bluuebunny/crossref_metadata_2025 huggingface dataset:
1395-P: The Role of Adipocyte Connexin-43 in Mediating Adipose Tissue Inflammation and Dysfunction in Obesity
Authors: RON SOPHIE , RON IDIT , RATHAUS MORAN , LIVNE RINAT , TIROSH AMIR
Published: May 2022

Obesity is a leading global health concern and a major risk factor for type-2 diabetes. Thus, identifying mechanisms linking obesity to insulin resistance is of great importance. Chronic low-grade inflammation of the adipose tissue (AT) observed in obesity has been shown as an important pathophysiological link between obesity and the development of systemic insulin resistance, by various mechanisms. Gap junction (GJ) intercellular-communication, primarily composed of connexin- (Cx) 43, has been recently reported to have immunomodulatory roles in various tissues, and increased Cx43 expression and GJ activity have been implicated in the response of various tissues to chronic stress conditions. We have therefore aimed to study the role of Cx43 in shaping the AT adaptive or maladaptive response to these cellular insults of obesity. In a diet-induced obesity mouse model we demonstrated an increase in Cx43 expression in the intact AT, which could be primarily attributed to increased Cx43 expression in adipocytes. In high fat diet-fed adipocyte-specific Cx43 knock-out mice (AdCx43KO) , we observed increased infiltration of macrophages to the AT as compared to wild-type (WT) mice. This was accompanied by morphologic changes in the AT, with increased variability in adipocyte size. While AdCx43KO mice did not significantly differ from WT mice in body weight, adiposity or food intake, the AdCx43KO mice demonstrated reduced whole-body insulin sensitivity assessed by insulin tolerance test. Mechanistically, we observed that adipocytes can interact directly with neighboring adipocytes as well as with macrophages via Cx43-composed gap junctions, an interaction that is likely to be key in attenuation of the tissue inflammatory response. Our results suggest an immunomodulatory and metabolic role for adipocyte-Cx43 in obesity.

Disclosure

S.Ron: None. I.Ron: None. M.Rathaus: None. R.Livne: None. A.Tirosh: Advisory Panel; Abbott Diagnostics, AstraZeneca, Boehringer Ingelheim International GmbH, Merck & Co., Inc., Novo Nordisk, Sanofi, Consultant; Bayer AG, DreaMed Diabetes, Ltd., Research Support; Medtronic, Speaker's Bureau; Eli Lilly and Company.

Funding

Israel Science Foundation (922/17)
1413-P: Semaglutide 2.4 mg Improved Glucose Metabolism and Reverted Prediabetes to Normoglycemia in Adults with Overweight/Obesity vs. Liraglutide 3.0 mg (STEP 8)
Authors: ROSENSTOCK JULIO , GARVEY W. TIMOTHY , GOLDMAN BRYAN , KHALID USMAN , SØRRIG RASMUS , RUBINO DOMENICA
Published: May 2022

The STEP 8 trial randomized 338 adults with overweight/obesity to weekly semaglutide 2.4 mg or placebo (PBO) , or daily liraglutide 3.0 mg or PBO, plus lifestyle intervention for 68 wks (3:1:3:1) ; PBO data not shown. Body weight (BW) change was greater with semaglutide vs. liraglutide (-15.8% vs. -6.4%, respectively; p<0.0001) . This exploratory analysis assessed effects on glucose metabolism and glycemic status (normoglycemia; prediabetes; T2D) at wk 68 in participants (pts) with prediabetes at baseline (BL; 43 semaglutide/45 liraglutide; mean HbA1c 5.9/5.8%; fasting plasma glucose [FPG] 102/100 mg/dL; HOMA-IR 4.9/3.7; BW 104/1kg) . Analyses were not adjusted for multiplicity. At wk 68, 89.5% of pts were normoglycemic with semaglutide vs. 64.9% with liraglutide (p=0.01) (Figure) ; in these pts, 76.5% had weight loss ≥10% with semaglutide vs. 33.3% with liraglutide. One pt in each group developed T2D (Figure) . From BL to wk 68, semaglutide led to greater reductions vs. liraglutide in HbA1c (difference: -0.22 %; p=0.0002) , FPG (difference: -6.2 mg/dL; p=0.02) , and HOMA-IR (relative % difference: -28%; p=0.02) .

In conclusion, both semaglutide 2.4 mg and liraglutide 3.0 mg had clinically beneficial effects on glucose metabolism and glycemic status in pts with overweight/obesity and prediabetes, though the effect of semaglutide was greater vs. liraglutide.

Disclosure

J.Rosenstock: Consultant; AstraZeneca, Other Relationship; Applied Therapeutics, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Hanmi Pharm. Co., Ltd., Intarcia Therapeutics, Inc., Novo Nordisk, Oramed Pharmaceuticals, Sanofi, Zealand Pharma A/S, Research Support; Genentech, Inc., Merck & Co., Inc., Metacrine, Inc., Novartis Pharmaceuticals Corporation, Pfizer Inc., vTv Therapeutics. W.Garvey: Other Relationship; Boehringer Ingelheim International GmbH, Eli Lilly and Company, Epitomee, JAZZ Pharmaceuticals, Novo Nordisk, Novo Nordisk, Pfizer Inc., Pfizer Inc. B.Goldman: Employee; Novo Nordisk A/S, Stock/Shareholder; Novo Nordisk A/S. U.Khalid: None. R.Sørrig: Employee; Novo Nordisk A/S. D.Rubino: Advisory Panel; Novo Nordisk, Other Relationship; Endocrine Society, PeerView, WebMD LLC, Research Support; Boehringer Ingelheim International GmbH, Novo Nordisk, Obesinov, SARL, Speaker's Bureau; Novo Nordisk, Stock/Shareholder; Novo Nordisk.

Funding

Funded by Novo Nordisk A/S
1416-P: UK First National Health Service (NHS) Endobarrier (EB) Service for Uncontrolled Diabesity: Four-Year Outcomes for All 62 Treated Patients
Authors: RYDER ROBERT E.J. , GUPTA PIYA SEN , YADAGIRI MAHENDER , BURBRIDGE WINNIFRED (WYN) , IRWIN SUSAN P. , BASHIR TAHIRA , GANDHI HARDEEP , WYRES MELANIE C. , ALLDEN RACHAEL , BLEASDALE JOHN , FOGDEN EDWARD , ANDERSON MARK
Published: May 2022

EB, a 60cm endoscopically inserted proximal intestinal liner, reduces weight and HbA1c over 1 year. In the 1st NHS EB service, we provided EB to patients with sub optimally controlled diabesity and monitored outcomes in a registry. 44/62 (71%) completing 3 years post EB removal (age 51.3±7.8 years, 54% male, diabetes duration 14.56 (8-21) years, BMI 41.7±7.2 kg/m2) attended follow-up. During EB treatment, mean ± SD HbA1c fell by 1.9±1.8 %, from 9.1±1.8 to 7.2±1.0 % (p<0.001) , weight by 17.3±9.0 kg from 122.8 ± 29.8 to 105.4±30.6 kg (<0.001) , systolic BP from 138.8±14.3 to 125.9±14.9 mmHg (<0.001) , serum alanine aminotransferase (liver fat marker) from 30.4±17.2 to 19.1±11.2 U/L (p<0.001) . Median (IQR) total daily insulin dose reduced from 1 (50-171) to 30 (0-63) units (n=29, p<0.001) ; 10/29 (34%) insulin treated patients discontinued insulin. 3-years post EB removal 33/44 (75%) maintained most of the improvement achieved with EB whilst 11/44 (25%) reverted to baseline (figure) . Of those deteriorating, all had depression and/or bereavement and/or major health problems/disability. 10/62 (16%) had early EB removal for adverse events or symptoms; all fully recovered after removal and most derived significant benefit. Our data demonstrates EB as highly effective in patients with refractory diabesity, with maintenance of significant improvement 3 years after removal in 75%

Disclosure

R.E.J. Ryder: None. P. Sen Gupta: None. M. Yadagiri: None. W. Burbridge: None. S.P. Irwin: None. H. Gandhi: None. M.C. Wyres: None. J. Bleasdale: None. E. Fogden: None. M. Anderson: None.
142-LB: Cognitive Screening among Older Adults with Diabetes
Authors: GUPTA DEEPASHREE , WILHALME HOLLY , SAUDER GABRIELA , MOIN TANNAZ
Published: May 2022

Background: Diabetes affects approximately 24.4% of older adults in the United States and increases the risk of cognitive dysfunction and future dementia. Patients with cognitive dysfunction may find it harder to manage their diabetes and have more complications such as hypoglycemia. Yearly cognitive screening is recommended for all older adults with diabetes but may not be widely conducted.

Methods: We performed cognitive screening with the Mini-Cog test for a subset of patients with diabetes seen in Endocrine Clinics in one large healthcare system in 2020-21. We used a retrospective chart review to compare patient screening results between Endocrine (Endo) , Geriatric (Geri) and multidisciplinary Endo-Geriatric practices (Geri-Endo) . We used Chi-square tests to examine differences between clinic cohorts and a multivariate logistic regression to predict risk of cognitive dysfunction. We adjusted for age, ethnicity, sex, type of diabetes, Sulfonylurea (SU) use, ACE inhibitors (ACEI) / Angiotensin Receptor Blocker (ARB) use, smoking history, statin use, and BMI.

Findings: Among 198 patients screened (Endo=80, Geri=32, Geri-Endo=86) , those seen in Geri-endo (40.7%) and Geri (31.3%) were more likely to have lower Mini-COG scores (lower scores suggest worse cognition) as compared to those in Endo (6.3%) (p<.0001) . Patients in Endo and Geri-Endo had longer duration of diabetes, were more likely to use insulin and had higher incidence of documented hypoglycemia. Higher age > 75 years (p=0.0105) , previous CV events (p=0.0006) and BMI <30 (p=0.0115) were significantly associated with lower Mini-COG scores.

Conclusion: Our study shows that cognitive screening can help identify older adults at risk for dementia in diabetes clinic. Patients with older age, previous CV events and lower BMI may be at higher risk for cognitive dysfunction. Older adults should be screened yearly for cognition as part of routine diabetes care.

Disclosure

D. Gupta: None. H. Wilhalme: None. G. Sauder: None. T. Moin: None.

Funding

NIH National Center for Advancing Translational Science (NCATS) , UCLA CTSI (UL1TR001881)
1440-P: Effects of Two Weight-Loss Medications, Liraglutide and Orlistat, on Liver Enzymes in Patients with Overweight or Obesity in a Real-World Setting
Authors: GORGOJO-MARTINEZ JUAN J. , CHICHARRO PABLO L. LOIS , FERREIRA-OCAMPO PABLO J. , DOEJO-MARCIALES SANDRA C. , BARRA-MALIG SOLANGE F. , ALMODOVAR-RUIZ FRANCISCA
Published: May 2022

The aim of this analysis of the XENSOR study was to assess the effect of 2 weight-loss drugs on hepatic serum biomarkers in patients with overweight or obesity and insufficient weight loss (WL) after a lifestyle modification program. A retrospective real-world study (RWS) was conducted comparing the effect of orlistat 120 mg tid and liraglutide (up to 3 mg daily) on liver enzymes in adult patients with BMI > 30 kg/m² (or > 27 kg/m² with at least a weight-related comorbidity) . The primary endpoint was to assess in a multivariate model the differences in ALT and GGT reductions between both drugs over a median follow-up period of 7 months. A secondary endpoint was to study the impact of the amount of WL on both liver enzymes. 400 patients in the group of orlistat (BMI 41.4 kg/m2, ALT 30.1 U/L, GGT 35.7 U/L) , and 100 in the group of liraglutide (BMI 39.7 kg/m2, ALT 27.3 U/L, GGT 29.1 U/L) , were included. WL with liraglutide (−7.7 kg) was significantly greater in comparison with orlistat (−3.3 kg) , p <0.0001. Treatment with both drugs significantly reduced ALT (orlistat −3.8 U/L; liraglutide −5.7 U/L, both p<0.05) . ALT reduction was significantly higher with liraglutide vs. orlistat (adjusted mean difference −3.4 U/L (95% CI −6.5 to −0.2 U/L) . A significant decrease in GGT levels was observed with liraglutide (−5.8 U/L, p<0.0001) , but not with orlistat. Adjusted mean differences in GGT changes between both drugs were not statistically significant. 46% of patients with high ALT levels lowered this enzyme into the normal range at the end of the follow-up, without significant differences between both drugs. When patients were split into 3 groups according to their WL (<5%, 5-10% and >10%) , ALT and GGT reductions were significantly higher in patients with WL>10%.

In summary, in this RWS ALT reductions were significantly greater with liraglutide in comparison with orlistat. The effects of both drugs on ALT and GGT were higher in those patients with clinically relevant WL (>10%) .

Disclosure

J.J.Gorgojo-martinez: Consultant; Mundipharma, Research Support; Novo Nordisk, Speaker's Bureau; Abbott Diabetes, AstraZeneca, Boehringer Ingelheim International GmbH, Lilly Diabetes, Merck Sharp & Dohme Corp., Mundipharma, Novo Nordisk. P.L.Lois chicharro: None. P.J.Ferreira-ocampo: None. S.C.Doejo-marciales: None. S.F.Barra-malig: None. F.Almodovar-ruiz: None.
1451-P: Peri-Islet Fibrosis in Cystic Fibrosis Is Associated with Altered Alpha-Cell Phenotype
Authors: KATTNER NICOLE , AL-SELWI YARA , TINIAKOS DINA , KLÖPPEL GÜNTER , SHAW JAMES A.
Published: May 2022

Cystic fibrosis (CF) is caused by CF transmembrane conductance regulator mutations leading to impaired epithelial cell anion transport and thickened pancreatic exocrine secretions. CF-related diabetes commonly ensues through impact of pancreatic exocrine pathology on islet cell mass and function although specific mechanisms remain unclear. Paraffin-embedded blocks obtained post mortem from pancreatic body in 9 donors with CF (6 (67%) male; age: 0-27 years) were studied. Following haematoxylin and eosin staining, CF changes were scored as early, advanced or end-stage and islet microenvironment was evaluated for presence of surrounding fibrosis and adipocyte replacement. Four donors were categorised as ‘early’ pathology with 0 (1) % (median (IQR)) of islets surrounded by uninterrupted fibrosis and 0 (0) % by adipocyte replacement; 1 donor was categorised as ‘advanced’ with 100% fibrosis, 0% adipocyte replacement; and 4 donors were categorised as ‘end-stage’ with 65 (46) % fibrosis, 12 (32) % adipocyte replacement. Presence of glucagon/vimentin co-expressing cells was demonstrated by immunofluorescence staining / confocal microscopy in 1 (25%) donor with ‘early’ CF pathology and in all with more advanced exocrine changes. All islets with altered alpha-cell phenotype had peri-islet fibrosis. In pancreata with <50% fibrosis (11±17% (mean±SD) ; n=6) altered alpha-cell phenotype was seen in 11±14% of islets vs. pancreata with >50% fibrosis (93±7%; n=3) with altered alpha-cell phenotype in 65±10% islets (p<0.001) . Analysis of a chronic pancreatitis resection with incrementally increasing fibrosis from head to tail showed parallel increase in islet proportion with altered alpha-cell phenotype (head: 28%; body: 80%; tail: 78%) . We have demonstrated that peri-islet fibrosis is associated with aberrant islet glucagon/vimentin co-expression providing further evidence for exocrine-endocrine signalling in human pancreatic pathology.

Disclosure

N.Kattner: None. Y.Al-selwi: None. D.Tiniakos: None. G.Klöppel: None. J.A.Shaw: None.

Funding

Cystic Fibrosis Trust (SRC 019)
1474-P: Testosterone Amplifies Glucose-Stimulated Insulin Secretion from Male ß-Cells via Soluble Adenylate Cyclase
Authors: QADIR MIRZA MUHAMMAD FAHD , XU WEIWEI , NASTESKA DANIELA , BHONDELEY MANIKA , MOTA DE SA PAULA , EVANS CHARLES R. , BUCK JOCHEN , LEVIN LONNY , BURANT CHARLES , HODSON DAVID , MAUVAIS-JARVIS FRANCK
Published: May 2022

Male mice with elimination of the androgen receptor (AR) in islet β cells (βARKO) exhibit blunted glucose-stimulated insulin secretion (GSIS) , leading to hypoinsulinemia and hyperglycemia. We showed that dihydrotestosterone (DHT) activation of an extranuclear AR in β cells potentiates GSIS by amplifying the insulinotropic action of islet-derived and exogenous GLP-1 via GLP-1R in mouse and human islets. Using electron microscopy, we observe that DHT increases the size of granules and the frequency of multivesicular granules, suggesting that DHT promotes multivesicular exocytosis. Using live β cell models expressing organelle-specific cAMP sensors and novel allosteric inhibitors in human islets, we show that DHT enhances the ability of GLP-1 to produce cAMP at the plasma membrane and endosomes and enhances islet GSIS via the bicarbonate (HCO3 -) sensor, soluble adenylate cyclase (sAC) . The end-products of glucose metabolism in the TCA cycle are CO2 and H2O, which is followed by rapid conversion of CO2 to HCO3 - and H+ by carbonic anhydrase (CA) . Therefore, CO2 production is translated to HCO3 - and specifically sensed by sAC. Bioenergetics analysis of DHT-stimulated respiration in mouse islets using a Seahorse Analyser revealed that DHT decreases glycolytic acidification and increases mitochondrial acidification (H+ efflux rate) , suggesting that DHT increases mitochondrial production of CO2 followed by conversion to HCO3 - and H+, allowing HCO3 - to stimulate sAC. We discuss experiments of single-cell transcriptomics, proteomics and metabolomics dissecting the impact of DHT on human islets glycolysis and mitochondrial CO2 production. Consistent with DHT increasing production of CO2 followed by conversion to HCO3 - by CA and activation of sAC, in cultured human male islets, the CA inhibitor acetazolamide blunted DHT-enhancement of GSIS. Together these experiments provide novel insights into the role of DHT action via an extranuclear AR in enhancing GSIS.

Disclosure

M.Qadir: None. D.Hodson: Other Relationship; Celtarys Research. F.Mauvais-jarvis: Consultant; Mithra Pharmaceutical. W.Xu: None. D.Nasteska: None. M.Bhondeley: None. P.Mota de sa: None. C.R.Evans: None. J.Buck: None. L.Levin: None. C.Burant: None.

Funding

National Institutes of Health (DK107444 and DK074970) U.S. Department of Veterans Affairs Merit Award (BX003725)
15-OR: Mitochondrial NRF2 in ß-cell Oxidative Stress Response
Authors: NOVAK ALISSA , CROWDER JUSTIN , MARASCO MICHELLE , LINNEMANN AMELIA K.
Published: May 2022

β-cells are highly metabolic and vulnerable to a variety of exogenous stressors, including excess glucose and proinflammatory cytokines. Many of these stressors elicit the accumulation of reactive oxygen species (ROS) , which can cause oxidative damage and lead to cell death. To restore homeostasis in the face of accumulating ROS, all cells, including β-cells, are equipped with adaptive stress response mechanisms such as the antioxidant response and autophagy. The antioxidant response is primarily driven by nuclear translocation of the transcription factor, NRF2. However, recently we observed a pool of NRF2 in the mitochondria that is associated with both the antioxidant response and a selective form of autophagy, known as mitophagy. We therefore hypothesized that NRF2 mediates coupling of mitophagy with the antioxidant response in the β-cell and is essential for restoration of β-cell homeostasis under conditions of increased ROS. We first asked whether loss of murine NRF2 was associated with increased susceptibility to β-cell damage induced by the toxic glucose analog, streptozotocin (STZ) . Multiple low dose STZ injections (45mg/kg for 5 consecutive days) rendered whole-body NRF2 knockout mice diabetic more quickly than wild type controls. To study the role of β-cell NRF2 in autophagy/antioxidant response coupling, we then generated a β-cell specific NRF2 knockout mouse (NRF2Δβ) by breeding NRF2 floxed mice with mice bearing the Ins1Cre/wt allele. NRF2Δβ mice appear to be phenotypically normal until at least 20 weeks of age and do not develop spontaneous diabetes. We then set out to determine how loss of β-cell NRF2 affects the mitochondria by both immunofluorescence analysis and transmission electron microscopy of isolated pancreata. We observed rounded β-cell mitochondrial morphology in the absence of NRF2. Collectively, our data are consistent with a role for NRF2 in β-cell mitochondrial function and suggest that NRF2 is required for proper response to β-cell damage.

Disclosure

A.Novak: n/a. J.Crowder: None. M.Marasco: Other Relationship; Eli Lilly and Company. A.K.Linnemann: Consultant; Janssen Research & Development, LLC.

Funding

NIDDK (R01DK124380)
159-LB: The Lipidomic Signature of Prediabetes among the Oldest Old—The Long Life Family Study
Authors: MILJKOVIC IVA , CVEJKUS RYAN , AN PING , THYAGARAJAN BHARAT , MENGEL-FROM JONAS , FEITOSA MARY , BENBOURENANE ANISA N. , ZMUDA JOSEPH M.
Published: May 2022

The epidemiology of prediabetes and its metabolomic features remain poorly characterized in the oldest old. Therefore, we characterized the serum lipidome in 635 nonagenarians and centenarians without type 2 diabetes (T2D) from the Long Life Family Study, a multicenter study of 583 families with a clustering of exceptional longevity. Despite a low T2D prevalence and incidence in the LLFS cohort, 361 (56.9%) of participants aged 90+ years had prediabetes at study entry based on either glycated hemoglobin or fasting serum glucose levels. We used LC/MS to initially assess 193 lipid metabolites in fasting serum in those with and without prediabetes. We tested the association of log transformed lipid metabolites with prediabetes using logistic regression with the Benjamini-Hochberg false-discovery rate method (FDR<0.05) , while adjusting for age, sex, fasting time, field center, and familial relatedness. 17 out of 193 annotated lipid metabolites were associated with prevalent prediabetes, with the strongest association found for phosphatidylethanolamine (PE 40:7) , which remained significant after additional adjustment for BMI (odds ratio (OR) =2.83; 95% confidence interval (CI) : 1.7-4.7) . Phosphatidylethanolamine might have proinflammatory properties and act as a positive modulator of membrane disruption induced by islet amyloid polypeptide, an amyloidogenic protein involved in T2D. Our data suggest that among the oldest old with exceptional longevity prediabetes burden is high, and that the phosphatidylethanolamine pathway might have an important role in the development of prediabetes. Future untargeted metabolomic analyses will investigate potential biomarkers that predict the transition from prediabetes to diabetes among the oldest old in this unique family study of healthy aging and exceptional longevity.

Disclosure

I. Miljkovic: None. R. Cvejkus: None. P. An: None. B. Thyagarajan: n/a. J. Mengel-from: None. M. Feitosa: None. A. N. Benbourenane: None. J. M. Zmuda: None.

Funding

National Institute of Health (U01AG023746)
159-OR: Endocrine and Metabolic Counterregulation to Standardized Postprandial Hypoglycemia after Bariatric Surgery
Authors: TRIPYLA AFRODITI , HERZIG DAVID , BRANCHAT GEMMA REVERTER , PAVAN JACOPO , SCHIAVON MICHELE , EUGSTER PHILIPPE J. , GROUZMANN ERIC , NAKAS CHRISTOS T. , VALÉRIE SAUVINET , ZEHETNER JOERG , GIACHINO DANIEL , DEL FAVERO SIMONE , THOMAS ANDREAS , THEVIS MARIO , MAN CHIARA DALLA , BALLY LIA
Published: May 2022

Aim: To compare endocrine and metabolic counterregulation to postprandial hypoglycemia in patients with postbariatric hypoglycemia (PBH) after gastric bypass (GB) vs. surgical and non-surgical controls.

Methods: Thirty-two adults (age 42.9±12.8 years, 13% male, BMI 28.2±4.3kg/m2) of 4 matched groups (PBH patients, GB, sleeve gastrectomy [SG] and non-surgical controls [CON]) ingested 15g glucose with subsequent induction of hypoglycemia (target 2.5mM) at t150-170 min using a constant insulin infusion with controller-guided iv dextrose adjustment. Primary outcome was mean glucagon during hypoglycemia (t150-170) . Further outcomes were responses of catecholamines, cortisol, growth hormone, pancreatic polypeptide (PP) , glucagon-like peptide 1, endogenous glucose production (EGP) and exogenous insulin. Values are shown as mean±SD.

Results: Plasma glucose and insulin at t150-170 were 2.6±0.2mM and 891±382pM across all groups. Glucagon was significantly lower in all surgical groups (PBH: 12.0±8.2pM, GB: 13.4±5.7pM and SG: 11.0±6.8pM vs. CON: 23.0±6.2pM, p=0.005) . PP was significantly lower in PBH vs. CON (28.1±49.6pM vs. 146.0±93.5pM, p=0.01) . EGP did not differ significantly between groups, nor did any other hormones.

Conclusion: Our results suggest diminished glucagon response to postprandial hypoglycemia after GB and SG compared to CON, with additionally lower PP values in PBH.

Disclosure

A.Tripyla: None. J.Zehetner: None. D.Giachino: None. S.Del favero: Research Support; Dexcom, Inc. A.Thomas: None. M.Thevis: None. C.Dalla man: Research Support; Becton, Dickinson and Company, Sanofi-Aventis Deutschland GmbH. L.Bally: None. D.Herzig: None. G.Reverter branchat: Employee; Fresenius Medical Care. J.Pavan: None. M.Schiavon: None. P.J.Eugster: None. E.Grouzmann: None. C.T.Nakas: None. S.Valérie: None.

Funding

Swiss National Science Foundation (PCEGP3_186978)
191-LB: Influenza Replication Dependent on Host Cell Pulmonary Metabolism
Authors: ROCHOWSKI MATTHEW , ALLEN SHAWN M.F. , CAMPOLO ALLISON , LACOMBE VERONIQUE
Published: May 2022

Diabetes is an independent risk factor for severe respiratory infections, including influenza (IAV) . We hypothesize that hyperglycemia predisposes diabetic subjects to excess glucose in the airway, allowing for greater IAV replication. To test this hypothesis, type 1 (T1Dx) and type 2 (T2Dx) diabetic mouse models were intranasally infected with influenza. Subsets of T1Dx and T2Dx mice were treated with insulin or metformin, respectively, to restore euglycemia. Glucose concentrations of the bronchoalveolar lavage fluid (BALF) were measured with a glucose oxidase assay. Human bronchial epithelial cells (HBECs) were incubated with varying glucose concentrations, 2-deoxyglucose, AMPK modulators, or insulin, then infected. Viral loads were determined by immunofluorescence and/or qrtPCR for the viral protein hemagglutinin. In vivo, diabetic and infected mice demonstrated increased glucose concentrations in BALF. Viral load was significantly greater in lung homogenates of both T1Dx and T2Dx mice, which was rescued by insulin and metformin treatment. In vitro, HBECs incubated in higher [glucose] had a significantly greater percentage of cells infected than those incubated in normal [glucose]. Conversely, cells treated with 2-deoxyglucose demonstrated reduced IAV replication. Activation of glycolysis using Metformin or AICAR (i.e. AMPK activators) increased IAV replication in a time-dependent manner, while inhibition of AMPK decreased IAV replication. Finally, insulin treatment modulated IAV replication in a similar time-dependent manner. These novel findings suggest that: 1) hyperglycemia increases BALF [glucose] and 2) influenza viral replication is dependent on host-cell glycolysis. Better understanding of the mechanisms altering glucose metabolism during IAV infection may lead to the discovery of novel therapeutic targets for diabetic patients infected with IAV.

Disclosure

M. Rochowski: None. S. M. F. Allen: None. A. Campolo: Employee; Alcon Research, LLC. V. Lacombe: None.

Funding

Centers of Biomedical Research Excellence
204-OR: The Chd4 Helicase Modulates ß-Cell Function In Vivo
Authors: DAVIDSON REBECCA K. , CASEY NOLAN , KANOJIA SUKRATI , SPAETH JASON
Published: May 2022

During diabetes progression, a subset of failing β-cells incurs a loss of transcription factors (TFs) such as Pdx1, which drives expression of numerous β-cell functional genes including Ins2, MafA, and Slc2a2. Like other TFs, Pdx1 requires the recruitment of transcriptional coregulators to modulate its transcriptional activity. It has been established that the Pdx1-bound Swi/Snf complex plays a critical role in maintaining β-cell function in part by recruiting Pdx1 to the insulin gene and facilitating its transcription. Furthermore, interactions between Pdx1 and the Swi/Snf ATPase subunit Brg1 are reduced in type 2 diabetes (T2D) β-cells. Recently, through an IP-Mass Spec approach, Pdx1 has been shown to interact with numerous other coregulatory complexes, including the Chd4 ATPase subunit of the Nucleosome Remodeling and Deacetylase complex. Here, we confirmed Pdx1 and Chd4 interact in mouse and human β-cell lines and primary β-cells using co-immunoprecipitation and proximity ligation assays (PLA) . Tissue harvested from mice injected with glucose had more β-cell nuclei containing multiple PLA signals than tissue from fasted mice. Conversely, tissue from high-fat diet mice and T2D human donors had fewer β-cell nuclei containing multiple PLA signals than tissue from control mice and nondiabetes human donors. To investigate the role of Chd4 on mature β-cell function, we generated a tamoxifen inducible, β-cell-specific Chd4 knockout mouse model (Chd4 Δβ ) . Four weeks following Chd4 removal, Chd4 Δβ mutants were glucose intolerant and had elevated ad libitum fed blood glucose levels and impaired insulin secretion from isolated islets. Bulk mRNA-sequencing from flow-sorted lineage labeled Chd4Δβ β-cells identified 1093 upregulated genes (eg Adcy1 and Rapgef3) and 438 downregulated genes (eg MafA, Robo2, and Slc2a2) . Future efforts are focused on defining the mechanisms by which Chd4 controls gene expression in the β-cell, with a current focus on how Chd4 modulates chromatin accessibility and TF recruitment.

Disclosure

R.K.Davidson: None. N.Casey: None. S.Kanojia: None. J.Spaeth: None.

Funding

National Institutes of Health (K01DK115633) Showalter Trust Award
219-OR: Intranasal Insulin Enhanced Resting-State Functional Connectivity of Medio-Prefrontal Cortex in Type 2 Diabetes: A Substudy of the Memory Advancement with Type 2 Diabetes (MemAID) Randomized Control Trial
Authors: ZHANG ZONGPAI , NOVAK VERA , NOVAK PETER , MANTZOROS CHRISTOS , NGO LONG H. , LIOUTAS VASILEIOS , DAI WEIYING
Published: May 2022

Type 2 diabetes mellitus (T2DM) is associated with impairment of resting-state functional connectivity (rsFC) and cognition. Intranasal insulin (INI) has emerged as potential treatment for T2DM-related cognitive decline. INI treatment has increased perfusion and low-frequency fluctuations in medio-prefrontal cortex (mPFC) . We aimed to evaluate the effect of INI treatment on mPFC rsFC.MemAIDs trial consisted of 24-week of treatment with 40 IU of intranasal insulin (Novolin® R, off label use) or placebo (sterile saline) once daily and 24-week follow-up period. Of 244 randomized participants (115 T2DM and 129 controls, > 50 years old) , T2DM participants (eight INI, three placebo, > 50 years old) completed the baseline and end-of-treatment blood oxygen level dependent (BOLD) fMRI scans.INI treatment increased resting state mPFC-postcentral rsFC (p=0.005) and decreased mPFC-cerebellum rsFC (p<0.001) . The mPFC rsFC changes after INI treatment were different from those from placebo (Figure 1) .INI treatment can modulate resting state connectivity between pre-frontal and sensorimotor cortex and cerebellum, indicating its potential effects on decision making and goal-oriented behaviors. These findings warrant further investigation.

Disclosure

Z.Zhang: None. V.Novak: None. P.Novak: Stock/Shareholder; Moderna, Inc., Novavax, Pfizer Inc., Prothena. C.Mantzoros: Advisory Panel; Novo Nordisk. L.H.Ngo: None. V.Lioutas: None. W.Dai: None.

Funding

Funded by NIH-NIDDK (1R01DK103902) , FDA (IND#107690) , Novo Nordisk, Inc. (ISS-001063) , Medtronic Inc. (NERP15-0310) and registered on www.clinicaltrials.gov (NCT02415556.)
227-OR: Disruption of Adipocyte YAP Improves Glucose Homeostasis in Mice and Decreases Adipose Tissue Fibrosis
Authors: HAN DANIEL JONG JIN , ASLAM RUKHSANA , OJHA TANVI , YUEN DARREN , LUK CYNTHIA T.
Published: May 2022

Adipose tissue is a dynamic metabolic organ that is essential for the regulation of whole-body glucose homeostasis. Dysfunctional adipose tissue hypertrophy with obesity is associated with glucose intolerance and insulin resistance underlying type 2 diabetes. Yes-associated protein 1 (YAP) is a transcription cofactor important in the Hippo signaling pathway. However, the role of YAP in adipose tissue and glucose homeostasis is unknown. To understand the role of YAP in vivo under metabolic stress conditions, we assessed how increased weight and insulin resistance impact YAP protein levels. We found that YAP protein levels increase in adipose tissue in humans with type 2 diabetes and mouse models. This suggests that YAP signaling may contribute to adipocyte dysfunction and insulin resistance under metabolic stress conditions. To further investigate the role of YAP in adipose tissue and glucose homeostasis in vivo, we developed adipose tissue-specific YAP knockout (YAP-KO) mice and placed them on either chow or high fat diet (HFD) for 12-14 weeks. On an HFD, adipose tissue YAP-KO mice show improved glucose tolerance compared to controls. Perigonadal fat pad weight was also decreased in knockout animals, with smaller adipocyte size. Adipose tissue fibrosis and gene expression associated with fibrosis was decreased in vivo and in 3T3-L1 cells treated with a YAP inhibitor. Together, these data indicate that YAP is increased in adipose tissue with weight gain and insulin resistance. Disruption of YAP in adipocytes improves glucose tolerance and adipose tissue fibrosis; suggesting that YAP plays an important role in adipose tissue and modulating glucose homeostasis under metabolic stress conditions.

Disclosure

D.Han: None. R.Aslam: None. T.Ojha: n/a. D.Yuen: Consultant; Fibrocor Therapeutics. C.T.Luk: None.

Funding

NSERC Discovery Grant (RGPIN-2018-05671) BBDC Gales Family Charitable Foundation Pilot and Feasibility GrantCanadian Institutes of Health Research Project Grant (PJT 168996)
24-LB: Guanfacine Normalizes Presynaptic Alpha-2 Adrenoceptors Enrichment and Ameliorates Neuropathic Pain in Type 1 Diabetic Rats
Authors: MUNAWAR NEHA , AL MADHOUN ASHRAF , NADER JOELLE , AL-ALI WALEED , MASOCHA WILLIAS , AL-MULLA FAHD , BITAR MILAD S.
Published: May 2022

Diabetes is associated with several complications, including neuropathic pain, which is challenging to manage with currently available drugs. Descending noradrenergic neurons possess anti-nociceptive activity; however their involvement in diabetic neuropathic pain remains to be explored. To infer the regulatory role of this system, we examined in the pons, a part of the brainstem, lumbar nerves of the spinal cord and dorsal root ganglia of streptozotocin (STZ) -treated rats, a model for type 1 diabetes (T1D) , the localization (immunofluorescence) and the mRNA (qRT-PCR) and protein (Western blotting) expression of alpha-2A adrenoceptor (ADRA2A) . The data revealed that presynaptic SNAP-25 labeled ADRA2A in the central and peripheral nervous system of STZ diabetic rats were up-regulated both at the mRNA and protein levels. Interestingly, the levels of PSD-95 labeled postsynaptic neuronal ADRA2A remain unaltered as a function of diabetes. These biochemical abnormalities in the noradrenergic system of diabetic animals were associated with increased pain sensitivity as typified by the presence of hyperalgesia and cold/mechanical allodynia. The latter pain-related behaviors were assessed using Hargreaves apparatus, cold-plate and dynamic plantar aesthesiometer. Chronically administered guanfacine, a selective ADRA2A agonist, to diabetic animals downregulated the upregulation of neuronal presynaptic ADRA2A and ameliorated the hyperalgesia and the cold/mechanical allodynia in these animals. Together, these findings demonstrate that guanfacine may function as a potent analgesic and highlight ADRA2A, a key component of the descending neuronal autoinhibitory pathway as a potential therapeutic target in the treatment of diabetic neuropathic pain.

Disclosure

N. Munawar: None. A. Al madhoun: None. J. Nader: None. W. Al-ali: None. W. Masocha: None. F. Al-mulla: None. M. S. Bitar: None.

Funding

Grant YM05/19 from Kuwait University Research Sector.
247-LB: Pancreatic Elastase Regulates ß-Cell Viability by Impairing the Mechanosignaling Pathway
Authors: BASILE GIORGIO , VETERE AMEDEO , HU JIANG , IJADUOLA OLUWASEUN , DE JESUS DARIO F. , FUKUDA KAZUKI , ELTONY AMIRA M. , YUN ANDY , CHOUDHARY AMIT , WAGNER BRIDGET , KULKARNI ROHIT
Published: May 2022

The discovery of SerpinB1 as an inducer of human β-cell proliferation pointed to acinar cell-derived proteases such as pancreatic elastase (PE) being involved in regulating β-cell viability. Indeed, we observed that expression of a major form of PE, namely CELA3B, was increased in acinar cells from donors with type 2 diabetes (T2D) compared to samples from non-diabetic (ND) donors. Surprisingly, CELA3B protein levels were higher in T2D vs. ND islets, although its gene expression levels were virtually undetectable. We confirmed that increasing doses of exogenous PE is able to decrease viability of EndoC-βH3 (βH3) cells in vitro. To evaluate the effects of PE inhibition on β-cell survival, we used telaprevir, an FDA-approved drug, that we identified as a potent PE blocker using a High-Throughput Screening assays with an IC50 of 15.6 nM to inhibit PE activity. Consistent with the mitogenic effects of SerpinB1 on β-cells, telaprevir also increased human β-cell viability in in vitro and in vivo systems. By performing phospho-antibody microarrays and single-cell RNA-Seq in human islets treated with vehicle or telaprevir, we identified the mechano-signaling pathway, which integrates the signals generated by the extracellular matrix (ECM) , as a candidate pathway linking PE inhibition to β-cell viability. We validated the results by treating EndoC-βH1 (βH1) cells lacking focal adhesion kinase (siFAK) with PE +/- telaprevir. We observed that PE impaired the mechano-signaling cascade, and these effects were reversed by telaprevir in control cells, but not in siFAK cells. Furthermore, using the novel approach of Brillouin microscopy we observed that PE increased the intracellular stiffness in βH1 cells, as a readout of cytoskeleton arrangements, whereas telaprevir blunted these effects. These data point to novel pathways underlying the acinar-β-cell crosstalk that could be harnessed for therapeutic approaches to resolve diabetes in patients.

Disclosure

G. Basile: None. B. Wagner: Research Support; Japan Tobacco Inc., Novo Nordisk Foundation, Ono Pharmaceutical Co., Ltd., Stock/Shareholder; Moderna, Inc., Pfizer Inc. R. Kulkarni: Advisory Panel; Biomea, Novo Nordisk, REDD Pharma, Consultant; Relay Therapeutics, Research Support; Inversago, SerPlus. A. Vetere: None. J. Hu: None. O. Ijaduola: None. D. F. De jesus: n/a. K. Fukuda: None. A. M. Eltony: None. A. Yun: None. A. Choudhary: None.

Funding

JDRF: 3-PDF-2020-935-A-NNIH: U01 DK123717NIH: UC4 DK116255
247-OR: Maternal Prepregnancy Obesity and Gestational Diabetes Are Associated with Epigenetic Gestational Aging in the Placenta
Authors: HUFF KATELYN , ROELL KYLE , BULKA CATHERINE , BOYLE KRISTEN E. , BRETON CARRIE , BURT AMBER , DABELEA DANA , KAHN LINDA , KARAGAS MARGARET R. , MARSIT CARMEN , NIEMIEC SIERRA , VOLK HEATHER , O'SHEA THOMAS M. , FRY REBECCA
Published: May 2022

Pregnancy conditions like maternal pre-pregnancy obesity and gestational diabetes (GDM) have been linked with the development of placenta, a critical organ required for maternal-fetal interface during gestation, and can lead to adverse health outcomes in both mother and fetus. However, the molecular signatures associated with these conditions are not fully understood. Epigenetic clocks, which are based upon DNA methylation of age-related CpGs, are thought to better reflect biological age compared to chronological age and have been used in different tissues as potential biomarkers. In the present study, we hypothesized that altered placental epigenetic gestational aging is associated with pre-pregnancy obesity and GDM. We explored these relationships by leveraging data from three diverse cohorts in the Environmental influences on Child Health Outcomes (ECHO) program (overall N=830) . First, we investigated associations between maternal pre-pregnancy obesity and GDM and estimated placental epigenetic gestational age acceleration (eGAA) . Next, we examined associations between these two conditions and methylation of each CpG site within the robust placental clock (558 CpGs) . Significant associations were not observed between pre-pregnancy obesity and GDM and eGAA. Interestingly, pre-pregnancy obesity was associated with methylation levels at 28 placental clock CpGs (q<0.1) . These sites were annotated to 17 unique genes that enrich processes like circadian entrainment and cellular response to glucagon stimulus (GNAS, GNG2) . Additionally, GDM was associated with methylation levels at 22 CpGs that mapped to 14 genes related to apoptosis (TNFRSF10B, TRADD) and neurological development (CACNA1G, ATXN1) . Overall, findings suggest that maternal pre-pregnancy obesity and GDM are associated with CpG methylation of the placental epigenetic clock, revealing potential markers vital for healthy pregnancy and fetal development.

Disclosure

K.Huff: None. C.Marsit: None. S.Niemiec: None. H.Volk: None. T.M.O'shea: None. R.Fry: None. K.Roell: None. C.Bulka: None. K.E.Boyle: None. C.Breton: None. A.Burt: None. D.Dabelea: None. L.Kahn: None. M.R.Karagas: None.

Funding

National Institutes of Health (UH3OD023348)
250-LB: Tead1—A Regulator of Adult ß-Cell Proliferation and Function
Authors: LEE JEONGKYUNG , LIU RUYA , LI FENG , NEGI VINNY , JAGANNATHAN RAJAGANAPATHI , HUISING MARK , MA KE , SHIH BEN , MOULIK MOUSUMI , YECHOOR VIJAY
Published: May 2022

The transcriptional factor, Tead1, mediates the transcriptional output of the Hippo pathway. Tead1 activates the transcription of downstream genes including many cell cycle promoting genes that lead to cell proliferation in many tissues, but its role in β-cells is unknown. To test if Tead1 is required for β-cell proliferation and function, we deleted Tead1 in β-cell. Deletion of Tead1 in β-cells was confirmed in these β-cell Tead1 KO (β-Tead1-/-) , by a significant decrease in transcript and protein from whole islet lysates. The β-Tead1-/- mice had normal body weight but developed significantly higher fasting and fed glucose levels starting at 5 weeks of age and progressed to frank diabetes by 8 weeks, with fasting blood glucose >300 mg/dl, accompanied by hypoinsulinemia, as compared to floxed control mice. 8wk β-Tead1-/- mice display severe glucose intolerance due to an abrogation of GSIS during GTT, with a loss of first phase insulin secretion. Pancreatic insulin content was decreased by ∼50%, due to a decrease in expression of mature β-cell genes including, Pdx1, Nkx6.1 and MafA, all of which, we show to be direct transcriptional targets of Tead1. Tead1 also binds to the proximal promoter at the Ink4a locus to regulate p16/p19, critical cell cycle inhibitors, and this loss of activation in Tead1-deficient β-cells leads to an enhanced entry into cell cycle. Furthermore, to test if Tead1 has a similar regulatory role in human β-cells, we deleted TEAD1 knockout in human iPS cell-derived β-cells using CRISPR/Cas9 system. Tead1 deficient iPS cell-derived β-cells displayed a significant decrease in vitro GSIS. Our results demonstrate Tead1 to be a transcriptional switch required for β-cells to maintain mature function and remain in proliferative quiescence. This novel regulatory pathway could be targeted for β-cell replacement therapy to achieve β-cell proliferation without a loss of mature function.

Disclosure

J. Lee: None. V. Yechoor: n/a. R. Liu: None. F. Li: None. V. Negi: None. R. Jagannathan: None. M. Huising: Consultant; Crinetics Pharmaceuticals, Inc., Research Support; Crinetics Pharmaceuticals, Inc. K. Ma: None. B. Shih: None. M. Moulik: None.

Funding

VA-MERIT Grant (I01BX002678)
251-LB: Tracking Beta-Cell Regeneration in Human Pancreatic Slices using Adenovirus Transduction
Authors: ALVAREZ-CUBELA SILVIA , KLEIN DAGMAR , ALVER CHARLES G. , MUHAMMAD FAHD QADIR MIRZA , MATEUS GONCALVES LUCIANA , QURESHI FARHAN , ALMACA JOANA , FRAKER CHRISTOPHER , CAICEDO ALEJANDRO , PUGLIESE ALBERTO , RICORDI CAMILLO , OLTRA ELISA , PUGLIESE ALBERTO , AGARWAL ASHUTOSH G. , PASTORI RICARDO , DOMÍNGUEZ-BENDALA JUAN
Published: May 2022

Introduction: The study of pancreatic regeneration would benefit greatly from the design and validation of robust human-based models. Human Pancreatic Slices (HPSs) are thin organotypic sections of live pancreatic tissue. The sectioning method preserves the overall histological structure of the organ, maintaining the integrity of the extracellular matrix and the natural interaction between the endocrine and exocrine compartments, as well as the local neural, vascular and immune milieu. Conditions for the long-term culture of HPSs, recently reported by our team, have enabled the real-time analysis of beta-cell neogenesis using adenoviral (AV) co-transduction of a red-green reporter and an insulin tracer in human pancreatic slices. However, the ability of these cells to respond to glucose was not established at that time.

Methods: To determine whether new INS+ cells respond to glucose, we have designed an AV in which INS-dependent recombination leads to the expression of a blue marker (moxBFP) and a Calcium Imaging Reporter (gcAMP6s, green) whose intensity is proportional to glucose-dependent INS secretion. This allowed us to monitor glucose-stimulated calcium influx in the newly created cells. To track the generation of new beta cells with an even higher degree of resolution we subsequently placed the transduced slices in a custom-made microfluid chip.

Conclusions: This setting further allows for longitudinal functional analyses in precise and highly controlled experimental conditions. Our ability to study regeneration in a clinically meaningful model represents a groundbreaking advance that may fast-track the screening and preclinical development of therapeutic agents.

Disclosure

S. Alvarez-cubela: n/a. A. Pugliese: Advisory Panel; Provention Bio, Inc., Consultant; Quell Tx. C. Ricordi: Advisory Panel; Vertex Pharmaceuticals Incorporated. E. Oltra: None. A. Pugliese: Advisory Panel; Provention Bio, Inc., Consultant; Quell Tx. A. G. Agarwal: None. R. Pastori: None. J. Domínguez-bendala: None. D. Klein: None. C. G. Alver: None. M. Qadir: None. L. Mateus goncalves: None. F. Qureshi: None. J. Almaca: None. C. Fraker: None. A. Caicedo: None.

Funding

National Institutes of Health (RO1DK130846UO1DK12393)
252-OR: Maternal Obesity and Diabetes during Pregnancy and Quantitative Checklist for Autism in Toddlers (QCHAT) Scores in Offspring
Authors: CARTER SARAH A. , LIN JANE C. , CHOW TING , MARTINEZ MAYRA P. , ALVES JASMIN M. , FELDMAN R. KLARA , QIU CHUNYUAN , PAGE KATHLEEN A. , XIANG ANNY
Published: May 2022

In utero exposure to maternal obesity and diabetes has been associated with risk of autism spectrum disorders (ASD) in children. As early identification and support improves long-term outcomes in children with ASD, a 10-point Quantitative Checklist for Autism in Toddlers (QCHAT) screening questionnaire was developed and validated for clinical screening between 18-24 months at well-child visits. This study assessed associations between maternal metabolic exposures and offspring performance on early QCHAT screening. Data included 65699 singletons born in a single healthcare system, with QCHAT administered toddler well-child visits. Demographic and social data, maternal pregnancy obesity (BMI≥30 kg/m2) , pre-existing type 1/type 2 (T1D/T2D) , and gestational diabetes (GDM) status, and child’s QCHAT score were extracted from electronic medical records. Negative binomial regression was used to assess associations between maternal obesity and diabetes and QCHAT scores as a continuous measure, adjusting for covariates. Analyses included and excluded children with ASD diagnoses and were run with and without mutual adjustment for each metabolic condition. In this cohort, 18923 (28.8%) of children were exposed to maternal obesity, 2564 (3.9%) to T1/T2, 1946 (3.0%) to GDM diagnosed at ≤26 weeks, and 3290 (5.0%) GDM diagnosed >26 weeks. Of the total cohort, 1622 (2.5%) were later diagnosed with ASD. Maternal obesity (Incidence Rate Ratio: 1.12, 95% CI: 1.09,1.14) , T1D/T2D (1.13, 1.07-1.20) , and GDM ≤26 weeks (1.12, 1.05-1.19) were associated with one-unit increases in QCHAT scores. No associations were reported for GDM >26 weeks. Similar patterns were observed after excluding children with ASD diagnoses. Mutual adjustment did not change results. These data suggest maternal metabolic conditions during pregnancy are associated with increased social and communication impairment on early autism screening tools, even in typically developing children.

Disclosure

S.A.Carter: None. J.C.Lin: None. T.Chow: None. M.P.Martinez: None. J.M.Alves: None. R.Feldman: None. C.Qiu: None. K.A.Page: None. A.Xiang: None.