bluuebunny/crossref_metadata_2025_split huggingface dataset:
121-OR: Activation of FoxO1 Induces Akt Phosphorylation and Mimics the Metabolic Functions of Insulin without Proliferative Effects on 3T3-L1 Adipocytes
Authors: OHNO TOMOHIRO , ONO HIRAKU , TAKEDA KENJI , YOKOTE KOUTARO 
Published: June 2021

Objective: Insulin promotes glucose uptake, lipogenesis, cell proliferation and cell growth mainly via Akt phosphorylation. FoxO1 is a substrate of Akt, which is inactivated when phosphorylated by Akt. Via this mechanism, FoxO1 antagonizes insulin. On the other hand, constitutive activation of FoxO1 reportedly enhances Akt phosphorylation. Thus, FoxO1 activation, via Akt phosphorylation, may mimic the functions of insulin.
Methods: We overexpressed constitutively-active FoxO1 (CA-FoxO1) and FoxO1Δ256, a dominant-negative mutant, in 3T3-L1 adipocytes by adenoviruses and investigated the effects of these proteins on insulin’s functions.
Results: CA-FoxO1 enhanced Akt phosphorylation, glucose uptake (275%, P&lt;0.01), GLUT4 translocation and lipogenesis (176%, P&lt;0.05) in the non-insulin stimulated state, while DNA synthesis was decreased (44%, P&lt;0.01) in the insulin-stimulated state. Surprisingly, FoxO1Δ256 also enhanced Akt phosphorylation, glucose uptake (154%, P&lt;0.05) and lipogenesis (268%, P&lt;0.01) in the basal state. In contrast to CA-FoxO1, FoxO1Δ256 increased DNA synthesis (196%, P&lt;0.01). FoxO1Δ256 did not exert a dominant negative action even on gene expressions related to adipocyte differentiation, de novo lipogenesis and cell cycle regulation. Akt phosphorylation was more strongly induced by CA-FoxO1 than by 3 nM insulin, while PI 3-kinase activation by FoxO1 was weaker than that induced by 3 nM insulin, indicating the mechanism of Akt phosphorylation by FoxO1 to mainly be downstream from PI 3-kinase. Expressions of IR-β, p110α, Rictor and TRB3 were not significantly affected.
Conclusion: Activation of FoxO1 selectively mimics the metabolic functions of insulin, without its proliferative effects. Contrary to previously reported results, FoxO1Δ256 does not always exert a dominant negative action.

Disclosure
T. Ohno: None. H. Ono: None. K. Takeda: None. K. Yokote: None.
122-OR: Insulin and C-Peptide in Human Choroid Plexus of Type 1 Diabetes Mellitus
Authors: LIU QING-RONG , ZHU MIN , MAZUCANTI CAIO , MARENCO STEFANO , EGAN JOSEPHINE M. 
Published: June 2021

The choroid plexus (CP) produces cerebrospinal fluid (CSF) and forms a blood-CSF barrier. We already reported that mouse CP epithelial (CPE) cells synthesize and secrete insulin in response to serotonin but not glucose. We now report that insulin expression is present in human postmortem CP samples of humans - nondiabetic, T2DM, and T1DM - by RT-qPCR, in situ hybridization and LC-MS/MS based Selected Reaction Monitoring assay. Human CPE contains all the INS splicing machinery and the major INS splicing variants that are present in islets. Known β-cell specific transcription factors are also expressed in CPE, except for PDX1 and PAX4, which are absent. The SRM assay detected INS A-chain, B-chain, and C-peptide at similar levels in nondiabetic, T2DM, and T1DM islets and CP postmortem samples. Islet amyloid polypeptide (IAPP) expression was not detected in CPE in contrast to human islets where INS and IAPP mRNA, as expected, colocalize in β-cells. Furthermore, INS colocalizes with transthyretin (TTR) mRNA in CPE cells but in islets, TTR is expressed in α-cells and not β-cells. We found less or no expression of T1DM autoantigens in CPE compared to β-cells, with the exception of ICA1 (islet cell autoantigen1), which has similar expression in both tissues.
In conclusion, human CPE expresses and synthesizes insulin, it is protected from autoimmune destruction and from IAPP amyloidogenic fibril formation.

Disclosure
Q. Liu: None. M. Zhu: None. C. Mazucanti: None. S. Marenco: None. J. M. Egan: None.

Funding
National Institute on Aging
1220-P: Treatment with Testosterone Undecanoate Injections (TU) Results in Sustained Weight Loss in Men with Hypogonadism and Type 2 Diabetes: Update of Real-World Data from a Urological Practice
Authors: HAIDER AHMAD , HAIDER KARIM S. , SAAD FARID 
Published: June 2021

Background: The prevalence of hypogonadism is particularly high in men with obesity and T2DM.
Methods: Of 361 men with hypogonadism and T2DM, 293 (81.2%) were obese, 61 (16.9%) overweight and 7 (1.9%) had normal weight. 183 received TU 1000 mg/12 weeks (T-group) following an initial 6-week interval, 178 opted against treatment (CTRL). Changes over time between groups were compared and adjusted for age, weight, waist circumference, fasting glucose, blood pressure, lipids and quality of life to account for baseline differences between groups.
Results: Mean baseline age: 61.8±5.3 years, mean (median) follow-up: 8.7±3.1 (10) years. T-group: weight decreased by 22.5±0.5 kg (unadjusted numbers in brackets) (from 114.1±14.0 to 89.5±7.9) at 12 years. CTRL: weight increased by 8.5±0.5 (from 102.8±14.0 to 103.9±10.1) (p&lt;0.0001 for both). T-group: waist circumference decreased by 13.6±0.4 cm (from 117.1±14.4 to 98.9±4.9). CTRL: WC increased by 8.5±0.4 cm (from 116.9±14.0 to 118.2±8.5) (p&lt;0.0001 for both). T-group: BMI decreased by 7.4±0.2 kg/m² (from 36.5±4.5 to 28.9±2.6). CTRL: BMI increased by 2.8±0.2 kg/m² (from 33.4±5.3 to 33.2±3.6) (p&lt;0.0001 for both). There was a weight loss of 19.7±0.4% in the T-group and weight gain of 9.1±0.4% in CTRL (p&lt;0.0001 for both). Waist:height ratio decreased by 0.08±0.00 (from 0.66±0.08 to 0.56±0.03) in the T-group and increased by 0.05±0.00 (from 0.66±0.08 to 0.67±0.05) in CTRL (p&lt;0.0001 for both). Visceral adiposity index (VAI) decreased by 3.1±0.2 (from 5.8±3.1 to 1.9±0.5) in the T-group and increased by 3.7±0.2 (from 5.1±2.5 to 8.1±3.7) in CTRL (p&lt;0.0001 for both). Since injections were administered in the office and documented, there was a 100% adherence to testosterone therapy.
Conclusions: Treatment with testosterone undecanoate injections results in profound weight loss that is maintained long-term with continued testosterone therapy.

Disclosure
A. Haider: Other Relationship; Self; Bayer AG. K. S. Haider: Other Relationship; Self; Bayer AG. F. Saad: Consultant; Self; Bayer AG, Stock/Shareholder; Self; AbbVie Inc., Bayer AG, Novo Nordisk.

Funding
Bayer AG
1232-P: A Total Weight Loss of 25% Shows Better Predictivity in Evaluating the Efficiency of Bariatric Surgery
Authors: TU YINFANG , PAN YUNHUI , HAN JUNFENG , PAN JIEMIN , BAO YUQIAN , YU HAOYONG 
Published: June 2021

Objectives: The need for a unified definition of weight loss (WL) after bariatric surgery has recently been highlighted. We aimed to evaluate the reliability of two clinically common WL indications including percentage of total WL (%TWL) and percentage of excess WL (%EWL) through comparing their performances in predicting metabolic syndrome (MetS) remission 1 year after bariatric surgery.
Methods: A total of 430 individuals with obesity who underwent bariatric surgery were enrolled. Participants were evaluated for changes in anthropometric parameters, metabolic indexes, MetS components and medications before and 1 year after surgery. MetS was defined using the criteria of the National Cholesterol Education Program Adult Treatment Panel III criteria for Asian-Americans.
Results: The prevalence of MetS is 92.3% (397) at baseline. One year after bariatric surgery, 337 individuals (84.9%) were in MetS remission. The multivariate adjusted odds ratios (ORs) were 1.16 (95% confidence interval [CI] 1.10-1.22) for each 1% increase in %TWL for MetS remission and 1.18 (95% CI 1.11-1.25) for each 5% increase in %EWL. This association with MetS remission remained statistically significant for %TWL after additional adjustment for %EWL (P for trend 0.029), and disappeared for %EWL. Receiver operating curve (ROC) analyses showed that the %TWL was more predictive than the %EWL (AUC%TWL vs. AUC %EWL, 0.749 vs. 0.700, p=0.023). The Youden index indicated that the optimal %TWL cut-off point to identify MetS remission was 25%.
Conclusions: We recommend that good responders to bariatric surgery should be defined as those exhibiting % TWL ≥25%. Clinical Trial Registration: Evaluation of the effectiveness and safety of metabolic surgery for obesity: a clinical cohort study; Chinese Clinical Trial Registration:ChiCTR1900028513.

Disclosure
Y. Tu: None. Y. Pan: None. J. Han: None. J. Pan: None. Y. Bao: None. H. Yu: None.

Funding
National Key Research and Development Project of China (2016YFA0502003); National Natural Science Foundation of China (81670791); Municipal Natural Science Foundation of Shanghai (17ZR1421200); Shanghai Key Clinical Center for Metabolic Disease (2017ZZ01013)
1233-P: Relationship between Taste Recognition Thresholds and Obesity in Type 2 Diabetes
Authors: SAKAI CHIEKO 
Published: June 2021

Objective: An increased taste threshold can often lead to lifestyle diseases such as obesity. However, little knowledge is available on whether the coexistence of obesity and diabetes affects taste recognition. Therefore, this study investigated the relationship between degrees of obesity and taste recognition thresholds for the 5 basic tastes (sweet, salty, sour, bitter, and umami) in patients with type 2 diabetes.
Methods: The study was performed in 30 outpatients (22 men and 8 women) with type 2 diabetes who were not receiving pharmacotherapy (age: 59.1 ± 13.0 years, BMI: 25.2 ± 3.8 kg/m2, HbA1c: 7.0 ± 1.2%, fasting plasma glucose: 137 ± 40 mg/dl). Taste recognition thresholds for the 5 basic tastes were evaluated with the filter paper disk method. The Mann-Whitney U test and Spearman’s test were used for analysis.
Results: Taste thresholds measured by perception levels in patients with type 2 diabetes were as follows: sweetness, 3.5 ± 1.5; saltiness, 2.6 ± 0.9; sourness, 3.2 ± 1.3; bitterness, 2.7 ± 1.3; and umami, 4.8 ± 1.3. The recognition thresholds for sweetness and sourness were significantly higher than in healthy people. (sweetness, 0.001; sourness, 0.007; both P &lt; .05). The correlation coefficients between BMI and the 5 taste thresholds were not significant (sweetness, -0.075; saltiness, 0.060; sourness, -0.074; bitterness, -0.025; and umami, 0.211).
Conclusion: Although patients with type 2 diabetes who are not receiving pharmacotherapy have higher taste thresholds for sweetness and sourness than healthy people, these differences were not associated with the degree of obesity.

Disclosure
C. Sakai: None.
124-OR: Repetitive Ca2+ Waves Emanate from a Stable Leader Cell in Mouse Islets
Authors: CHABOSSEAU PAULINE L. , MARTINEZ-SANCHEZ AIDA , LECLERC ISABELLE , SALEM VICTORIA , RUTTER GUY A. 
Published: June 2021

Pancreatic beta cells form a highly connected network within the islet, with transcriptomic and functional heterogeneity existing between individual cells. A sub-population of leader cells has been proposed to act as functional hubs, controlling islet responses to glucose and other secretagogues. Using high speed confocal Ca2+ imaging, we aimed here to examine (a) the role of these cells in wave propagation and (b) their stability over time.
Methods: Islets were isolated from C57BL6 male mice expressing the recombinant Ca2+ probe GCaMP6f selectively in the beta cell (Ins1Cre:GCaMP6f/f). Islets were then infected with an adenovirus expressing photo-activatable- (PA) mCherry. Ca2+ imaging was performed across a single islet plane in modified KREBS-Ringer medium at 11mM glucose using a Zeiss LSM 810 laser-scanning microscope and data acquisition at 5 Hz. Cells were photopainted by excitation of PA-mCherry at 405nm.
Results: Leader cells were identified as those exhibiting the first detectable increase in Ca2+ signal during subsequent Ca2+ waves. The great majority of islets (85%, n=26, three independent experiments) displayed repetitive Ca2+ waves (1.10±0.26 min-1), and 82.9±5.0% of waves propagated directly from an identifiable leader cell. There was an average of 2.5±0.4 leader cells per islet with 62.9±0.70% of waves emanating from a “principal” leader cell. Ca2+ imaging experiments performed 16 h after the original session revealed that 37±0.08% (n=4) of waves emanated from the same, now photopainted, leader cell.
Conclusions: These data suggest that (a) a complex, non-binary, heterogeneity exists between leader and follower beta cells, (b) leader behaviour is an intrinsic phenotype of a sub-population of cells with a half-life in this role of ∼12 h.

Disclosure
P. L. Chabosseau: None. A. Martinez-sanchez: None. I. Leclerc: Consultant; Spouse/Partner; Sun Pharmaceutical Industries Ltd. V. Salem: None. G. A. Rutter: Advisory Panel; Self; Sun Pharmaceutical Industries Ltd.
1240-P: Changes in Protein Expression during Islet Regeneration in a Cystic Fibrosis-Related Diabetes (CFRD) Ferret Model
Authors: ISMAIL HEBA M. , MIRMIRA RAGHAVENDRA G. , EVANS-MOLINA CARMELLA , BONE ROBERT N. 
Published: June 2021

Diabetes is a common complication among persons with cystic fibrosis (CF), affecting nearly 20% of adolescents and 50% of all adults with CF. The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) knockout (KO) ferret is a well-characterized animal model of CF that exhibits distinct periods of dysglycemia and pancreatic remodeling that are similar to patterns observed in humans. After the first month of life (normoglycemia/Phase I), hyperglycemia develops during the second month of life (Phase II) with reduced islet mass, followed by a period of near normoglycemia (Phase III) during which the islet mass regenerates, followed by persistent hyperglycemia/CFRD at the 4th month of life (Phase IV). While dynamic changes in islet hormone mRNA levels have been described across these phases, changes in protein expression patterns have not been characterized. To this end, immunofluorescence was performed to characterize islet hormone expression patterns in age-matched CFTR KO and wild type (WT) ferrets, focusing on the regenerative Phase III. The relative intensity of immunofluorescent staining per islet area was calculated. Compared to WT ferrets, the relative staining intensity of insulin and glucagon increased (4.687 vs. 3.228, p&lt;0.05 and 0.827 vs. 0.238, p&lt;0.0001, respectively ), while the intensity of proinsulin decreased (1.061 vs. 1.455, p&lt;0.05) in CF ferret sections. Our findings suggest increased efficiency of proinsulin processing and a potential compensatory upregulation of insulin expression during this phase with the surprising finding of increased glucagon expression. Additional studies are underway to further characterize changes in islet hormone expression across each of the four described phases as well as potential cellular dedifferentiation patterns.

Disclosure
H. M. Ismail: None. R. G. Mirmira: Advisory Panel; Self; Hibercell Inc., Sigilon Therapeutics, Inc., Veralox Therapeutics, Employee; Spouse/Partner; Beta Bionics, Inc. C. Evans-molina: Advisory Panel; Self; Provention Bio, Inc., Consultant; Self; Dompe, Other Relationship; Self; Bristol-Myers Squibb Company, Nimbus Pharmaceuticals, Pfizer Inc. R. N. Bone: None.
1244-P: The Protein Phosphatase PPm1k Regulates Ribosomal Protein S6 Phosphorylation in Beta Cells
Authors: DELEYE YANN , HERRING JACOB , HESS KAVAN H. , LEININGER BRENNAN , MCGARRAH ROBERT W. , TESSEM JEFFERY S. , WHITE PHILLIP J. 
Published: June 2021

Phosphorylation of ribosomal protein S6 (RPS6) on ser235/236 and ser240/244 in the pancreatic beta cell increases cell size and insulin content and influences systemic glucose homeostasis. Our phosphoproteomic analysis identified the protein phosphatase, PPm1K, as a novel regulator of RPS6 phosphorylation. We find that beta cells account for most PPm1K expression within pancreatic islets and that exposure of INS1 832/13 beta cells to increasing concentrations of glucose results in suppression of PPm1K mRNA expression. siRNA mediated knockdown of PPm1K in INS1 832/13 cells, to levels achieved by glucose, increased insulin content and secretion in basal and glucose stimulated conditions that was accompanied by robust phosphorylation of RPS6 on ser235/236 and ser240/244. PPm1K knockdown did not increase phosphorylation of p70 S6 Kinase (S6K) or the mammalian target of rapamycin (mTOR) upstream of RPS6 suggesting that PPm1K regulates RPS6 phosphorylation in an mTOR-S6K independent manner. Accordingly, treatment with the mTORC1 inhibitor, Rapamycin, lowered S6K and RPS6 phosphorylation but did not override the effect of PPm1K knockdown to increase RPS6 phosphorylation compared to control siRNA treated cells. Lowering phosphorylation of PPm1K’s classical target, ser293 of the branched-chain keto acid dehydrogenase (BCKDH) with the small molecule, BT2, also did not impact the effect of PPm1K knockdown to increase RPS6 phosphorylation. Thus PPm1K exerts its effects on RPS6 phosphorylation in a BCKDH and mTOR/S6K independent manner. The effects of siRNA-mediated PPm1K knockdown on RPS6 phosphorylation were accompanied by a 1.5 fold increase in protein translation and the induction of an unfolded protein response as determined by an increase in XBP-1 splicing, and BiP expression. Together our data identify a new role for the phosphatase PPm1K as a glucose sensitive modulator of RPS6 phosphorylation and protein translation that impacts beta cell insulin content and secretion.

Disclosure
Y. Deleye: None. J. Herring: None. K. H. Hess: None. B. Leininger: None. R. W. Mcgarrah: None. J. S. Tessem: None. P. J. White: None.

Funding
American Diabetes Association/Pathway to Stop Diabetes (1-16-INI-17 to P.J.W.)
1255-P: The Role of BAI3 Adhesion G-Protein Coupled Receptor in Insulin Secretion and Glucose Tolerance
Authors: ALSHARIF HAIFA , RAHMAN MOSTAFIZUR , PATHAK ASMITA , TROMBLEY JOHN E. , BHATNAGAR SUSHANT 
Published: June 2021

Insulin secretion from beta-cells of pancreatic islets is essential for regulating whole-blood glucose homeostasis. Impaired insulin secretion is an early event in the development of obesity-linked type 2 diabetes (T2D). However, underlying molecular mechanisms that cause a reduction in insulin secretion are not completely understood. We recently identified that brain angiogenesis inhibitor-3 (BAI3) adhesion G-protein coupled receptor is expressed in beta-cells, and when activated by its native ligand, complement-1q like-3 (C1ql3), blunts insulin secretion in response to cyclic adenosine monophosphate (cAMP) in an autocrine/paracrine manner. The expression of BAI3 is conserved in human islets, yet the role of islet BAI3 in whole-body glucose homeostasis remains uncharacterized. We generated C57BL6/J mice that are homozygous for the loss of the BAI3 gene by using CRISPR technology. Our data show that the BAI3-/- mice have reduced body weight and impaired glucose tolerance compared to the wild-type control (WT) mice. These mice have elevated in vivo insulin secretion after 10 min of glucose challenge and an increased ex vivo islet insulin secretion in response to stimulation by 40 mM KCL and 11 mM glucose with no effect observed at 2.8 mM glucose. Concomitantly, the insulin action is decreased in the BAI3-/- mice. These outcomes show that increased insulin secretion from beta-cells due to the loss of the BAI3 gene contributes to reduced insulin sensitivity and glucose tolerance. The expression of BAI3 is increased by obesity, indicating that the activation of BAI3 signaling may contribute to reduced insulin secretion, causing beta-cell dysfunction in T2D. In sum, we have established an important role of an understudied BAI3 GPCR signaling in inhibiting insulin secretion, potentially identifying an attractive target for novel T2D therapeutics.

Disclosure
H. Alsharif: None. M. Rahman: None. A. Pathak: None. J. E. Trombley: None. S. Bhatnagar: None.

Funding
National Institutes of Health (R01DK120684-01)
1268-PUB: The Relationship of Time-in-Range (TIR) and HbA1c and the Factors That Contribute to a Better TIR
Authors: GONG LI , XU WEN , LIN BEISI , LIU ZHIGU , YANG DAIZHI , WANG CHAOFAN , YAO BIN 
Published: June 2021

Background: HbA1c, the current gold standard treatment target for patients with diabetes, cannot reveal the glucose fluctuation. Therefore, new metrics need to be incorporated with HbA1c to better understand the dynamic nature of glucose. As the use of continuous glucose monitoring (CGM) increases, there is a need to better understand key metrics of time in range 70-180 mg/dL (TIR).
Methods: Patients with T2DM underwent a 48-h to 72-h CGM were included and divided into two groups: low TIR group (LTIR, TIR&lt;70%) and high TIR group (HTIR, TIR≥70%). The characteristics between the two groups were compared. The relationship of HbA1c, glucose variability, time above range (TAR), time below range (TBR) and TIR were analyzed. And the contributions of BG and PPG increments to TIR were investigated.
Results: A total of 110 patients were enrolled. The age, BMI, duration of disease, treatment types and the index of glucose variability were not statistically significant (P &gt; 0.05) between the LTIR group (n=11) and the HTIR group (n=99). HbA1c (7.96±1.08% vs. 6.69±1.23%, p =0.002) and TAR (61.59±7.88% vs. 90.59±7.73%, p&lt;0.001) were lower in the HTIR group than that in the LTIR group, but TBR did not differ between groups. We further evaluated the contributions of BG and PPG to TIR, It showed that the relative contributions of BG in the LTIR group and the HTIR group were(47%vs33 %, p = 0.045), while the relative contributions of PPG were (53% vs. 67%, p = 0.045)in the two groups, respectively.
Conclusions: TIR is correlated with HbA1c and TAR, but not TBR, indicating the need to control the hyperglycemia for a better TIR. Furthermore, BG seems to a predominant contributor to a better TIR, and obviously exceed PPG in the T2DM patients with TIR &gt;70%.

Disclosure
L. Gong: None. W. Xu: None. B. Lin: None. Z. Liu: None. D. Yang: None. C. Wang: None. B. Yao: None.

Funding
National Key Research and Development Program of China (2016YFC1304801)