Nondiabetic Research Articles

Topical Reconstituted High-Density Lipoproteins Elicit Anti-Inflammatory Effects in Diabetic Wounds.

Objective: Reconstituted high-density lipoproteins (rHDL) improve wound healing in diabetes. We aimed to determine if rHDL elicit anti-inflammatory effects in diabetic wounds, as a mechanism to explain their wound healing benefits. Approach: Diabetes was induced using streptozotocin in C57Bl6/J mice. Two full-thickness wounds were placed on the subflanks of diabetic and nondiabetic (ND) mice. Phosphate-buffered saline (PBS) or rHDL (50 µg/wound/day) were applied topically. Wound closure was assessed daily. Inflammatory gene transcripts were measured by qPCR and proteins by Western blotting and enzyme-linked immunosorbent assay in wounds collected at baseline, 24 h, and 3 days postwounding. Wound macrophages were assessed by flow cytometry 7 days postwounding. The fate of fluorescent 3,3-dioctadecyloxacarbocyanine, perchlorate (DiO)-labeled rHDL was tracked by flow cytometry, fluorescent imaging, and microscopy. Results: In diabetic mice, rHDL increased wound closure rates at days 6 (+288%, p < 0.01) and 7 (+639%, p < 0.0001) postwounding, compared with PBS controls. After 3 days, rHDL-treated diabetic wounds had lower Rela (-65%) and C-C motif chemokine ligand 2 (Ccl2) (-59%) mRNA levels and CCL2 protein (29%) than PBS controls, p < 0.05 for all. Wound macrophage content was higher in diabetic than ND wounds, but rHDL did not change macrophage content or polarity. DiO-rHDL were taken up by key wound cells including fibroblasts, macrophages, keratinocytes and endothelial cells, and retained in wounds for at least 48 h. Innovation: rHDL exerts anti-inflammatory effects in diabetic wounds early postwounding, which may contribute to its wound healing properties. Conclusion: The anti-inflammatory properties of rHDL in diabetic wounds present topical rHDL as a novel treatment option for improving healing in patients with diabetic foot ulcers.

6524 Sex-Specific Single Cell Genetic Architecture Of Human Islets With And Without Type 2 Diabetes

Abstract Disclosure: M. Qadir: None. R.M. Elgamal: None. P. Kudtarkar: None. K.J. Gaulton: None. F. Mauvais-Jarvis: None. Type 2 diabetes (T2D) is a heterogeneous disease and biological sex affects its pathogenesis including the development of adiposity, insulin resistance, and β cell failure. For example, ketosis-prone diabetes is a phenotypically-defined form of T2D with acute β cell failure and severe insulin deficiency, predominantly observed in men. However, what mechanisms drive male predominance in β cell failure is unknown. To fully understand the sex-based pathogenesis of islet failure in diabetes, the study of sex differences in human islet biology and dysfunction is essential. To decipher sex differences in human islets genetic architecture and function, we performed an orthogonal series of experiments from 15 islets of non-diabetic (ND) donors, using single cell RNAseq (scRNAseq) and single nucleus assay for transposase-accessible chromatin sequencing (snATACseq). We also leveraged 37 ND and T2D donors for scRNAseq data from the human pancreas analysis program (HPAP) dataset. We studied over 200,000 single cells spanning the accessible genome and transcriptome. It is the largest study of sex-based single-cell genomics and transcriptomics of human islets to date. We observe that in cultured human ND islets, in the absence of the in vivo hormonal environment, islet cell transcriptome, and genome accessibility across coding regions are predominantly limited to sex chromosome genes. Of particular interest are sex differences in gene accessibility and expression of the X-linked KDM6A and Y-linked KDM5D demethylase genes in female (F) and male (M) islet cells respectively. Furthermore, analysis of T2D islets revealed a greater number of autosomal differentially expressed genes (DEGs) in F (721 upregulated and 1164 downregulated, FDR &amp;lt; 0.01) compared to M β cells (111 upregulated and 99 downregulated, FDR &amp;lt; 0.01). M β cell DEGs over-representation analysis revealed suppressed insulin secretion and processing pathways, while F β cell DEGs exhibited suppression of oxidative phosphorylation and electron transport chain pathways. Thus, although, sex differences in gene expression and accessibility of cultured ND human islets are restricted to sex chromosome genes, during the transition from ND to T2D, major sex differences in autosomal DEGs appear, with a greater number of autosomal DEGs in F compared to M T2D islets. Presentation: 6/3/2024

A-035 Profiling of circulating-microRNAs' signatures in Pre-diabetic versus Diabetic patients

Abstract Background MicroRNAs (miRs) are small non-coding RNAs involved in the process of gene regulation, and hence affect various diseases including metabolic pathways in diabetes. Given their key roles, they are promising candidates as potential biomarkers or better understanding of disease etiology. In this study, our aim is to identify miRs signatures of plasma circulating-miRs related to glycemic control, vitamin D deficiency and platelet activation in diabetic, pre-diabetic versus non-diabetic patients. Methods Plasma samples were collected from 24 patients (both sexes) in total (eight non-diabetic (ND) control, twelve diabetic (DM), and four pre-diabetic (PD)), classified based on their glycemic control (HbA1c levels). miRs were extracted using the miRNeasy Serum/Plasma Advanced kit of Qiagen, run and analyzed on Next Generation Sequencing (NGS). Results Differential expression analysis revealed miRs expression variations between diabetic to pre-diabetic and non-diabetic groups. Expression levels of 131 miRs varied significantly between DM to ND group (92 down- and 39 up- regulated), and 141 miRs in PD compared to ND (97 down- and 39 up- regulated). Although DM and PD had similar differential expression, three miRs (has-miR4776-5p, -6778-3p, and 5002-3p) were over-expressed in PD with very low p-value. Differential expression comparisons accounting sex were performed showing significant differences between males to females that will be presented here. Conclusions Our findings supports the important involvement of circulating miRs in the regulation of glucose homeostasis and pathogenesis of diabetes.Further investigation of candidate miRs could identify promising new biomarkers for the early diagnosis and prevention of Diabetes and related health complications.

Signalment and laboratory findings in cats with diabetes mellitus in Germany: a retrospective review of laboratory submissions of 129,505 cats.

The aims of this study were to compare signalment and laboratory parameters between diabetic (D) and non-diabetic (ND) cats and poorly-controlled diabetic (PD) and well-controlled diabetic (WD) cats in Germany. Laboratory data from Antech Lab Germany between 2015 and 2018 were retrospectively analysed. Age, sex, red blood cell count (RBC), creatinine (CREA), alkaline phosphatase (AP), alanine aminotransferase (ALT), bilirubin (BILI), cholesterol (CHOL), triglycerides (TRI), glucose (GLU) and total thyroxine (TT4) were compared between D (fructosamine ⩾340 µmol/l) and ND cats, and PD (fructosamine >500 µmol/l) and WD (fructosamine 340-500 µmol/l) cats. The proportion of cats with anaemia (RBC ⩽4.21 ×1012/l), CREA >250 µmol/l, ALT >455 U/l, AP >315 U/l, BILI ⩾35 µmol/l and TT4 > reference interval (RI) was compared between PD and WD cats. Data are presented as median and interquartile range (IQR) and analysed using non-parametric tests. Significance was P<0.05, and effect size was assessed by Cramér V or r. In total, 129,505 cats were included (D: n = 9334 [prevalence 7.2%], WD: n = 5670/9334 [60.7%]). The median age of D and ND cats was 12 years (IQR D 9-14; ND 9-15); there was no difference in sex. A significant difference was found between groups (D vs ND; PD vs WD) for all parameters studied. Considering the effect sizes and medians outside the RI, the only relevant difference was higher CHOL, TRI, AP and GLU in PD compared with WD (CHOL: PD 7.46 [5.85-9.32] vs WD 5.44 [4.32-6.97] mmol/l, P<0.001, r = 0.39; TRI: PD 1.44 [0.84-3.66] vs WD 0.78 [0.5-1.35] mmol/l, P <0.001, r = 0.35; AP: PD 66 [47-92] vs WD 35 [23-59] U/l, P <0.001, r = 0.39; GLU: PD 23.7 [20.15-27.3] vs WD 6.89 [5-11.31] mmol/l, P <0.001, r = 0.69). Laboratory changes in diabetic cats were mild and mainly associated with lipid derangements.

Single cell regulatory architecture of human pancreatic islets suggests sex differences in β cell function and the pathogenesis of type 2 diabetes.

Type 2 and type 1 diabetes (T2D, T1D) exhibit sex differences in insulin secretion, the mechanisms of which are unknown. We examined sex differences in human pancreatic islets from 52 donors with and without T2D combining single cell RNA-seq (scRNA-seq), single nucleus assay for transposase-accessible chromatin sequencing (snATAC-seq), hormone secretion, and bioenergetics. In nondiabetic (ND) donors, sex differences in islet cells gene accessibility and expression predominantly involved sex chromosomes. Islets from T2D donors exhibited similar sex differences in sex chromosomes differentially expressed genes (DEGs), but also exhibited sex differences in autosomal genes. Comparing β cells from T2D vs. ND donors, gene enrichment of female β cells showed suppression in mitochondrial respiration, while male β cells exhibited suppressed insulin secretion. Thus, although sex differences in gene accessibility and expression of ND β cells predominantly affect sex chromosomes, the transition to T2D reveals sex differences in autosomes highlighting mitochondrial failure in females.

Regulation of endocrine cell alternative splicing revealed by single-cell RNA sequencing in type 2 diabetes pathogenesis

The prevalent RNA alternative splicing (AS) contributes to molecular diversity, which has been demonstrated in cellular function regulation and disease pathogenesis. However, the contribution of AS in pancreatic islets during diabetes progression remains unclear. Here, we reanalyze the full-length single-cell RNA sequencing data from the deposited database to investigate AS regulation across human pancreatic endocrine cell types in non-diabetic (ND) and type 2 diabetic (T2D) individuals. Our analysis demonstrates the significant association between transcriptomic AS profiles and cell-type-specificity, which could be applied to distinguish the clustering of major endocrine cell types. Moreover, AS profiles are enabled to clearly define the mature subset of β-cells in healthy controls, which is completely lost in T2D. Further analysis reveals that RNA-binding proteins (RBPs), heterogeneous nuclear ribonucleoproteins (hnRNPs) and FXR1 family proteins are predicted to induce the functional impairment of β-cells through regulating AS profiles. Finally, trajectory analysis of endocrine cells suggests the β-cell identity shift through dedifferentiation and transdifferentiation of β-cells during the progression of T2D. Together, our study provides a mechanism for regulating β-cell functions and suggests the significant contribution of AS program during diabetes pathogenesis.

Islet-antigen reactive B cells display a unique phenotype and BCR repertoire in autoantibody positive and recent-onset type 1 diabetes patients.

Autoreactive B cells play an important but ill-defined role in autoimmune type 1 diabetes (T1D). To better understand their contribution, we performed single cell gene and BCR-seq analysis on pancreatic islet antigen-reactive (IAR) B cells from the peripheral blood of nondiabetic (ND), autoantibody positive prediabetic (AAB), and recent-onset T1D individuals. We found that the frequency of IAR B cells was increased in AAB and T1D. IAR B cells from these donors had altered expression of B cell signaling, pro-inflammatory, infection, and antigen processing and presentation genes. Both AAB and T1D donors demonstrated a significant increase in certain heavy and light chain V genes, and these V genes were enriched in islet-reactivity. Public clones of IAR B cells were restricted almost entirely to AAB and T1D donors. IAR B cells were clonally expanded in the autoimmune donors, particularly the AAB group. Notably, a substantial fraction of IAR B cells in AAB and T1D donors appeared to be polyreactive, which was corroborated by analysis of recombinant monoclonal antibodies. These results expand our understanding of autoreactive B cell activation during T1D and identify unique BCR repertoire changes that may serve as biomarkers for increased disease risk.

Single cell regulatory architecture of human pancreatic islets suggests sex differences in β cell function and the pathogenesis of type 2 diabetes.

Biological sex affects the pathogenesis of type 2 and type 1 diabetes (T2D, T1D) including the development of β cell failure observed more often in males. The mechanisms that drive sex differences in β cell failure is unknown. Studying sex differences in islet regulation and function represent a unique avenue to understand the sex-specific heterogeneity in β cell failure in diabetes. Here, we examined sex and race differences in human pancreatic islets from up to 52 donors with and without T2D (including 37 donors from the Human Pancreas Analysis Program [HPAP] dataset) using an orthogonal series of experiments including single cell RNA-seq (scRNA-seq), single nucleus assay for transposase-accessible chromatin sequencing (snATAC-seq), dynamic hormone secretion, and bioenergetics. In cultured islets from nondiabetic (ND) donors, in the absence of the in vivo hormonal environment, sex differences in islet cell type gene accessibility and expression predominantly involved sex chromosomes. Of particular interest were sex differences in the X-linked KDM6A and Y-linked KDM5D chromatin remodelers in female and male islet cells respectively. Islets from T2D donors exhibited similar sex differences in differentially expressed genes (DEGs) from sex chromosomes. However, in contrast to islets from ND donors, islets from T2D donors exhibited major sex differences in DEGs from autosomes. Comparing β cells from T2D and ND donors revealed that females had more DEGs from autosomes compared to male β cells. Gene set enrichment analysis of female β cell DEGs showed a suppression of oxidative phosphorylation and electron transport chain pathways, while male β cell had suppressed insulin secretion pathways. Thus, although sex-specific differences in gene accessibility and expression of cultured ND human islets predominantly affect sex chromosome genes, major differences in autosomal gene expression between sexes appear during the transition to T2D and which highlight mitochondrial failure in female β cells.

Defining the miRnome of Saphenous Vein Smooth Muscle Cells from Patients with Type 2 Diabetes Mellitus

Type 2 diabetes mellitus (T2DM) patients suffer premature development of cardiovascular disease and commonly require cardiac revascularization using the autologous saphenous vein (SV). Smooth muscle cells (SMCs) are the principal cell type within the vascular wall and are dysfunctional in T2DM SV-SMCs, yet the mechanisms underpinning this are incompletely understood. The purpose of this study was to interrogate differential microRNA (miRNA) expression in SV-SMCs to enhance our understanding of T2DM SV-SMC phenotypic change. miRNA expression in primary human SV-SMCs from T2DM and non-diabetic (ND) donors was determined using an array (n = 6 each of ND and T2DM SV-SMCs). Differentially expressed miRNAs were ranked, and functional annotation of the 30 most differentially expressed miRNAs using DAVID and KEGG analysis revealed pathways related to SMC phenotype, including proliferation, migration, cytokine production and cell signaling. After selecting miRNAs known to be involved in SMC phenotypic regulation, miR-17, miR-29b-2, miR-31, miR-130b and miR-491 were further validated using qRT-PCR (n = 5 each of ND and T2DM SV-SMC), with miR-29b-2 subsequently being removed from further investigation. Potential mRNA targets were identified using mirDIP. Predicted target analysis highlighted likely dysregulation in transcription, epigenetic regulation, cell survival, intracellular signaling and cytoskeletal regulation, all of which are known to be dysfunctional in T2DM SV-SMCs. In conclusion, this paper identified four miRNAs that are dysregulated in T2DM SV-SMCs and are implicated in functional changes in the behavior of these cells. This provides a step forward in our understanding of the molecular and epigenetic regulation of vascular dysfunction in T2DM.

Abstract 1028: Cytotoxic CD4+ T Cells: Unraveling The Residual Risk In Lipid Lowering For Type 2 Diabetes Patients

Type 2 Diabetes (T2D) significantly increases the risk of cardiovascular diseases (CVDs), marked by severe plaque pathology resistant to standard cholesterol-lowering treatments. While the roles of the innate immune system and macrophages in CVDs are well-established, T cell alterations, particularly involving cytotoxic CD4 T cells (CD4 CTLs), remain unexplored in human atherosclerotic plaques. In our study, we aimed to identify the phenotype and function of CD4 CTLs in blood &amp; plaque tissue, in the context of T2D. Using unbiased single-cell RNA sequencing (scRNA-seq) analysis, we examined over 70,000 immune cells from 22 plaques of patients undergoing carotid endarterectomy (T2D: n=12, Non-Diabetic (ND): n=10), &amp; peripheral blood mononuclear cells (PBMCs) from the CHORD study (NCT04369664) (T2D: n=12, ND: n=4). scRNA-seq analysis revealed a diverse population of CD4 T cells, like naïve, central memory (CM), effector memory (EM), tissue resident memory (Trm), T regulatory (Tregs), &amp; two clusters of cytotoxic (CTL) T cells expressing canonical cytotoxicity-related genes. Cluster 5 expressed GZMB, GNLY, PRF1 , whereas Cluster 10 expressed GZMA, GZMK . Cell-cell communication analysis demonstrated that both CTL clusters received signaling from dendritic cells and macrophages, including TREM2+ macrophages. CTL T cells reciprocally communicated with clusters of EM and EMRA CD8 T cells, underscoring their central role in plaque cell-cell communication. Notably, Cluster 5, one of these two CTL clusters, was expanded in T2D plaque samples (p=0.032) and expressed genes involved in chemotaxis, IL-2 signaling, &amp; transcription factors (e.g., RUNX3, TBX21 ) involved in T cell activation. The equivalent of CD4 CTL phenotype was found to be expanded in T2D individuals compared to controls, &amp; these elevated levels persisted in T2D patients following intensive lipid-lowering therapy, indicating resistance to the treatment regimen. In summary, our study emphasizes the crucial role of CD4 CTLs in the immune dynamics of atherosclerotic plaques. The observed resistance of CTLs to lipid-lowering interventions underscores the necessity for targeted therapeutic strategies to address the distinctive immune changes in T2D-associated cardiovascular diseases.

Abstract 3140: Upregulation Of Mirna-409-3p Contributes To Endothelial Cell Dysfunction In Patients With Diabetes

Endothelial cell (EC) dysfunction is prevalent in individuals with insulin resistance and diabetes, characterized by diminished phosphorylation of endothelial nitric oxide synthase (p-eNOS) and reduced nitric oxide (NO) production in the endothelium. Despite its common occurrence, the fundamental mechanism underlying EC dysfunction remains incompletely understood. Dysregulation of microRNAs (miRNAs) has been implicated in EC biology and metabolic disorders, yet studies exploring the functional relevance of these miRNAs are limited. In this investigation, venous EC samples were collected from patients with type 2 diabetes mellitus (T2DM) and non-diabetic (ND) individuals (n=20/group). Subsequent miRNA sequencing revealed 37 upregulated and 21 downregulated miRNAs (P&lt;0.05, L2FC&gt;1). Notably, increased expression of miR-409-3p (L2FC&gt;2) emerged as a potential target associated with EC dysfunction. Validation experiments conducted on human aortic EC (HAEC) exposed to high glucose palmitate (HGPAL) confirmed a significant upregulation of miR-409-3p (P&lt;0.001, L2FC&gt;2). In the same experimental setup, insulin-induced p-eNOS was decreased following HGPAL treatment. However, inhibition of miR-409-3p restored p-eNOS levels (18% to 20%), surpassing controls (21%), by enhancing insulin action in EC. Further investigations involving knockdown and overexpression of miR-409-3p demonstrated its negative regulation of p-eNOS in EC. The molecular mechanism uncovered indicates that miR-409-3p modulates p-eNOS signaling through JNK activity, as confirmed by fluorescence microscopy in patient ECs and by western blot in HAECs. In summary, this study identifies an altered miRNA repertoire in EC derived from T2DM patients, suggesting miR-409-3p as a key player in regulating insulin resistance and EC function. Ongoing study looks is delving into the transcriptional targets of miR-409-3p and their functional implications in both T2DM patients and cultured EC. These findings will shed light on the intricate interplay between miRNAs and EC dysfunction, offering potential avenues for therapeutic exploration in the realm of diabetes-associated vascular complications.

Neuroprotective Potential of Hygrophila auriculata Targeting Oxidative Stress-Mediated Deficits in Streptozotocin-Induced Sciatic Nerve Injury

Abstract Objective Diabetic neuropathy, a microvascular complication of diabetes, affects 50% of individuals. Addressing this challenge is challenging due to its poorly understood origin and existing therapeutic approaches. This study used a methanolic extract from Hygrophila auriculata (MEHA) to treat oxidative stress-induced sciatic nerve injury in diabetic rats. Materials and Methods A study was conducted to assess the nociceptive reflex after a single streptozotocin (STZ) (45 mg/kg intraperitoneal.) injection. The rats were divided into six groups (n = 6 rats per group). Group I nondiabetic (ND) rats received oral gavage of 1% carboxymethyl cellulose (CMC). The diabetic rats in groups II to VI were given 1% CMC, 100, 200, and 400 mg/kg of MEHA, and 180 mg/kg of metformin (MET). The freshly prepared 1% (w/v) CMC suspension of both MEHA and MET was administered over a 4-week period, commencing from the 28th day through the 56th day post-STZ injection. The impact of STZ-induced sciatic nerve injury was analyzed through the estimation of serum glucose and glycohemoglobin levels, paw withdrawal and tail-flick latencies, oxidative stress markers, and neural histoarchitecture. Results Diabetic (STZ) control group II showed significantly altered serum glucose and glycohemoglobin levels, a reduced paw withdrawal threshold, and reduced paw withdrawal and tail-flick latencies in contrast to ND group I. Furthermore, increased oxidative stress in the sciatic nerve correlates with a reduced nociceptive threshold and disrupted neural histoarchitecture in diabetic rats. These behavioral, biochemical, and molecular changes were markedly and dose-dependently reduced by MEHA and MET treatments. Conclusion The antioxidant efficacy of MEHA modulated oxidative stress in STZ-sensitized diabetic rats and corrected neuropathic pain by attenuating hyperglycemia.

Hepatic and renal effects of oral stingless bee honey in a streptozotocin-induced diabetic rat model.

Diabetes is known damage the liver and kidney, leading to hepatic dysfunction and kidney failure. Honey is believed to help in lowering the blood glucose levels of diabetic patients and reducing diabetic complications. However, the effect of stingless bee honey (SBH) administration in relieving liver and kidney damage in diabetes has not been well-studied. To investigate the effect of SBH administration on the kidney and liver of streptozotocin-induced (STZ; 55 mg/kg) diabetic Sprague Dawley rats. The rats were grouped as follows (n = 6 per group): non-diabetic (ND), untreated diabetic (UNT), metformin-treated (MET), and SBH+metformin-treated (SBME) groups. After successful diabetic induction, ND and UNT rats were given normal saline, whereas the treatment groups received SBH (2.0 g/kg and/or metformin (250 mg/kg) for 12 d. Serum biochemical parameters and histological changes using hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) staining were evaluated. On H&E and PAS staining, the ND group showed normal architecture and cellularity of Bowman's capsule and tubules, whereas the UNT and MET groups had an increased glomerular cellularity and thickened basement membrane. The SBH-treated group showed a decrease in hydropic changes and mild cellularity of the glomerulus vs the ND group based on H&E staining, but the two were similar on PAS staining. Likewise, the SBME-treated group had an increase in cellularity of the glomerulus on H&E staining, but it was comparable to the SBH and ND groups on PAS staining. UNT diabetic rats had tubular hydropic tubules, which were smaller than other groups. Reduced fatty vacuole formation and dilated blood sinusoids in liver tissue were seen in the SBH group. Conversely, the UNT group had high glucose levels, which subsequently increased MDA levels, ultimately leading to liver damage. SBH treatment reduced this damage, as evidenced by having the lowest fasting glucose, serum alanine transaminase, aspartate transaminase, and alkaline phosphatase levels compared to other groups, although the levels of liver enzymes were not statistically significant. The cellularity of the Bowman's capsule, as well as histological alteration of kidney tubules, glomerular membranes, and liver tissues in diabetic rats after oral SBH resembled those of ND rats. Therefore, SBH exhibited a protective hepatorenal effect in a diabetic rat model.

Mechanism exploration and biomarker identification of glycemic deterioration in patients with diseases of the exocrine pancreas

The damage to the endocrine pancreas among patients with diseases of the exocrine pancreas (DP) leads to reduced glycemic deterioration, ultimately resulting in diabetes of the exocrine pancreas (DEP). The present research aims to investigate the mechanism responsible for glycemic deterioration in DP patients, and to identify useful biomarkers, with the ultimate goal of enhancing clinical practice awareness. Gene expression profiles of patients with DP in this study were acquired from the Gene Expression Omnibus database. The original study defines DP patients to belong in one of three categories: non-diabetic (ND), impaired glucose tolerance (IGT) and DEP, which correspond to normoglycemia, early and late glycemic deterioration, respectively. After ensuring quality control, the discovery cohort included 8 ND, 20 IGT, and 12 DEP, while the validation cohort included 27 ND, 15 IGT, and 20 DEP. Gene set enrichment analysis (GSEA) employed differentially expressed genes (DEGs), while immunocyte infiltration was determined using single sample gene set enrichment analysis (ssGSEA). Additionally, correlation analysis was conducted to establish the link between clinical characteristics and immunocyte infiltration. The least absolute shrinkage and selection operator regression and random forest combined to identify biomarkers indicating glycemic deterioration in DP patients. These biomarkers were further validated through independent cohorts and animal experiments. With glycemic deterioration, biological processes in the pancreatic islets such as nutrient metabolism and complex immune responses are disrupted in DP patients. The expression of ACOT4, B2M, and ACKR2 was upregulated, whereas the expression of CACNA1F was downregulated. Immunocyte infiltration in the islet microenvironment showed a significant positive correlation with the age, body mass index (BMI), HbA1c and glycemia at the 2-h of patients. It was a crucial factor in glycemic deterioration. Additionally, B2M demonstrated a significant positive correlation with immunocyte infiltration and clinical features. Quantitative real-time PCR (qRT-PCR) and western blotting confirmed the upregulation in B2M. Immunofluorescent staining suggested the alteration of B2M was mainly in the alpha cells and beta cells. Overall, the study showed that gradually increased immunocyte infiltration was a significant contributor to glycemic deterioration in patients with DP, and it also highlighted B2M as a biomarker.

Identification of claudin-2 as a promising biomarker for early diagnosis of pre-diabetes.

Introduction: Pre-diabetes, a high-risk metabolic state, is situated between normal glucose homeostasis and diabetes. Early identification of pre-diabetes offers opportunities for intervention and diabetes reversal, highlighting the crucial need to investigate reliable biomarkers for this condition. Methods: We conducted an in-depth bioinformatics analysis of clinical samples from non-diabetic (ND), impaired glucose tolerance (IGT), and type 2 diabetes mellitus (T2DM) categories within the GSE164416 dataset. Thereafter the HFD and STZ treated mice were used for validation. Results: This analysis identified several codifferentially expressed genes (Co-DEGs) for IGT and T2DM, including CFB, TSHR, VNN2, APOC1, CLDN2, SLPI, LCN2, CXCL17, FAIM2, and REG3A. Validation of these genes and the determination of ROC curves were performed using the GSE76895 dataset. Thereafter, CLDN2 was selected for further verification. Gene expression analysis and immunofluorescence analysis revealed a significant upregulation of CLDN2 expression in the pancreas islets of mice in the high-fat diet and T2DM groups compared to the control group. Similarly, serum level of CLDN2 in patients with IGT and T2DM were significantly higher than those in the healthy group. Discussion: These results suggest that CLDN2 can serve as a novel biomarker for pre-diabetes, providing a new direction for future research in the prevention of type 2 diabetes.

Insulin Therapy on Bone Macroscopic, Microarchitecture, and MechanicalProperties of Tibia in Diabetic Rats.

This study evaluated tibia's macroscopic structure, mechanical properties, and bone microarchitecture in rats with type 1 diabetes mellitus (T1DM). Eighteen animals were divided into three groups (n=6): Non-diabetic (ND), diabetic (D), and diabetic+insulin (DI). T1DM was induced by streptozotocin; insulin was administered daily (4IU). The animals were euthanized 35 days after induction. The tibiae were removed and analyzed using macroscopic, micro-computed tomography (micro-CT) and three-point bending. The macroscopic analysis measured proximal-distal length (PD), antero-posterior thickness (AP) of proximal (AP-P) and distal (AP-D) epiphysis, and lateral-medial thickness (LM) of proximal (LM-P) and distal (LM-D) epiphysis. Micro-CT analysis closed porosity, tissue mineral density, and cortical thickness. The three-point bending test measured maximum strength, energy, and stiffness. The macroscopic analysis showed that D presented smaller measures of length and thickness (AP and AP-P) than ND and DI. More extensive measurements were observed of LM and AP-D thickness in DI than in D. In micro-CT, DI showed larger cortical thickness than D. Mechanical analysis showed lower strength in D than in other groups. T1DM reduces bone growth and mechanical strength. Insulin therapy in diabetic rats improved bone growth and fracture resistance, making diabetic bone similar to normoglycemic animals.

Influence of Diabetes Mellitus and Universal Adhesive Application Mode on the Bond Strength of Composite Resin to Dentine.

The aim of the current study was to evaluate the influence of diabetes mellitus and the mode of applying a universal adhesive on the shear bond strength of composite resin to dentin. Forty teeth were extracted from diabetic individuals who had been living with diabetes for a period of 5 to 15 years. These forty teeth were divided into two groups: twenty molars from patients with type 1 diabetes and twenty molars from patients with type 2 diabetes. The remaining twenty sound human molars were collected from non-diabetic patients. The collected teeth were assigned into 3 groups, and each group was further divided into 2 subgroups. Group A (n = 20): non-diabetic (ND) where sub group IA: Universal adhesive applied in the total-etch mode (n = 10) and sub group IIA: Universal adhesive applied in the self-etch mode (n = 10). Group B (n = 20): diabetic type 1 (D1) where sub group IB: Universal adhesive applied in the total-etch mode (n = 10) and sub group IIB: Universal adhesive applied in the self-etch mode (n = 10). Group C (n = 20): diabetic type 2 (D2). Where Sub group IC: Universal adhesive applied in the total-etch mode (n = 10) and sub group IIC: Universal adhesive applied in the self-etch mode (n = 10). A Teflon mold measuring 3mm in diameter was attached to the dentin surface, used to build Filtek Z550 to a height of 3mm. The specimens were fixed to the universal testing to measure shear bond strength. There was a statistically significant difference of Mean ±SD of shear bond strength among the three tested groups. In group A, the Mean ±SD were (21.710 ± 0.638), it was decreased in group B to (14.626 ± 0.726) and group C to (17.740 ± 0.668). Subgroup I had lower shear bond strength values than subgroup II in all tested groups. The difference between each subgroup in groups A, B, and C was significant. 1. Diabetes mellitus has an adverse effect on the shear bond strength of composite to dentine. 2. Type 1 diabetes mellitus significantly reduces the shear bond strength of composite resin to dentin. 3. Shear bond strength of the universal adhesive was higher when applied to dentin using the self-etch mode, as compared to the total etch mode, in all groups, regardless of whether the participants had diabetes or not. Key words:Diabetes Mellitus, Universal Adhesive, Application Mode, Shear bond Strength, Dentine, Composite.

Evaluation of the Clinical Effect of Hyaluronic Acid Mouthwash on Palatal Secondary Wound Healing in Diabetic Rats

Objective: The objective of this study was to investigate clinical effect of topically administered hyaluronic acid (HA) mouthwash on healing of secondary palatal wound in diabetic (D) rats. &#x0D; Methods: 60 Wistar albino male rats were divided into D and non-diabetic (ND) groups. Diabetes was induced to 30 randomly selected rats by initially administering 110 mg/kg of nicotinamide intraperitoneally, followed by 15 min of intraperitoneal injection of 65 mg/kg of streptozotocin solution. 5 mm excisional wounds were made in the centre of the palate. After that, 6 animals from each group were sacrificed. Then, both groups were subdivided into two groups: 0.12% HA mouthwash and saline (S) (n=12 per group), depending on the agent to be administered to the wound area (WA). On days 7 and 14, six rats from each group were sacrificed, and the WAs were measured through photographic measurements utilizing Image J software.&#x0D; Results: The WA decreased with time in each group (p

Impact of RYGB surgery on plasma immunoglobulins: association between blood pressure and glucose levels six months after surgery.

We aimed to study levels of natural antibodies in plasma, and their associations to clinical and fecal biomarkers, before and 6 months after Roux-en-Y gastric bypass (RYGB) surgery. Thirty individuals with obesity [16 type 2 diabetic, 14 non-diabetic (ND)] had RYGB surgery. Total plasma IgA, IgG and IgM antibody levels and specific antibodies to oxidized low-density lipoprotein (oxLDL), malondialdehyde-acetaldehyde adducts, Porphyromonas gingivalis gingipain A hemagglutinin domain (Rgp44), and phosphocholine were measured using chemiluminescence immunoassay. Associations between plasma and fecal antibodies as well as clinical markers were analyzed. RYGB surgery reduced blood pressure, and the glycemic state was improved. A higher level of diastolic blood pressure was associated with lower plasma antibodies to oxLDL after surgery. Also, lower level of glucose markers associated with lower level of plasma antibodies to bacterial virulence factors. Antibodies to oxLDL decreased after surgery, and positive association between active serum lipopolysaccharide and specific oxLDL antibodies was detected. Total IgG levels decreased after surgery, but only in ND individuals. Reduced level of total plasma IgG, improved state of hypertension and hyperglycemia and their associations with decreased levels of specific antibodies in plasma, suggest an improved state of systemic inflammation after RYGB surgery.

Comparison of central corneal thickness and endothelial cell count among diabetic retinopathy and non-diabetic cases

: Diabetes is associated with alterations in the structure and functions of corneal endothelial cells. Current research has been carried out to compare the central corneal thickness and endothelial cell density in patients with diabetic retinopathy (DR) and non-diabetic (ND) individuals. : A Cross Sectional comparative research was carried out in a tertiary eye care hospital, over the period of two years from April 2021 to March 2023. Among 160 study participants, 80 cases with diabetic retinopathy (DR) and 80 non-diabetic (ND) cases as age matched control have been selected. Thorough Ophthalmic assessment was done. We have included only retinopathy cases as they have longer duration of diabetis and poorer metabolic control, for better comparison of the endothelium parameters with non-diabetics. Specular microscopy has been done in all cases for endothelial cell count evaluation and thickness of cornea has been estimated by Pachymeter. Statistical analysis was carried out by students‘t’ test by comparing the variables between two groups. : P-value was not significant for the mean age and sex distribution in the two groups. Mean endothelial cell density was lesser (2512.12±260.23cells/mm2) in DR than in ND group (2699.10±95.68cells/mm2). Mean central corneal thickness was greater (522.65±36.56μm) in DR than in ND group (486.50±18.67μm) (P&amp;#60;0.05). Also, the Co-efficient of variation percentage was more whereas the percentage of hexagonality was found to be statistically less in DR than ND group. Among DR patients, endothelial cell density was significantly reduced and central corneal thickness considerably raised when compared with ND. Our results also suggest that poor metabolic control and advanced Diabetic Retinopathy are risk factors for developing keratopathy.