bluuebunny/crossref_metadata_2025_split huggingface dataset:
1757-P: Ginsenoside Rb2 Alleviates Obesity by Activation of Brown Fat and Induction of Browning of White Fat
Authors: GU XUEJIANG 
Published: June 2019

Ginsenoside Rb2 (Rb2), the most abundant saponin contained in Panax ginseng, has been used to treat variety of metabolic diseases. However, its effects in obesity and potential mechanism are not well understood. In the present study, we investigate the metabolic performances with Rb2 supplement in diet induced obese (DIO) mice, focusing on assessing the effects and mechanisms of Rb2 on brown and beige fat functions. Our results demonstrated that Rb2 effectively reduced body weight, improved insulin sensitivity, as well as induced energy expenditure in DIO mice. Histological and gene analysis revealed that Rb2 induced activation of brown fat and browning of white fat as shown by reduced lipid droplets, increased UCP1 staining and increased thermogenic and mitochondrial genes, which could be recapitulated in 3T3-L1, C3H10T1/2 and primary adipocytes. In addition, Rb2 induces AMPK phosphorylation both in vitro and in vivo and these effects were shown to be dependent on AMPK since its inhibitor blocked Rb2 induced expressions of Pgc1α and Ucp1. Overall the present study revealed that Rb2 activates brown fat and induces browning of white fat, which increases energy expenditure and thermogenesis and consequently ameliorates obesity and metabolic disorders, suggesting Rb2 to be a promising beneficial compound treating obesity.

Disclosure
X. Gu: None.
1759-P: Effects of High-Fat Feeding on Adipose Tissue Matrisome Composition in Female Mice
Authors: BAILEY JENNIFER L. , ELKS CARRIE M. 
Published: June 2019

Alterations in adipose tissue (AT) extracellular matrix (ECM) organization and/or composition accompany several pathologies, including obesity and diabetes. The need to better understand overall AT ECM composition and structure is essential to unraveling the pathogenesis of obesity, especially when considering that a given tissue ‘matrisome’ (a protein signature comprised of core ECM proteins plus ECM-associated proteins) can consist of several hundred proteins. Recent methods have exploited the insolubility of ECM proteins to allow their enrichment from decellularized tissues and subsequent proteomic analysis, but to our knowledge these methods have not been performed in AT. In this study, we used this methodology to characterize the AT matrisome in female mice fed control or high-fat diets. Female C57BL/6J mice were fed control (n=6) or high-fat (n=6) diets for 8 weeks. Body weights were obtained weekly and body composition was analyzed biweekly via NMR. At study end, AT samples were decellularized and enriched for ECM proteins using a commercially available kit. The ECM-enriched fractions were digested, deglycosylated, fractionated into peptides, LC-MS/MS performed, and raw data analyzed using Proteome Discoverer v2.2. Our data reveal that high-fat feeding does not significantly affect AT collagen levels in female mice, but does significantly increase glycoprotein levels (spondin 1, SPARC, adiponectin). Conversely, a significant decrease in the proteoglycan, PRG3, was also observed in AT with high-fat feeding. Several additional ECM-associated proteins, which have undefined roles AT, were differentially regulated by high-fat diet feeding in female mice. Current studies are focused on determining how these proteins influence AT metabolism.

Disclosure
J.L. Bailey: None. C.M. Elks: None.

Funding
National Institutes of Health
1760-P: Tetrahydrobiopterin Regulates Developmental Differentiation of Brown Adipose Tissue
Authors: MINAMINO HIROTO , FUJITA YOSHIHITO , OGURI YASUO , GOTO TSUYOSHI , OHASHI AKIKO , FURUYA FUTOSHI , ISOMURA NOZOMI , KAWARASAKI SATOKO , KAWADA TERUO , HASEGAWA HIROYUKI , INAGAKI NOBUYA 
Published: June 2019

Brown adipose tissue (BAT) is a key organ that produces heat and increases energy expenditure; its regulatory factors could be clinically useful in the treatment of obesity and diabetes. Tetrahydrobiopterin (BH4) is an essential co-factor of tyrosine hydroxylase and nitric oxide synthase. Our group recently showed that BH4 activates BAT function and regulates systemic energy metabolism using Hph-1 mice, a mouse model of BH4 deficiency in which GTP-cyclohydrolase I (GTPCH I), a rate-limiting enzyme of BH4, synthesis is deficient. Interestingly, we found that dysplasia of BAT already has been confirmed in the neonatal period. We investigated the role of BH4 in the development and differentiation of BAT using the mouse model of BH4 deficiency. Compared with control mice, Hph-1 neonatal mice exhibited lower weight of BAT as well as attenuated thermogenesis-related gene expressions (UCP1, Dio2, Pgc1a). We also evaluated the role of BH4 in brown adipocyte differentiation using brown pre-adipocytes isolated from control and Hph-1 mice. Compared with control mice, brown pre-adipocytes from Hph-1 mice reduced differentiation capacity measured by accumulated lipid droplets and thermogenesis-related gene expressions. In addition, inhibition of GTPCH I by 2,4-Diamino-6-Hydroxypyrimidine (DAHP) reduced differentiation capacity in brown pre-adipocytes. These data strongly suggest that BH4 is required for brown adipocyte differentiation. Furthermore, supplementation of BH4 in brown pre-adipocytes from Hph-1 mice dose-dependently increased accumulation of lipid droplets, thermogenesis-related gene expressions, and nitric oxide production. Taken together, BH4 plays an integral role in developmental differentiation of BAT, and has therapeutic potential for metabolic disorders such as obesity and diabetes.

Disclosure
H. Minamino: None. Y. Fujita: None. Y. Oguri: None. T. Goto: None. A. Ohashi: None. F. Furuya: None. N. Isomura: None. S. Kawarasaki: None. T. Kawada: None. H. Hasegawa: None. N. Inagaki: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Japan Tobacco Inc., Kyowa Hakko Kirin Co., Ltd., Merck Sharp & Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Ono Pharmaceutical Co., Ltd., Sanofi, Sumitomo Dainippon Pharma Co., Ltd., Takeda Pharmaceutical Company Limited.
177-OR: Genome-Wide Association Study of Lipid Traits in Youth with Type 2 Diabetes: The ProDiGY (Progress in Diabetes Genetics in Youth) Consortium
Authors: CHEN LING , SRINIVASAN SHYLAJA , SANTORO NICOLA , TODD JENNIFER , DIVERS JASMIN , SHAH AMY SANGHAVI , GIDDING SAMUEL , BURKE BRIAN K. , HAYMOND MOREY W. , LANGE LESLIE , MARCOVINA SANTICA M. , CHAIT ALAN , FLANNICK JASON , CAPRIO SONIA , FLOREZ JOSE C. 
Published: June 2019

Type 2 diabetes (T2D) in youth is associated with a pro-atherogenic lipid profile characterized by high triglycerides (TG), high low-density lipoprotein cholesterol (LDL-c), and low high-density lipoprotein cholesterol (HDL-c). Genome wide association studies (GWAS) in adults have discovered more than 100 variants associated with lipid traits, but the association of genetic variants to dyslipidemia in youth with T2D remains unknown. Therefore, we genotyped 206,928 variants and imputed 17,642,824 variants in 1,076 youth with T2D from the Treatment Options for type 2 diabetes in Adolescents and Youth (TODAY) and SEARCH for Diabetes in Youth (SEARCH) as part of ProDiGY, a collaboration with type 2 diabetes Genetic Exploration by Next-generation sequencing in multi-Ethnic Samples (T2D-GENES). Using a linear mixed model, we performed association testing for TG, LDL-c and HDL-c adjusted for age, sex, body mass index (BMI) Z-score, HbA1c, and genetic relationship matrix within each sub-study followed by meta-analyses for each lipid trait. We identified a novel variant between a deletion on chromosome 3 (3:67817380_AT/A_Deletion:RP11-81N13.1) that was associated with TG levels at genome wide level of significance (P-value=2.26×10-8, β=0.21±0.09). We also identified a genome-wide significant variant rs247617 (P-value=5.12×10-9, β=2.52±0.43) between HERFUD1 and CETP for association with HDL-c. This variant has been previously reported to be associated with HDL-c in adults in the Global Lipids Genetics Consortium (GLGC). We did not find any genome-wide significant variants in the LDL-c analysis.
In summary, our genetic analyses of lipid traits in youth with T2D identified one novel and one previously known locus. Further multi-phenotype analysis may uncover additional loci associated with lipid traits in youth with T2D.

Disclosure
L. Chen: None. S. Srinivasan: None. N. Santoro: None. J. Todd: None. J. Divers: None. A.S. Shah: None. S. Gidding: Research Support; Self; Color Genomics. B.K. Burke: None. M.W. Haymond: Advisory Panel; Self; Daiichi Sankyo Company, Limited, Zealand Pharma A/S. Stock/Shareholder; Self; Xeris Pharmaceuticals, Inc. Other Relationship; Self; AstraZeneca. L. Lange: None. S.M. Marcovina: None. A. Chait: Consultant; Self; MedImmune. J. Flannick: Consultant; Self; Decibel Therapeutics. S. Caprio: None. J.C. Florez: None.

Funding
National Institutes of Health
1771-P: Upregulation of RETN, TNFα, and CCL2 mRNAs in Adipocytes by Intermittent Hypoxia and Its MicroRNA-Mediated Mechanism
Authors: UCHIYAMA TOMOKO , ITAYA-HIRONAKA ASAKO , YAMAUCHI AKIYO , MAKINO MAI , SAKURAMOTO-TSUCHIDA SUMIYO , SHOBATAKE RYOGO , OTA HIROYO , TAKEDA MAIKO , OHBAYASHI CHIHO , TAKASAWA SHIN 
Published: June 2019

Sleep apnea syndrome (SAS), characterized by recurrent episodes of oxygen desaturation and reoxygenation (intermittent hypoxia [IH]), is a risk factor for insulin resistance/type 2 diabetes. IH is considered to cause adipose tissue inflammation, leading to insulin resistance. In this study, we exposed mouse 3T3-L1 and human SW872 adipocytes to experimental IH (1% O2, 5% CO2 [5 min], 21% O2, 5% CO2 [10 min]) or normoxia (21% O2, 5% CO2) for 24 h, and analyzed mRNA expression of adipokines (leptin, adiponectin, resistin [RETN], tumor necrosis factor α [TNFα], IL-6, and C-C motif chemokine ligand 2 [CCL2]). We found that the mRNA levels of RETN, TNFα and CCL2 in SW872 cells (P=0.0257, P=0.0312, and P&lt;0.0001, respectively) and 3T3-L1 adipocytes (P=0.0498, P=0.0237, and P=0.0002, respectively) were significantly increased by IH. RETN, TNFα, and CCL2 in IH-treated cell medium were also increased (P=0.0256, P=0.0215, and P&lt;0.0001, respectively). We next prepared reporter plasmids containing human RETN (-979∼+20), TNFα (-966∼+19) and CCL2 (-3455∼+25) upstream of luciferase reporter gene, and transfected them into SW872 cells. The promoter activities of these genes were not increased by IH. Target mRNA search of microRNA (miR)s using MicroRNA.org program revealed that all the mRNAs have a potential target sequence for miR-452. The miR-452 level of IH-treated cells was significantly decreased than that of normoxia-treated cells (P=0.0458). To clarify the role of miR-452, miR-452 mimic and non-specific control RNA (miR-452 mimic NC) were introduced into SW872 cells, and the IH-induced up-regulation of RETN, TNFα, and CCL2 was abolished by introduction of miR-452 mimic but not by miR-452 mimic NC. These results indicate that IH stress down-regulates the miR-452 in adipocytes, resulting in the increased levels of RETN, TNFα, and CCL2 mRNA via the inhibition of the miR-452-mediated mRNA degradation, leading SAS patients to insulin resistance.

Disclosure
T. Uchiyama: None. A. Itaya-Hironaka: None. A. Yamauchi: None. M. Makino: None. S. Sakuramoto-Tsuchida: None. R. Shobatake: None. H. Ota: None. M. Takeda: None. C. Ohbayashi: None. S. Takasawa: None.
1773-P: Beta-Catenin Is a Key Player in Insulin-Stimulated Skeletal Muscle Glucose Transport
Authors: MASSON STEWART W.C. , SHEPHERD PETER R. , MERRY TROY L. 
Published: June 2019

Recent work has identified β-catenin as a potential regulator of vesicle trafficking within various tissues including synapses, pancreatic β-cells, and adipocytes. The trafficking of GLUT4 vesicles to the cell membrane is essential for insulin-stimulated glucose uptake. Here we provide evidence for a role of β-catenin in regulating skeletal muscle glucose transport. In in vivo models, β-catenin is sensitive to insulin, which through PI3 kinase and Akt triggers phosphorylation of β-catenin at serine 552 (S552). Substituting S552 to alanine impairs glucose transport in L6-GLUT4-myc cells suggesting S552 is key in β-catenin-regulated glucose transport. Furthermore, inhibition of β-catenin in ex vivo models and cell culture impairs insulin-stimulated glucose transport. Using transcriptional inhibitors we demonstrate that β-catenin’s regulation of glucose transport is at least in part independent of its role in gene regulation. Finally, we investigated whether diet induced insulin-resistance alters insulin stimulated β-cateninS552 phosphorylation in vivo. Based on β-catenin’s structure and role in other cellular processes we believe that β-catenin regulates glucose transport via interactions with cytoskeletal proteins which ultimately regulate vesicle movement.

Disclosure
S.W.C. Masson: None. P.R. Shepherd: None. T.L. Merry: None.

Funding
Royal Society of New Zealand; Maurice Wilkins Centre for Molecular Biodiscovery
1778-P: Delivery of the Insulin Gene Using an Integrating Adeno-Associated Viral Vector (AAV) to Diabetic NOD Mice
Authors: LA QUE T. , REN BINHAI , LOGAN GRANT J. , CUNNINGHAM SHARON C. , KHANDEKAR NEETA , NASSIF NAJAH T. , O'BRIEN BRONWYN A. , ALEXANDER IAN E. , SIMPSON ANN M. 
Published: June 2019

Previously we introduced furin-cleavable human insulin (INSFUR) to the livers in several animal models using a lentiviral vector. Reversal of diabetes with normal glucose tolerance and pancreatic transdifferentiation was seen. The aim of the present study was to ascertain if we could similarly reverse diabetes in the non-obese diabetic (NOD) mouse using a clinically-applicable AAV system to deliver INSFUR ± the β-cell transcription factor (TF) Pdx1 to the livers of the diabetic mice. The traditional AAV8 that allows predominantly episomal expression, and the hybrid AAV8/piggyBac that results in transgene integration into the host genome were used. Weight and blood glucose levels were monitored. Intraperitoneal glucose tolerance tests (IPGTTs) were performed. Immunofluorescence was carried out to monitor transduction utilising marker genes Venus and mCherry. RT-PCR was used to identify expression of pancreatic genes. Diabetic mice that received intraperitoneal (IP) injections of AAV8-INSFUR-mCherry (2.3x 1010-4.5x1010vg) became hypoglycaemic with abnormal responses to IPGTTs. No β-cell TF could be detected in the livers. Reversal of diabetes could not be achieved in the mice (n=8) that received combinations of AAV8-INSFUR-venus (5x1010 or 7.5x1010vg) and AAV8-Pdx1-mCherry (7.5x1010 vg). Both INSFUR and Pdx1 were detected using RT-PCR and immunofluorescence showed the expression of both marker genes. IPGTTs were abnormal; however, the β-cell TF MafA was detected by RT-PCR. Euglycaemia and normal IPGTTs were achieved in mice (n=4) that received AAV8/piggyBac-INSFUR-mCherry (transposase 4x1010 vg and transposon 3.5x1010 vg). The INSFUR gene and the β-cell TFs NKx6.1 and MafA were detected in the livers, however wider evidence of pancreatic transdifferentiation was not seen. This is the first in vivo study using the hybrid AAV8/piggyBac system to treat type 1 diabetes (T1D). The AAV system, with further development, may hold promise for gene therapy of T1D.

Disclosure
Q.T. La: None. B. Ren: None. G.J. Logan: None. S.C. Cunningham: None. N. Khandekar: None. N.T. Nassif: None. B.A. O'Brien: None. I.E. Alexander: Other Relationship; Self; LogicBio. A.M. Simpson: Consultant; Self; Pharmacyte Biotech.
178-LB: Use of Random Glucose Followed by HbA1c Improves the Accuracy of Opportunistic Screening
Authors: LEGVOLD BRIAN T. , STAIMEZ LISA R. , LI DONG , PHILLIPS LAWRENCE S. , RHEE MARY K. 
Published: June 2019

The diagnostic accuracy of HbA1c to identify diabetes and dysglycemia as found in OGTTs is limited. Since opportunistic screening at health visits is cost-effective, and glucose is commonly included in metabolic panels obtained at such visits, we asked if use of glucose followed by HbA1c could improve the accuracy of screening. With the Screening for Impaired Glucose Tolerance (SIGT) study dataset of 1,573 adults without known diabetes, we used Receiver Operating Characteristic (ROC) analysis and OGTTs with ADA diagnostic criteria as a "standard" to evaluate screening; random plasma and capillary glucose (RPG and RCG) were measured on a separate day. Subjects were 58% female and 58% black, with mean age 47.9 year, BMI 30.3 kg/m2 and HbA1c 5.4%. Using OGTTs, 51.5% had normal glucose metabolism, 33.3% prediabetes (PreDM), 4.6% type 2 diabetes (DM), and 37.9% dysglycemia (PreDM or DM). For HbA1c to identify DM by ADA OGTT criteria, the ROC area under the curve (AUC) was 0.818 in all 1573 subjects. However, using an RPG cutoff of 100 mg/dl, the ROC was increased to 0.857 in the 576 with RPG &gt;100 mg/dl but only 0.603 in the 997 with RPG &lt;100 mg/dl (both p&lt;0.001 vs. all subjects). There was similar improvement in identification of DM in the 756 subjects with RCG &gt;100 mg/dl (ROC 0.841, p&lt;0.001), and parallel improvement in identification of dysglycemia (ROC 0.671 for all subjects, vs. 0.740 and 0.741 for those with RPG or RCG &gt;110 mg/dl, respectively, both p&lt;0.001). Results were similar with RCG in a separate dataset (n=1,037). For identification of DM among those with RPG &gt;100 mg/dl in the SIGT dataset, specificity and sensitivity were 79% and 74%, respectively, for HbA1c &gt;5.7%, 84% and 69%, for HbA1c &gt;5.8%, and 90% and 64% for HbA1c &gt;5.9%.
Conclusions: Use of random glucose followed by HbA1c improves the accuracy and efficiency of screening, identifying both individuals who should and should not have an OGTT. Such a strategy might improve recognition of diabetes and prediabetes, permitting initiation of preventive management.

Disclosure
B.T. Legvold: None. L.R. Staimez: None. D. Li: None. L.S. Phillips: Advisory Panel; Self; Janssen Pharmaceuticals, Inc. Research Support; Self; AbbVie Inc., GlaxoSmithKline plc., Kowa Pharmaceutical Europe Co. Ltd., Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Pfizer Inc. Stock/Shareholder; Self; Diasyst Inc. Other Relationship; Self; Diasyst Inc., Janssen Pharmaceuticals, Inc. M.K. Rhee: None.

Funding
U.S. Department of Veterans Affairs; National Institutes of Health
178-OR: Long-Term Treatment with Injectable Testosterone Undecanoate (TU) Improves Lipid Profile in Men with Hypogonadism and Type 2 Diabetes in Comparison to an Untreated Control Group: Ten-Year Data from a Urological Registry Study
Authors: SAAD FARID , HAIDER KARIM S. , HAIDER AHMAD 
Published: June 2019

Background: In the literature, effects of testosterone on lipid pattern in men with hypogonadism are inconsistent.
Methods: Of 324 men with hypogonadism and T2DM in a registry, 152 received TU 1000 mg/12 weeks (T-group), 172 opted against treatment (CTRL). Mean changes over time between groups were compared by mixed effects model for repeated measures with random effect for intercept and fixed effects for time, group and their interaction. Changes were adjusted for age, weight, waist circumference, fasting glucose, blood pressure (BP), lipids and quality of life to account for baseline differences between the two groups.
Results: Mean age: 62.7±5.1 years.
Follow-up: Mean: 7.9, median: 9 years. All values are in mmol/L ±SE. Total cholesterol decreased by 2.8±0.1 at 10 years (T-group) and increased by 1.0±0.1 (CTRL). Estimated adjusted difference between groups: 3.9 (p&lt;0.0001 for all). HDL increased by 0.3±0.0 (T-group) and decreased by 0.3±0.0 (CTRL), difference between groups: 0.6 (p&lt;0.0001 for all). LDL decreased by 1.8±0.1 (T-group) and increased by 0.8±0.1 (CTRL), difference between groups: 2.7 (p&lt;0.0001 for all). Triglyceride decreased by 1.1±0.1 (T-group) and increased by 0.4±0.0 (CTRL), difference between groups: 1.6 (p&lt;0.0001 for all). Non-HDL decreased by 4.6±0.3 (T-group) and increased by 3.8±0.3 (CTRL), difference between groups: 8.4 (p&lt;0.0001 for all). Remnant cholesterol decreased by 1.3±0.1 (T-group) and increased by 0.4±0.1 (CTRL), difference between groups: 1.8 (p&lt;0.0001 for all). The lipid accumulation product (LAP, cm*mmol/L) decreased by 85.0±3.7 (T-group) and increased by 40.3±3.2 (CTRL), difference between groups: 125.3 (p&lt;0.0001 for all).
Conclusions: Long-term TTh with TU in men with hypogonadism and T2DM sustainably improved lipid profile which worsened in untreated controls.

Disclosure
F. Saad: Employee; Self; Bayer AG. Stock/Shareholder; Self; AbbVie Inc., Amgen Inc., Bayer AG. K.S. Haider: Speaker's Bureau; Self; Bayer AG. A. Haider: Research Support; Self; Bayer AG. Speaker's Bureau; Self; Bayer AG.

Funding
Bayer AG
1788-P: The Natural Product ZYM01 Exerts Antidiabetic and Islet-Protective Effects in Diabetic Mice and Promotes Glucagon-Like Peptide-1 Secretion
Authors: SUN YILI , LI JINGYA , LI JIA 
Published: June 2019

Over the last few years, incretin-based therapies have emerged as important agents in the treatment of type 2 diabetes (T2D). These agents exert their effect via the incretin system, specifically targeting the receptor for the incretin hormone glucagon-like peptide 1 (GLP-1). Two classes of incretin-based therapies are available: GLP-1 receptor agonists and dipeptidyl peptidase-4 inhibitors. However, there are currently no clinical drugs that directly increase endogenous GLP-1 secretion. Although several nutrients induce GLP-1 secretion, there is little evidence to suggest that non-nutritive compounds directly increase GLP-1 secretion. In the present study, we demonstrated that the natural product ZYM01 from the Bougainvillea glabraChoisy (which was used for hypoglycemic by the Dai ethnic group in China) had a good hypoglycemic effect and a significant islet protection function in diabetic mouse model. ZYM01 treatment (50 mg•kg-1 •day-1) decreased plasma levels of glucose, body weight and food intake; ameliorated OGT and plasma lipid profiles; augmented plasma insulin levels; alleviated islets pathological morphology; and reduced liver lipid accumulation in db/db diabetic mouse model. Furthermore, ZYM01 enhanced GLP-1 release in C57BL/6 mice and exerted an acute hypoglycemic effect through the secretion of GLP-1 in vivo since the exendin 9-39, a GLP-1R antagonist could completely block the acute hypoglycemic effect of ZYM01. Also, in cultured STC-1 cells, ZYM01 increased intracellular calcium and promoted GLP-1 secretion in a dose- dependent manner accompanied by elevated levels of phosphorylated Erk signaling.
In summary, our study found that ZYM01 exerted the hypoglycaemic and islets protective effects, which were associated with an enhanced release of GLP-1 mediated by the activation of the Erk signaling. These findings firmly identified ZYM01 as a new viable therapeutic option for diabetes control.

Disclosure
Y. Sun: None. J. Li: None. J. Li: None.