bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Hyperinsulinemic Isoglycemic Hyperlactemic Clamp
Authors: ZARETSKY DMITRY , YOUNG ANDREW A. 
Published: June 2018

The Cori Cycle (glucose-lactate interchange) dominates carbohydrate fluxes. Glucose-to-lactate flux follows peripheral disposal; lactate-to-glucose occurs during gluconeogenesis. As glucose flux can be assessed by the change in an exogenous glucose infusion rate (GIR) required to maintain a constant concentration (glucose clamp), so too can lactate flux be assessed from an exogenous lactate infusion rate (LIR) required to maintain isolactemia. Simultaneous assessment of both fluxes will enable estimation of lactate-derived gluconeogenesis (GNG), and its contribution to the rate of glucose appearance (Ra) without using radiolabeled tracers, the mainstay of flux measurement. Anesthetized Sprague-Dawley rats were intubated, catheterized via artery and vein, and partially pancreatectomized to eliminate endogenous glucagon secretion. Establishment of a hyperinsulinemic (100mU/kg/h), hyperglycemic (7mM) clamp was followed by a hyperlactemic (4mM) clamp via independent variable infusions of glucose/Na-lactate in response to 5-min measures of plasma concentrations (YSI 2950D Analyzer with glucose/lactate electrodes). Hyperlactemia (∼10x resting) rendered it the dominant gluconeogenic substrate. After at least 30 min of isolactemia, glucagon was infused for 60 min (0, 1, 3, 10 or 50 µg/kg/h) to enable dose/concentration response analysis. Glucagon administration dose-dependently reduced the GIR and increased the LIR necessary to maintain isoglycemia/isolactemia (respective ED50’s 1.33, 0.98 µg/kg/h). δ-GIR of up to -12.2 mmol/kg/h potentially represented increased glucose Ra. δ-LIR of up to +8.6 mmol/kg/h potentially represented GNG. Since 2 lactate moieties are required per glucosyl, GNG contributed ∼35% of glucagon-stimulated glucose Ra (∼65% being glycogenolysis).
In conclusion, the hyperinsulinemic isoglycemic hyperlactemic clamp enables a quantitative assessment of hepatic processes, complementing that of peripheral processes revealed with the glucose clamp.

Disclosure
D. Zaretsky: Employee; Self; Intarcia Therapeutics, Inc. A.A. Young: Employee; Self; Intarcia Therapeutics, Inc.. Stock/Shareholder; Self; GlaxoSmithKline plc..
Autoimmunity in Patients with Type 2 Diabetes in Newfoundland and Labrador
Authors: IQBAL SAHAR J. 
Published: June 2018

Latent Autoimmune Diabetes of the Adult (LADA) is a slowly progressive form of diabetes associated with autoantibody positivity. LADA shares characteristics with type 1 and 2 diabetes (T1D, T2D) but is often misdiagnosed as T2D. Epidemiologic data shows patients with LADA may account for 2 to 12% of all cases of diabetes.
The aim of the study was to collect information on prevalence of autoimmunity in patients with T2D and describe their characteristics in Newfoundland and Labrador, Canada.
A cross- sectional study design was used to collect data in 140 patients. The frequency of GAD 65 antibody, ICA, Insulin and C-peptide levels was examined by radioimmunoassay and enzyme-linked immunosorbent assays. We collected data on height, weight, Body Mass Index (BMI) and tested for Thyroid Stimulating Hormone and Glycosylated Hemoglobin levels. Data collected included age at diagnosis, family history and number of co-morbidities. Adults 18 years and above with T2D were included. Pregnant and lactating females were excluded. SPSS version 24 was used to analyze data.
Results obtained showed 13 (8 males and 5 females, p 0 .181) of the 132 patients tested were GAD 65 positive thus placing prevalence of LADA in NL at 9.7%. Baseline characteristics were similar in both GAD 65 positive and negative patients. None of the patients tested were positive for ICA antibodies thereby improving our confidence that the sample did not contain any patients with T1D. Prevalence of GAD 65 was higher in the patients requiring Insulin whereas the C-peptide levels were lower. BMI was high in both GAD 65 positive and negative group with data trending towards higher BMI in the GAD 65 positive group.
GAD 65 positivity was associated with older age group, higher BMI which is different from reported literature indicating that older age and obesity have a predictive value in diagnosis of T2D but not in LADA.

Disclosure
S.J. Iqbal: None.
Role of ROCK1 in Pancreatic ß Cells
Authors: SUNG BYUNG-JUN , HUR KYU YEON , KIM JAE HYEON , KIM YOUNG-BUM , LEE MOON-KYU 
Published: June 2018

Insulin signaling in pancreatic β-cell is critical for normal insulin secretion. ROCK1 plays an important role in the regulation of insulin and leptin signaling in the context of glucose and energy metabolism. However, the role of ROCK1 in pancreatic β-cell remains unknown. The current study was to determine whether deleting ROCK1 from β-cell affects insulin secretion and glucose metabolism in mice and cultured cell lines. Here we show that β-cell-specific deletion of ROCK1 results in a significant decrease insulin secretion in mice fed a normal chow diet, leading to hyperglycemia. These effects were independent of changes in adiposity. Consistent with this, in vitro studies with INS-1 and MIN6 cells indicated that inhibition of ROCK1 decreased insulin secretion in response to glucose without changes in β-cell proliferation and apoptosis. In addition, β-cell-ROCK1 deficient mice displayed glucose intolerance, as evidenced by the fact that the area under the glucose curve was increased 53% in β-cell-ROCK1 deficient mice compared with control mice. However, neither body weight, food intake nor insulin sensitivity was changed in theses mice. Upon insulin stimulation, proximal insulin signaling components, including p-IRS2, p-PI3K and p-Akt were impaired when ROCK1 expression was inhibited in INS-1 cells, suggesting a role for ROCK1 in insulin signaling. These data indicate that activation of ROCK1 in pancreatic β-cell is required for the maintenance of normal insulin secretion. Thus, pancreatic ROCK1 might be a key regulator of glucose homeostasis.

Disclosure
B. Sung: None. K. Hur: None. J. Kim: None. Y. Kim: None.M. Lee: None.
The Simultaneous Changes of Endogenous Glucose Production, Postprandial Glucagon, and Fasting Glutamine during Weight Loss in Type 2 Diabetes
Authors: OTTEN JULIA , STOMBY ANDREAS , WALING MARIA , CHORELL ELIN , RYBERG MATS , SVENSSON MICHAEL B. , HOLST JENS J. , OLSSON TOMMY 
Published: June 2018

In type 2 diabetes, high blood glucose levels are associated with increased glucagon levels and insufficient suppression of endogenous glucose production (EGP). Notably, glutamine, a gluconeogenic amino acid, is decreased compared to healthy individuals.
We randomized 32 overweight patients with type 2 diabetes to either a Paleolithic diet (PD) or a Paleolithic diet combined with supervised exercise (PD-EX). Study duration was 12 weeks. Subjects conducted a solid mixed meal test: glucagon, insulin and glucose were measured at 0, 30, 60, 120 and 180 min with calculation of the total area under the curve for the response. On another study day, fasting glutamine was measured with GC-MS and suppression of EGP was examined with the hyperinsulinemic euglycemic clamp technique with [6,6-2H2]glucose as a tracer and with an insulin infusion of 40 mIU/m2/min.
Median weight loss was 7 kg in both study groups. Fasting glucose and fasting insulin decrease significantly only in the PD-EX group. Postprandial glucose decreased by 13% in the PD group (P&lt;0.05) and by 26% in the PD-EX group (P&lt;0.01), without difference between groups. Fasting glucagon tended to decrease in both study groups. Postprandial glucagon decreased by 22% in the PD-group (P&lt;0.01) and by 21% in the PD-EX group (P=0.13). Fasting glutamine increased non-significantly in both groups. Suppression of EGP increased by 26% in the PD group (P&lt;0.05) and by 10% in the PD-EX group (P=0.75). During the intervention, the increased suppression of EGP was associated with a) reduction of postprandial glucagon levels (rS=-0.65, P&lt;0.01), b) decreasing fasting glucagon (rS=-0.51, P&lt;0.05) and c) the increasing fasting glutamine levels (rS=0.51, P&lt;0.05).
We conclude that glucagon, an activator of EGP, decreases in concert with improved suppression of EGP during weight loss in type 2 diabetes patients. An increase in plasma glutamine may be due to decreased gluconeogenesis.

Disclosure
J. Otten: None. A. Stomby: None. M. Waling: None. E. Chorell: None. M. Ryberg: None. M.B. Svensson: None. J.J. Holst: Advisory Panel; Self; Novo Nordisk A/S. Board Member; Self; Zealand Pharma A/S. Speaker's Bureau; Self; Merck Sharp & Dohme Corp., AstraZeneca. Research Support; Self; Danish Diabetes Academy, Novo Nordisk Foundation. Other Relationship; Self; Antag Therapeutics. Other Relationship; Spouse/Partner; Antag Therapeutics. T. Olsson: None.
GLP-1 Secretion Is Increased upon Blockade of the Somatostatin Receptor Subtype 2 and 5 Resulting in GLP-1 Receptor-Mediated Lowering of Blood Glucose in Mice
Authors: JEPSEN SARA L. , WEWER ALBRECHTSEN NICOLAI J. , PEDERSEN JENS , ENGELSTOFT MAJA S. , DEACON CAROLYN F. , HOLST JENS J. 
Published: June 2018

Somatostatin (SS) is a hormone inhibiting the secretion of the incretin hormone, glucagon-like peptide-1 (GLP-1) by binding to 5 SS receptors (SSTr). However, which SSTrs involved is still unclear. We hypothesized that antagonizing the SSTrs on enteroendocrine L-cells may increase secretion of GLP-1 and via GLP-1 receptor mediated mechanism lead to lowering of blood glucose. To investigate the level of the SSTrs we did expression analysis on L-cells isolated from throughout the murine intestine (n=3). We perfused the proximal small intestine from mice (n=6-7) to investigate the impact of SSTr2 and 5 on GLP-1 secretion using intra-arterial administration of specific antagonists. Lastly we performed oral glucose tolerance tests (OGTT) on mice (n=5-8) after giving 1mg/ml SSTr2 or 5 antagonist with and without the GLP-1 receptor antagonist exendin(9-39) to investigate the role of these receptors on the glucose profile and to elucidate if their effect was mediated via GLP-1. The expression levels of the SSTrs showed that SSTr2 and especially 5 had the highest expression on L-cells throughout the intestine. The SSTr2 antagonist resulted in a increase in GLP-1 secretion from the perfused intestine (100nM, 4.96pM±0.56; 1µM, 14.55pM±1.4; 10µM, 16.46pM±1.18pM; all p&lt;0.0001). The same doses of SSTr5 antagonist gave a more potent GLP-1 secretion in this setup (100nM, 5.82pM±0.7 p=0.0004; 1µM, 19.79pM±1.2 p&lt;0.0001; 10µM, 48pM±2.6 p&lt;0.0001). Both SSTr2 and SSTr5 antagonists significantly reduced the blood glucose during an OGTT compared to the control (p=0.00and p=0.0072 respectively). This reduction in blood glucose was significantly increased when adding exendin(9-39) together with either the SSTr2 or 5 antagonist (p&lt;0.0001 for SSTr2, p=0.0036 for SSTr5). Our results demonstrate that inhibiting the SSTr2 and SSTr5 increases GLP-1 secretion which in turn may cause the lowering in blood glucose during an OGTT.

Disclosure
S.L. Jepsen: None. N.J. Wewer Albrechtsen: Speaker's Bureau; Self; Merck Sharp & Dohme Corp.. Other Relationship; Self; Mercodia, Alpco. J. Pedersen: None. M.S. Engelstoft: None. C.F. Deacon: Other Relationship; Self; Merck & Co., Inc., Eli Lilly and Company. Employee; Spouse/Partner; Merck Sharp & Dohme Corp.. Stock/Shareholder; Spouse/Partner; Merck & Co., Inc. J.J. Holst: Advisory Panel; Self; Novo Nordisk A/S. Board Member; Self; Zealand Pharma A/S. Speaker's Bureau; Self; Merck Sharp & Dohme Corp., AstraZeneca. Research Support; Self; Danish Diabetes Academy, Novo Nordisk Foundation. Other Relationship; Self; Antag Therapeutics. Other Relationship; Spouse/Partner; Antag Therapeutics.
In Insulin-Resistant Subjects, Islet Functional Changes Might Represent an Attempt to Increase the Incretin Effect
Authors: MEZZA TERESA , MOFFA SIMONA , SORICE GIAN PIO , ASSUNTA CEFALO CHIARA MARIA , CINTI FRANCESCA , MARI ANDREA , GIACCARI ANDREA 
Published: June 2018

Nondiabetic insulin-resistant subjects (InsRes) display an increased α-cell mass, suggesting a key role of α-cells in the compensatory mechanisms to cope with insulin resistance states. The mechanism contributing to greater GLP-1 secretion detected in healthy patients following pancreatoduodenectomy (PD), is not fully understood and suggests other potential sources of the incretin hormone, such as α-cells. To investigate the dynamic changes of in-vivo GLP-1 secretion, insulin secretion (IS), and incretin effect (IE), 16 nondiabetic patients (10 F/6 M, 51±15 years.) scheduled for PD underwent a mixed meal test (MMT), a 2-h hyperglycemic clamp (HC), and a hyperinsulinemic euglycemic clamp (HEC) before and after surgery. β-cell glucose sensitivity (GS) was calculated as the ratio of IS and glucose increments, both during HC and MMT. The IE was estimated as the ratio of GS during the HC and MMT. InsRes displayed a reduction in the 1st IS at baseline (p=0.04), which did not decrease following PD. Both groups experienced a reduction of 2nd phase ISR (p&lt;0.for both groups) and arginine-induced IS (p&lt;0.for both groups) after surgery, with no difference observed in the reduction delta; while a significant reduction in basal ISR and GS was observed only in InsRes after surgery (respectively p=0.03 and p=0.06). Following PD, GLP-1 secretion significantly increased in both groups (P=0.and P=0.02 respectively), but the percentage of increase was greater in InsRes compared to insulin-sensitive subjects (InsSen) (respectively 54.4% vs. 36.2%). Further, the IE was significantly decreased in the InsSen (p=0.05), whereas no changes were observed in InsRes. Our data suggest that acute islet mass reduction leads to different functional effects in InsRes compared to InsSen. Impaired β-cell GS and reduced 1stIS are relatively compensated by preserved IE following PD in InsRes. This may be driven by an increased α-mass in InsRes, that might be the source of the increased GLP-1 secretion.

Disclosure
T. Mezza: None. S. Moffa: None. G. Sorice: None. C. Cefalo: None. F. Cinti: None. A. Mari: None. A. Giaccari: Advisory Panel; Self; Sanofi-Aventis. Speaker's Bureau; Self; AstraZeneca, Merck Sharp & Dohme Corp., Amgen Inc..
The Family Coriobacteriaceae Is a Potential Contributor to the Beneficial Effects of Roux-en-Y Gastric Bypass on Type 2 Diabetes
Authors: HU CHENG , JIA WEIPING , LIU HAIJUN 
Published: June 2018

Background: Changes in gut microbiota induced by bariatric surgery have been associated with metabolic benefits.
Objectives: Our aim was to identify specific gut microbiota that may contribute to the improvement of type 2 diabetes (T2D) after RYGB.Setting: Laboratories of Shanghai Diabetes Institute and Shanghai Sixth People’s Hospital.
Methods: Diabetic rats induced via a high-fat diet and low-dose streptozotocin administration were randomized to RYGB or sham surgery (SHAM), and stool samples were collected at baseline and at postoperative week 8. The faecal microbiota was profiled using 16S ribosomal RNA (rRNA) gene sequencing. Additionally, we performed a case-control study of the gut microbial community profiles of T2DM patients compared to those of healthy individuals (CONT) via 16S rRNA gene sequencing of mucosal-luminal interface samples collected from the ascending colon during colonoscopy.
Results: RYGB significantly reduced the body weight and improved glucose tolerance and insulin sensitivity in diabetic rats. Principal coordinate analysis showed that RYGB caused marked alterations in the gut microbiota. The RYGB group was postoperatively enriched for Bacteroidetes, Proteobacteria, Fusobacteria and Actinobacteria; whereas the SHAM group was enriched for Firmicutes and Verrucomicrobia. Based on the gut microbial patterns in the T2D patients, we found that the family Coriobacteriaceae within Actinobacteria might contribute to the beneficial effects of RYGB on T2D.
Conclusions: RYGB significantly improves glucose metabolism and alters the gut microbiota. Moreover, the family Coriobacteriaceae may partly mediate the beneficial effects of RYGB on T2D and thus possibly contribute to the development of novel bacteria-based therapeutic approaches.

Disclosure
C. Hu: None. W. Jia: None. H. Liu: None.
The Regulation of BRD7 in the Insulin Signaling Pathway
Authors: PARK SANG , HAN YOUNGAH , SALAZAR HERNANDEZ MARIO A. 
Published: June 2018

Elevated endoplasmic reticulum (ER) stress plays crucial roles in the development of insulin resistance, type 2 diabetes, and obesity. X-Box binding protein 1 (XBP1) is one of the master regulators of ER folding function and is involved in the regulation of ER stress. Haploinsufficiency of XBP1 leads to the development of obesity, insulin resistance, and type 2 diabetes even in mice on a background that is resistant to the development of obesity. To date, our understanding of the regulation of XBP1 in metabolic processes is still limited. In our previous work, we demonstrated that bromodomain containing protein 7 (BRD7) regulates glucose homeostasis by increasing the nuclear translocation of XBP1s through its interaction with the regulatory subunits of phosphatidyl inositol 3-kinase (PI3K), p85s. We also showed that BRD7 protein levels are significantly reduced in the liver of genetically obese ob/ob and high fat diet (HFD)-induced obese mouse models, and over-expression of BRD7 in the liver of ob/ob and HFD fed mice ameliorates hyperglycemia and glucose intolerance through increasing the activity of XBP1s. In our current unpublished work, we show that BRD7 heterozygous knockout mice are prone to developing obesity when they are challenged with a HFD. They display increased hepatic triglyceride levels with moderately severe steatosis. In addition, we generated a liver-specific BRD7 knockout mouse model (LBKO) to examine the effect of BRD7 specifically in the liver. LBKO mice fed on a HFD exhibit increased body weights compared to their wild type control mice. Downregulation of BRD7 in the liver led to lack of nuclear XBP1s in the liver in both the knockout mouse model and adenovirus-mediated knockdown system. Our results provide the first evidence that BRD7, which was originally reported as a tumor suppressor, is involved in the development of obesity as well as in the regulation of unfolded protein response (UPR) and glucose metabolism.

Disclosure
S. Park: None. Y. Han: None. M.A. Salazar Hernandez: None.
Berberine Hydrochloride Protects Cytokine-Induced Inflammation through Multiple Pathways
Authors: POUDEL ANIL , ZHOU JOSEPH , MASSEY SCOTT , LI LIXIN 
Published: June 2018

Berberine hydrochloride (BBR) has been reported to improve insulin resistance. However, the underlying mechanism remains largely elusive. We therefore investigate the anti-inflammatory role of BBR and the cell signaling pathways that involved.
C2C12 myoblasts were treated with IL-1β (10ng/mL) alone or in combination with TNF-α (50 ng/mL) in the presence or absence of BBR (10 µM) for 24 hours. Western blot analysis was used to assess the protein level. In vivo study is conducted using C57BL/6 mice fed with high fat high sucrose (HFHS) diet to induce insulin resistance. Skeletal muscle was collected after 4 weeks treatment with either 5 mg/kg BBR or vehicle.
BBR reduced activity of iNOS, and stress-related kinases, including p38 MAPK, NF-kB, SAP/JNK, MMK4 and C-JUN that was induced by cytokines in C2C12 myoblast. Furthermore, BBR reversed cytokine mediated suppression of AMPK-α, augmented activity of SIRT-1 and PGC-1α. Antioxidant enzyme or transcription factor, nuclear factor like 2 (NRF-2) were also elevated after BBR treatment. In addition, BBR improved insulin signaling in C2C12 myoblast. HFHS diet induced iNOS expression was reversed by BBR in skeletal muscle. Augmentation of the activity of uncoupled protein-3 (UCP-3) and PPARα in HFHS diet skeletal muscle was observed.
Our results demonstrate that BBR ameliorates cytokine induced inflammation and oxidative stress. BBR improves mitochondrial function, promotes insulin sensitivity. The anti-inflammatory effect of BBR in skeletal muscle may mediated through multiple pathways including activation of AMPKα-SIRT-1-PGC-1α pathway, and inhibition of MMK4- SAP/JUNK-C-JUN pathway. BBR is a potential medication in treating obesity induced insulin resistance.

Disclosure
A. Poudel: None. J. Zhou: None. S. Massey: None. L. Li: None.
Endothelial ULK1 Regulates High-Fat Diet–Induced Obesity via FGF-21
Authors: LI MANNA , QIAN MING , XU JIAN 
Published: June 2018

Fibroblast grow factor (FGF) 21 is a hormone with huge potential for pharmaceutical development for treatment of metabolic disease. The role of endothelial cell (EC)-derived FGF-21 in obesity remains largely unknown, although we detected lower EC-FGF-21 levels in visceral fat of Ob/Ob and high-fat diet (HFD)-fed mice than the control lean mice. This study was to identify the role and mechanism of EC-FGF-21 in HFD-induced obesity using cell culture and mouse models. Mimicking HFD, long-term incubation with palmitate (100 µM, 16h) reduced FGF-21 levels in HUVEC that were co-cultured with 3T3-L1-adipocytes. The reduction was prevented in ULK1-siRNA-treated HUVEC, via ATF4-promoted mRNA expression and increased protein stability of FGF-21. Thus, FGF-21-induced expression of FGF-R1c, β-Klotho, UCP1 and GLUT1 was enhanced in co-cultured adipocytes. To confirm these findings in vivo, we used a Tie2/Cre-lox system to generate mice with EC-knockout of FGF-21 (FKO), ULK1 (UKO), or both (DKO). In 8-to-10-week-old male mice (n=15-20/group) fed an HFD (TestDiet: 60% energy from fat) for 3 months, wild type (WT, C57BL/6J) mice developed obesity and insulin resistance, which was worsened in FKO mice. In contrast, UKO mice gained less body weight and visceral fat mass and had improved glucose tolerance, without significant differences in food and water intake. Levels of adipose EC-FGF-21 was normalized in UKO mice, as determined by immunohistochemistry of FGF-21 and EC markers (CD31 and lectin). Expression of adipose FGF-R1c, β-Klotho, UCP1, and GLUT1 was restored in UKO, but not WT, mice. Concomitant with UCP1 induction, the visceral fat showed beige-cell-like deposits with fewer expanded adipocytes. However, this favorable phenotype was absent in DKO mice.
In summary, EC-ULK1 mediated HFD-induced obesity and insulin resistance through loss of EC-FGF-21, at least in part, by blunting local FGF-21-signaling in adipose tissue. Therapeutic targeting of the EC-ULK1-FGF-21 pathway may be a new treatment for obesity and insulin resistance.

Disclosure
M. Li: None. M. Qian: None. J. Xu: None.