bluuebunny/crossref_metadata_2025_split huggingface dataset:
214-OR: Effects of Fluid Rehydration Strategy on Correction of Acidosis and Electrolyte Abnormalities in Children with Diabetic Ketoacidosis
Authors: REWERS ARLETA , KUPPERMANN NATHAN DAVIS , STONER MICHAEL J. , GARRO ARIS , QUAYLE KIMBERLY , BENNETT JONATHAN E. , MYERS SAGE , MCMANEMY JULIE K. , NIGROVIC LISE E. , TRAINOR JENNIFER L. , TZIMENATOS LEAH , KWOK MARIA , OLSEN CODY S. , CASPER THERON C. , GHETTI SIMONA , GLASER NICOLE 
Published: June 2019

Background: Fluid replacement to correct acidosis and electrolyte abnormalities is the cornerstone of treatment of diabetic ketoacidosis (DKA). We evaluated the rates of change in pH, pCO2, anion gap, Na, K, and Cl in children randomized to four DKA fluid treatment protocols.
Methods: From a large study conducted by The Pediatric Emergency Care Applied Research Network we used data from 714 children &lt;18 y old with moderate or severe DKA. In a 2-by-2 factorial design, we evaluated the effects of intravenous rehydration with 0.9% vs. 0.45% NaCl and fast vs. slow rate of fluid administration, after an initial 0.9% NaCl bolus. The fast arm assumed 10% dehydration - half replaced in 12 h + maintenance fluids, the other half replaced over subsequent 24 h + maintenance fluids. The slow arm assumed 5% dehydration replaced evenly over 48 hours + maintenance fluids. We tested for differences in the rate of change of laboratory values between treatments using stratified Van Elteren and Cochran Mantel Haenszel tests.
Results: The median age, sex, and baseline laboratory characteristic of the patients did not differ by treatment arm. The rate of change of pH did not differ by the arm, however the rate of rise of pCO2 was more rapid in the fast vs. slow fluid arms during the initial 4 h. The rate of decrease in anion gap was more rapid in the fast vs. slow arms during the initial 4 h and 8 h (p&lt;0.01 for all). Glucose-corrected Na remained steady for subjects assigned to 0.9% NaCl vs. those assigned to 0.45% NaCl whose glucose-corrected Na decreased at 4 and 8 h (p&lt;0.01). K levels decreased and Cl increased more rapidly with 0.9% vs. 0.45% NaCl (p&lt;0.01). Hyperchloremic acidosis developed in 46.1% of patients in the fast arms vs. 35.2% in the slow arms (p&lt;0.01).
Conclusions: In children treated for DKA, faster fluid administration led to more rapid normalization of anion gap and pCO2 than slower fluid rates. However, more rapid fluid rates were associated with increased frequency of hyperchloremic acidosis.

Disclosure
A. Rewers: None. N.D. Kuppermann: Other Relationship; Self; InsuCalc. M.J. Stoner: None. A. Garro: None. K. Quayle: None. J.E. Bennett: None. S. Myers: None. J.K. McManemy: None. L.E. Nigrovic: None. J.L. Trainor: None. L. Tzimenatos: None. M. Kwok: None. C.S. Olsen: None. T.C. Casper: None. S. Ghetti: None. N. Glaser: Consultant; Self; Bristol-Myers Squibb Company. Other Relationship; Self; Insucalc.

Funding
Eunice Kennedy Shriver National Institute of Child Health and Human Development
2140-P: Pancreatic Islet and Exocrine Tissue Innervation Is Not Altered in Type 1 Diabetes
Authors: RICHARDSON TIFFANY M. , HALIYUR RACHANA , ARAMANDLA RADHIKA , BOTTINO RITA , POWERS ALVIN C. , BRISSOVA MARCELA 
Published: June 2019

Impaired counterregulatory response of glucagon to hypoglycemia is a common complication of type 1 diabetes (T1D). One proposed contributor to abnormal glucagon secretion is decreased sympathetic innervation in T1D islets. To systematically assess the innervation of human pancreatic tissues, we examined samples from donors with recent-onset T1D (&lt;10 years, age 13-45 years, n = 4), longstanding T1D (&gt;10 years, age 27-63 years, n = 3), and nondiabetic controls (age 10-52 years, n = 4). Islet and exocrine tissue innervation was visualized by pan-neuronal marker tubulin β-3 and analyzed by a 2-D morphometry and 3-D rendering. By quantifying the nerve fiber length (1.9±0.4 nm/μm2) and density (646±94 fibers/mm2), we found that innervation in control human islets was nearly 20-fold less than previously reported in mouse islets. In contrast to mouse, human exocrine tissue was far more innervated than islets with fiber length of 6.8±0.4 nm/μm2(p&lt;0.05), and density of 1336±50 fibers/mm2(p&lt;0.05). The 3-D analysis showed that nerve fibers follow extracellular matrix (ECM) formed around acini and from there, they extend further into islets along the ECM made by islet vasculature. Although T1D islets were primarily composed of α cells, the islet area was similar to controls (control: 15.7±1.5 mm2, recent-onset T1D: 16.5±1.4 mm2, long-standing T1D: 15.3±1.5 mm2; p&gt;0.05), and so was the 3-D arrangement of nerve fibers in exocrine tissue and islets. Regardless of T1D duration, the islet fiber length (recent-onset: 1.5±0.2 nm/μm2; long-standing: 1.0±0.2 nm/μm2) and density (recent-onset: 623±61 fibers/mm2; longstanding: 486±49 fibers/mm2), as well as exocrine fiber length (recent-onset: 5.2±.5 nm/μm2; longstanding: 5.8±0.4 nm/μm2) and density (recent-onset: 1119±77 fibers/mm2; longstanding: 1537±72 fibers/mm2) did not differ from controls (p&gt;0.05). These data indicate that neuronal patterning of human pancreas is significantly different from that of mouse and islet innervation does not appear to be altered in T1D.

Disclosure
T.M. Richardson: None. R. Haliyur: None. R. Aramandla: None. R. Bottino: Research Support; Self; Imagine Pharma. A.C. Powers: None. M. Brissova: None.
2149-P: Mitochondrial Akt Modulated Human Embryonic Stem Cell Differentiation Lineages—Analysis with Single Cell RNA Sequencing
Authors: LEE HSIAOCHEN , CHEN YU-HAN , LIN HUGO YOU-HSIEN , TA ALBERT , WANG PING H. , CHEN YUMAY , ANDERSEN BOGI 
Published: June 2019

How signaling pathways control embryonic stem cell (ESC) differentiation is poorly understood. Recent studies suggested mitochondria metabolism may play a role in the regulation of stem cell differentiation. In this study, we found a novel role for mitochondrial Akt1 (mito-Akt) that modulated human ESC (hESC) differentiation lineage. We observed that Akt was activated and translocated into mitochondria in hESC upon growth factor stimulation. To determine the effect of mito-Akt on hESC differentiation, hESC were transduced with His-tagged mitochondrial-targeting dominant negative Akt1/GFP (Ad-mdnAkt) or GFP adenoviral vector (Ad-GFP). The transgene was expressed in &gt;90% cells, and mitochondria-targeting was verified. Ad-mdnAkt altered hESC respiration, as measured with Seahorse extracellular flux analyzer, and mitochondria proton leak was increased. After 21 days of differentiation in vitro, H&E stain showed Ad-mdnAkt transduced cells formed more organized cell clusters than control ones. Bulk RNA was extracted for qRT-PCR with primer array of representative marker genes, and the results suggested that marker genes for mesoderm and endoderm were up-regulated whereas marker genes for ectoderm were down-regulated when mito-Akt was inhibited by Ad-mdnAkt in hESCs. To further dissect how mito-Akt modulates cell lineage, single-cell RNA seq was analyzed with 10X Genomics platform. Among the 24 clusters identified by t-SNE plot, Ad-mdnAkt cells were over-represented in 5 clusters and under-represented in 4 clusters when compared to the controls. Interestingly, hESCs with Ad-mdnAkt favored lineages toward hematological system and lymphoid tissue development instead of pancreatic progenitor cells.
In summary, mito-Akt is a novel modulator of stem cell differentiation, altering mito-Akt signaling during the early phase of hESC differentiation may impact later embryonic development.

Disclosure
H. Lee: None. Y. Chen: None. H.Y. Lin: None. A. Ta: None. P.H. Wang: None. Y. Chen: None. B. Andersen: None.
2154-P: Mice Fed a Low Protein Diet In Utero Show Decreased Apelin Receptor Presence in Ins+Glut2Lo Cells during Pregnancy Associated with Lower ß-Cell Mass
Authors: AZIMI HOUSHANG , SZLAPINSKI SANDRA , HILL DAVID J. 
Published: June 2019

The apelinergic system, consisting of the apelin receptor (APJ) and its cognate ligand apelin, is active in pancreas, skeletal muscle, adipose and other tissues. Apelin/APJ are implicated in energy metabolism as apelin is secreted by adipocytes and may bind APJ on β-cell membranes to reduce glucose-stimulated insulin secretion. Apelin-13 administration has been shown to increase islet cell mass and β-cell insulin content in mice with T1DM and can increase insulin sensitivity in obese, hyperinsulinaemic mice. The metabolic stress of pregnancy is normally accommodated by β-cell mass (BCM) expansion and increased glucose-stimulated insulin secretion. A failure to increase BCM can contribute to the risk of gestational diabetes. Exposure of mice to a low protein diet (LP) in utero can limit expansion of BCM during subsequent pregnancy. As previously shown, insulin-expressing but glucose-transporter-2-low (Ins+Glut2Lo) progenitor cells contribute to gestational BCM expansion. Thus, we investigated how APJ presence on Ins+Glut2Lo cells might differ in islets of pregnant C57Bl/6 mice previously exposed to LP vs. control (C) diet. F1 female offspring exposed to LP or C diet in utero and postnatal development were time mated and pancreata collected at gestational days (GD) 9 and 12. Insulin, Glut2, and APJ were visualized by immunohistochemistry with respect to β-cell localization in small endocrine clusters (1-5 β-cells) and in islets (≥ 6 β-cells). The percentage of Ins+Glut2LoAPJ+ cells relative to all Ins+ cells was significantly decreased in LP-exposed mice vs. C at GD12 (C 5.7±1.0%, LP 2.7±0.1%, p&lt;0.05) when Ins+Glut2Lo cell expansion is maximal, but not at GD9 (C 4.9±1.6%, LP 3.4±0.9%). Significant differences were not found in % Ins+Glut2LoAPJ+ cells relative to extra-islet clusters or islets between LP vs. C at GD 9 and 12. Results suggest that APJ presence may contribute to BCM expansion during pregnancy mediated by Ins+Glut2Lo cells.

Disclosure
H. Azimi: None. S. Szlapinski: None. D.J. Hill: None.
2167-P: Gamma-Hydroxybutyrate Prevents Lipotoxicity-Induced ß-Cell Dysfunction
Authors: NGA YEUNG LUCY SHU , IVOVIC ALEKSANDAR , TAN YAO FANG , BATCHULUUN BATTSETSEG , MAMELAK MORTIMER , GIACCA ADRIA 
Published: June 2019

Prolonged exposure to high levels of fatty acids impairs the insulin secretory capacity of pancreatic β-cells through mechanisms involving oxidative stress (lipotoxicity). Beta-cell lipotoxicity is thought to play an important role in the β-cell failure of type 2 diabetes. Antioxidants can protect β-cells from lipotoxicity by decreasing oxidative stress. Gammahydroxybutyrate (GHB), an endogenous neurotransmitter currently safely used as a treatment for narcolepsy, is also found in cells as an energy source and antioxidant. Previous studies have shown that GHB can protect mice from streptozotocin and alloxan induced diabetes; both compounds are generators of oxidative stress and yield models of type 1 diabetes. However, the effect of GHB in models of type 2 diabetes has not been investigated. We hypothesized that GHB can protect β-cells from lipotoxicity.
We first investigated the effectiveness of GHB in preventing β-cell dysfunction in high-fat treatment (palmitate) in an in vitro model with freshly isolated mouse islets. Beta-cell function was determined by glucose stimulated insulin secretion assays. Islets incubated with GHB were protected from β-cell lipotoxicity that resulted from the concurrent 48h exposure to palmitate. The antioxidant effects of GHB in lipotoxicity were further investigated by measuring reactive oxygen species (ROS) levels. GHB prevented the increase in ROS that was observed with the palmitate treatment while GHB by itself had no effects. While further studies need to be done to elucidate possible mechanisms of action of GHB beyond its antioxidant effect as well as GHB effectiveness in vivo, our findings suggest that GHB or potentially GHB derivatives may be of interest for their β-cell protective properties.

Disclosure
L. Yeung: None. A. Ivovic: None. Y. Tan: None. B. Batchuluun: None. M. Mamelak: None. A. Giacca: Research Support; Self; Jazz Pharmaceuticals.

Funding
Jazz Pharmaceuticals plc
217-LB: Associations between Multimorbidity and HbA1c in People with Type 2 Diabetes in Australian Family Practice
Authors: CHIANG JASON I. , FURLER JOHN , JANI BHAUTESH D. , MAIR FRANCES S. , THURAISINGAM SHARMALA , NICHOLL BARBARA I. , MANSKI-NANKERVIS JO-ANNE E. 
Published: June 2019

Introduction: Multimorbidity (MM) is common in people with T2D (PwT2D) who in Australia, receive medical care predominantly in family practice (FP). We explored the association between MM condition count and HbA1c in PwT2D attending Australian FP, using electronic health record (EHR) data.
Methods: Cross-sectional study of 69,718 PwT2D who had a FP encounter between 2013-15 captured in the NPS MedicineInsight dataset (EHR data from &gt;500 family practices and 3.5 million patients across Australia). MM condition count was based on coded and free text conditions in the reason for visit, prescription or medical history fields. Primary outcome: HbA1c. Multivariable mixed-effects linear regression examined associations between MM counts (total, concordant (T2D-related) and discordant (unrelated)) and HbA1c (%).
Results: We found statistically significant associations between HbA1c and total and discordant MM counts. Mean HbA1c (95% CI) was lower for those having 1, 2, 3 and 4+ total conditions compared to T2D only, -0.065 (-0.089, -0.041), -0.097 (-0.128, -0.066), -0.089 (-0.133, -0.046), and -0.097 (-0.146, -0.047) respectively, and for discordant conditions -0.085 (-0.108, -0.061), -0.100 (-0.134, -0.065), -0.141 (-0.195, -0.087), and -0.204 (-0.276, -0.133). There was no statistically significant association between HbA1c and increasing concordant count.
Conclusions: While EHR counts of MM and discordant conditions are associated with lower HbA1c, this is not likely to be clinically significant, suggesting the impact of MM on HbA1c remains unclear. The NPS dataset uses routine data entered by clinicians for the purpose of patient care, rather than epidemiological studies and may thus underestimate the number of conditions experienced by PwT2D. While clinicians need to consider the MM burden of PwT2D, the NPS MedicineInsight dataset needs further validation, eg through combining with other data sources, in the study of MM.

Disclosure
J.I. Chiang: None. J. Furler: Research Support; Self; Abbott, Sanofi. B.D. Jani: None. F.S. Mair: None. S. Thuraisingam: None. B.I. Nicholl: None. J.E. Manski-Nankervis: Research Support; Self; Australian National Health and Medical Research Council, Boehringer Ingelheim International GmbH, Diabetes Australia, Eli Lilly and Company. Speaker’s Bureau; Self; RACGP.

Funding
Royal Australian College of General Practitioners
2173-P: Effects of AMP-Activated Protein Kinase Activation on Insulin Secretion in Mice
Authors: NGUYEN-TU MARIE-SOPHIE , HARRIS JOSEPH P. , MARTINEZ-SANCHEZ AIDA , LECLERC ISABELLE , RUTTER GUY A. 
Published: June 2019

AMP-activated protein kinase (AMPK) is an important cellular energy sensor in regulation of metabolic activity. AMPK activation improves glucose homeostasis by increasing muscle glucose uptake and reducing hepatic glucose production. Although the effects on glucose-stimulated insulin secretion in the pancreatic β cells are still controversial. Our aim was to investigate the regulation of insulin secretion in pancreatic β cells using specific pharmacological AMPK activators and in mouse models of constitutive activation. We used a chronic global AMPK (γ1D316A) and a pancreatic β cell specific (β-D316A) activation mouse model that express the AMPK γamma1 isoform with a D316A mutation under the control of β-actin and ins1 promoter respectively. We tested how chronic AMPK activation impacted insulin secretion on C57BL6 mouse isolated islets using direct activators PF-06409577 (β1 isoform-selective) and RA089 (binds both β1 and β2 isoforms). Global activation of AMPK reduced insulin secretion from islets isolated from γ1D316A mice on chow and high-fat diet (2.4-fold, p&lt;0.05 and 1.9-fold p&lt;0.01, respectively). Dual AMPK activation of β1 and β2 (RA089) on control mouse islets enhanced in a dose-response manner insulin secretion in response to high glucose (17mM) and KCl after 30 minutes of incubation (1.7-fold p&lt;0.005 and 1.7-fold p&lt;0.005, respectively). The same effects were observed by activating AMPK β1 isoform with PF-06409577, increasing insulin release in response to high glucose and KCl (2-fold, p&lt;0.005 and 1.7-fold, p&lt;0.05). These findings suggest that the effects of AMPK on insulin secretion depends on its mode of activation. Acute pharmacological activation targetting β1 and/or β2 subunit increased insulin secretion whereas chronic global activation in mice reduced insulin release. Investigating our second model of chronic activation β-D316A and further studies of our models will help us decipher the role of AMPK in the pancreas.

Disclosure
M. Nguyen-Tu: None. J.P. Harris: None. A. Martinez-Sanchez: None. I. Leclerc: None. G.A. Rutter: Consultant; Self; Sun Pharma.
2175-P: Isoform-Specific Roles of Prolyl Hydroxylase Domain Proteins in Regulating Beta-Cell Function
Authors: HOANG MONICA , JANSSEN SARAH M. , JOSEPH JAMIE W. 
Published: June 2019

Pancreatic β-cells secrete insulin in a biphasic manner via KATP-channel dependent and independent pathways. The KATP-channel independent pathway produces a rise in the NADPH/NADP+ ratio and cytosolic α-ketoglutarate (α-KG). Prolyl hydroxylase domain proteins (PHDs) belong to the α-KG-dependent dioxygenase superfamily and regulates stability of hypoxia-inducible factor α (HIF-α). Prolyl hydroxylation of HIF-α for ubiquitin-mediated proteasomal degradation requires sufficient levels of oxygen, iron and α-KG. We have previously shown pharmacological inhibition of PHDs reduces glucose-stimulated insulin secretion (GSIS), glucose utilization, and mitochondrial function in clonal β-cells. Glucose-induced changes in cytosolic α-KG may regulate PHD and thus β-cell function. In mouse pancreatic β-cells, PHD isoforms express unique subcellular localization: PHD1 is expressed exclusively in the cytosol, PHD2 is expressed in the cytosol and nucleus, and PHD3 is mainly expressed in the nucleus. To further explore the isoform-specific roles of PHD, we employed a β-cell specific knockout (β-PHD KO) mouse model for each of the PHD isoforms. β-PHD1 KO, β-PHD2 KO and β-PHD3 KO. β-PHD KO mice show no significant in vivo defects associated with glucose tolerance and insulin resistance; however, β-PHD1 KO and β-PHD2 KO mice have significantly increased plasma insulin compared to wild type (WT) controls (p&lt;0.05). Consistent with our cell culture model of PHD knockdown, PHD1 and PHD3 regulate insulin secretion as both β-PHD1 KO and β-PHD3 KO mice show impaired GSIS. Only β-PHD1 KO mice have reduced beta-cell mass (p&lt;0.01).
In conclusion, there are significant and unique effects observed with isoform-specific suppression of PHD.

Disclosure
M. Hoang: None. S.M. Janssen: None. J.W. Joseph: None.

Funding
Natural Sciences and Engineering Research Council of Canada; Canadian Institutes of Health Research
218-OR: Does the Reparixin (CXCR1/2 Inhibitor) Improve Islet Cell Engraftment and Clinical Outcomes after Pancreatic Islet Allotransplantation in Patients with Type 1 Diabetes Mellitus? A Randomized, Double-Blind, Prospective Study
Authors: BACHUL PIOTR J. , GOLEBIEWSKA JUSTYNA E. , PARA MONICA , ANTIC FILIP , BASTO LINDSAY , PEREA LAURENCIA , GOLAB KAROLINA , WANG LING-JIA , TIBUDAN MARTIN L. , THOMAS CELESTE C. , WITKOWSK PIOTR 
Published: June 2019

We present results of the clinical study testing Reparixin- CXCR1/2 inhibitor for improved engraftment of islet allografts. Twelve patients were randomized blindly (2:1) to receive Reparixin (N=8) or placebo N=4) in addition to the same immunosuppression.
Results: Patient and donor characteristics as well as islet quality and quality did not differ significantly between the groups. Four (50%) patients from Reparixin vs. none (0%) from placebo group achieved persistent insulin independence after only one islet infusion for over 2 years (2.2, &gt;4.5,&gt;5,&gt;5 years) (p=0.2). Islet engraftment index (IEI) measured on day 75 based on the ratio of area under the curve for serum c-peptide (AUC c-peptide) and glucose (AUC glucose) in 120 min mixed meal test standardized for islet mass in IEQ of the first transplant was higher in each of 5 individuals without early complications in Reparixin group: 5.92, 6.02, 7.04, 9.44, 11.99, than in the placebo group (0, 4.56, 5.02) (p=0.035). Islet function after engraftment measured by BETA-2 on day 75 was higher but did not reached statistical difference (p=0.07); BETA-2 was 15.5, 21, 22, 23, 23 in Reparixin vs. 0, 10, 19 in placebo group. Diabetic retinopathy improved in 2 and remained stable in all patients who achieved insulin independence and got worse in one patient who has never become insulin free. Diabetic neuropathy based on EMG and neurological exam improved in 2 patients and remained stable in remaining 8.
Conclusion: Half of the patients from Reparixin group achieved long- term (&gt;2 years) insulin independence after only one islet infusion, while none from placebo group had such outcome. Islet engraftment index was significantly higher in Reparixin group than in placebo, while assessed in patients without complications. Diabetic retinopathy and neuropathy remained stable or improved after successful IT.

Disclosure
P.J. Bachul: None. J.E. Golebiewska: None. M. Para: None. F. Antic: None. L. Basto: None. L. Perea: None. K. Golab: None. L. Wang: None. M.L. Tibudan: None. C.C. Thomas: None. P. Witkowsk: None.

Funding
Dompé Pharmaceuticals
2182-P: Protective Effects of Lin28a-RNA Binding Protein Against Glucotoxicity in Pancreatic Beta Cell
Authors: LEE KYEONG-MIN , HWANG YEOJIN , CHOI JUN-HYEOK 
Published: June 2019

In pancreatic beta cell, excessive and prolonged increase of glucose causes beta cell dysfunction which is characterized by impaired insulin synthesis and secretion, termed the glucotoxicity. Although it is known that both Lin28a and LIN28B promoted an insulin-sensitized state that resisted high fat diet-induced diabetes, other molecular mechanisms that protect from glucotoxicity are poorly defined. We investigated the effect of Lin28a overexpression on glucotoxicity-induced beta cell dysfunction. In this study, we found that high glucose leads to a decrease in Lin28a mRNA and protein expression, followed by a decrease in insulin gene expression and secretion in the rat insulinoma cell line, INS-1. In addition, PDX-1and BETA2 mRNA and protein expression were downregulated by high glucose in INS-1. Similarly, mRNA expression of Lin28a and insulin was decreased in pancreas of high fat diet induced diabetic mouse. Adenovirus mediated Lin28a overexpression in INS-1 reversed the glucotoxicity-induced downregulation of insulin, PDX-1 and BETA2 expression, and improved glucose stimulated insulin secretion (GSIS). In mouse islet, mRNA expressions of insulin, PDX-1 and ki67 and GSIS were decreased by glucotoxicity. Lin 28a overexpression in mouse islet partially restored their expression and GSIS. These results suggest that Lin28a protects pancreatic beta cells from glucotoxicity through regulation of insulin transcription factors.

Disclosure
K. Lee: None. Y. Hwang: None. J. Choi: None.

Funding
Daegu Gyeongbuk Institute of Science and Technology (18-LC-01, 2018010133)