAKENO AYUMU , NOTSU MASAKAZU , TANAKA SAYURI , SUGIMOTO TOSHITSUGU
Published: June 2019

Previous studies have shown that diabetes mellitus increases the risk of sarcopenia. We previously reported that higher serum advanced glycation end product (AGE) and lower serum insulin-like growth factor-I (IGF-I) were independently associated with muscle mass reduction in patients with type 2 diabetes. The present study examined effects of AGEs and IGF-I on myogenic differentiation and apoptosis in C2C12 cells. Real-time PCR and western blot analyses showed that AGE3 significantly decreased the mRNA and protein expressions of MyoD and Myogenin, whereas IGF-I significantly increased them and reversed the effects of AGE3. AGE3 significantly decreased endogenous IGF-I expression and suppressed IGF-I-induced Akt activation. High glucose (22 mM), but not AGE3, significantly increased expression of receptor for AGEs (RAGE), while IGF-I significantly decreased it. Moreover, DNA fragment ELISA showed that AGE2 and AGE3 significantly increased apoptosis of C2C12 cells, whereas IGF-I significantly suppressed the AGE2- and AGE3-induced apoptosis. To investigate whether high glucose enhances the AGE3’ effects, the effects of co-treatment with high glucose and AGE3 were examined. Although high glucose alone did not affect the apoptosis of C2C12 cells, it enhanced AGE3-induced apoptosis. IGF-I significantly reversed the effects of co-incubation of high glucose and AGEs on the expression of MyoD and Myogenin as well as the apoptosis. These findings indicate that AGE3 inhibits myogenic differentiation and increases apoptosis in C2C12 cells, and that high glucose increases RAGE and enhances the AGE3-induced apoptosis, suggesting that AGEs and high glucose might contribute to the reduction of muscle mass and function. Moreover, IGF-I reversed the detrimental effects of AGEs and high glucose in myoblastic cells; thus, IGF-I-Akt signal could be a therapeutic target of DM-induced sarcopenia.

Disclosure

I. Kanazawa: None. N. Adachi: None. K. Tanaka: None. A. Takeno: None. M. Notsu: None. S. Tanaka: None. T. Sugimoto: None.
1947-P: Comparative Effects of PCSK9 Inhibitor and High-Dose Atorvastatin on Mitochondria of Red Muscle Fibers in Obese Female Rats
Authors: THONUSIN CHANISA , PALEE SIRIPONG , ARUNSAK BUSARIN , AMPUT PATCHAREEYA , PRATCHAYASAKUL WASANA , CHATTIPAKORN NIPON , CHATTIPAKORN SIRIPORN C.
Published: June 2019

Obesity and dyslipidemia cause mitochondrial dysfunction of skeletal muscle. Low dose atorvastatin impairs mitochondrial function of white muscle fibers but does not damage that of red muscle fibers. However, the effects of PCSK9 inhibitors and high dose atorvastatin on mitochondrial function of red muscle fibers have not been identified. Twenty-four female rats (180-200 g) were divided into 2 groups. Six rats were fed with normal diet (ND) and 18 rats were fed with high-fat diet (HFD) for 15 weeks. At week 12, ND-fed rats were treated with vehicle (NDV), while HFD-fed rats were equally subdivided and treated with either vehicle (HFV), 4 mg/kg/day of PCSK9 inhibitor (HFP), or 40 mg/kg/day of atorvastatin (HFA) for another 3 weeks. HFV rats developed obesity and dyslipidemia, as indicated by increased weight, visceral fat, and total cholesterol. Treatment with atorvastatin and PCSK9 inhibitor improved obesity and dyslipidemia. Increased lipid peroxidation (MDA), mitochondrial fission (pDrp1/Drp1), mitochondrial ROS production, and mitochondrial membrane depolarization were found in soleus muscle of HFV rats. Treatment with atorvastatin and PCSK9 inhibitor similarly improved lipid peroxidation and mitochondrial fission. However, only PCSK9 inhibitor improved mitochondrial ROS production and mitochondrial membrane depolarization. Our findings suggest that obesity increases lipid peroxidation, mitochondrial fission, mitochondrial ROS production, and mitochondrial membrane depolarization in red muscle fibers. PCSK9 inhibitor attenuates those deleterious effects of obesity. High dose atorvastatin improves lipid peroxidation and mitochondrial fission, but does not attenuate obesity-induced mitochondrial ROS production and mitochondrial membrane depolarization in red muscle fibers. The results imply that high dose atorvastatin causes adverse effects on red muscle fibers despite its benefit on the improvement of obesity and dyslipidemia.

Disclosure

C. Thonusin: None. S. Palee: None. B. Arunsak: None. P. Amput: None. W. Pratchayasakul: None. N. Chattipakorn: None. S.C. Chattipakorn: None.

Funding

Thailand Research Fund
1948-P: Secretion of the Short-Form Gastric Inhibitory Polypeptide in Nondiabetic and Diabetic Subjects
Authors: TAKEDA YASUTAKA , FUJITA YUKIHIRO , YANAGIMACHI TSUYOSHI , MARUYAMA NOBUHIRO , BESSHO RYOICHI , SAKAGAMI HIDEMITSU , HANEDA MASAKAZU , OTA TSUGUHITO
Published: June 2019

We previously reported that the short-form gastric inhibitory polypeptide (GIP) 1-30 is released from islet alpha cells and promotes insulin secretion in a paracrine manner in vitro. However, the role played by GIP 1-30 in glucose metabolism in vivo remains unclear. We developed an enzyme-linked immunosorbent assay (ELISA) specific for GIP 1-30 and measured GIP 1-30 secretion in nondiabetic subjects (ND, n = 8) and patients with type 2 diabetes (T2D, n = 9). We developed a sandwich ELISA by combining a novel antibody to the GIP C-terminus with the N-terminal anti-GIP 1-42 antibody that is already available. We explored cross-reactivities with incretins and glucagon-related peptides. We next subjected ND and T2D subjects to the cookie meal test (CMT: carbohydrates 75 g, lipids 28.5 g, proteins 8.5 g) and measured GIP 1-30 blood levels. Absorbance increased in a dose-dependent manner on addition of the GIP 1-30 amide but not GIP 1-42, GLP-1, or glucagon. Post-CMT loading, GIP 1-30 concentrations increased in both ND and T2D subjects; however, in ND subjects, the increases were much lower than those of GIP 1-42 (GIP 1-30, before loading: 1.4 ± 0.5 pmol/L, after: 1.9 ± 0.6 pmol/L; GIP 1-42, before: 8.2 ± 0.1 pmol/L, after: 204.0 ± 35.2 pmol/L, P<0.05). In T2D patients, the GIP 1-30 levels, as assessed by the areas under the curves, tended to be lower than those in ND patients. The DPP-4 inhibitor significantly increased both GIP 1-30 and GIP 1-42 secretion in T2D patients.

In conclusion, we developed a novel ELISA that is highly specific for GIP 1-30, the secretion of which was promoted by a mixed meal to a blood level lower than that of GIP 1-42. As is also true of the incretins, GIP 1-30 levels increased upon administration of DPP-4 inhibitor; GIP 1-30 secretion may differ between ND and T2D subjects.

Disclosure

Y. Takeda: None. Y. Fujita: None. T. Yanagimachi: None. N. Maruyama: Employee; Self; Immuno-Biological Laboratoties Co.,Ltd. R. Bessho: None. H. Sakagami: None. M. Haneda: None. T. Ota: None.

Funding

Japan Society for the Promotion of Science
1961-P: Identification of a Novel Brown/Beige Fat-Derived Adipokine
Authors: CHOI CHAN HEE J. , CRANE AUDREY A. , COHEN PAUL
Published: June 2019

Fat cells or adipocytes play a central role in obesity, diabetes, and metabolic disease. White adipocytes store excess energy in the form of triglycerol, and brown adipocytes dissipate chemical energy into heat. In addition to these bioenergetics functions, adipocytes also have important endocrine functions. While several adipocyte-derived secreted factors, or adipokines, have been shown to have important physiological roles, bioinformatic analyses suggest that adipocytes may secrete more than 1,000 unique polypeptides. The vast majority of these gene products remain uncharacterized, representing a major opportunity for discovering new biology with translational potential. To that end, we have employed chemical and genetic tools to label and enrich circulating proteins in a cell-selective manner. Using bio-orthogonal non-canonical amino acid tagging (BONCAT) along with mass spectrometry we have characterized the secretome of white, brown, and beige adipocytes. We have identified more than 800 unique proteins, with approximately 300 showing fat cell type specific enrichment. Our studies have focused on a brown and beige fat specific protein encoded by a previously uncharacterized open reading frame (BAT-ORF). The mRNA encoding this protein is expressed in UCP1+thermogenic adipocytes, induced by cold and repressed by warm temperature, and upregulated following thiazolidinedione treatment. We have found that the BAT-ORF gene encodes two isoforms, only one of which is secreted. We have generated mice with genetic deletion of each isoform using CRISPR-Cas9. Preliminary data suggests these mice have decreased energy expenditure. Complete characterization of these mice is underway and data will be presented. We will also present a method using genetically engineered mouse models that allows us to specifically label and enrich secreted proteins in the blood in a cell-type specific fashion.

Disclosure

C.J. Choi: None. A.A. Crane: None. P. Cohen: None.

Funding

American Diabetes Association/Pathway to Stop Diabetes (1-17-ACE-17 to P.C.)
1987-P: Nampt Deletion Changes Metabolism in Adipose Tissue and Prevents Diet-Induced Obesity
Authors: OKABE KEISUKE , NAWAZ ALLAH , FUJISAKA SHIHO , YAGI KUNIMASA , KADO TOMONOBU , USUI ISAO , NAKAGAWA TAKASHI , TOBE KAZUYUKI
Published: June 2019

Nampt is the rate-limiting enzyme in salvage pathway in which NAD is mainly synthesized in mammalian cells. NAD plays very important role in various biological processes, such as energy production, energy consumption, aging, cell death, and DNA damage repair as a coenzyme of redox reactions or as a substrate of Sirtuins or Poly (ADP-ribose) polymerases. Besides, Nampt is highly expressed in mature adipocyte and secreted into extracellular. The extracellular Nampt is known to regulate systemic metabolic state. In short, Nampt works as a metabolic sensor and NAD integrates systemic metabolism. Recently, metabolism is thought to be closely related to cell differentiation. However, how NAD metabolism affects to adipogenesis remains unknown. In this study, we generated the adipose tissue specific Nampt knock out mice, Adiponectin-Cre Nampt flox/flox (KO) and conducted targeted-metabolomic analysis to investigate the metabolic effect, employing liquid-chromatograph mass spectrometry and gas-chromatograph mass spectrometry. Interestingly, KO showed resistant against diet-induced obesity (DIO), fed high fat high sucrose diet. Body weight and both inguinal and epididymal fat mass were significantly reduced in KO compared to wild type control mice. On the other hand, we revealed that Nampt-mediated NAD synthesis was up-regulated during differentiation of 3T3-L1 preadipocytes and the inhibition of Nampt significantly suppressed the differentiation. Our metabolomic analysis and ChIP-qPCR assay showed NAD contributed to the differentiation through the metabolic reprogramming and the subsequent epigenetic alteration to activate the transcription of adipogenic genes. This result indicated Nampt-mediated NAD synthesis regulated differentiation of preadipocytes to mature adipocytes. Together, our data suggested that adipose tissue specific Nampt deletion resulted in the resistance against DIO and it could be partly attributed to the inhibited differentiation of preadipocytes.

Disclosure

K. Okabe: None. A. Nawaz: Research Support; Spouse/Partner; Astellas Pharma Inc., Astellas Pharma Inc., AstraZeneca, Daiichi Sankyo Company, Limited, Eli Lilly and Company. Research Support; Self; Japan Society for the Promotion of Science. Research Support; Spouse/Partner; Merck Sharp & Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Novo Nordisk A/S, Ono Pharmaceutical Co., Ltd., Pfizer Inc., Sanofi, Shionogi & Co., Ltd., Takeda Pharmaceutical Company Limited. S. Fujisaka: None. K. Yagi: None. T. Kado: None. I. Usui: None. T. Nakagawa: None. K. Tobe: Advisory Panel; Self; Novo Nordisk A/S. Board Member; Self; Japan Diabetes Society. Research Support; Self; Bristol-Myers Squibb Company.

Funding

Japan Society for the Promotion of Science
1998-P: Progressive Liver Dysfunction Resulted from a High-Fat, High-Fructose Diet in Fatzo Mice
Authors: WANG XIAOLI , YE WEIWEI , CHEN YANG , ZHANG XIAOYU , WANG YIXIN , XIAO YONG-FU
Published: June 2019

The FATZO mouse is a polygenic model with an intact leptin system and was developed by cross breeding of C57BL6/J (C57) with AKR/J. FATZO mice have been used in many dysmetabolic studies. The epidemic health issue of nonalcoholic fatty liver disease (NAFLD, including NASH) is the organ dysmetabolic manifestation. This study was to investigate a high fat/fructose diet (HFFD) on liver function in FATZO mice. The body weights of FATZO (n=30) at age of 9 to 10 weeks were significantly heavier than those of age-matched C57 (n=19) mice (39±0.4g vs. 23±0.2g, p<0.001). The serum glucose levels in FATZO were higher than those in C57 (212±5mg/dL vs. 131±7mg/dL). The body weights were gradually increased during 5-month feeding with HFFD or 5C08 (Figure 1). FATZO+HFFD mice significantly elevated serum AST, ALT and ALP, compared with FATZO+5C08, C57+HFFD and C57+5C08 mice. These elevations in FATZO+HFFD mice reached peak in 3-month feeding and then maintained at the high levels (Figure 1). Serum TC, HDL and LDL (except TG) were also significantly elevated in FATZO+HFFD mice. Interestingly, serum glucose and HbA1c levels were significantly reduced in FATZO+HFFD mice (from 216±7mg/dL and 6.2±0.1% at the baseline to 169±5mg/dL and 4.3±0.1% after 5-month feeding, respectively). Our data demonstrate that the HFFD progressively caused liver dysfunction in FATZO, but not C57. FATZO + the HFFD could thus be a very valuable model for NAFLD/NASH research.

Disclosure

X. Wang: None. W. Ye: None. Y. Chen: None. X. Zhang: None. Y. Wang: None. Y. Xiao: Employee; Self; Crown Bioscience Inc.
201-OR: ADA Presidents' Select Abstract: Shared Medical Appointments (SMA) Improve Psychosocial Outcomes in Adolescents with Type 1 Diabetes (T1D)
Authors: RAYMOND JENNIFER , REID MARK W. , BERGET CARI , CAIN CINDY L. , ANDERSON BARBARA , KLINGENSMITH GEORGEANNA J.
Published: June 2019

Background: T1D care delivery via SMA is associated with increased attendance and satisfaction in adolescents, but the impact of the model on clinical and psychosocial outcomes remains unclear.

Objective: Evaluate the efficacy of SMA for improving psychosocial outcomes in adolescents with T1D.

Methods: Participants in the prospective Team Clinic study were randomized to attend SMA or individual appointments (usual care). They completed the Children’s Depression Inventory (CDI), Problem Areas in Diabetes (PAID), Diabetes Family Conflict Scale (DFCS), and Diabetes Strengths and Resilience (D-STAR) measures at baseline and after one year.

Results: Eighty-six adolescents (51% female; 74% white; 10% Latinx) completed at least one study visit. Team Clinic participants (n = 44) attended more visits than patients in usual care (n = 42; p < 0.0001), and they attended all four study visits more frequently (66%) than those in usual care (33%). Although all adolescents reported nearly equal levels of depressive symptoms, emotional distress, and family conflict associated with diabetes care at baseline, significant differences emerged over the course of the study. Specifically, adolescents in Team Clinic reported reduced levels of conflict with family, while those in usual care reported slightly increased levels of conflict (p=0.03). Team Clinic adolescents also reported reduced depressive symptoms when compared to usual care, including more positive mood (p=0.001), greater self-esteem (p=0.03), improved effectiveness in completing daily tasks (p=0.03), and reduced anhedonia (p=0.04). Team Clinic adolescents also reported improved resilience and reduced emotional distress in diabetes care compared to controls, but these results were only marginally significant.

Conclusions: SMA may support adolescents with T1D in reducing familial conflict associated with diabetes care, while providing a decrease in comorbid depressive symptoms.

Disclosure

J. Raymond: Other Relationship; Self; Insulet Corporation. M.W. Reid: None. C. Berget: Consultant; Self; Insulet Corporation, Medtronic. C.L. Cain: None. B. Anderson: None. G.J. Klingensmith: Consultant; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Takeda Pharmaceutical Company Limited. Research Support; Self; Novo Nordisk Foundation. Stock/Shareholder; Spouse/Partner; Dexcom, Inc.

Funding

The Leona M. and Harry B. Helmsley Charitable Trust
2014-P: The Effect of High-Fat Diet on Adipose Tissue Glycerol Metabolism Is Influenced by Sex and GLP-1 Agonist Treatment
Authors: IENA FRANCESCO M. , VEGGER JENS BAY , THOMSEN JESPER SKOVHUS , BRÜEL ANNEMARIE , LEBECK JANNE
Published: June 2019

During fasting triglyceride (TG) in adipose tissue (AT) is hydrolyzed into fatty acids and glycerol. Glycerol exits AT through the glycerol channel aquaporin 7 (AQP7). AQP7 KO mice have increased activity of AT glycerol kinase (GlyK), increased storage of TG, and become obese indicating that changes in AT glycerol metabolism influences TG storage. Moreover, in humans an association between low expression of AQP7 and obesity has been found in women only. This study aims to investigate whether high fat diet (HFD) influences AT glycerol metabolism in a sex-specific manner and whether the glycerol metabolism is influenced by GLP-1 agonist treatment. Female and male C57BL/6J mice (10 weeks) were fed a control diet or HFD for 12 or 24 weeks. The GLP-1 agonist Liraglutide was administered (SC, 1 mg/kg/day) to a subgroup of control and HFD fed female mice during the last 12 of the 24 weeks. Body weight (BW) and blood glucose (BG) were monitored and the expression of AQP7, GlyK, and lipolytic enzymes were investigated using immunoblotting of perigonadal AT and analyzed using Two-Way ANOVA. After 12 weeks, HFD increased BW by 34% in female and by 49% in male mice. BG levels were significantly increased in male mice only. After 24 weeks, HFD increased BW by 76% in female and by 49% in male mice and BG levels was significantly increased in both sexes. Liraglutide decreased both BW and BG levels compared to untreated controls. HFD increased the expression of AQP7 in female mice only and after 24 weeks of HFD this was paralleled by a more than 4-fold increase in GlyK and a 50% reduction in adipose triglyceride lipase abundance (ATGL). Liraglutide treatment normalized the expression of both AQP7 and GlyK. In male mice the abundance of GlyK was increased after 12 and 24 weeks of HFD.

In conclusion, HFD influences the expression of AQP7 in AT in a sex-specific manner. The increase in GlyK suggests an increased utilization of glycerol for TG synthesis in both sexes. Liraglutide treatment influences AT glycerol metabolism.

Disclosure

F.M. Iena: None. J. Vegger: None. J. Thomsen: None. A. Brüel: None. J. Lebeck: None.

Funding

Aarhus University
2015-P: 14-3-3zeta Is Required for PKA-Dependent Lipolysis
Authors: OPPONG ABEL , MUGABO YVES , LIM GARETH E.
Published: June 2019

The molecular scaffold, 14-3-3?, was previously found to be essential for visceral adipogenesis, but its contributions to the function of mature adipocytes is not known. As it can regulate the activities of metabolic effectors, we hypothesized that 14-3-3? also has essential roles in adipocyte function.

3T3-L1 adipocytes and mouse models were used to study if 14-3-3? regulates lipolysis. Depletion of 14-3-3? by siRNA abrogated glycerol and free fatty acid (FFA) release from 3T3-L1 cells treated with Isoproterenol (ISO, 1 μM), Forskolin (FSK, 10 μM), and dibutyryl cAMP (1 mM). In contrast, over-expression of 14-3-3? potentiated ISO-mediated FFA release. Knockdown of 14-3-3? did not affect cAMP generation in ISO- and FSK-treated 3T3-L1 cells, but mRNA levels of lipases (Atgl, Hsl, and Magl) and Pparg were reduced, suggesting a loss of adipocyte identity. Decreased activation and total expression of PKA substrates, including Hsl and CREB, were detected in 14-3-3?-depleted 3T3-L1 cells. Taken together, these data suggest that 14-3-3? is necessary for lipolysis from 3T3-L1 adipocytes.

To understand adipocyte-specific roles of 14-3-3?, tamoxifen (TMX)-inducible, adipocyte-specific 14-3-3? knockout (adi14-3-3?KO) mice were used. Four weeks after TMX exposure (5 days, 50 mg/kg), no effects on body weight were found. After an overnight fast, adi14-3-3?KO mice displayed impaired lipolysis following i.p CL-316,243 (1 mg/kg) injections. In contrast, transgenic over-expression of 14-3-3? did not affect lipolysis. Adi14-3-3?KO mice also displayed glucose intolerance following i.p. glucose (2 g/kg). Real-time PCR confirmed significant reductions in Atgl, Hsl, and Pparg mRNA levels in adi14-3-3?KO mice, suggesting impaired adipocyte function.

Collectively, these results demonstrate essential functions of 14-3-3? in facilitating lipolysis and, potentially, adipocyte maturity. Future studies are aimed at understanding how 14-3-3? regulates other aspects of adipocyte function, including diet-induced expansion of fat mass.

Disclosure

A. Oppong: None. Y. Mugabo: None. G.E. Lim: None.

Funding

Canadian Institutes of Health Research
2019-P: Deletion of Hepatic SIRT1 Enhances the Effect of Ketogenic Diets on Ameliorating Obesity and Metabolic Disorders
Authors: GUO WANRONG , XU FEN , LI GUANGRONG , LIANG HUA , CAI MENGYIN
Published: June 2019

A ketogenic diet, one kind of carbohydrate-restricted diets, is considered to reduce body weight and improve metabolic disorders. However, the mechanism of how a ketogenic diet provides such metabolic benefits is unclear. SIRT1, a NAD-dependent deacetylase, plays an important role in lipid metabolism. We investigated the role of hepatic SIRT1 in the beneficial effect of ketogenic diets on improving obesity and metabolic disorder. In this study, eight-week-old male liver-specific SIRT1 deletion (SIRT1 LKO) mice and control mice were subjected to a high fat diet (HFD) or an intermittent ketogenic diet (iKD), ketogenic diet alternated biweekly with HFD, for 12 weeks. The results showed that the body weight of mice fed on iKD was significantly lower than mice fed on HFD, even though higher energy intake was observed when mice were fed on a ketogenic diet. Interestingly, SIRT1 LKO mice on HFD were slightly heavier than control mice on HFD, but SIRT1 LKO mice on iKD were thinner than control mice on iKD. Lower fasting blood glucose was observed after every 2-week ketogenic diets intervention, and there was no significant difference between SIRT1 LKO mice and control mice. Compared to HFD, iKD significantly improved insulin resistance. Notably, iKD improved the glucose metabolism of SIRT1 LKO mice, but such improvement was not observed in control mice. These results indicate that an intermittent ketogenic diet improves metabolic disorders induced by HFD and has further beneficial effect on obesity and metabolic disorder in the absent of hepatic SIRT1. The underlying mechanism will be further studied.

Disclosure

W. Guo: None. F. Xu: None. G. Li: None. H. Liang: None. M. Cai: None.
203-LB: Comparison of Health-Related Quality of Life Between Elderly Men and Women with Diabetes in Korea
Authors: KANG HYUN SOOK
Published: June 2019

Objective: This study aimed to identify the differences in health-related quality of life between elderly men and women with diabetes in Korea.

Methods: Health-related quality of life was measured using the Korean version of the EQ-5D (EuroQol 5-dimension). Using a descriptive study, this study incorporated a secondary analysis of the Korean version of the EQ-5D data from the 10th wave of the Korea Health Panel Survey in 2015.

Results: Total 1011 patients (aged ≥65 years), including 446 men and 565 women. The elderly Korean men with diabetes scored significantly higher on their quality of life than the elderly women with diabetes (t=-6.083, p=.000). The elderly women with diabetes had more problems in all EuroQol ED-5Q dimensions than did the elderly Korean men with diabetes. Subjective health status (t=-5.063, p=.000) and peer-to-peer health status (t=-5.073, p=.000) were also perceived to be worse among women compared with those among men. There were significant differences in education (X2=232.888, p=.000), marital status (X2=123.090, p=.000), economic activity (X2=185.585, p=.000), drinking status (X2=106.444, p=.000), physical activity (X2=35.581, p=.000), and visual acuity (X2=7.024, p=.030) among the elderly men and women with diabetes.

Conclusion: Based on these results, there is a need for specific health care programs targeting elderly women with diabetes that may help improve their health-related quality of life.

Disclosure

H. Kang: None.
2033-P: APX-115, a Pan-NADPH Oxidase (NOX) Inhibitor, Ameliorates Diabetic Nephropathy in NOX5 Transgenic Mice
Authors: LEE EUNSOO , CHUNG CHOON H. , HUH JI HYE , LEE SUN HEE
Published: June 2019

Several studies verified that NOXs which is comprise isoform NOX1 to 5, Duox 1 and 2, trigger diabetic nephropathy incidences in diabetes. Recently, it has been studied that the expression of NOX5 is increased in the patients with diabetic nephropathy over other isoforms and the importance of NOX5 in the development of therapeutic agents has been suggested. Previous study investigated that APX-115 has the effect of pan-NOX inhibition in type 2 diabetic mice.

However, since the NOX5 gene is absent in the mouse, we evaluated the pan-NOX inhibitory of APX-115 in Nox5 transgenic mouse. Wild type and renal podocyte specifically NOX5 transgenic mice (NOX5 pod+) were fed high fat diet (60% kcal fat) and treated APX-115 (60mg/kg) by oral gavage for 14 weeks.

APX-115 significantly improved pancreatic beta cell function with decreased fasting blood glucose and increased insulin levels. And serum total cholesterol, triglycerides and urinary albumin/creatinine levels were also significantly decreased by APX-115 treatment. In NOX5 pod+ mice kidney, increased NOX5 mRNA expression, increased desmin and reduced podocin protein expressions were also significantly recovered by APX-115. Moreover, APX-115 inhibited inflammatory related protein expressions such as TRAF6.

Therefore, these data suggest that APX-115 might be a promising therapy for diabetic nephropathy by an efficacy as a pan-NOX inhibitor including NOX5 inhibition and anti-inflammatory effects.

Disclosure

E. Lee: None. C.H. Chung: None. J. Huh: None. S. Lee: None.
2053-P: Low AMY1 Gene Copy Number Is Associated with Increased Childhood Obesity in Alabama
Authors: KURAPATY VENKATAPOORNA CHANDRA MOHAN , AYINE PRISCILLA , PARRA EMILY P. , KOENIGS TAYLOR , SANDEY MANINDER , JEGANATHAN RAMESH B. , THANGIAH GEETHA
Published: June 2019

Salivary amylase (AMY1) is the most abundant enzyme in human saliva, responsible for the hydrolysis of α-1,4 glycosidic linkages that aids in the digestion of dietary starch. Recently few studies have shown that copy number of AMY1 is associated with obesity, however, the data varies with different location. One-third of children are overweight/obese in Alabama. In this study, we specifically aim to determine the relationship between the copy number of AMY1 gene and obesity measurements in elementary school children from Alabama. Sixty elementary school children (30 normal weight and 30 overweight/obese) aged between 6 to 10 years participated in this study. Anthropometric measurements were measured using WHO recommendations. Saliva was collected, genomic DNA was extracted and the copy number of the AMY1 gene was estimated by digital PCR. Linear regression analysis was used to investigate the association between AMY1 copy number and obesity measurements. The mean AMY1 copy number was found to be significantly decreased in obese compared to normal weight children. The results showed a significant inverse association between obesity measurements and AMY1 copy number. Gender-specific analysis showed boys had a stronger association between low AMY1 copy number and obesity compared to girls. Our findings suggest that low AMY1 copy number plays a major role in childhood obesity in Alabama elementary school children and the effect is more prominent in boys compared to girls.

Disclosure

C. Kurapaty Venkatapoorna: None. P. Ayine: None. E.P. Parra: None. T. Koenigs: None. M. Sandey: None. R.B. Jeganathan: None. G. Thangiah: None.

Funding

National Institute of Food and Agriculture
2056-P: Psychiatric Profile and Quality of Life in Subjects with Excess Weight Treated with Transcranial Direct Current Stimulation Combined with Hypocaloric Diet
Authors: NATIVIDADE GABRIELLA RICHTER , ARAUJO CARINA DE , FITZ RAQUEL C. , OSÓRIO AMANDA F. , MERELLO PAULA NUNES , SCHESTATSKY PEDRO , GERCHMAN FERNANDO
Published: June 2019

Transcranial direct current stimulation (tDCS) of the right dorsolateral prefrontal cortex (DLPFC) may reduce appetite and caloric intake. However, it is used as an adjunct treatment for depression by stimulation of the left DLPFC. As a result, its safety in mental health and its impact on quality of life of subjects with excess weight needs to be addressed.

Objective: To assess the effect of repeated tDCS of the right DLPFC on mood, daytime sleepiness, anxiety and quality of life in subjects with excessive weight on a hypocaloric diet.

Methods: We randomly assigned 28 participants to receive 20 sessions of active or sham tDCS for 20 consecutive weekdays. Anode and cathode electrodes were respectively placed on the right and left DLPFC. The Beck Depression Inventory (BDI), a daytime sleepiness questionnaire (DSQ), State-Trait Anxiety Inventory-State (STAI-S) and the 36-Item Short Form Health Survey (SF-36) were administered before (t0) and after (t20) the intervention period.

Results: There were no statistically significant changes in BDI, DSQ, STAI-S and SF-36 subscales between the groups throughout the study (Figure 1A and 1B).

Conclusion: The absence of significant changes in the scales suggests that tDCS is not associated with impairment in mental health and quality of life in this population. ClinicalTrial.gov: NCT02683902.

Disclosure

G.R. Natividade: None. C. de Araujo: None. R.C. Fitz: None. A.F. Osório: None. P.N. Merello: None. P. Schestatsky: None. F. Gerchman: Speaker's Bureau; Self; Novo Nordisk Inc. Other Relationship; Self; Sanofi-Aventis.

Funding

Hospital de Clínicas de Porto Alegre (FIPE15-0119)
2060-P: Serum Wnt Inhibitor DKK1 Is Associated with Adiposity and Insulin Resistance in Nondiabetic North Americans
Authors: ALI HIRA , OCZYPOK ELIZABETH , MILJKOVIC IVA , CVEJKUS RYAN , ZMUDA JOSEPH M. , DELANY JAMES , KERSHAW ERIN E.
Published: June 2019

Background: The Wnt pathway is known to influence body composition during early human development by regulating bone formation, myogenesis and adipogenesis. We have recently shown that DKK1 is related to measures of adiposity in nondiabetic African ancestry males. Whether these relationships are true across sexes and/or in other human populations remains unknown. In this study we evaluated the relationship of DKK1 with adiposity in a sample of male and female, mixed race, nondiabetic North American individuals.

Methods: Fasting serum DKK1 levels were measured using ELISAs in 201 nondiabetic subjects (age range 30.7-60 years, BMI range 19.9-64.0 kg/m2) recruited from the general population via television advertisement and mass mailing. The subjects were predominantly female (85%) and Caucasian (63%). Anthropometrics were obtained and markers of inflammation and glucose metabolism were measured from fasting serum samples. Spearman and partial Spearman correlation analysis was used to determine the association of DKK1 with measures of adiposity and insulin resistance.

Results: Circulating DKK1 was positively associated with BMI (r=0.30, p<0.001) and waist circumference (r=0.25, p<0.001), independent of age, race and sex. In addition, DKK1 was positively associated with fasting serum insulin level (r=0.18, p<0.01) and HOMA-IR (r=0.19, p<0.01) in age, sex and race adjusted models. However, these associations were no longer significant after adjustment for BMI.

Conclusion: This is the first study looking at correlation of DKK1 with adiposity in North American nondiabetic individuals. Our findings suggest that the link between DKK1 and insulin resistance maybe mediated indirectly through total and regional adiposity and that DKK1 could be a potential early biomarker for diabetes in high risk individuals. These findings support our findings in African ancestry males and add to the evidence favoring potential role of Wnt modulators in adiposity regulation in humans.

Disclosure

H. Ali: None. E. Oczypok: None. I. Miljkovic: None. R. Cvejkus: None. J.M. Zmuda: None. J. DeLany: None. E.E. Kershaw: Research Support; Self; Regeneron Pharmaceuticals.

Funding

National Institutes of Health (T32DK007252); Endocrine Fellows Foundation
2081-P: Obesity and Bariatric Surgery Trends in Type 1 Diabetes
Authors: UWAIFO GABRIEL I.
Published: June 2019

The prevalence of obesity and its complications and comorbidities continue to increase. It is well established that obesity is a major etiologic factor in the onset of type 2 diabetes (T2DM) and that bariatric surgery (BS) is an effective management strategy for both weight and glycemic control in obese T2DM patients. The relationship between obesity and type 1 diabetes (T1DM) is less clear though recent reports suggest a recent surge in the prevalence of obesity among T1DM patients. To explore the prevalence of obesity in T1DM and the impact of bariatric surgery in these patients a 1 year (2018) retrospective chart review of T1DM patients was done and the identified patients were compared to T2DM patients seen over the same time period in the Ochsner clinical network. Of 93,367 subjects with T2DM seen over the 1 year period, 62, 034 (66.4%) are obese and of these 18,670 (30%) are morbidly obese. The T1DM patients over the same period are 7,143 of which 3,728 (52%) are obese and 969 (26%) are morbidly obese. Over this period while 1,818 Obese T2DM patients (2.9% of the cohort) had BS only 6 obese T1DM patients (0.16%) of the cohort had BS. This group consists of mostly Caucasian (5/6) women (5/6) with 3 having gastric bypass and 4 having sleeve gastrectomy. While the prevalence of cardiometabolic comorbidities (hypertension, dyslipidemia, ESRD, CAD, and CVD) were essentially similar in both cohorts of obese diabetics, the effects of bariatric surgery on weight, HBA1c, blood pressure and lipids were more variable in the T1DM patient than the T2DM patients. Obesity is presently nearly as prevalent in T1DM as T2DM. Morbid obesity is just as prevalent in both diabetes cohorts. While BS remains underutilized as a management strategy in obese T2DM it remains a rarity in obese T1DM and the little data available suggests that its impact on cardiometabolic surrogates may be more variable in this unique cohort. More study is needed of the potential utility and impact of BS in weight management of obese T1DM.

Disclosure

G.I. Uwaifo: None.
2089-P: Regional Renal Hemodynamics and Fatty Acid Uptake: Effects of Obesity and Weight Loss
Authors: REBELOS ELENI , DADSON PRINCE , OIKONEN VESA , IIDA HIDEHIRO , HANNUKAINEN JARNA C. , IOZZO PATRICIA , FERRANNINI ELE , NUUTILA PIRJO
Published: June 2019

Objective: Changes of obesity on renal perfusion and metabolism are poorly understood. Our aims were to evaluate whether obesity increases renal regional hemodynamics and metabolism in vivo, and to determine the effects of weight loss.

Methods: 23 morbidly obese and 15 age- and sex-matched nonobese controls were recruited. Renal perfusion and fatty acid uptake (FAU) were measured using PET-CT scanning and [15O]- H2O and [18F]fluoro-6-thia-heptadecanoic acid ([18F]THA), respectively. Kidney volumes and radiodensity were measured by CT and cardiac output by MRI. Intraglomerular pressure (Pglo) was estimated by Gomez formula. Obese subjects were re-studied six months after bariatric surgery.

Results: As compared to controls, obese subjects had larger renal volume (340[118] vs. 256[63] ml, median[interquartile range]) and lower renal radiodensity (15.3[13.1] vs. 31.5[9.8] units, p≤0.05 for both), suggesting accumulation of water and/or lipid. Both cardiac output and glomerular filtration rate (eGFR) were increased by ∼25% in the obese. While there were no differences in total renal perfusion between the two groups, Pglo was higher in the obese (61±5 mmHg vs. 55±3 of controls, p<0.001). In both groups, regional blood flow was 50% higher in the cortex than medulla; in either region, FAU was higher in the obese (6.3±1.8 vs. 4.1±1.5 µmol.min-1.100g-1, and 7.5±2.2 vs. 4.6±2.1, both p≤0.05), mostly as a consequence of higher plasma FFA levels. Post-surgery (weight loss = 26±8 kg), renal volume, GFR, and Pglo all decreased and renal radiodensity increased to levels similar to controls. On the contrary, cortical and medullary FAU was not normalized.

Conclusions: In obesity, renal hemodynamics are indicative of an incomplete protection against hyperperfusion and renal fatty acid uptake is increased, driven by higher substrate availability. Surgically-induced major weight loss reverses the hemodynamic but not the metabolic changes.

Disclosure

E. Rebelos: None. P. Dadson: None. V. Oikonen: None. H. Iida: None. J.C. Hannukainen: None. P. Iozzo: None. E. Ferrannini: None. P. Nuutila: Research Support; Self; AstraZeneca.

Funding

Academy of Finland; University of Turku; Turku University Hospital
21-OR: Final Results of a Phase 2, Randomized, Double-Blind, Placebo-Controlled Clinical Trial of the MetAP2 Inhibitor ZGN-1061 in Patients with Type 2 Diabetes
Authors: KIM TERRI , ZHUANG DONGLIANG , HAUGEN TONYA L. , KIM DENNIS D. , TAYLOR KRISTIN
Published: June 2019

ZGN-1061 is a second-generation methionine aminopeptidase 2 (MetAP2) inhibitor in development for treatment of T2D. This clinical trial evaluated ZGN-1061 (0.05, 0.3, 0.9, or 1.8 mg) vs. placebo (PBO) administered SC every 3 days for 12 weeks on change from baseline in A1C in individuals with T2D at multiple sites in Australia/New Zealand. Stable noninsulin diabetes therapy was permitted. Baseline (ITT population N=175): 52% female, 77% white, (mean±SD) age 54±8 y, A1C 8.6±1.1%, BMI 37±7 kg/m2, T2D duration 8±6 y. Interim analysis results (0.05, 0.3, 0.9 mg ZGN-1061) were presented previously. Here, we report additional results with 0.9 and 1.8 mg ZGN-1061. The LS mean±SE A1C change for PBO was 0.2±0.1% (N=42) vs. -0.4±0.1% (N=44) and -0.9±0.2% (N=23) for 0.9 and 1.8 mg ZGN-1061, respectively. The LS mean±SE difference vs. PBO was -0.6±0.2% and 1.1±0.2% for 0.9 mg (p=0.0003) and 1.8 mg (p<0.0001). The LS mean±SE weight change for PBO was -0.6±0.3 kg vs. -1.4±0.3 kg (p=ns) and -2.8±0.5 kg (p<0.001) for 0.9 and 1.8 mg ZGN-1061. Dose-dependent fasting and postprandial glucose reductions were observed for ZGN-1061 (p<0.05 vs. PBO); β-cell function and insulin sensitivity were improved with 1.8 mg ZGN-1061 (p<0.05 vs. PBO). ZGN-1061 also improved triglycerides, hsCRP, adiponectin, and leptin (p<0.05 vs. PBO). Across all ZGN-1061 treatment groups, adverse events (AEs) were primarily mild/moderate without any notable trends. The completion rate was 93% (163/175). Three subjects in ZGN-1061 groups withdrew due to AEs (sensory disturbance, 0.05 mg; upper abdominal pain, 0.9 mg; injection site urticaria, 1.8 mg). There were no reports of venous thrombosis or changes in thrombosis markers indicative of a thrombosis event. ZGN-1061 (1.8 mg) demonstrated statistically significant and clinically meaningful efficacy; the largest reductions in A1C and weight were observed at 12 weeks with no evidence of a waning effect and no notable safety signals.

Disclosure

T. Kim: Employee; Spouse/Partner; Celgene Corporation. Employee; Self; Zafgen, Inc. D. Zhuang: Employee; Self; Zafgen, Inc. T.L. Haugen: Employee; Self; Zafgen, Inc. D.D. Kim: Employee; Self; Zafgen, Inc. K. Taylor: Employee; Self; Zafgen, Inc.
211-LB: Developing "Gold Standard" Diagnoses for Type 1 and Type 2 Diabetes in Adults from Electronic Health Record Data
Authors: SUE KIRKMAN M. , NOONEY JENNIFER G. , BENOIT STEPHEN R. , BERGAMO KATHERINE A. , BULLARD KAI M. , CAMPIONE JOANNE R. , MARDON RUSS , PFAFF EMILY , ROLKA DEBORAH B. , SAYDAH SHARON
Published: June 2019

Type 1 (T1D) and type 2 (T2D) diabetes are distinct diseases, but most federal surveys used to monitor diabetes prevalence have not included questions on diabetes type. To design and validate survey questions that will distinguish between T1D and T2D in adults, we developed a survey sample in whom we first determined a "gold standard" for diabetes type from review of electronic health record (EHR) data. From >50,000 adult patients in the UNC Health Care System with diabetes, we assembled a sample of 2479 patients using a "strawman" definition based on diabetes-related diagnostic codes, laboratory results, and drugs. We stratified the sample using age, sex, race/ethnicity and crude diabetes type, oversampling T1D. For visits between 10/2014 and 9/2017, we collected clinical and demographic data including body mass index, diagnosis codes, medications, family history, and laboratory tests. We also abstracted chart notes, using trained personnel, to find additional information including diabetes duration and historical treatments. We created two models to determine diabetes type. The first was a decision tree, in which higher nodes sorted straightforward cases; lower nodes included more specific factors for those not categorized in higher nodes. The second was a weighted model, in which factors drawn from clinical or epidemiologic literature weighed weakly, moderately, or strongly in favor of or against each type of diabetes. The two models were 89% congruent for T1D and 96% congruent for T2D. An endocrinologist reviewed and determined type for the 10% of patients for which results of the two models were not congruent or for which neither was able to determine type. Overall, 70% of our sample has T2D, 27% has T1D, and 3% has indeterminate or other diabetes. We will use this sample to validate survey questions to determine type of diabetes. Such "gold standard" determinations may support validation of new and improved methods for measuring diabetes type among the adult population.

Disclosure

M. Kirkman: Research Support; Self; Bayer AG, Novo Nordisk A/S, Theracos, Inc. J.G. Nooney: None. S.R. Benoit: None. K.A. Bergamo: None. K.M. Bullard: None. J.R. Campione: None. R. Mardon: None. E. Pfaff: None. D.B. Rolka: None. S. Saydah: None.

Funding

Centers for Disease Control and Prevention
2156-P: Development of the Immune System in the Normal Rat Pancreas
Authors: GOLDEN THEA , WORTHEN G. SCOTT , SIMMONS REBECCA A.
Published: June 2019

The role of resident immune cells in organogenesis has not been extensively studied in pancreas. We hypothesized that during the critical stage of ß-cell remodeling during the newborn period, immune cell populations would also change. Using flow cytometry and immunohistochemistry, we identified immune cell populations in the endocrine (islets) and exocrine portion of the pancreas at postnatal day 1, 7 and 14. When corrected for the expected increase in pancreatic weight, immune cell populations increase in the exocrine pancreas in males and females from P1 to P7 but not further at P14. Interestingly, in islets the number of immune cells further increases in males from P7 to P14. Macrophages, T cells, B cells, granulocytes and NK cells are present in islets and exocrine pancreas. Macrophages are the dominant population in islets (M: 47%, F: 47%) and exocrine (M:30%, F: 34%) pancreas. In islets, M2 macrophages are favored at P1. M1 macrophages dramatically decrease with age. In exocrine pancreas, M1 macrophages are initially favored, but with age the M2 population increases. T cells are also present in islets (M:6%, F:6%) and exocrine (M:8%, F: 6%) pancreas. Cd3+ Cd4- Cd8- is the dominant T cell population at P1 but decreases with age replaced by Cd3+ Cd4+ Cd8- cells. In islets from male, but not females, the number of T cells, specifically Cd4+ T cells, increases with age. B cells (Cd45Ra+) are present in islets (M:20%, F:19%) and exocrine (M:20%, F:25%) pancreas and numbers do not change with age. Neutrophils (HIS48hi Cd68) are also present in islets (M:6%, F:6%) and exocrine (M:13%, F: 10%) pancreas. NK cells are a minor cell population present at about 1% of total immune cells in male and female pancreas at P1. Interestingly, NK cells increase with age in females in islets and exocrine pancreas. These data demonstrate significant sex specific changes in the pancreas during the critical period of postnatal pancreas development suggesting a role for immune cells in normal postnatal development and function in the endocrine and exocrine pancreas.

Disclosure

T. Golden: None. G. Worthen: None. R.A. Simmons: None.

Funding

National Institutes of Health