Nondiabetic Research Articles

Antidiabetic and Toxicological Effects of the Tea Infusion of Summer Collection from Annona cherimola Miller Leaves

Annona cherimola Miller (Ac) is a plant used in Mexican traditional medicine for the treatment of diabetes. In this work, the tea infusion extracts obtained from 1.5 g of leaf powder from Ac collected in May (AcMa), June (AcJun), July (AcJul), and August (AcAu) were evaluated on streptozocin-induced diabetic (STID) mice and for subchronic toxicity in STID and non-diabetic (ND) mice. In addition, extracts were subjected to high-performance liquid chromatography with diode array detection (HPLC-DAD). Results showed that the tea infusion extract of the sample collected in August (AcAu) exhibited the most significant antihyperglycemic activity during all acute assays. The analysis of the extracts (AcMa, AcJu, AcJul, and AcAu) by HPLC-DAD revealed that flavonoid glycosides, rutin, narcissin, and nicotiflorin were the major components. In addition, the sample AcAu contained the best concentration of flavonoids. In the case of subchronic oral toxicity, the AcAu sample did not cause mortality in STID mice, and histopathological analysis revealed significant improvement in the changes associated with diabetes in the liver and kidneys. These findings suggest that the Ac leaves collected in August may be a source of flavonoids such as rutin, with antidiabetic potential. In addition, these findings support the use of Ac to treat diabetes in traditional medicine.

Abstract 15087: Chronic Inhibition of NADPH Oxidase Improves Coronary Endothelial Function in Mice With Diabetes

Introduction:: Endothelial dysfunction is the leading cause for diabetes (DM) related coronary heart disease. Diabetes is associated with NADPH Oxidase (Nox) overproduction and related ROS amplification in endothelial cells and coronary vasculature, which may contribute to coronary endothelial dysfunction. Small conductance calcium-activated-potassium (SK) channels also play a key role in diabetes-induced coronary endothelial dysfunction . Hypothesis: Chronic inhibition of Nox may improve coronary endothelial function/coronary arteriolar relaxation, and SK activator-induced vasodilation in mice with or without diabetes. Methods: Wild type (control), non-diabetic (ND) mice and genetically modified, obesity/type-2 diabetic mice (BKS.Cg- Dock7 m +/+ Lepr db /J) with or with treatment with Nox inhibitor apocynin (n = 4-6/group) were used in the study. In the Nox inhibitor treated group, apocynin (30mg/kg/day, i.p) was applied to the diabetic mice for 4 weeks. Small coronary arteries (70-120μM) isolated for harvested hearts were pre-contracted and examined in the presence or absence of the vasodilatory agents, SNP, ADP, and NS309, respectively. Results: After 4-week treatment with apocynin, endothelium-NO-dependent relaxation responses to ADP, and endothelium-dependent, SK channel activator NS309 in the diabetic mice were significantly increased when compared with that in untreated diabetic mice, respectively (# p<0.05 vs ND Figure ). There were no significant differences in responses to the endothelium-independent vasodilator SNP (P>0.05) among the three groups. Conclusions: Chronic inhibition of Nox appears to improve coronary endothelial function/coronary relaxation and SK channel activity in mice with diabetes.

The human batokine EPDR1 regulates β-cell metabolism and function

ObjectiveEpendymin-Related Protein 1 (EPDR1) was recently identified as a secreted human batokine regulating mitochondrial respiration linked to thermogenesis in brown fat. Despite that EPDR1 is expressed in human pancreatic β-cells and that glucose-stimulated mitochondrial metabolism is critical for stimulus-secretion coupling in β-cells, the role of EPDR1 in β-cell metabolism and function has not been investigated. MethodsEPDR1 mRNA levels in human pancreatic islets from non-diabetic (ND) and type 2 diabetes (T2D) subjects were assessed. Human islets, EndoC-βH1 and INS1 832/13 cells were transfected with scramble (control) and EPDR1 siRNAs (EPDR1-KD) or treated with human EPDR1 protein, and glucose-stimulated insulin secretion (GSIS) assessed by ELISA. Mitochondrial metabolism was investigated by extracellular flux analyzer, confocal microscopy and mass spectrometry-based metabolomics analysis. ResultsEPDR1 mRNA expression was upregulated in human islets from T2D and obese donors and positively correlated to BMI of donors. In T2D donors, EPDR1 mRNA levels negatively correlated with HbA1c and positively correlated with GSIS. EPDR1 silencing in human islets and β-cell lines reduced GSIS whereas treatment with human EPDR1 protein increased GSIS. Epdr1 silencing in INS1 832/13 cells reduced glucose- and pyruvate- but not K+-stimulated insulin secretion. Metabolomics analysis in Epdr1-KD INS1 832/13 cells suggests diversion of glucose-derived pyruvate to lactate production and decreased malate-aspartate shuttle and the tricarboxylic acid (TCA) cycle activity. The glucose-stimulated rise in mitochondrial respiration and ATP/ADP-ratio was impaired in Epdr1-deficient cells. ConclusionThese results suggests that to maintain glucose homeostasis in obese people, upregulation of EPDR1 may improve β-cell function via channelling glycolysis-derived pyruvate to the mitochondrial TCA cycle.

Identification of novel microRNAs associated with type 2 diabetes by an integrative bioinformatic analysis

To identify potential miRNAs involved in the development/progression of type 2 diabetes (T2D) we performed data screening and integration of microRNA (miRNA) and messenger RNA (mRNA) expression profiles available in public data repositories. We retrieved two independent sequencing datasets: GSE52314 (small RNA-Seq) and GSE50244 (RNA-Seq). Then, we identified Differentially Expressed Genes (DEGs) and Differentially Expressed miRNAs (DEMs) from non-diabetic (ND) vs. people with T2D. The integrative analysis of identified DEGs and DEMs revealed 303 interactions involving 33 miRNAs and 256 genes. Functional enrichment analysis showed these interactions could play a functional role in carbohydrate, lipid, and nucleotide impaired metabolism. Some interactions we identified could also be relevant at an early stage of T2D. We also performed a similar integration analysis using a single study from a rat model, including both RNA-Seq and small RNA-Seq in the same pancreatic tissue: CRA000791. Interestingly, in this analysis one identified miRNA was also identified in the integrative analysis in human transcriptome: hsa-miR-27a-5p. Moreover, functional enrichment analysis showed that the target genes are mainly enriched in biological processes and pathways related to lipid and nucleotide metabolism.The miRNAs and interactions we identified could be relevant to gene expression regulation in T2D. Although further validation is necessary for extensive use in clinics, they provide new and useful knowledge to develop potential novel strategies for early diagnosis, prevention, and treatment of T2D.

A single-cell human islet interactome atlas identifies disrupted autocrine and paracrine communications in type 2 diabetes.

A sensible control of hormone secretion from pancreatic islets requires concerted inter-cellular communications, but a comprehensive picture of the whole islet interactome is presently missing. Single-cell transcriptomics allows to overcome this and we used here a single-cell dataset from type 2 diabetic (T2D) and non-diabetic (ND) donors to leverage islet interaction networks. The single-cell dataset contains 3046 cells classified in 7 cell types. The interactions across cell types in T2D and ND were obtained and resulting networks analysed to identify high-centrality genes and altered interactions in T2D. The T2D interactome displayed a higher number of interactions (10 787) than ND (9707); 1289 interactions involved beta cells (1147 in ND). High-centrality genes included EGFR, FGFR1 and FGFR2, important for cell survival and proliferation. In conclusion, this analysis represents the first in silico model of the human islet interactome, enabling the identification of signatures potentially relevant for T2D pathophysiology.

Bidirectional Relationship between Oral Health and Diabetes Mellitus: A Knowledge, Attitude, and Practice Survey

Background: Diabetes mellitus is a chronic disease, known to be associated with several oral complications. A less well-known, but proven bidirectional relationship exists between diabetes and oral health, where one affects the other. Previous studies have shown a lack of awareness about this two-way relationship. This Knowledge, Attitude, and Practice (KAP) study was undertaken to identify the gaps in awareness about diabetes mellitus and its association with oral health. Objectives: The objective of this study was to evaluate and compare the KAPs of diabetics and nondiabetics (ND) regarding the bidirectional relationship between diabetes and oral health. Methodology: This study was conducted at a tertiary dental care institute wherein 75 diabetics, both Type I and II and 75 ND were enrolled. A questionnaire was administered by the investigators, in which responses to various questions regarding their demographics, KAPs in the context of oral health and diabetes were collected. The Chi-Square test was utilized for statistical analysis. Results: 27.3% of respondents had no knowledge about the bidirectional relationship between diabetes and oral health. Although the majority (69.3%) considered regular dental check-ups to be important for good oral health, they were found to be lacking in their practices, where 51.3% followed once a day brushing routine and 71.3% did not know about flossing or interdental cleaning. Conclusion: The study highlights the gaps in KAPs of people regarding the bidirectional relationship between diabetes and oral health and thus emphasizes the need for educational interventions at the community level.

Total fecal IgA levels increase and natural IgM antibodies decrease after gastric bypass surgery.

Obesity is associated with low-grade inflammation and increased systemic oxidative stress. Roux-en-Y gastric bypass (RYGB) surgery is known to ameliorate the obesity-induced metabolic dysfunctions. We aimed to study the levels of natural antibodies in feces, before and 6 months after RYGB surgery in obese individuals with and without type 2 diabetes (T2D). Sixteen individuals with T2D and 14 non-diabetic (ND) individuals were operated. Total IgA, IgG and IgM antibody levels and specific antibodies to oxidized low-density lipoprotein (oxLDL), malondialdehyde-acetaldehyde adducts (MAA adducts), Porphyromonas gingivalis gingipain A hemagglutinin domain (Rgp44) and phosphocholine (PCho) were measured using chemiluminescence immunoassay. Total fecal IgA was elevated, while total IgM and IgG were not affected by the surgery. Fecal natural IgM specific to oxLDL decreased significantly in both T2D and ND individuals, while fecal IgM to Rgp44 and PCho decreased significantly in T2D individuals. A decrease in IgG to MAA-LDL, Rgp44 and PCho was detected. RYGB surgery increases the levels of total fecal IgA and decreases fecal natural IgG and IgM antibodies specific to oxLDL. Natural antibodies and IgA are important in maintaining the normal gut homeostasis and first-line defense against microbes, and their production is markedly altered with RYGB surgery.

Type 2 diabetes disrupts circadian orchestration of lipid metabolism and membrane fluidity in human pancreatic islets.

Recent evidence suggests that circadian clocks ensure temporal orchestration of lipid homeostasis and play a role in pathophysiology of metabolic diseases in humans, including type 2 diabetes (T2D). Nevertheless, circadian regulation of lipid metabolism in human pancreatic islets has not been explored. Employing lipidomic analyses, we conducted temporal profiling in human pancreatic islets derived from 10 nondiabetic (ND) and 6 T2D donors. Among 329 detected lipid species across 8 major lipid classes, 5% exhibited circadian rhythmicity in ND human islets synchronized in vitro. Two-time point-based lipidomic analyses in T2D human islets revealed global and temporal alterations in phospho- and sphingolipids. Key enzymes regulating turnover of sphingolipids were rhythmically expressed in ND islets and exhibited altered levels in ND islets bearing disrupted clocks and in T2D islets. Strikingly, cellular membrane fluidity, measured by a Nile Red derivative NR12S, was reduced in plasma membrane of T2D diabetic human islets, in ND donors' islets with disrupted circadian clockwork, or treated with sphingolipid pathway modulators. Moreover, inhibiting the glycosphingolipid biosynthesis led to strong reduction of insulin secretion triggered by glucose or KCl, whereas inhibiting earlier steps of de novo ceramide synthesis resulted in milder inhibitory effect on insulin secretion by ND islets. Our data suggest that circadian clocks operative in human pancreatic islets are required for temporal orchestration of lipid homeostasis, and that perturbation of temporal regulation of the islet lipid metabolism upon T2D leads to altered insulin secretion and membrane fluidity. These phenotypes were recapitulated in ND islets bearing disrupted clocks.

Detection of Antiviral Tissue Responses and Increased Cell Stress in the Pancreatic Islets of Newly Diagnosed Type 1 Diabetes Patients: Results From the DiViD Study.

Aims/hypothesisThe Diabetes Virus Detection (DiViD) study has suggested the presence of low-grade enteroviral infection in pancreatic tissue collected from six of six live adult patients newly diagnosed with type 1 diabetes. The present study aimed to compare the gene and protein expression of selected virally induced pathogen recognition receptors and interferon stimulated genes in islets from these newly diagnosed type 1 diabetes (DiViD) subjects vs age-matched non-diabetic (ND) controls.MethodsRNA was extracted from laser-captured islets and Affymetrix Human Gene 2.0 ST arrays used to obtain gene expression profiles. Lists of differentially expressed genes were subjected to a data-mining pipeline searching for enrichment of canonical pathways, KEGG pathways, Gene Ontologies, transcription factor binding sites and other upstream regulators. In addition, the presence and localisation of specific viral response proteins (PKR, MxA and MDA5) were examined by combined immunofluorescent labelling in sections of pancreatic tissue.ResultsThe data analysis and data mining process revealed a significant enrichment of gene ontologies covering viral reproduction and infectious cycles; peptide translation, elongation and initiation, as well as oxidoreductase activity. Enrichment was identified in the KEGG pathways for oxidative phosphorylation; ribosomal and metabolic activity; antigen processing and presentation and in canonical pathways for mitochondrial dysfunction, oxidative phosphorylation and EIF2 signaling. Protein Kinase R (PKR) expression did not differ between newly diagnosed type 1 diabetes and ND islets at the level of total RNA, but a small subset of β-cells displayed markedly increased PKR protein levels. These PKR+ β-cells correspond to those previously shown to contain the viral protein, VP1. RNA encoding MDA5 was increased significantly in newly diagnosed type 1 diabetes islets, and immunostaining of MDA5 protein was seen in α- and certain β-cells in both newly diagnosed type 1 diabetes and ND islets, but the expression was increased in β-cells in type 1 diabetes. In addition, an uncharacterised subset of synaptophysin positive, but islet hormone negative, cells expressed intense MDA5 staining and these were more prevalent in DiViD cases. MxA RNA was upregulated in newly diagnosed type 1 diabetes vs ND islets and MxA protein was detected exclusively in newly diagnosed type 1 diabetes β-cells.Conclusion/interpretationThe gene expression signatures reveal that pathways associated with cellular stress and increased immunological activity are enhanced in islets from newly diagnosed type 1 diabetes patients compared to controls. The increases in viral response proteins seen in β-cells in newly diagnosed type 1 diabetes provide clear evidence for the activation of IFN signalling pathways. As such, these data strengthen the hypothesis that an enteroviral infection of islet β-cells contributes to the pathogenesis of type 1 diabetes.

Research on Potential Network Markers and Signaling Pathways in Type 2 Diabetes Based on Conditional Cell-Specific Network.

Traditional methods concerning type 2 diabetes (T2D) are limited to grouped cells instead of each single cell, and thus the heterogeneity of single cells is erased. Therefore, it is still challenging to study T2D based on a single-cell and network perspective. In this study, we construct a conditional cell-specific network (CCSN) for each single cell for the GSE86469 dataset which is a single-cell transcriptional set from nondiabetic (ND) and T2D human islet samples, and obtain a conditional network degree matrix (CNDM). Since beta cells are the key cells leading to T2D, we search for hub genes in CCSN of beta cells and find that ATP6AP2 is essential for regulation and storage of insulin, and the renin-angiotensin system involving ATP6AP2 is related to most pathological processes leading to diabetic nephropathy. The communication between beta cells and other endocrine cells is performed and three gene pairs with obvious interaction are found. In addition, different expression genes (DEGs) are found based on CNDM and the gene expression matrix (GEM), respectively. Finally, ‘dark’ genes are identified, and enrichment analysis shows that NFATC2 is involved in the VEGF signaling pathway and indirectly affects the production of Prostacyclin (PGI2), which may be a potential biomarker for diabetic nephropathy.

Comparison of Blood Pressure Variability between 24 h Ambulatory Monitoring and Office Blood Pressure in Diabetics and Nondiabetic Patients: A Cross-Sectional Study.

Background Evidence regarding blood pressure (BP) variability (BPV) and its independent association with adverse outcomes has grown. Diabetic patients might have increased BPV, but there is still an evidence gap regarding relation between BPV and type 2 diabetes beyond mean values of BP. Objective To examine the relationship between 24 h ambulatory BP monitoring (ABPM, short-term variability) and visit-to-visit in-office BPV (OBP, long-term variability), in diabetics (D) and nondiabetics (ND), and to explore BPV relation with estimated glomerular filtration rate (eGFR), and pulse wave velocity (PWV) as indicators of target organ lesion. Materials and Methods We conducted a single-center cross-sectional study in an outpatient BP unit, including adult patients consecutively admitted from 1999 to 2019. Multivariate was performed to compare BPV between D and ND adjusted for clinical variables. Pearson's correlation was performed to evaluate relation of BPV with eGFR and PWV. Results A total of 1123 patients with ABPM and OBP measurements were included. Values of eGFR and PWV were worse in D than in ND. Measurements of OBPV did not differ between groups. Of ABPM BPV, the coefficient of variation and standard deviation for daytime systolic BP were higher in D compared to ND, but only in ND did BPV correlated with both eGFR and PWV. Conclusion We found that diabetes is associated with higher variability of daytime BP than nondiabetics along with worse damage of vascular and renal function. However, in contrast to nondiabetics, in diabetics eGFR and PWV may not be dependent on BP variability, suggesting that other mechanisms might explain more rigorously the greater damage of target organ lesion markers.

The role of pancreas to improve hyperglycemia in STZ-induced diabetic rats by thiamine disulfide

BackgroundThe present study investigated the effect of thiamine disulfide (TD) on the pancreas in terms of hyperglycemia improvement and insulin sensitivity increase in diabetic male rats. We also aimed to study the function of Pdx1 (pancreatic and duodenal homeobox 1) and Glut2 (glucose transporter 2) genes in pancreatic tissue.MethodsType 1 diabetes was induced through injection of 60 mg/kg streptozotocin (STZ). The diabetic rats were divided into four groups, namely diabetic control (DC), diabetic treated with thiamine disulfide (D-TD), diabetic treated with insulin (D-insulin), and diabetic treated with TD and insulin (D-insulin+TD). The non-diabetic (NDC) and diabetic groups received a normal diet (14 weeks). Blood glucose level and body weight were measured weekly; insulin tolerance test (ITT) and glucagon tolerance test (GTT) were performed in the last month of the study. The level of serum insulin and glucagon were measured monthly and a hyperglycemic clamp (Insulin Infusion rate (IIR)) was done for all the groups. Pancreas tissue was isolated so that Pdx1and Glut2 genes expression could be measured.ResultsWe observed that TD therapy decreased blood glucose level, ITT, and serum glucagon levels in comparison with those of the DC group; it also increased serum insulin levels, IIR, and expression of Pdx1 and Glut2 genes in comparison with those of the DC group.ConclusionAdministration of TD could improve hyperglycemia in type 1 diabetic animals through improved pancreas function. Therefore, not only does TD have a significant effect on controlling and reducing hyperglycemia in diabetes, but it also has the potential to decrease the dose of insulin administration.

247-LB: Pancreatic Elastase Regulates ß-Cell Viability by Impairing the Mechanosignaling Pathway

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

Transient silencing of TGF‐β1 reverses myelopoietic bias in older diabetic CD34+ cells

Adult circulating CD34+ stem progenitor cells (HSPCs) participate in vascular repair by stimulating angiogenic functions of the endothelium via paracrine mechanisms. Aging and diabetes are associated with reduced number and impaired vasculogenic potential of CD34+cells. Previous studies showed that diabetic dysfunction was largely due to the paracrine pro‐inflammatory and anti‐angiogenic switch in the phenotype of CD34+ cells. Furthermore, diabetic cells have a higher potential to generate pro‐inflammatory monocyte‐macrophages. We have recently shown that older‐diabetic (DB) cells have increased expression of transforming growth factor β1 (TGF‐β1) compared to the age‐matched nondiabetic (ND) cells. Transient silencing of this pleiotropic regulator restored reparative functions such as migration, proliferation, and nitric oxide generation.The current study tested the hypothesis that TGF‐β1‐silencing reverses myelopoietic bias and paracrine pro‐inflammatory phenotype in CD34+ cells derived from diabetic older adults. Expression of alarmins and receptor for advanced glycation end‐products (RAGE) were evaluated.CD34+ cells were isolated from either male or female non‐diabetic (ND) (n=31) or diabetic (DB), both type 1 and type 2, (n=39) subjects. Phosphorodiamidate morpholino oligomers (PMO) were used for TGF‐β1‐silencing. Circulating CD34+cells and monocyte‐macrophages were enumerated by flow cytometry in both groups. Gene‐expression assays were carried out by qPCR. Myelopoiesis was determined by CFU‐GM assay followed by characterization of monocytes and macrophages by flow cytometry.Circulating CD34+ cells (p<0.05, n=31) and classical monocytes (anti‐inflammatory, CD14++CD16‐) were lower (p<0.05, n=16) whereas non‐classical (pro‐inflammatory, CD14lowCD16++) monocytes (P<0.05, n=16) were higher in DB compared to the ND group. However, no differences were observed in the intermediate monocyte and macrophage populations in circulation between the groups. DB‐CD34+ cells generated a higher number of colonies in the CFU‐GM assay than ND cells. Nonclassical monocytes were higher in DB‐CFUs (p<0.05 vs ND‐CFUs, n=4) with no difference in classical or intermediate monocytes. A higher number of pro‐inflammatory (CCR2+CD68+) (p<0.05, n=4) and lower number of anti‐inflammatory (CX3CR1+CD206+) macrophages (p<0.05, n=4) were detected in DB‐CFUs compared to that observed in ND‐CFUs. This imbalance was not observed in DB‐cells that were modified with TGF β1‐PMO. Gene expression of S100A8/S100A9, S100A10, S100A14, S100P, HMGb1 and RAGE were higher (p<0.05, n=5) in DB CD34+ cells compared to the ND. These changes were reversed by TGFβ1‐PMO. RAGE‐antagonist recapitulated the effects of TGFβ1‐PMO in assays for myelopoiesis.This study suggests that silencing of TGF‐β1 is a promising approach for enhancing the revascularization potential of CD34+cells from diabetic older adults by reversing the paracrine pro‐inflammatory functions that would negatively impact vascular repair.

IDF21-0682 Wrist circumference as a marker for abdominal obesity correlates with waist circumference, independent of diabetes

Background: Contemporary literature highlights the importance of wrist circumference as an easy-to-use marker of general and abdominal obesity. Aim: To examine whether Wrist Circumference (WrC) correlates with Waist Circumference (WC) and BMI among diabetics and non-diabetics. Method: We measured WrC and WC in a heavy volume outpatient department of a specialised endocrine centre with and without diabetes over three months (July–September 2021). Patients with known medical conditions, that were confounding and influencing anthropometric parameters like body weight and BMI, including thyroid related disorders, genetic disorders were excluded. A dedicated nurse assistant was trained to eliminate observer bias. The measurements were by standard protocols using calibrated instruments. Unpaired t test, Fisher's exact test and Pearson r were used for statistical analysis. Results: We evaluated overall 1089 individuals (264 males, 825 females), which comprised diabetics (T2DM) n = 551, and Non-Diabetics (ND) n = 538. Mean age (years) in overall cohort (n = 1089) was 46 (±16, 95% CI 45–47), in T2DM (50.8 ± 12.7, 95% CI 49.8–51.9) which was more than ND (41 ± 16.6, 95% CI 39.6–42.4), p < 0.0001. Mean body weight (kg) in overall cohort was 72 (±15, 95% CI 71–73); in T2DM (72.7 ± 14.1, 95% CI 71.5–73.9) it was more than ND (70.5 ± 15.4, 95% CI 69.2–71.9), p = 0.016. Mean BMI (kg/m2) in overall cohort was 28 (±5.3, 95% CI 28–29); in T2DM (28.3 ± 4.9, 95% CI 27.8–28.7) which was comparable with ND (28.3 ± 5.5, 95% CI 27.8–28.7), p = 0.95 ns. Mean WC (cm) in overall cohort was 93 (±10, 95% CI 93–94); in T2DM (94.9 ± 10.3, 95% CI 94–95.7) it was more than ND (91.9 ± 9.6, 95% CI 91.1–92.8), p < 0.0001. Mean WrC (cm) in overall cohort was 16 (±1.3, 95% CI 16–16); in T2DM (16.2 ± 1.3, 95% CI 16.1–16.3) it was more than ND (15.8 ± 1.1, 95% CI 15.7–15.9), p < 0.0001. There was significant correlation between WrC and WC across overall cohort (r 0.49, 95% CI 0.45–0.54 p < 0.0001), in T2DM (r 0.46, 95% CI 0.39–0.53 p < 0.0001), in ND (r 0.51, 95% CI 0.44–0.57 p < 0.0001). Significant correlation between BMI and WrC across overall cohort (r 0.5, 95% CI 0.46–0.55 p < 0.0001) in T2DM (r 0.52, 95% CI 0.46–0.58 p < 0.0001), in ND (r 0.50, 95% CI 0.44–0.56 p < 0.0001). Significant correlation between BMI and WC across overall cohort (r 0.53, 95% CI 0.49–0.57 p < 0.0001) in T2DM (r 0.55, 95% CI 0.48–0.6 p < 0.0001), in ND (r 0.54, 95% CI 0.48–0.59 p < 0.0001). Discussion: The significant positive correlation of wrist circumference with waist circumference and BMI enables wrist circumference to be considered as a new measure of anthropometric assessment, irrespective of the presence or absence of diabetes. It is easy, quick, and convenient to measure than waist circumference, especially in daily clinical practice. Our findings need corroboration with larger epidemiological studies, involving varied multi-ethnic population.

SDF-1α loaded bioengineered human amniotic membrane-derived scaffold transplantation in combination with hyperbaric oxygen improved diabetic wound healing

Based on its multifactorial nature, successful treatment of diabetic wounds requires combinatorial approach. In this regard, we hypothesized that engraftment of a bioengineered micro-porous three-dimensional human amniotic membrane-scaffold (HAMS) loaded by SDF-1α (SHAMS) in combination with hyperbaric oxygen (HBO), throughout mobilization and recruitment of endothelial progenitor cells (EPCs), could accelerate wound healing in rats with type 1 diabetes mellitus. To test this hypothesis, 30 days after inducting diabetes, an ischemic wound was created in rat skin and treatments were performed for 21 days. In addition to wounded non-diabetic (ND) group, diabetic animals were randomly divided into non-treated (NT-D), HBO-treated (HBO-D), HBO-treated plus HAMS transplantation (HBO+HAMS-D) or HBO-treated in combination with SHAMS transplantation (HBO+SHAMS-D) groups. Our results on post-wounding days 7, 14 and 21 showed that the wound closure, volume of new dermis and epidermis, numerical density of basal cells of epidermis, fibroblasts and blood vessels, number of proliferating cells, deposition of collagen and biomechanical properties of healed wound were considerably higher in both HBO+HAMS-D and HBO+SHAMS-D groups in comparison to those of the NT-D and HBO-D groups, and were the highest in HBO+SHAMS-D ones. The transcripts for Vegf, bFgf, and Tgf-β genes were significantly upregulated in all treatment regimens compared to NT-D group and were the highest for HBO+SHAMS-D group. This is while expression of Tnf-α and Il-1β as well as cell density of neutrophil and macrophage decreased more significantly in HBO+SHAMS-D group as compared with NT-D or HBO-D groups. Overall, it was found that using both HAMS transplantation and HBO treatment has more impact on diabetic wound healing. Moreover, SDF-1α loading on HAMS could transiently improve the wound healing process, as compared with the HBO+HAMS-D group on day 7 only.

Transforming growth factor-β1/Thrombospondin-1/CD47 axis mediates dysfunction in CD34+ cells derived from diabetic older adults

Aging with diabetes is associated with impaired vasoprotective functions and decreased nitric oxide (NO) generation in CD34+ cells. Transforming growth factor- β1 (TGF-β1) is known to regulate hematopoietic functions. This study tested the hypothesis that transforming growth factor- β1 (TGF-β1) is upregulated in diabetic CD34+ cells and impairs NO generation via thrombospondin-1 (TSP-1)/CD47/NO pathway. CD34+ cells from nondiabetic (ND) (n=58) or diabetic older adults (DB) (both type 1 and type 2) (n=62) were isolated from peripheral blood. TGF-β1 was silenced by using an antisense delivered as phosphorodiamidate morpholino oligomer (PMO-TGF-β1). Migration and proliferation in response to stromal-derived factor-1α (SDF-1α) were evaluated. NO generation and eNOS phosphorylation were determined by flow cytometry. CD34+ cells from older, but not younger, diabetics have higher expression of TGF-β1 compared to that observed in cells derived from healthy individuals (P<0.05, n=14). TSP-1 expression was higher (n=11) in DB compared to ND cells. TGFβ1-PMO decreased the secretion of TGF-β1, which was accompanied with decreased TSP-1 expression. Impaired proliferation, migration and NO generation in response to SDF-1α in DB cells were reversed by TGF-β1-PMO (n=6). TSP-1 inhibited migration and proliferation of nondiabetic CD34+ cells that was reversed by CD47-siRNA, which also restored these responses in diabetic CD34+ cells. TSP-1 opposed SDF-1α-induced eNOS phosphorylation at Ser1177 that was reversed by CD47-siRNA. These results infer that increased TGF-β1 expression in CD34+ cells induces dysfunction in CD34+ cells from diabetic older adults via TSP-1/CD47-dependent inhibition of NO generation.

Abstract P1-04-12: Pathway analysis of immune checkpoint gene regulation as altered in Type 2 diabetes: Implications for breast cancer patients treated with checkpoint inhibitors

Abstract Background: Mediators of immune exhaustion are an active area of breast cancer research. Type II diabetes mellitus (T2D), the most common metabolic disorder, both increases breast cancer incidence and decreases survival. Despite this clear link, impacts of the T2D immune phenotype on cancer remain poorly understood. We have previously demonstrated that the chronic inflammatory state of T2D leads to immune exhaustion, but how the T2D breast microenvironment interacts with T cells is unclear. Exosomes are crucial components of intercellular communication and are associated with increased breast cancer aggressiveness. Given this function, we hypothesized that T2D adipocyte-derived exosomes drive T cell exhaustion. Here, we characterize expression patterns and regulatory networks driving T2D immune phenotypes. Methods: Exosomes were first isolated from culture media of mature human primary breast adipocytes from, either insulin-sensitive (IS), or rendered insulin-resistant (IR) by ex vivo TNFα-treatment. Human primary peripheral blood mononuclear cells (PBMCs) from nondiabetic (ND) and T2D donors were stimulated ex vivo with plate bound anti-CD3 (5 ug/ml) and soluble CD28 (2 ug/ml) for 48 hours and treated with exosomes. Different small molecule inhibitors of the bromo and extraterminal (BET) protein family, including the pan-BET inhibitor JQ1 and the BRD4-selective PROTAC degrader MZ-1, were used to identify BET protein-regulated targets. We also used the AMP-activating protein kinase (AMPK) inhibitor Compound-C to identify the role of this pathway in activating this major epigenetic player. Multicolor flow cytometry was subsequently performed with an LSRII cytometer to assess expression of inhibitory receptors PD-1, CTLA-4, TIM-3 and TIGIT on immune subsets. Events for live cells were analyzed in FlowJo. Cytokines were collected from conditioned media and analyzed via Th17 cytokine staining panel. Results: We observed that exosomes derived from IR breast adipocytes increase expression of immune exhaustion markers in CD4+ and CD8+ T cells, compared to IS or ND matched controls. Additionally, we define signal transduction among BET proteins, AMPK signaling, and immune checkpoint expression. Lastly, we identify changes in cytokine profile between IS and IR treated groups. Taken together, our findings suggest a network of immune regulation imparted by exosomes. Impacts: Metabolic health does not inform the current standard of care in breast medical oncology, which contributes to a large, underserved population in which treatment plans are not well established or optimized for their comorbidities. Our findings offer a deeper understanding of immune checkpoint regulation in T2D and suggest new insights into treatment of diabetic breast cancer patients. Citation Format: Christina S. Ennis, Pablo LLevenes, Manohar Kolla, Naser Jafari, Anna C Belkina, Gerald V. Denis. Pathway analysis of immune checkpoint gene regulation as altered in Type 2 diabetes: Implications for breast cancer patients treated with checkpoint inhibitors [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-04-12.

The Effects of Passive Simulated Jogging on Parameters of Explosive Handgrip in Nondiabetics and Type 2 Diabetics: A Single Arm Study

Aims Type 2 diabetes (T2D) is associated with sarcopenia and decreased muscle strength. Explosive and isometric voluntary handgrip strengths (EHGS and HGS) are frequently utilized methods to ascertain health status and a marker of overall muscle strength. We have previously shown that a portable, motorized device, which produces effortless, rapid stepping in place (passive simulated jogging device (JD)), improves glucose homeostasis. This study quantitatively evaluated the effects of JD in modifying parameters of the modified EHGS curve in T2D and nondiabetic (ND) subjects. Methods Twenty-one adult participants (11 ND and 10 T2D) (mean age: 41.3 ± 13.5 yr) performed a modified explosive handgrip strength (EHGS) test on study day 1 followed by daily use of JD (90 min per day) for 7 days. The EHGS was repeated after 3 and 7 days' use of JD (JD3 and JD7) and 3 days after completion of JD (Carryover). EHGS curves were analyzed for the following: maximal peak force value (MAX); rate of force development at 25%,75%, and 90% of maximum force; and maximum force (RFD25%, RFD75%, RFD90%, and RFDmax); time to 90%, 75%, and 25% of maximal force (t90, t75, t25) and time to maximal force (tmax); and the integrated area under the curve for force vs. time until task failure (iAUCTF); and fatigue resistance times at 50% and 25% of maximal force (FR50 and FR25) and fatigue resistance time to task failure (FRTF). Results At baseline, T2D had lower MAX compared to ND. There were no differences at baseline for force development time or fatigue resistance time between T2D and ND. In both T2D and ND, 7 days of JD increased FR25 and FRTF and iAUCTF compared to baseline. Conclusion JD for at least 7 days prior to EHGS increased time to task failure (fatigue resistance) and iAUCTF of the force-time curve. JD is a reasonable intervention to decrease sedentary behavior and improve muscle fatigue resistance under various clinical and nonclinical scenarios. This trial is registered with NCT03550105 (08-06-2018).

Compensatory intestinal antibody response against pro-inflammatory microbiota after bariatric surgery

ABSTRACT Obesity and type 2 diabetes (T2D) are growing burdens for individuals and the health-care system. Bariatric surgery is an efficient, but drastic treatment to reduce body weight, normalize glucose values, and reduce low-grade inflammation. The gut microbiome, which is in part controlled by intestinal antibodies, such as IgA, is involved in the development of both conditions. Knowledge of the effect of bariatric surgery on systemic and intestinal antibody response is limited. Here, we determined the fecal antibody and gut microbiome response in 40 T2D and non-diabetic (ND) obese individuals that underwent bariatric surgery (N = 40). Body weight, fasting glucose concentrations and inflammatory parameters decreased after bariatric surgery, whereas pro-inflammatory bacterial species such as lipopolysaccharide (LPS), and flagellin increased in the feces. Simultaneously, concentrations of LPS- and flagellin-specific intestinal IgA levels increased with the majority of pro-inflammatory bacteria coated with IgA after surgery. Finally, serum antibodies decreased in both groups, along with a lower inflammatory tone. We conclude that intestinal rearrangement by bariatric surgery leads to expansion of typical pro-inflammatory bacteria, which may be compensated by an improved antibody response. Although further evidence and mechanistic insights are needed, we postulate that this apparent compensatory antibody response might help to reduce systemic inflammation by neutralizing intestinal immunogenic components and thereby enhance intestinal barrier function after bariatric surgery.