bluuebunny/crossref_metadata_2025_split huggingface dataset:
1252-P: Alteration of Alternative Splicing of RYR2 Modulates Insulin Biosynthesis in Pancreatic ß-Cells
Authors: MAKINO MAI , ITAYA-HIRONAKA ASAKO , YAMAUCHI AKIYO , SAKURAMOTO-TSUCHIDA SUMIYO , TAKEDA YOSHINORI , UCHIYAMA TOMOKO , TAKASAWA SHIN 
Published: June 2021

In pancreatic β cells, intracellular Ca2+ is essential for insulin biosynthesis. Ryanodine receptors (Ryrs) mediate Ca2+ mobilization from the endoplasmic reticulum. Previously we found that type 2 Ryr (Ryr2; originally called cardiac-type) of three isoforms from separate genes was expressed in pancreatic β cells and that Ryr2 mRNA expressed in the pancreatic β cells was deficient in the region of exon 75, unlike that expressed in the heart. This alternative splicing occurred tissue-specifically, which is presumably due to a unique splice donor site “gg” in intron 75 of Ryr2 gene. In order to clarify the role of the tissue-specific splice variant of Ryr2 on insulin biosynthesis, we constructed the homozygous genome-modified pancreatic β cells which express cardiac RYR2 by changing the splice donor “gg” of RYR2 gene to “gt” in human 1.1B4 pancreatic β cells using the CRISPR/Cas9 system. Insulin content in the culture medium and insulin mRNA level were measured by ELISA and real-time RT-PCR, respectively. The results showed that both insulin and insulin mRNA levels were significantly lower in the genome-modified cells than those in parental 1.1B4 cells. We examined the effect of complementation of Ryr2 by introducing the rat cardiac-type or islet-type Ryr2 expression vector into each cell. Real-time RT-PCR analysis showed that the expression of insulin mRNA was significantly increased in the cells transfected with the islet-type Ryr2 expression vector, whereas that in the cells transfected with the cardiac-type Ryr2 expression vector was not increased. Furthermore, the result of luciferase assay showed that the promoter activity of insulin gene was not decreased in the genome-modified cells as compared with the parental 1.1B4 cells.
In conclusion, those results indicate that exon 75-deficient Ryr2 could be important in the post-transcriptional step of insulin biosynthesis via Ca2+ signaling in pancreatic β cells.

Disclosure
M. Makino: None. A. Itaya-hironaka: None. A. Yamauchi: None. S. Sakuramoto-tsuchida: None. Y. Takeda: None. T. Uchiyama: None. S. Takasawa: None.

Funding
Japan Society for the Promotion of Science (JP19K16543)
1254-P: ß-Cell Cdk1 Deletion Reveals a- and ß-Cell Compensatory Mechanisms for Hypoplasia
Authors: SDAO SOPHIA , ADAMS MELISSA , FOSTER HANNAH R. , BLUM BARAK , MERRINS MATTHEW J. 
Published: June 2021

Diabetes is characterized by a loss of functional β-cell mass. With current rodent models of chemically-induced diabetes such as streptozotocin, it is difficult to distinguish the consequence of β-cell loss from the decline in individual β-cell function. Here, we generated a mouse model of severe β-cell mass loss by deleting β-cell CDK1 within the first week of life (Cdk1Ucn3Cre, Cdk1-floxed:Ucn3-Cre). At 4 weeks of age, islet mass in Cdk1Ucn3Cre islets is comparable to controls, although islet architecture is disrupted with α-cells having formed more heterotypic contacts with β-cells. By 12 weeks of age, Cdk1Ucn3Cre mice exhibit significantly reduced β-cell mass as well as α-cell mass, but maintain glucose tolerance and ex vivo insulin secretion in response to glucose, amino acids, and the gut incretin GIP. We therefore hypothesized that functional compensations in Cdk1Ucn3Cre islets arise from both β- and α-cells, which are insulinotropic in the fed state. Interestingly, α-cell glucagon secretion from Cdk1Ucn3Cre islets exhibits hypersensitivity to suppression by glucose, including when glucagonotropic amino acids are present. In parallel, we observed a reduction in amino acid-stimulated cAMP in β‑cells from Cdk1Ucn3Cre islets. We also found a left-shift in the glucose dependence of β-cell calcium oscillations in Cdk1Ucn3Cre islets, identifying β-cell recruitment as one mechanism offsetting the loss of α-cell paracrine signaling. Importantly, GIP elicited a much larger glucagon response from glucose- and amino-acid stimulated Cdk1Ucn3Cre islets, implying that α-cell compensation contributes significantly to the maintained insulin secretory output in vivo. In sum, the Cdk1Ucn3Cre model reveals the interdependence of α- and β-cell mass, and function, and will be useful to define the temporal sequence of islet compensation for chronic loss of β-cell mass under stressed conditions.

Disclosure
S. Sdao: None. M. Adams: None. H. R. Foster: None. B. Blum: None. M. J. Merrins: None.
1257-P: LncRNA Kcnq1ot1 Affects Islet ß-Cell Proliferation and Insulin Secretion
Authors: CHEN YALAN , LI YANLI , LIU ZIYU , XU WEN , LIN BEISI , SU YANNA , YAN JINHUA , LI WANGEN 
Published: June 2021

Background: Accumulating evidences indicated that long noncoding RNA (LncRNA) were involved in the regulation of diverse biological functions, including the islet function. In our previous study, relatively high expression of LncRNA Kcnq1ot1 was found in the pancreas of normal mice. This study was designed to clarify the role of LncRNA Kcnq1ot1 in pancreatic β cells.
Methods: The expressions of LncRNA Kcnq1ot1 in different tissues of normal adult male C57BL/6J mice were detected by qRT-PCR. siRNA was used to downregulate Kcnq1ot1 in MIN6 β cells. The proliferation and apoptosis of β cells were measured by cell counting kit-8 (CCK-8) assay and flow cytometry. Glucose stimulated insulin secretion (GSIS) assay was used to determine the effect of Kcnq1ot1 on insulin secretion of β cells. Moreover, the expressions of cell cycle and insulin related genes (including Ccnd1, Ccnd2, Ins1 and Ins2) were assessed by qRT-PCR. The LncRNA Kcnq1ot1 siRNA and scrambled siRNA were injected into the tail vein of male C57BL/6J, then, the blood glucose and serum insulin levels were measured at 72h after injection.
Results: The expression of Kcnq1ot1 was much more abundant in islets of normal male C57BL/6J mice than other tissues. The CCK8 and mRNA expression of Ccnd1 and Ccnd2 showed that the proliferation of Min6 cells was significantly decreased with the downregulation of kcnq1ot1 at 48h and 72h. However, flow cytometry indicated that inhibition of Kcnq1ot1 had no effects on β cells apoptosis. Impaired β-cell function measured by GSIS was found in kcnq1ot1 silenced Min6 cells, along with decreased mRNA levels of INS1 and INS2. Higher glucose and lower serum insulin were detected in the LncRNA Kcnq1ot1 siRNA-treated mice when compared to control group.
Conclusions: LncRNA Kcnq1ot1 has multiple effects on β-cell function, including enhanced proliferation and insulin secretion, which suggest Kcnq1ot1 as a new biomarker of islet function.

Disclosure
Y. Chen: None. Y. Li: None. Z. Liu: None. W. Xu: None. B. Lin: None. Y. Su: None. J. Yan: None. W. Li: None.

Funding
National Natural Science Foundation of China (81800682)
126-LB: Evaluation of the Efficacy of the Therapeutic Agent Cytoreg in Patients with Diabetic Foot: A Clinical Humanitarian Protocol
Authors: CARRILLO DARIO A. , VILLASANA KATIUSCA , TORRES GEIZON , POZO LEWIS J. , JIMENEZ WILLIAM J. , BAUTA WILLIAM E. , DUGARTE LEONARDO 
Published: June 2021

Diabetic foot wound complications, including bacterial infections, ulcerations, and gangrene, are a major cause of hospitalizations and account for 85% of diabetic amputations. The investigational therapeutic agent Cytoreg®, an aqueous mixture of hydrofluoric, hydrochloric, sulfuric, phosphoric, phosphoric, citric, and oxalic acids, was recently shown to increase arterial oxygen levels in an orally administered Wistar rat model. It previously demonstrated bactericidal effect. Under a compassionate use protocol, we tested oral and topical treatment with diluted Cytoreg in patients with diabetic foot ulcers in a 30-day study in Venezuela. The total study population was ten patients under informed consent. On average, all patients received oral Cytoreg daily for 30 days. Five patients received oral Cytoreg plus topical wound cleansing with Cytoreg in isotonic saline. In addition to standard clinical observations, wound healing was monitored using the St. Elian’s checklist system. Four of five patients in the oral plus topical group had complete closure of their wounds after thirty days. Two of the patients in the oral group had complete wound closure. There was no significant change in oxygen saturation in the affected tissues, nor in insulin and glycemia levels. However, there were significant increases in arterial hemoglobin and arterial oxygen-pO2- were observed. Significant decreases HbA1c, aspartate aminotransferase, glutamic-pyruvic transaminase, creatinine, and urea levels were also observed. The overall results are encouraging and justify an expanded study in the treatment of diabetic foot wounds.

Disclosure
D. A. Carrillo: Research Support; Self; Cytorex Biosciences Inc. K. Villasana: None. G. Torres: Research Support; Self; Cytorex Bioscience Inc. L. J. Pozo: None. W. J. Jimenez: Board Member; Self; Cytorex Biosciences Inc. W. E. Bauta: Advisory Panel; Self; Cytorex Biosciences, Vascular Perfusion Solutions, Employee; Self; bioAffinity Technologies. L. Dugarte: None.
1265-PUB: Basic Carbohydrate Counting in Patients with Type 2 Diabetes: Improvement in Glycemic Control and Effects on Body Compositions
Authors: TANIGUCHI MARIKO , MORITA RYUHEI , HARADA REIKO , YOSHINO FUYUMI , MUKAI YUMI , IKEDA MIWA , HARADA KAZUE , YOSHIYAMA MAO , NAKANO HIROMI , MAEDA KANAKO 
Published: June 2021

Objective: This study aims to show the usefulness of basic carbohydrate counting (BCC) in the glycemic control of patients with type 2 diabetes (T2DM) and to investigate the effects of BCC on body compositions.
Subjects and Methods: We included T2DM 20 patients in no insulin therapy with stable dosage of antidiabetic medication 8 weeks before and during the study period. We provided the meal planning according to the indicated sugar mass, which was set to 50% of their energy amount per day. HbA1c levels, daily carbohydrate intake, and body compositions (measured with InBody 770) were collected before and 12 ± 4 weeks after BCC.
Results: Before/after BCC of their daily carbohydrate intake and HbA1c levels were 210.7 ± 42.1/171.5 ± 34.9 g (p = 0.0018), and 8.1 ± 1.1/7.4 ± 0.9 % (p = 0.0002), respectively. To examine the effects of BCC on their body compositions, 16 patients with improved HbA1c levels were selected for further analysis. BCC improved their body weight (before/after BCC, 64.4 ± 17.6/63.0 ± 16.6 kg, p = 0.005), BMI (25.7 ± 5.3/25.2 ± 5.0 kg/m2, p = 0.004), body fat mass (21.8 ± 9.5/20.1 ± 8.7 kg, p = 0.014), and abdominal circumference (95.0 ± 13.6/92.9 ± 13.2 cm, p = 0.0004). There was no significant difference in overall SMI, but the muscle mass of right arm, left arm, and trunk significantly decreased while the muscle mass of legs was not affected by BCC.
Discussion and Conclusion: Our findings demonstrate that BCC is an effective strategy for improving the glycemic control of T2DM patients. The patients with improved HbA1c levels also showed significant reduction in their body weight, BMI, body fat mass, and abdominal circumference. While preserving the muscle mass of their legs, the muscle mass reduction in their arms and trunks was observed. Combining with resistant training of arms and trunk, BCC may make more positive effects on T2DM patients.

Disclosure
M. Taniguchi: None. K. Maeda: None. R. Morita: None. R. Harada: None. F. Yoshino: None. Y. Mukai: None. M. Ikeda: None. K. Harada: None. M. Yoshiyama: None. H. Nakano: None.
1267-PUB: Sustained Reduction of A1C through Use of AI Digital Coach and Consumer-Friendly Glucose Measurement
Authors: BALDER TIM 
Published: June 2021

Management of diabetes has always depended upon sustained self-management by the person with diabetes. Data continues to show that people do a poor job of diabetes management. Many telehealth solutions achieve short-term, but not sustained results. This investigated platform implements two unique elements to improve the ability to achieve sustained self-management. One element is an Artificial Intelligence (AI) digital coach that exists in the mobile app--keeping the right behaviors in front of people with diabetes and encouraging continual engagement. The second element is an integrated glucose meter with a dramatically more friendly user experience--defined by smaller size, discreet (no visible assembly), and less painful. The platform has been commercialized for more than 12 months. A clinical registry has been established and currently includes 50 commercial users of the platform and more than 25,000 glucose tests are included. Demographics include both type 1 and type 2, 42% male, 58% female, with ages ranging from 24 to 69 years old. With use of the platform, Figure 1 shows a significant decrease in A1C, as calculated from glucose measurements, of 1.4 points (p = 0.0001) in early use to a sustained reduction of 0.9 points (p=0.0001). These results may indicate that easier visibility of results causes immediate reduction of glucose and that this easier visibility combined with the AI coach may sustain results. These results potentially demonstrate that a simpler experience for self-management will improve sustained diabetes management results. Future studies of the registry data will include improved compliance toward ADA guidelines and glucose time in range.

Disclosure
T. Balder: Advisory Panel; Self; POPS Diabetes Care.
1279-PUB: Postpartum Screening for Diabetes in Women with a History of Gestational Diabetes
Authors: RAKHRA VARUNDEEP , RILEY MARCELLA , JORDAN SYMONE , BULCHANDANI SHEETAL , ALLSWORTH JENIFER , DREES BETTY 
Published: June 2021

Background: Gestational diabetes (GDM) is glucose intolerance detected for the first time during pregnancy. Women with a history of GDM have a 7-fold higher lifetime risk of developing type 2 diabetes (DMII) compared to women without a history of GDM. Women with a history of GDM should be screened for DMII 6 to 12 weeks postpartum with a 2-hour 75-g oral glucose tolerance test (OGTT) and then again one year after delivery. The long-term goal of this project is to improve screening for DMII in women with a history of GDM. This report establishes the prevalence of GDM, determines the postpartum screening rate, and assesses for barriers to screening at an urban safety-net hospital. This information will be used to design interventions to improve postpartum screening.
Methods: We conducted a retrospective chart review of women who had a delivery at Truman Medical Center between January 1, 2017 to December 31, 2019 and had a diagnosis of GDM. Charts were reviewed for demographic data, management of GDM, postpartum appointments, and screening for DMII.
Results: There were 9569 deliveries during the three-year period. Of the total deliveries, 537 (5.6%) had a diagnosis of GDM. Of the patients with GDM, 426 (79%) had a postpartum visit scheduled, 354 (66%) attended a 6-week postpartum appointment, 219 (41%) had the recommended test ordered, and 49 (9%) completed the test. The data revealed a significant association with ethnicity, language, insurance type, and site of prenatal care with attending the postpartum appointment (p&lt;0.05) as well as with insurance status on completing the screening test (p&lt;0.05).
Conclusion: There is gap in the number of women with GDM and those who are screened for DMII in the postpartum period. There appears to be a disconnect in the ordering of the recommended 2-hour OGTT and the completion of the test. The baseline data supports a multi-prong approach including patient education, system changes, and advocacy for extended insurance benefits to cover screening.

Disclosure
V. Rakhra: None. M. Riley: None. S. Jordan: None. S. Bulchandani: None. J. Allsworth: None. B. Drees: None.
144-LB: Collaborative Telehealth Approach Where Traditional Care Fails Diabetes Care Network
Authors: CLARK KRISTY , LARSON MEG J. , LUTZ-MCCAIN STACEY J. , BANDI ARCHANA 
Published: June 2021

Clinical inertia is the failure to initiate or intensify treatment in a timely manner in people with type-2 diabetes mellitus (T2DM) in primary care (PC) where collaborative care can be an approach to overcome inertia. Diabetes care network (DCN) is a telehealth based collaborative care model to address clinical inertia in the PC environment. In DCN, initial care was delivered via E-Consult by hub diabetes team for Veterans with A1c of 9% or higher and the longitudinal collaborative care delivered by PC liaison with weekly team huddle. To study the difference in the clinical outcomes with DCN vs. PC practices, we compared the DCN cohort (97.7% male, 90.8% white, with a mean age 67.2 (8.9), with a PC cohort (100% male, 94.9% white, mean age: 68 year (10.5), with A1C &gt;9%).
Methods: Means (SD), frequencies and percentages were presented. The DCN cohort had a significant decline in the baseline A1C of 10.2% (1.4), to 8.1% (0.99), 7.6% (0.96), 7.5 % (0.86) at 3, 6, and 12 months while A1c in PC cohort stayed poor with baseline A1C of 10.1% (0.89), to 10.2% (1.69), 9.7% (1.74) and, 9.5% (1.83) at 3, 6, and 12 months. Patients who achieved A1c less than 8% in DCN cohort were 38 (43.6%), 56 (64%), and 56 (64%) at 3, 6, and 12 months and were 1(1.7%), 5(8.5%), and 6(10.2%) at 3, 6, and 12 months in PC cohort. An A1c of less than 7% was achieved in DCN cohort in 10 (11.4%), 21 (24.1%), and 23 (26.4%) patients at 3, 6, and 12 months and only 1(1.7%), 0(0%), and 0(0%) patients achieved A1c less than 7% at 3, 6, and 12 months in PC cohort. Our study shows participation in the DCN telehealth program for 1 year was associated with significant improvement in A1c. This improvement was not seen within the PC with traditional care practices. Thus, addressing clinical inertia in PC will need a paradigm shift in current practices. The proposed collaborative approach of DCN can overcome clinical inertia in PC and improve care for people with T2D by supporting PC access to specialty care expertise, decreasing the burden of diabetes care for patients and PC providers.

Disclosure
K. Clark: None. M. J. Larson: None. S. J. Lutz-McCain: None. A. Bandi: None.
146-OR: A New Type 2 Diabetes Microsimulation Model to Estimate Long-Term Health Outcomes, Costs, and Cost-Effectiveness
Authors: HOERGER THOMAS J. , HILSCHER RAINER , NEUWAHL SIMON , CHENG YILING J. , BENOIT STEPHEN R. , SHAO HUI , LAXY MICHAEL , YANG WENYA , CINTINA INNA , KAUFMANN MATTHEW , CHEN HAIYING , ANDERSON ANDREA M. , STAIMEZ LISA R. , NARAYAN K.M. VENKAT , ZHANG PING 
Published: June 2021

We developed a new microsimulation model to estimate health effects, costs, and cost-effectiveness of public health interventions for type 2 diabetes. The model combined risk equations for complications and mortality, risk factor progression equations, and patient utility equations based on the U.S. ACCORD and Look AHEAD studies with a cost equation estimated from a large panel dataset of privately insured U.S. adults. The model features a web-based interface that allows users to change model parameters. The model includes interventions for glycemic, blood pressure, and cholesterol control; smoking cessation; and a generic intervention that users can modify to represent control of multiple risk factors.
To demonstrate how the model estimates cost-effectiveness, we analyzed an intervention to reduce HbA1c from 9% to 7%. We estimated the long-run health effects, costs, and quality-adjusted life-years (QALYs) for a simulated cohort of 10,000 U.S. adults with type 2 diabetes. On average, the simulated cohort of U.S. patients was projected to have 20.16 remaining life-years (from a mean age of 61), incur $187,435 in discounted medical costs, and experience 8.89 discounted QALYs. Results were most sensitive to age and duration of diabetes at baseline and the discount factor. The intervention to reduce HbA1c produced $3,151 in incremental costs and gained 0.4 QALYs, yielding an incremental cost-effectiveness ratio of $7,838 per QALY.
The model performed well in internal validation exercises. External validation results were more mixed, with the model better at predicting clinical trial results than at predicting observational study results.
As the model’s risk progression, patient utility, and costs are all derived using recent U.S. studies, our new model can more accurately project the long-run health impact, costs, and cost-effectiveness of interventions for type 2 diabetes in the United States than existing models.

Disclosure
T. J. Hoerger: None. M. Kaufmann: None. H. Chen: None. A. M. Anderson: None. L. R. Staimez: None. K. Narayan: None. P. Zhang: None. R. Hilscher: None. S. Neuwahl: None. Y. J. Cheng: None. S. R. Benoit: None. H. Shao: Research Support; Self; Sanofi. M. Laxy: None. W. Yang: None. I. Cintina: None.

Funding
Centers for Disease Control and Prevention (200-2016-92270)
154-OR: People Who Get More Hours of Sleep Have Lower Mean Glucose: Analysis of Continuous Glucose Monitoring and Fitbit Data in a Free-Living Population without Diabetes
Authors: CRAWFORD MARGARET A. , PICKUS SARAH K. , HAMES KAZANNA C. , JOHNSON MATTHEW , KLEIN ILENE J. , NORMAN GREGORY J. 
Published: June 2021

Background: Sleep disorders are associated with having or developing type 2 diabetes, increased insulin resistance, and higher BMI. The relationship between sleep duration and continuous glucose monitoring (CGM) metrics has not been well studied. We examined the relationship between CGM-based metrics and sleep duration in a population without diabetes.
Methods: Glucose metrics from a 10-day blinded CGM (Dexcom G6) wear and sleep metrics from a concurrent unblinded Fitbit wear were collected. Participants (mean (SD) age= 49.1 (10) years, BMI= 29.5 (8) kg/m2, 90% female, 14% African American) were recruited from an employee health plan in the Southeastern US. Forty-nine individuals without diabetes or prediabetes were included in this analysis. A linear regression model adjusting for age and BMI was used to evaluate the relationship between mean daytime glucose and nightly hours of sleep.
Results: Participants recorded a median (IQR) 6.4 (6.1, 7.1) nightly hours of sleep, 6416 (5112, 8071) daily steps, and 12.7 (12.1, 13.5) daily sedentary hours. Mean (SD) mean daytime glucose was 112.7 (11.5) mg/dL and coefficient of variation was 15.4 (3) %. Mean daytime glucose was significantly and inversely associated with nightly hours of sleep (Beta= -3.1, p &lt;0.05).
Conclusion: In this population without diabetes, each additional one hour of sleep was associated with 3.1 mg/dL lower mean daytime glucose. This finding aligns with existing research on the protective effects of sleep on glycemic health and suggests it may be beneficial for people at risk for developing diabetes to improve their sleep habits. Longitudinal research where behavioral data is collected along with CGM can provide further insights into correlates of developing prediabetes and type 2 diabetes.

Disclosure
M. A. Crawford: Employee; Self; Dexcom, Inc. S. K. Pickus: Employee; Self; Dexcom, Inc. K. C. Hames: Employee; Self; Dexcom, Inc. M. Johnson: Employee; Self; Dexcom, Inc. I. J. Klein: None. G. J. Norman: Employee; Self; Dexcom, Inc.