bluuebunny/crossref_metadata_2025_split huggingface dataset:
Renal C3 Production Drives Acute Kidney Injury and Acute Kidney Disease in Diabetics with Sepsis
Authors: FRYDRYCH LYNN M. , BIAN GUOWU , WARD PETER A. , BITZER MARKUS , DELANO MATTHEW 
Published: June 2018

Introduction: Type 2 diabetics (T2D) suffer the greatest sepsis mortality due to the development of acute kidney injury (AKI), which progresses to acute kidney disease (AKD), renal failure, and death. Systemic inflammation and complement production are known to provoke AKI during sepsis. We hypothesize that sepsis induced renal inflammation and complement production drives progression of AKI to AKD in T2D.
Methods: C3-/-, TLR2-/-, TNF-/-, C57BL/6, and Diet Induced Obese (T2D) 30 week old mice underwent sepsis from cecal ligation and puncture (CLPLD20) or sham procedure. At serial time points, plasma and kidneys were harvested (n=5 mice/group). Multiplex RT-PCR arrays were used to evaluate toll like receptors (TLR), complement, and immune signaling cascades. TNF, BUN, creatinine, and NGAL were evaluated by ELISA.
Results: In lean mice, renal NGAL, representing AKI, peaked at 1 day and returned to baseline 7 days after CLP. In T2D mice, NGAL persisted past 7 days and peaked after 14 days, indicating transition to AKD (p&lt;0.001). Renal C3 mRNA levels were eight-fold higher 1 day after CLP in lean and T2D mice compared to sham (p&lt;0.001). In lean mice, C3 levels returned to baseline at 7 days. C3 levels in T2D mice peaked at 2 weeks and persisted. Renal in-situ hybridization showed significantly more C3 in septic T2D mice compared to lean mice. RT-PCR analysis showed significantly more renal TLR2, TNF, and C3 mRNA in T2D mice compared to lean mice. C3, TLR2, and TNF null mice each produced less renal C3 and NGAL, demonstrating less AKI and AKD (p&lt;0.001). In T2D mice, TNF inhibition blocked C3 and NGAL production, representing prevention of AKI and AKD after sepsis.
Conclusion: Lean mice resolve sepsis induced AKI by 7 days. In T2D mice, AKI persists past 7 days and progresses to AKD. The T2D AKI to AKD progression is dependent on local renal C3 production through TLR2 and TNF signaling. TNF inhibition is a novel strategy to prevent AKI progression to AKD, preserve renal function, and improve T2D sepsis mortality.

Disclosure
L.M. Frydrych: None. G. Bian: None. P.A. Ward: None. M. Bitzer: None. M. Delano: None.
Phosphoregulation Profiles of Transcription Factors during Early Stage of Thermogenesis in Brown Fat
Authors: GAO XIAOJING 
Published: June 2018

Brown adipose receives extensive attention for dissipating stored chemical energy in heat via nonshivering thermogenesis. However, whether and how the dynamics of key transcription factors (TFs) phosphorylation contributes to initial activation of thermogenesis remains unknown. Here, we employed the stat-of-the-art data-independent acquisition (DIA) mass spectrometry to explore the global and temporal phosphorylation profiles of signaling networks to elucidate mechanism responsible for the TF activation for thermogenesis gene transcription. The nucleus was isolated before and after stimulation for 5, 10, 20, 30, 40, 45, 50, 55, 60, 70 and 80 min with NE. Consequently, 401 transcription regulators bearing 1007 phosphosites were identified with high reproducibility across 12 time points. Firstly, the UCP1 transcriptional level was measured, which began at 30min, then rose until 70min to the peakedness. The global phosphoproteome displayed dramatic changes early to 20-30min, which could be classified into two dynamic clusters, namely UCP1 pre-transcription phase and transcription phase. The former changed from 5min, and reached the extremum at 20-40min, which functioned before UCP1 action. Some well- characterized transcriptional regulators of UCP1 were founded to be phosphorylated, such as PRDM16 and RIP140. Alternatively, other phosphorylation events of TFs such as Jun and ATF2 began from 30min, and maintained until 70min, almost synchronizing with UCP1 transcription. Intriguingly, we found THRAP3 posed multiple phosphosites but involving in different regulation patterns. This might indicate that THRAP3 had distinct transcriptional regulation and functions upon thermogenesis. We, for the first time, uncovered temporal phosphoregulation profiles of TFs, which might be potentially required for initial activation of thermogenesis in brown fat. A new sight was provided to increase energy expenditure for treatment of obesity and other metabolic syndrome.

Disclosure
X. Gao: None.
Fat Shadows from DEXA for Documentation of Fat Distribution in Patients with Lipodystrophy
Authors: MERAL RASIMCAN , RYAN BENJAMIN J. , HOROWITZ JEFFREY F. , ORAL ELIF A. 
Published: June 2018

Lipodystrophy (LD) syndromes are a heterogeneous group of disorders causing atypical diabetes, associated with selective absence of fat with diagnosis mostly depending on the clinical acumen of the physician. With niche therapies either approved or under investigation, development of objective diagnostic tools for these syndromes are urgently needed. Here we describe a new method using the built-in features of the enCore software v14.10 to render out non-fat tissues from Dual Energy X-ray Absorptiometry (DEXA) scans to derive a “fat-shadow” (Figure). We evaluated fat-shadows from 58 LD (50F:8M) patients and 91 non-LD patients (51F:40M). Among the LD, 3 had Generalized LD, 2 had Acquired Partial LD, and 53 had Familial Partial LD. Controls consisted of a mixed population of lean (12F:14M) or obese patients (39F:26M) with or without metabolic disease. There was substantial overlap in % body fat between the LD and non-LD groups (interquartile range: total, 22.2-39.8% vs. 33.6-45.9%; trunk, 27.0-46.8% vs. 39.1-50.2%; legs, 16.6-30.6% vs. 27.7-42.6%). However, fat-shadows obtained from patients with LD stood out due to the paucity of fat signal either in a generalized or regional manner compared to controls. Overall, fat-shadows provided sufficient qualitative information to infer clinical phenotype. We propose that these fat-shadows could be used for accurate documentation of fat distribution in LD.

Disclosure
R. Meral: None. B.J. Ryan: None. J.F. Horowitz: None. E.A. Oral: Advisory Panel; Self; Aegerion Pharmaceuticals. Research Support; Self; Aegerion Pharmaceuticals, Akcea Therapeutics. Advisory Panel; Self; Akcea Therapeutics. Research Support; Self; Ionis Pharmaceuticals, Inc., AstraZeneca. Other Relationship; Self; Aegerion Pharmaceuticals.
National Trends in Type 2 Diabetes Treatment—Comparing Older and Younger Adults
Authors: PILLA SCOTT J. , SEGAL JODI B. , CALEB ALEXANDER G. , BOYD CYNTHIA , MARUTHUR NISA 
Published: June 2018

With expanding therapeutic options for type 2 diabetes, the treatment of older adults requires special care to balance safety and complexity with glucose control. We compared national trends in type 2 diabetes treatment among older (≥65 years) and younger adults (30-64 years) using the 2006-2015 National Ambulatory Medical Care Survey (NAMCS), an annual probability sample of visits to U.S. outpatient providers. We included all visits of patients with type 2 diabetes using ≥1 diabetes medication. Analyses were weighted to yield nationally-representative estimates. In the most recently available data, 2014-2015, there were 23.3 million and 20.1 million annual treated diabetes visits for older and younger adults, respectively. The most frequently used medications in older and younger adult visits were metformin (50.4% vs. 62.3% respectively, p&lt;0.001), sulfonylureas (31.1% vs. 26.7%, p=0.301), and long-acting insulins (30.2% vs. 22.4%, p=0.017). GLP-1 receptor agonists were used in fewer older than younger adult visits (2.9% vs. 6.2%, p=0.004); DPP-4 inhibitor use was similar (12.9% vs. 11.1%). In the past decade, mean diabetes medication counts increased in both older and younger adult visits (1.47 to 1.54, p-trend=0.046; and 1.53 to 1.67, p-trend 0.052, respectively). Between 2006 and 2015, the proportion of visits with metformin, DPP-4 inhibitors, and insulin increased for both older and younger adults; sulfonylurea and thiazolidinedione use decreased (p&lt;0.05 for all trends). Long-acting insulin use increased markedly in older adults, particularly between 2010-2015 where it rose from 12.5% to 30.2% of older adult visits. In younger adult visits, the increase was modest, from 17.2% to 22.4%, a slower rate of increase than in older adults (p-interaction &lt;0.001).
In conclusion, the outpatient treatment of type 2 diabetes differs between older and younger adults driven in part by a marked increase in the use of long-acting insulin among older adults, which may have implications for risk of hypoglycemia.

Disclosure
S.J. Pilla: None. J.B. Segal: None. G. Alexander: Other Relationship; Self; See disclosure. C. Boyd: None. N. Maruthur: None.
Multitrait Genome-Wide Association Analysis of Macronutrient Intake Identifies 12 Novel Loci for Dietary Intake and Unravels Genetic Overlap with Lifestyle Traits and Cardiometabolic Diseases
Authors: DASHTI HASSAN S. , MERINO JORDI , LANE JACQUELINE M. , FLOREZ JOSE C. , RUTTER MARTIN K. , SAXENA RICHA 
Published: June 2018

Suboptimal diets are associated with increased risk of type 2 diabetes, cardiovascular disease and cancer. Genome-wide association studies (GWAS) for individual macronutrients have facilitated the discovery of two relevant loci (FGF-21, FTO), however a single-nutrient approach may preclude the discovery of additional relevant loci. Here, we conducted a multi-trait macronutrient GWAS for carbohydrate, fat, and protein in the UK Biobank to factor in the significant correlation among macronutrient intake. In the UK Biobank, dietary data were collected from over 210,000 individuals aged 37-74 using the Oxford WebQ, a web-based 24 hour diet recall. For multi-trait GWAS, we combined GWAS association evidence of ∼11m imputed variants in participants of European ancestry (n =192,005) for percent of carbohydrate, fat, and protein adjusted for age and sex by applying the CPASSOC package. We identified 14 independent genome-wide significant (Pυ x 10-8) loci for macronutrient intake. We discovered loci associated with lipid metabolism (APOE, GCKR, SLC39A8, PPP1R3B, MLXIPL), alcohol intake (ADH1B, KLB), and new candidate genes (CCDC171, PSMD7, ARL17B, METTL4), and replicated previously associated loci with dietary intake (FGF-21 and FTO). Of the 14 signals, 12 replicated (P003C;0.0036) in multi-trait analyses using previous GWAS on dietary intake from the CHARGE Consortium (n =91,114). Tissue enrichment analysis across 53 tissues in GTEx revealed enrichment of gene expression in the cerebellum. Genome-wide genetic correlations across 234 traits with publicly available GWAS data indicated shared biological links between dietary intake and lifestyle factors (smoking and education) as well as type 2 diabetes and coronary artery disease (P&lt;2.14x10-4). The present findings expand our understanding of the genetic architecture of dietary intake and the shared genetic links with related traits.

Disclosure
H.S. Dashti: None. J. Merino: None. J.M. Lane: None. J.C. Florez: Consultant; Self; Intarcia Therapeutics, Inc.. Consultant; Spouse/Partner; Santen. M.K. Rutter: Consultant; Self; Novo Nordisk Inc.. Speaker's Bureau; Self; Novo Nordisk Inc.. Consultant; Self; Ascensia Diabetes Care, Roche Diabetes Care Health and Digital Solutions. R. Saxena: None.
DBC1, Deleted in Breast Cancer 1, Is a Nuclear STAT5A-Interacting Protein in Adipocytes, but Its Primary Effects on GLUT4 Gene Expression and Lipolysis Are STAT5 Independent
Authors: ABLE ASHLEY , RICHARD ALLISON J. , STEPHENS JACQUELINE M. 
Published: June 2018

STAT5A (signal transducer and activator of transcription 5A) is a transcription factor that plays a role in adipocyte development and function. Although the role of STAT5A in fat cell development is well characterized, its specific roles in mature fat cells are still largely unknown. Moreover, the potential role of STAT5A in metabolic diseases, in particular type 2 diabetes, remains unclear. To further understand the function of STAT5A in adipocytes, we employed a non-biased co-immunoprecipitation and mass-spectrometry-based approach to identify novel STAT5A-interacting proteins. One of the proteins we identified was DBC1 (deleted in breast cancer 1; also known as CCAR2). DBC1 is typically localized in the nucleus and has been primarily studied in tumor cells. However, the functions of DBC1 in adipocytes are relatively unknown. Using mouse fat cells, we confirmed that there is a physical association between endogenous STAT5A and DBC1 proteins under physiological conditions in the nucleus that is not dependent upon STAT5 tyrosine phosphorylation. Knockdown of DBC1 in 3T3-L1 adipocytes using siRNA did not affect the expression of several STAT5A target genes including socs3, cish, bcl-6, socs2, and igf-1. However, we did observe increased levels glycerol and free fatty acids released from adipocytes with reduced DBC1 following TNF (tumor necrosis factor) α stimulation. Although loss of DBC1 did not alter STAT5 transcriptional activity for the genes examined, it appears to modulate TNFα-mediated lipolysis in a STAT5-independent manner. In addition, DBC1 knockdown increased GLUT4 expression in murine fat cells but did not have a profound effect on TNFα-mediated changes in gene expression. On-going studies are being performed to elucidate the function of the DBC1/STAT5A interaction and to clarify the impact of DBC1 on lipolysis in adipocytes.

Disclosure
A. Able: None. A.J. Richard: None. J.M. Stephens: None.
Adipocyte Gamma-Secretase Inhibition May Alter IL-6 Expression and Trafficking
Authors: SPARLING DAVID , MCCULLOUGH NILE 
Published: June 2018

Adipocyte-mediated inflammatory signaling has been proposed to alter adipose physiology in obesity. Activation of adipocytes can increase secretion of inflammatory cytokines such as IL-6. IL-6 secretion is regulated through changes in expression of both IL-6 itself, as well as its specific SNARE trafficking machinery. Recent data suggests that the γ-secretase enzyme complex is involved in both adipose insulin sensitivity and immune system regulation. We therefore hypothesized that adipocyte IL-6 secretion could be altered by γ-secretase inhibition. To explore this hypothesis, 3T3-L1 adipocytes were stimulated with lipopolysaccharide (LPS) in the absence or presence of a γ-secretase inhibitor. Changes in gene expression of inflammatory cytokines and SNARE complex components, including IL-6, MCP1, Stx6, Vti1b, SCAMP5, VAMP3, Stx4, and SNAP23 were examined by qRT-PCR, and immunofluorescence (IF) was used to determine adipocyte IL-6 expression and localization. IL-6 secretion was analyzed by ELISA. Transcription of IL-6 and several of the SNARE complex proteins were not significantly affected by γ-secretase inhibition. However, total cellular IL-6 increased by LPS stimulation by IF, which could be blocked by pre-treatment with a γ-secretase inhibitor. IL-6-associated SNARE component Vti1b mRNA levels were specifically affected by overnight γ-secretase inhibition. LPS stimulation increased Vti1b transcription 3-fold, which was completely blocked by γ-secretase inhibition (p&lt;0.01, n=6 per condition). Increasing amounts of LPS (range 0 to 10 ng/mL) for 6 hours led to an expected increase in IL-6 secretion, which was significantly blocked by overnight pre-treatment with a γ-secretase inhibitor. LPS-stimulated IL-6 secretion could also be decreased by short-term (30 minute) γ-secretase blockade. Taken together, this data suggests that the γ-secretase enzyme complex may be involved in a variety of signaling cascades controlling both the production and secretion of IL-6 in adipocytes.

Disclosure
D. Sparling: None. N. McCullough: None.
Insulin Receptor–Mediated Signaling Regulates Pluripotency Markers and Lineage Differentiation
Authors: GUPTA MANOJ K. , DE JESUS DARIO F. , KAHRAMAN SEVIM , SHAMSI FARNAZ , HU JIANG , TSENG YU-HUA , KULKARNI ROHIT 
Published: June 2018

Insulin receptor (IR)-mediated signaling is involved in the regulation of pluripotent stem cells, however its direct effects on regulating the maintenance of pluripotency and lineage development are not fully understood. To this end, we generated IR knock-out (IRKO) mouse induced pluripotent stem cells (miPSCs) from E14.5 mouse embryonic fibroblasts (MEFs) of global IRKO mice using a cocktail of four reprogramming factors (Oct4, Sox2, Klf4 and cMyc). iPSCs were maintained in a 2-inhibitor (2i) feeder-free system. Interestingly, expression of pluripotency markers including Klf4, Lin28a, Tbx3 and cMyc were upregulated while abundance of Oct4 and Nanog were enhanced by 4-fold and 3-fold respectively in IRKO iPSCs. Analyses of signaling pathways demonstrated downregulation of phospho-STAT3, pmTor and pErk, and an increase in the total mTor and Erk proteins in IRKO iPSCs. Stimulation with leukemia inhibitory factor (LIF) showed a 3-fold decrease of phospho-ERK in IRKO iPSCs (p&lt;0.vs. C, n=3 clones/group). On the contrary, Erk phosphorylation was increased during in-vitro spontaneous differentiation of iPSCs lacking IR. Lineage specific directed differentiation of the iPSCs revealed that cells lacking IR showed enhanced expression of neuronal lineage markers (Pax6, Tubb3, Asci and Oligo2) while exhibiting a decrease in adipocyte (Fas, Acc, Pparg, Fabp4, C/ebpa and Fsp27) and pancreatic beta cell markers (Ngn3, Isl1 and Sox9) (p&lt;0.05, n=3 clones/group). Further molecular characterization by phosphoproteomics confirmed the novel IR-mediated regulation of the global pluripotency network and several key proteins involved in various aspects of growth and embryonic development ((p&lt;0.05, n=3 clones/group). Thus, IRKO iPSCs provide an opportunity to explore the crosstalk of insulin receptor signaling with key signaling pathways required for the maintenance of pluripotency and lineage determination.

Disclosure
M.K. Gupta: None. D.F. De Jesus: None. S. Kahraman: None. F. Shamsi: None. J. Hu: None. Y. Tseng: Other Relationship; Self; Chugai Pharmaceutical Co., Ltd.. Research Support; Self; MedImmune. R. Kulkarni: None.
Use of Oral Contraceptives at Child-Bearing Age Are Associated with the Prevalence of Diabetes in Postmenopausal Women
Authors: KIM SUNG-WOO , KIM REBECCA , JEON EONJU , LEE JI-HYUN , SHON HO-SANG 
Published: June 2018

Objective: The effect of past use of oral contraceptives (OCs) at child-bearing on the incidence of diabetes and insulin resistance (IR) after menopause is not clearly elucidated. This study aimed to evaluate the association of past use of OCs with the development of diabetes and IR in post-menopausal women.
Methods: This cross-sectional study was based on data from the Korea National Health and Nutrition Examination Survey from 2007 to 2012. Of the 50,4participants, 6554 post-menopausal women were selected and included in the analysis. The long-term effects of OCs use on the prevalence of diabetes in post-menopausal women were examined using multivariate logistic analysis. In addition, fasting glucose and insulin levels were measured in 3338 nondiabetic post-menopausal women, and the association between IR and OCs was examined by analysis of covariance.
Results: The prevalence of diabetes was significantly higher in post-menopausal participants who had taken OCs for more than 6 months than in those who had never taken OCs. The association remained significant after adjusting for multiple confounding factors (odd ratio, 1.379; 95% CI, 1.115-1.707; P=0.003). The duration of OCs use was also positively associated with the prevalence of diabetes. Furthermore, taking OCs for more than 6 months led to a significant increase in fasting insulin levels and HOMA-IR in nondiabetic participants.
Conclusion: Past use of OCs for more than 6 months led to a significant increase in the prevalence of diabetes in post-menopausal women, and increased IR in nondiabetes participants. These results suggested that prolonged use of OCs at a reproductive age is an important risk factor for developing diabetes in post-menopausal women.

Disclosure
S. Kim: None. R. Kim: None. E. Jeon: None. J. Lee: None. H. Shon: None.
An Oral Glucose Load Decreases Post-Prandial Microparticles in Obese Adults With and Without Prediabetes
Authors: EICHNER NATALIE , GILBERTSON NICOLE M. , MUSANTE LUCA , SALVIA SABRINA LA , BARRETT EUGENE , WELTMAN ARTHUR , ERDBRUEGGER UTA , MALIN STEVEN K. 
Published: June 2018

Microparticles (MPs) are a novel mediator and/or biomarker of cardiovascular disease (CVD) and type 2 diabetes. A high fat meal can elevate MP release in relation to CVD risk, but the effects of carbohydrates are unknown. This is clinically relevant as low-fat diets are advised for glycemic control. We tested the hypothesis that a 75g oral glucose tolerance test (OGTT) would promote changes in MPs linked to CVD risk. Twenty-five obese adults (Age: 52.4±3.2y, BMI: 32.5±1.2kg/m2) were screened for prediabetes using ADA criteria (75g OGTT and/or HbA1c). Eight were normal glucose tolerant (NGT) and 17 had prediabetes (PD). Body composition (bioelectrical impedance) was measured and arterial stiffness (augmentation index; AI), insulin and glucose were collected at 0 and 2 hour of the OGTT to assess CVD risk. Annexin V+ (AV+) and Annexin V- (AV-) total MPs and platelet MPs (CD31+/CD41+; CD41+) were also collected at these times and analyzed from fresh plasma via imaging flow cytometry. There were no statistical differences in age, BMI, or body fat (all P&gt;0.21) between NGT and PD, although fasting and 2 hour glucose as well as insulin were higher in PD vs. NGT (P&lt;0.03). Glucose and insulin increased during the OGTT (both P≤0.005), while arterial stiffness at 2 hour decreased by 6.9% (P=0.06). Fasting MPs were not different between groups. Total MPs, AV+ CD41+ and AV+ CD31+/CD41+ MPs decreased after the OGTT (P≤0.04), while AV- CD41+ (P=0.08) and CD31+/CD41+ (P=0.10) trended. Attenuated OGTT insulin responses were related to lower post-prandial platelet AV- CD31+/CD41- (r=0.46, P=0.06) and AV- CD41+ (r=0.45, P=0.06) MPs, while arterial stiffness was associated with reduced total MPs (r=-0.49, P=0.03) and AV+CD41+ (r=-0.52, P=0.02). An oral glucose load lowers post-prandial total and platelet MPs in obese adults with NGT and PD. Further work is required to examine MP content in order to gain mechanistic insight for optimizing type 2 diabetes management.

Disclosure
N. Eichner: None. N.M. Gilbertson: None. L. Musante: None. S. La Salvia: None. E. Barrett: None. A. Weltman: None. U. Erdbruegger: None. S.K. Malin: None.
Generation of a Novel Gene Therapy Vector Encoding Beta-Cell–Specific Insulin for Diabetes Treatment
Authors: SAHIN ELIF O. , ERENDOR FULYA , BALCI MUSTAFA KEMAL , SANLIOGLU SALIH 
Published: June 2018

Insulin injection is the main treatment modality especially for patients with type 1 diabetes. Despite the development of several insulin analogs, recent meta-analysis indicated that rapid and long acting insulin analogs provide little benefit compared to conventional insulins regarding to glycemic control. Therefore, gene therapy methods are investigated to develop a novel treatment approach to reconstitute a natural endogenous insulin expression profile in diabetic patients. For this purpose, Multisite Gateway Technology was employed to generate a transfer vector carrying lentiviral backbone with minimal insulin promotor hooked up to proinsulin gene sequence. Following confirmation of the transfer plasmid by restriction enzyme digestion and DNA sequence analysis, high-titer LentiINS vectors at a concentration of 10e9 TU/ml were successfully produced by CaPO4 cotransfection of 293T cells with packaging and transfer plasmids. While transduction of NIT1 mouse pancreatic beta cell lines produced 3 fold increase in insulin gene expression compared to controls, no insulin expression was detected in 293T kidney cell line. These results indicate that we successfully constructed an HIV-based lentiviral gene therapy vector being capable of beta cell-specific insulin gene expression and secretion (TUBITAK 215S820).

Disclosure
E.O. Sahin: None. F. Erendor: None. M. Balci: None. S. Sanlioglu: None.