bluuebunny/crossref_metadata_2025_split huggingface dataset:
252-OR: Cardiorespiratory Fitness Predicts Lower Mortality Rates in Type 2 Diabetes
Authors: WILLS ANDREW C. , KNOWLER WILLIAM C. , CURTIS JEFFREY , OLAIYA MUIDEEN T. 
Published: June 2019

Look AHEAD was a randomized multi-center clinical trial comparing effects of an intensive lifestyle intervention (ILI) with diabetes support and education (DSE) on CVD events in 5145 adults aged 45-76 years who were overweight or obese and had type 2 diabetes. After 10 years of follow-up, the ILI had no significant effect on the primary outcome of fatal or non-fatal CVD events or on all-cause or CVD mortality rates. We now present a secondary analysis of the effects of cardiovascular fitness at baseline on mortality in 4865 participants. Fitness was defined as the metabolic equivalence (METs) level achieved during maximal workload on a treadmill. For all participants, the mean (SD) were 7.20 (1.97) METs. The all-cause death rate was 14% lower per 1 MET greater fitness at baseline, adjusted for age, sex, race/ethnicity, CVD history, BMI and treatment group (HR 0.86 95% CI 0.80 -0.92). In this model, CVD history (HR 1.82 95% CI 1.42-2.35) but not ILI (HR 0.87 95% CI 0.71-1.07) significantly predicted mortality. Similarly, CVD mortality was 28% lower per 1 MET (HR 0.72, 95% CI 0.62-0.83). Fitness was similarly protective from the other CVD outcomes. The Figure shows all-cause mortality rates modeled by history of CVD, treatment group, age 60 years, and white men not on beta blockers because they were the largest population in the dataset.
In conclusion, fitness measured by maximal workload on a treadmill was highly predictive of CVD and mortality.

Disclosure
A.C. Wills: None. W.C. Knowler: None. J. Curtis: None. M.T. Olaiya: None.

Funding
National Institutes of Health
257-LB: Long-Term Effects of Birth Mode on Gut Microbiome Colonization
Authors: NIES LAURA DE , BUSI SUSHEEL BHANU , HABIER JANINE , WAMPACH LINDA , HEINTZ-BUSCHART ANNA , DE BEAUFORT CARINE , WILMES PAUL 
Published: June 2019

Background: Caesarean section (CS) disrupts mother-to-neonate transmission of specific microbial strains, linked functional repertoires and immune system priming. We investigated whether the differences in microbial populations and functional potentials persist at 1 year of age.
Methodology: We performed high-resolution, quantitative metagenomic analyses of the gut microbiomes of 8 infants born by vaginal delivery (VD) (n=3) or by CS (n=5) at 1 year. Metagenomic sequencing reads were used to generate functional repertoire profiles based on the homology of the assembled genes/contigs with the KEGG database. Simultaneously, LPS from the infant stool samples was used to stimulate primary immune cells (dendritic cells) obtained from healthy human donors. An enzyme-linked immunosorbent assay for TNFα was used to determine concomitant levels of cytokines. Differential mOTU (metagenomic operational taxonomic unit) and KEGG pathway analyses were performed using the DESeq2 R package.
Results: Seven mOTUs including Anaerotruncus colihominis, [Ruminococcus obeum], and Subdoligranulum variabile demonstrated between 10- to 30- fold enrichment in the VD samples compared to the CS group (p &lt; 0.05). Although ordination analyses did not indicate significant differences between the groups, the functional repertoire analysis found several KEGG pathways differentially enriched when comparing VD and CS samples. The CS samples exhibited enrichments in CD226 antigen, cell adhesion and vancomycin resistance pathways, while the VD group had an increased abundance of carbon metabolism and epithelial cell invasion pathways. Previously increased TNFα levels observed in VD samples right after birth compared to the CS group, were not observed at 1 year.
Conclusion: After 1-year, persistent differences in gut mOTUs were observed between CS and VD born infants. Whether the CD226 pathway upregulation relates to human auto-immune disease, requires further analysis and validation.

Disclosure
L. de Nies: None. S. Busi: None. J. Habier: None. L. Wampach: None. A. Heintz-Buschart: None. C. de Beaufort: Consultant; Self; Medtronic. P. Wilmes: None.

Funding
Fondation Andr? et Henriette Losch; National Research Fund; University of Luxembourg; Integrated BioBank of Luxembourg
257-OR: High Glucose Levels and Risk of Vascular Diseases—Observational Studies and Mendelian Randomization Studies of the General Population
Authors: EMANUELSSON FRIDA , MAROTT SARAH , TYBJAERG-HANSEN ANNE , NORDESTGAARD BØRGE GRØNNE , BENN MARIANNE 
Published: June 2019

Aim: To investigate the observational and causal associations between glucose levels in the nondiabetic range and risk of vascular diseases in the general population.
Methods: We included 112,457 nondiabetic individuals from the Copenhagen City Heart Study and the Copenhagen General Population Study and assessed their prospective risk of retinopathy, neuropathy, diabetic nephropathy, peripheral arterial disease (PAD), and myocardial infarction (MI) as a function of increasing glucose levels. We used Mendelian randomization (MR) to assess a potential causal effect of high glucose levels on the vascular endpoints using genetic variants known to be associated with elevated glucose levels. We replicated our analyses in a 2-sample MR design using glucose data from the MAGIC consortium and vascular endpoint data from the UK Biobank.
Results: Observationally, glucose levels in the nondiabetic range and higher were associated with increased risks of retinopathy, neuropathy, diabetic nephropathy, PAD, and MI (p trend for all &lt;0.001). In genetic, causal analyses risk ratios for a 1 mmol/L higher glucose were 2.01 (95% confidence interval: 1.18-3.41) for retinopathy, 2.15 (1.38-3.35) for neuropathy, 1.58 (1.04-2.40) for diabetic nephropathy, 1.19 (0.90-1.58) for PAD, and 1.49 (1.02-2.17) for MI. Summary level data from the MAGIC consortium and the UK Biobank gave similar results.
Conclusion: In this general population, glucose levels in the nondiabetic range and higher were prospectively associated with a high risk of retinopathy, neuropathy, diabetic nephropathy, PAD, and MI. These associations were causal for retinopathy, neuropathy, diabetic nephropathy, and MI, but causality could not be confirmed for PAD. These findings suggest that elevated glucose levels in the nondiabetic range is an important risk factor for microvascular diseases.

Disclosure
F. Emanuelsson: None. S. Marott: None. A. Tybjaerg-Hansen: None. B.G. Nordestgaard: None. M. Benn: None.

Funding
Danish Council for Independent Research
26-OR: Digenic FGFR1/KLB Variants Associated with Endocrine Specific FGF-21 Signaling Defects and Extreme Insulin Resistance
Authors: STONE STEPHEN I. , WEGNER DANIEL J. , WAMBACH JENNIFER A. , COLE F. SESSIONS , ORNITZ DAVID M. , URANO FUMIHIKO 
Published: June 2019

Fibroblast growth factor 21 (FGF-21) is a hepatokine with multiple important metabolic functions. However, the role of FGF-21 in human insulin sensitivity remains unclear. We report an adolescent female with severe acanthosis nigricans and extremely elevated insulin levels (2446 uIU/mL). Due to her severe insulin resistance, we looked into the possibility that she might have a genetic syndrome of insulin resistance. Whole exome sequencing revealed that she carries 2 variants of unknown significance in genes critical for FGF-21 signaling, Fibroblast Growth Factor Receptor 1 (FGFR1) and beta-Klotho (KLB). FGFR1 and KLB are transmembrane co-factors that bind FGF-21 leading to downstream signaling. A bicistronic expression plasmid was created recapitulating the above variants found in our subject. These plasmids were transfected into L6 myoblasts, a cell line known to express low levels of endogenous FGF receptor. ERK phosphorylation (pERK), a major downstream signal of FGF-21, was measured after treatment with FGF-21. pERK was strongly upregulated in cells transfected with wild type FGFR1 and KLB (500,710 AU). However, variant FGFR1 and KLB greatly attenuated pERK (107,068 AU). Canonical FGF-2 treatment led to robust upregulation of pERK in both wild type and variant cell lines (374,968 AU and 441,654 AU respectively). Expression of Slc2a1 (Glut1) and Slc2a4 (Glut4) was also measured in response to FGF-21. Cells transfected with wild type FGFR1 and KLB led to a 1.7-fold increase in both Glut1 and Glut4 expression, whereas Glut1 and Glut4 expression was attenuated in cells transfected with the FGFR1 and KLB variants (1.2 and 0.7-fold respectively). These results suggest that the digenic FGFR1 and KLB variants found in our subject lead to an endocrine specific FGF-21 resistance with decreased GLUT1- and GLUT4-mediated glucose uptake. This helps explain our subject’s severe insulin resistance, and may represent a novel disorder related to FGF-21.

Disclosure
S.I. Stone: Research Support; Self; National Institute of Diabetes and Digestive and Kidney Diseases. Speaker's Bureau; Self; AbbVie Inc. D.J. Wegner: None. J.A. Wambach: None. F.S. Cole: None. D.M. Ornitz: None. F. Urano: Board Member; Self; Healthbeat. Research Support; Self; Aetas, Amylyx, JDRF, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health. Stock/Shareholder; Self; CytRx. Other Relationship; Self; Novus Biologicals.

Funding
National Institutes of Health (HL120002); Institute of Clinical and Translational Sciences/Children's Discovery Institute (UL1TR00448)
261-LB: Robotic Pancreas Kidney Transplantation for Type 1 Diabetic Patient with End-Stage Renal Disease: First Year Experience in Single Institute
Authors: DUK LEE SEUNG , CANTRELL LEIGH A. , OBERHOLZER JOSE 
Published: June 2019

Background: Simultaneous pancreas kidney transplantation for type 1 diabetic patients with ESRD is a highly successful, but also very invasive surgery. Robotic assistance facilitates a minimally invasiveness approach to this technically intricate surgery. The present study aims to evaluate the feasibility and outcomes of the first year`s experience in robotic pancreas kidney transplantation (RPKT) for type 1 diabetic patients with ESRD.
Patients and Methods: From October 2017 to October 2018, we enrolled consecutive cases of RPKT at the University of Virginia. Operative and clinical features including graft and patient survivals were retrospectively analyzed. The organs were introduced via 6 cm Pfannenstil incision.
Results: Six patients (3 male/3 female) successfully underwent RPKT. Median age was 32 years old (range: 30-46years). Median pre-transplant body mass index (BMI) was 25.5 mg/kg2 (Range: 20.6-33.4 mg/kg2). Operative time and estimated blood loss were 593 min (473-733) and 300cc (150-400), respectively. Total cold and warm ischemic time were 436 min (242-585) for pancreas/725min (403-835) for kidney and 43 min (34-60) for pancreas/56min (44-69) for kidney, respectively. We created 5 enteric and 1 bladder drainage for transplant pancreas. There were no conversions to open. No major complications during surgery were observed. We reported one case of delayed graft function of kidney and two cases of rejection with complete recovery. We found one patient with a minor trocar wound dehiscence and revision of ureterocystostomy for stricture. There was one pancreas graft loss at postoperative 7 days due to graft vein thrombosis. No patient death was noted.
Conclusion: RPKT for type 1 diabetic patients with ESRD appears to be a safe and effective surgical procedure with potential advantages of wound complication, quality of vascular anastomosis, and shorter recovery with minimal incision compared to conventional open surgery.

Disclosure
S. Lee: None. L.A. Cantrell: None. J. Oberholzer: Employee; Self; CellTrans. Research Support; Self; JDRF. Stock/Shareholder; Self; Sigilon Therapeutics.
262-LB: Human Pancreas-Derived Multipotent Stromal Cells Secrete Islet Regenerative Stimuli
Authors: COOPER TYLER , MA JUN , BELL GILLIAN I. , LAJOIE GILLES , HESS DAVID A. 
Published: June 2019

We recently demonstrated that intra-pancreatic (iPan) delivery of conditioned media (CM) generated by human bone marrow-derived multipotent stromal cells (hBM-MSC) can act as a biotherapeutic to reduce hyperglycemia in STZ-treated mice by stimulating endogenous islet regeneration. We have established analogous stromal cell lines derived from human islet cultures that grow adherent to plastic and possess multipotent mesodermal differentiation in vitro. Furthermore, CM generated by these human pancreas-derived MSC (hPanc-MSC) contained proangiogenic stimuli within extracellular vesicles (EVs) that accelerated blood vessel regeneration after injection into ischemic hindlimbs. Herein, we investigated whether iPan-injection of CM generated by hPanc-MSC could stimulate islet regeneration in STZ-treated mice. We hypothesized that islet regenerative stimuli would be harboured within EVs secreted by hPanc-MSC. iPan-injection of unfractioned CM (6µg) generated by hBM-MSC and hPanc-MSC equally reduced hyperglycemia in STZ-treated mice, compared to mice injected with media control (*p&lt;0.05). To determine whether islet regenerative stimuli was harboured within EVs, ultrafiltration was used to separate CM into EV enriched (EV+) or depleted (EV-) subfractions. Enrichment of EVs was validated by nanoscale flow cytometry and atomic force microscopy, and the EV+ and EV- hPANC-MSC subfractions were compared for protein content by label-free mass spectrometry. Expression of typical EV-associated proteins (CD9, CD81, CD63) were exclusive to the EV+ fraction and putative islet regenerative factors were detected in both the EV- and EV+ subfractions C. Mice iPan-injected with either EV+ or EV- CM sub-fractions showed significantly reduced hyperglycemia compared to unconditioned media controls (*p&lt;0.05). These initial studies provide a foundation to further explore the use the hPanc-MSC secretome or related small-molecule pharmaceuticals to enhance islet-regeneration in situ.

Disclosure
T. Cooper: None. J. Ma: None. G.I. Bell: None. G. Lajoie: None. D.A. Hess: None.
264-LB: Evaluation of Awareness about Insulin Injection Technique and Prevalence of Lipohypertrophy amongst Insulin-Taking Diabetics in Central India
Authors: SUNIL GUPTA SHLOK , CHAOJI SUNANDA , GUPTA SUNIL , GUPTA KAVITA , GATHE SACHIN 
Published: June 2019

Background: Lipohypertrophy (LH) at insulin injection sites is a preventable complication in patients of diabetes mellitus taking insulin injection therapy. We evaluated the prevalence, contributing risk factors, and consequences of LH along with awareness regarding correct insulin injection technique amongst these diabetic patients.
Methodology: It is a cross sectional observational questionnaire-based study conducted at a tertiary health centre in Nagpur, India. 31 type 1 diabetics (T1DM) and 43 type 2 diabetics (T2DM) who are on insulin injection therapy for over one year, were randomly selected and presence of LH was evaluated through visual and palpation techniques. Demography, anthropometry, awareness of correct insulin injection technique were noted using a validated questionnaire. Their effect on LH was determined along with its correlation with complains of unexplained hypoglycaemia (UH).
Results: The prevalence of LH amongst T1DM is 48.39% and amongst T2DM is 18.60%. Prevalence was significantly higher in T1DM than in T2DM (p=0.0133). Young age (p=0.0042), Male gender (p=0.0011), decreased duration of diabetes (p=0.0281), improper rotation of the correct sites (p=0.0032) significantly increased the prevalence of LH. Occurrence of UH were significantly higher in patients with LH (p=0.0078). Independent risk factors for LH amongst T2DM include less age (p=0.0358), Male gender (p=0.0048) and improper rotation of sites (p=0.0071). No independent risk factors were found significant for LH amongst T1DM as well as no significance was observed for prevalence of LH with respect to needle reuse, number of injections taken per day and total daily dosage of insulin, which may be regarded to low sample size.
Conclusion: Improper rotation of injection sites, Male gender, younger age, lesser duration of diabetes and T1DM contribute to a higher prevalence of LH, which in turn will increase the risk of UH.

Disclosure
S.S. Gupta: None. S. Chaoji: None. S. Gupta: None. K. Gupta: None. S. Gathe: None.
27-LB: Alteration of the Gut Microbiome by Resveratrol Is Associated with Improvement in Function of Kidney in db/db Mice
Authors: DAFA DING , YIBING LU , TINGTING CAI , XIAOLONG YE , JIANHUA MAO 
Published: June 2019

Oral administration of resveratrol is able to amelioration the progression of diabetic nephropathy, but the underlying cause is unknown. In this report, we demonstrated that resveratrol relieved a series of indicators of diabetic nephropathy. Oral administration of resveratrol was able to sequester negatively charged bacterial endotoxin in the gut, leading to 1) reduced plasma endotoxin levels, 2) resolved systemic inflammation and the Intestinal environment, and 3) lowered renal inflammation and improved renal function. Gut dysbiosis, characterized as an increment of Bacteroides, Alistipes, Turicibacter and Parabacteroides, and reduction of Enterococcus , was fully modulated by resveratrol, indicating that inflammatory-associated microbiome in the gut are critical for the metabolic endotoxemia. Furthermore, fecal bacteria transplant, derived from heathy resveratrol-treated mices, was sufficient to antagonize the renal dsyfunction and rebalance of gut microbiome in the recipient mice. These results indicate that resveratrol mediated changes in the gut microbiome may play an important role in the mechanism of action of resveratrol, which provide supporting evidence for the gut-kidney axis in DN.

Disclosure
D. Dafa: None. L. Yibing: None. C. Tingting: None. Y. Xiaolong: None. M. Jianhua: None.

Funding
Major Research and Development Project of Jiangsu (BE2016800); Nanjing Science and Technology Plan Project (BE201715015)
271-OR: Whole-Exome Sequence Identifies Potentially Functional Rare Variants in Bardet-Biedl Syndrome (BBS) Genes that Associate with Obesity in American Indians
Authors: DAY SAMANTHA E. , MULLER YUNHUA L. , KOBES SAYUKO , KIM HYE IN , VAN HOUT CRISTOPHER V. , GOSALIA NEHAL , YE BIN , SHULDINER ALAN R. , HANSON ROBERT L. , ALTOK CIGDEM KOROGLU , BOGARDUS CLIFTON , BAIER LESLIE J. 
Published: June 2019

American Indians suffer from a high prevalence of obesity. Although their obesity is typically polygenic in nature, we are systematically analyzing known monogenic obesity genes in a population-based sample of Pima Indians. Previously, we have reported that 3% of this population carries functional mutations in MC4R. The goal of the current study is to determine whether variation in any of the 21 known BBS genes also contribute to obesity in this population. Analysis of whole exome sequence data from 6,809 Pima Indians (44.8% male; mean±SD age 34.8 ± 16.0 years; mean±SD maximum body mass index (BMI) 36.2 ± 8.8 kg/m2; 31.9% had T2D) identified 1,726 variants in the 21 BBS genes, of which 273 variants had a Combined Annotation Dependent Depletion (CADD) score &gt;20 (top 1% most likely to be deleterious). Among these potentially deleterious variants, 32 were exclusive to, or enriched in, obese subjects (mean BMI for carriers &gt;40). Five of the 32 variants were in the BBS9 gene and these were prioritized for functional studies. BBS mutations may impact obesity via leptin signaling, where BBS gene dysfunction has led to increased food intake. Therefore, in vitro luciferase reporter assays targeting STAT3 transcriptional activity were used to assess alterations in leptin signaling by the BBS variants. Analysis of a Thr549Ile (rs59252892) in BBS9 (mean BMI of Ile allele carriers was 41.3 ± 12.4 kg/m2 vs. 36.4 ± 8.8 kg/m2 for non-carriers, P=0.0008) showed that the Ile allele had a 28% increase in leptin signaling activity compared to the Thr allele (P=0.006, 5 experiments). Moreover, western blot analysis showed that the Ile allele had a 19% increase in STAT3 phosphorylation compared to the Thr allele. Comparable in vitro results were observed with a S545G, but not a K810E or L665F in BBS9.
In summary, whole exome sequencing in Pima Indians has identified potentially functional variants in BBS9 that may increase risk for obesity through leptin signaling.

Disclosure
S.E. Day: None. Y.L. Muller: None. S. Kobes: None. H. Kim: Employee; Self; Regeneron Pharmaceuticals. C.V. Van Hout: Employee; Self; Regeneron Pharmaceuticals. N. Gosalia: Employee; Self; Regeneron Pharmaceuticals. B. Ye: Employee; Self; Regeneron Pharmaceuticals. A.R. Shuldiner: Employee; Self; Regeneron Pharmaceuticals. Stock/Shareholder; Self; Rhythm Pharmaceuticals, Inc. R.L. Hanson: None. C. Koroglu Altok: None. C. Bogardus: None. L.J. Baier: None.
272-LB: The Micropeptide ORF60 Modulates Insulin Sensitivity
Authors: GUPTA NIKHIL , SHI YU , LI PING , CAO HAIMING 
Published: June 2019

Although long non-coding RNAs (lncRNAs) are non-protein-coding transcripts by definition, recent studies have shown that a number of putative small open reading frames within lncRNAs are translated. The biological significance of these hidden polypeptides is still unclear, although studies have demonstrated functional significance for some polypeptides.
Recently, our lab discovered a novel micropeptide ORF60 encoded by a lncRNA. The micropeptide is conserved across species. Topology prediction indicates that the micropeptide is likely transmembrane. RNA transcript expression is nearly ubiquitous across tissues. In mice, ORF60 expression was found to be increased in the setting of high fat diet (HFD). Using CRISPR/Cas9 technology we developed ORF60 KO mice. In both mice feeding on normal chow diet and HFD, we observed impaired glucose tolerance in KO mice compared to wild type mice. ORF60 KO mice also demonstrated impaired response to insulin compared to WT mice on insulin tolerance testing (ITT), suggesting insulin resistance. Mouse tissue harvested under the condition of insulin infusion demonstrated insulin signaling attenuation in white adipose tissue (WAT) and brown adipose tissue (BAT). We are currently dissecting the molecular mechanism underlying ORF60’s influence on insulin signaling pathways.
In summary, our results demonstrate that the conserved micropeptide ORF60 regulates insulin sensitivity in WAT and BAT in mice. Micropeptides involved in diabetes may serve as novel drug targets.

Disclosure
N. Gupta: None. Y. Shi: None. P. Li: None. H. Cao: None.