Early Stages Of Diabetes Mellitus Research Articles

Early impaired insulin tolerance among Vietnamese diabetes with or without dyslipidemia.

This study aims to evaluate impaired insulin tolerance among Vietnamese diabetes with or without dyslipidemia. Diabetes mellitus (DM) remains the serious global health and social burden that has increased over the past few decades. It progresses silently to vascular injury and disability of injured vascular-perfused tissues/organs. Insulin intolerance and dyslipidemia exacerbate and accelerate the implications of DM. Thus, early detection and more evidence of early insulin intolerance and dyslipidemia is needed for proactive management. This cross-sectional descriptive study recruited 100 healthy control (HC) and 297 DM patients in Military Hospital 103 from 2021 to 2023. Patients with DM were subgrouped into lipid metabolism disorder (LMD, n = 98) and non-LMD (NLMD, n = 99). The biochemists' serum levels were measured automatically and the accuracy of the test result was strictly controlled. Insulin tolerance indices (HOMA2-IR, HOMA2-%S and HOMA2-%B) were compared between HC, DM with or without dyslipidemia as well as correlated with lipid ingredients (total Cholesterol, triglyceride, LDL-C and HDL-C). Among DM patients, HOMA2-IR was significantly high and HOMA2-%S and HOMA2-%B were significantly low. HOMA2-IR was higher and HOMA2-%S and HOMA2-%B were lower in DM with LMD than in DM without LMD. In addition, HOMA2-IR was positively correlated with serum cholesterol, triglyceride and LDL-C concentration, and negatively correlated to HDL-C concentration. In contrast, HOMA2-%S and HOMA2-%B was negatively correlated with serum cholesterol, triglyceride and LDL-C, and positively correlated with HDL-C. Impaired insulin intolerance occurred in early stage of DM, and more serious among DM with LMD, compared to DM with NLMD.

Orodispersible Dosage Forms with Rhinacanthin-Rich Extract as a Convenient Formulation Dedicated to Pediatric Patients.

Rhinacanthins, derived from Rhinacanthus nasutus, widely used in traditional medicine, exhibit antifungal, anticancer, antiviral, antibacterial, and antiplatelet aggregation effects. Recently, their anti-diabetic activity was confirmed, which makes them an interesting natural alternative in the therapy of the early stage of diabetes mellitus. The aim of this study was to demonstrate the possibility of formulating orodispersible tablets (ODTs) and orodispersible films (ODFs) containing rhinacanthin-rich extract (RRE). Tablets with 50 mg or 100 mg of RRE were produced by direct compression. ODFs were manufactured by casting of Lycoat RS 720 or polyvinyl alcohol solution with RRE and additional excipients. The mechanical properties and disintegration times of the prepared formulations were studied. The effectiveness of taste masking was analyzed with an electronic tongue system. Six months simplified stability studies were performed in conditions complying to ICH guidelines. Appropriate friability of ODTs was achieved, despite low tensile strength (0.45-0.62 MPa). All prepared ODFs successfully met the acceptance criteria regarding Young's modulus, tensile strength, and elongation at break. The observed variations in their mechanical properties were dependent on the type and quantity of polymers and plasticizers used. Disintegration time of ODTs ranged from 38.7 s to 54.2 s, while for ODFs from 24.2 to 40 s in the pharmacopoeial apparatus. Analyses made with the electronic tongue showed the significant taste-masking effect in both formulations. The addition of sucralose as a sweetener and menthol with mint flavor as a taste-masking agent was sufficient to mask an RRE's taste in the case of ODTs and ODFs. Stability studies of ODTs packed in the PVC/Alu blisters showed a decrease in the RRE content below 90% after 6 months. However, ODFs with PVA were physicochemically stable for 6 months while being stored in Alu/Alu sachets. Our study proved for the first time the possibility of the formulation of orodispersible dosage forms with RRE, characterized by good mechanical properties, disintegration time, and appropriate taste masking.

PET imaging for the early evaluation of ocular inflammation in diabetic rats by using [18F]-DPA-714

Ocular complications of diabetes mellitus (DM) are the leading cause of vision loss. Ocular inflammation often occurs in the early stage of DM; however, there are no proven quantitative methods to evaluate the inflammatory status of eyes in DM. The 18 kDa translocator protein (TSPO) is an evolutionarily conserved cholesterol binding protein localized in the outer mitochondrial membrane. It is a biomarker of activated microglia/macrophages; however, its role in ocular inflammation is unclear. In this study, fluorine-18-DPA-714 ([18F]-DPA-714) was evaluated as a specific TSPO probe by cell uptake, cell binding assays and micro positron emission tomography (microPET) imaging in both in vitro and in vivo models. Primary microglia/macrophages (PMs) extracted from the cornea, retina, choroid or sclera of neonatal rats with or without high glucose (50 mM) treatment were used as the in vitro model. Sprague-Dawley (SD) rats that received an intraperitoneal administration of streptozotocin (STZ, 60 mg/kg once) were used as the in vivo model. Increased cell uptake and high binding affinity of [18F]-DPA-714 were observed in primary PMs under hyperglycemic stress. These findings were consistent with cellular morphological changes, cell activation, and TSPO up-regulation. [18F]-DPA-714 PET imaging and biodistribution in the eyes of DM rats revealed that inflammation initiates in microglia/macrophages in the early stages (3 weeks and 6 weeks), corresponding with up-regulated TSPO levels. Thus, [18F]-DPA-714 microPET imaging may be an effective approach for the early evaluation of ocular inflammation in DM.

SUMOylation of GMFB regulates its stability and function in retinal pigment epithelial cells under hyperglycemia

BackgroundGlia maturation factor beta (GMFB) is a growth and differentiation factor that acts as an intracellular regulator of signal transduction pathways. The small ubiquitin-related modifier (SUMO) modification, SUMOylation, is a posttranslational modification (PTM) that plays a key role in protein subcellular localization, stability, transcription, and enzymatic activity. Recent studies have highlighted the importance of SUMOylation in the inflammation and progression of numerous diseases. However, the relationship between GMFB and SUMOylation is unclear. ResultsHere, we report for the first time that GMFB and SUMO1 are markedly increased in retinal pigment epithelial (RPE) cells at the early stage of diabetes mellitus (DM) under hyperglycemia. The GMFΒ protein could be mono-SUMOylated by SUMO1 at the K20, K35, K58 or K97 sites. SUMOylation of GMFB led to its increased protein stability and subcellular translocation. Furthermore, deSUMOylation of GMFΒ downregulates multiple signaling pathways, including the Jak-STAT signaling pathway, p38 pathway and NF-kappa B signaling pathway. ConclusionsThis work provides novel insight into the role of SUMOylated GMFB in RPE cells and provides a novel therapeutic target for diabetic retinopathy (DR).

Skin characterization of diabetes mellitus revealed by polarization-sensitive optical coherence tomography imaging.

Diabetes can lead to the glycation of proteins and dysfunction of skin collagen. Skin lesions are a prevalent clinical symptom of diabetes mellitus (DM). Early diagnosis and assessing the efficacy of treatment for DM are crucial for patient health management. However, performing a non-invasive skin assessment in the early stages of DM is challenging. By using the polarization-sensitive optical coherent tomography (PS-OCT) imaging technique, it is possible to noninvasively assess the skin changes caused by diabetes. The PS-OCT was used to monitor the polarization characteristics of mouse skin at different stages of diabetes. Based on a multi-layered adhesive tape model, we found that the polarization characteristics (retardation, optic axis, and polarization uniformity) were sensitive to the microstructure changes in the samples. Through this method, we observed significant changes in the polarization states of the skin as diabetes progressed. This was in line with the detected microstructure changes in skin collagen fibers using scanning electron microscopy. This study presents a highly useful approach for non-invasive skin assessment of diabetes.

Early left ventricular microvascular dysfunction in diabetic pigs: a longitudinal quantitative myocardial perfusion CMR study

BackgroundMicrovascular pathology is one of the main characteristics of diabetic cardiomyopathy; however, the early longitudinal course of diabetic microvascular dysfunction remains uncertain. This study aimed to investigate the early dynamic changes in left ventricular (LV) microvascular function in diabetic pig model using the cardiac magnetic resonance (CMR)-derived quantitative perfusion technique.MethodsTwelve pigs with streptozotocin-induced diabetes mellitus (DM) were included in this study, and longitudinal CMR scanning was performed before and 2, 6, 10, and 16 months after diabetic modeling. CMR-derived semiquantitative parameters (upslope, maximal signal intensity, perfusion index, and myocardial perfusion reserve index [MPRI]) and fully quantitative perfusion parameters (myocardial blood flow [MBF] and myocardial perfusion reserve [MPR]) were analyzed to evaluate longitudinal changes in LV myocardial microvascular function. Pearson correlation was used to analyze the relationship between LV structure and function and myocardial perfusion function.ResultsWith the progression of DM duration, the upslope at rest showed a gradually increasing trend (P = 0.029); however, the upslope at stress and MBF did not change significantly (P > 0.05). Regarding perfusion reserve function, both MPRI and MPR showed a decreasing trend with the progression of disease duration (MPRI, P = 0.001; MPR, P = 0.042), with high consistency (r = 0.551, P < 0.001). Furthermore, LV MPR is moderately associated with LV longitudinal strain (r = − 0.353, P = 0.022), LV remodeling index (r = − 0.312, P = 0.033), fasting blood glucose (r = − 0.313, P = 0.043), and HbA1c (r = − 0.309, P = 0.046). Microscopically, pathological results showed that collagen volume fraction increased gradually, whereas no significant decrease in microvascular density was observed with the progression of DM duration.ConclusionsMyocardial microvascular reserve function decreased gradually in the early stage of DM, which is related to both structural (but not reduced microvascular density) and functional abnormalities of microvessels, and is associated with increased blood glucose, reduced LV deformation, and myocardial remodeling.

O-Linked GlcNAcylation mediates the inhibition of proximal tubule (Na++K+)ATPase activity in the early stage of diabetes mellitus

BackgroundDiabetic kidney disease (DKD) is a severe complication of diabetes mellitus (DM). It has been proposed that modifications in the function of proximal tubule epithelial cells (PTECs) precede glomerular damage during the onset of DKD. This study aimed to identify modifications in renal sodium handling in the early stage of DM and its molecular mechanism. MethodsStreptozotocin (STZ)-induced diabetic BALB/c mice (STZ group) and LLC-PK1 cells, a model of PTECs, were used. All parameters were assessed in the 4th week after an initial injection of STZ. ResultsEarly stage of DKD was characterized by hyperfiltration and PTEC dysfunction. STZ group exhibited increased urinary sodium excretion due to impairment of tubular sodium reabsorption. This was correlated to a decrease in cortical (Na++K+)ATPase (NKA) α1 subunit expression and enzyme activity and an increase in O-GlcNAcylation. RNAseq analysis of patients with DKD revealed an increase in expression of the glutamine-fructose aminotransferase (GFAT) gene, a rate-limiting step of hexosamine biosynthetic pathway, and a decrease in NKA expression. Incubation of LLC-PK1 cells with 10 μM thiamet G, an inhibitor of O-GlcNAcase, reduced the expression and activity of NKA and increased O-GlcNAcylation. Furthermore, 6-diazo-5-oxo-L-norleucine (DON), a GFAT inhibitor, or dapagliflozin, an SGLT2 inhibitor, avoided the inhibitory effect of HG on expression and activity of NKA associated with the decrease in O-GlcNAcylation. ConclusionOur results show that the impairment of tubular sodium reabsorption, in the early stage of DM, is due to SGLT2-mediated HG influx in PTECs, increase in O-GlcNAcylation and reduction in NKA expression and activity.

Dynamic evolution of left ventricular strain and microvascular perfusion assessed by speckle tracking echocardiography and myocardial contrast echocardiography in diabetic rats: Effect of dapagliflozin.

This experimental study aimed to determine the dynamic changes in myocardial strain and microvascular perfusion in diabetic rats by comprehensive echocardiography while evaluating the effect of dapagliflozin (DAPA). Male Sprague-Dawley rats (n = 128) were randomly divided into four groups based on the presence or absence of a high-fat diet and streptozotocin-induced diabetes with or without DAPA treatment (n = 32/group). Serial conventional ultrasound, two-dimensional speckle tracking echocardiography (2D-STE) and myocardial contrast echocardiography (MCE) were performed at 2, 4, 6, and 8 weeks, and left ventricular global longitudinal strain (GLS), myocardial blood flow velocity (MBFV), myocardial blood flow (MBF), and myocardial blood volume (MBV) were determined. All animals were sacrificed immediately after the last echo measurement for histopathological assessment. Despite similar conventional Doppler-echo indexes among the groups at 2, 4, 6, and 8 weeks (p > 0.05), left ventricular GLS, MBFV, MBF, and MBV were decreased at 8 weeks in diabetic rats (p < 0.05) as detected by both 2D-STE and MCE. These indexes were significantly improved at 6 and 8 weeks after treatment with DAPA for diabetic rats (p < 0.05), reaching similar values observed in non-diabetic controls. DAPA treatment was associated with increased myocardial vacuolization and microvessel density and reduced interstitial fibrosis in diabetic rats. Combined 2D-STE and MCE is sensitive for detecting left ventricular deformity and impaired microvascular perfusion in prediabetes and the early stage of diabetes mellitus. DAPA exerts a beneficial effect on protecting myocardial perfusion in diabetic rats.

Correlation of dyslipidemia and athrogenic index of plasma with anthropometric measurements and clinical variables among diabetic patients in Dessie Comprehensive Specialized Hospital, Ethiopia, 2021

Background: Control of increased athrogenic index of plasma and lipid parameters in the early stage of diabetes mellitus helps diabetic patients to improve their quality of life and treatment outcomes. Limited studies were conducted on the assessment of dyslipidemia and its correlation with clinical and anthropometric variables among diabetes patients but no study was conducted on the correlation of the athrogenic index of plasma with anthropometric measurements among diabetes patients in this study area. The objective of this study was to assess the correlation of dyslipidemia and athrogenic index of plasma with anthropometric measurements and clinical variables among diabetic patients in Dessie Compressive Specialized Hospital, Northeast Ethiopia. Methods: Institution-based comparative cross-sectional study was conducted from August 2020 to June 2021. A total of 250 diabetic and healthy control respondents were included in the study with convenience sampling. Semi-structured questionnaire of a modified WHO stepwise Approach to Surveillance for chronic disease was used to collect data. Finally, descriptive statistics and correlation analysis were conducted to assess the correlation between variables. A p - value of less than 0.05 was declared as the level of significance. Results: Athrogenic Index of Plasma, Triglyceride to High-density Lipoprotein Ratio, Very-Low-Density Lipoprotein, systolic blood pressure, diastolic blood pressure, triglyceride, waist circumference, WHtR and BMI were statistically significantly higher among Type 2 DM groups. There was a significant positive linear correlation between triglycerides and waist circumference, between TG/HDL and WHtR, and between cholesterol and WHtR, but a significant negative linear correlation between HDL and waist circumference among the Type 2 DM group. Systolic blood pressure and pulse showed a significant positive linear correlation with WC, BMI, and WHtR among diabetics groups only. Our study showed that the pattern of lipid abnormalities observed among DM patients was high AIP in 68%, moderate AIP in 16% and all four groups of hyperlipidemia were found in 9% of diabetic patients. All lipid profiles showed a significant very strong positive linear correlation with AIP, but DHL has a significant very strong negative linear correlation with AIP among Type 2 DM groups. Conclusion: The proportions of high athrogenic index of plasma and lipid profile disorders were higher in DM patients compared to healthy controls. Dyslipidemia and a high athrogenic index of plasma had a considerable correlation with anthropometric measurements and clinical outcomes of Type 2 DM patients. DM patients who have a higher athrogenic index of plasma and higher lipid parameters should be strictly followed based on their anthropometric measurements.

Phosphoproteome reveals molecular mechanisms of aberrant rhythm in neurotransmitter-mediated islet hormone secretion in diabetic mice.

BackgroundAcetylcholine (ACh) and norepinephrine (NE) are representative neurotransmitters of parasympathetic and sympathetic nerves, respectively, that antagonize each other to coregulate internal body functions. This also includes the control of different kinds of hormone secretion from pancreatic islets. However, the molecular mechanisms have not been fully elucidated, and whether innervation in islets is abnormal in diabetes mellitus also remains unclear.Methods and resultsImmunofluorescence colocalization and islet perfusion were performed and the results demonstrated that ACh/NE and their receptors were highly expressed in islet and rapidly regulated different hormones secretion. Phosphorylation is considered an important posttranslational modification in islet innervation and it was identified by quantitative proteomic and phosphoproteomic analyses in this study. The phosphorylated islet proteins were found involved in many biological and pathological processes, such as synaptic signalling transduction, calcium channel opening and insulin signalling pathway. Then, the kinases were predicted by motif analysis and further screened and verified by kinase‐specific siRNAs in different islet cell lines (αTC1‐6, Min6 and TGP52). After functional verification, Ksr2 and Pkacb were considered the key kinases of ACh and NE in insulin secretion, and Cadps, Mlxipl and Pdcd4 were the substrates of these kinases measured by immunofluorescence co‐staining. Then, the decreased expression of receptors, kinases and substrates of ACh and NE were found in diabetic mice and the aberrant rhythm in insulin secretion could be improved by combined interventions on key receptors (M3 (pilocarpine) or α2a (guanfacine)) and kinases (Ksr2 or Pkacb).ConclusionsAbnormal innervation was closely associated with the degree of islet dysfunction in diabetic mice and the aberrant rhythm in insulin secretion could be ameliorated significantly after intervention with key receptors and kinases in the early stage of diabetes mellitus, which may provide a promising therapeutic strategy for diabetes mellitus in the future.

Molecular Mechanism of Pancreatic β-Cell Failure in Type 2 Diabetes Mellitus.

Various important transcription factors in the pancreas are involved in the process of pancreas development, the differentiation of endocrine progenitor cells into mature insulin-producing pancreatic β-cells and the preservation of mature β-cell function. However, when β-cells are continuously exposed to a high glucose concentration for a long period of time, the expression levels of several insulin gene transcription factors are substantially suppressed, which finally leads to pancreatic β-cell failure found in type 2 diabetes mellitus. Here we show the possible underlying pathway for β-cell failure. It is likely that reduced expression levels of MafA and PDX-1 and/or incretin receptor in β-cells are closely associated with β-cell failure in type 2 diabetes mellitus. Additionally, since incretin receptor expression is reduced in the advanced stage of diabetes mellitus, incretin-based medicines show more favorable effects against β-cell failure, especially in the early stage of diabetes mellitus compared to the advanced stage. On the other hand, many subjects have recently suffered from life-threatening coronavirus infection, and coronavirus infection has brought about a new and persistent pandemic. Additionally, the spread of coronavirus infection has led to various limitations on the activities of daily life and has restricted economic development worldwide. It has been reported recently that SARS-CoV-2 directly infects β-cells through neuropilin-1, leading to apoptotic β-cell death and a reduction in insulin secretion. In this review article, we feature a possible molecular mechanism for pancreatic β-cell failure, which is often observed in type 2 diabetes mellitus. Finally, we are hopeful that coronavirus infection will decline and normal daily life will soon resume all over the world.

The role of neutrophilic granulocytes in the development of acute lung injury in experimental diabetes mellitus

Diabetes mellitus takes one of the first places in the structure of endocrine diseases. Among the complications of diabetes are sufficiently described nephro- and retinopathy, neuropathy, damage to the cardiovascular system. However, changes in the respiratory system and, in particular, the state of the endothelium of the pulmonary hemocapillaries and the morphofunctional state of neutrophilic granulocytes remain poorly understood. The aim of this study was to determine the role of neutrophilic granulocytes in the pathogenesis of acute lung injury in experimental diabetes mellitus. The experiments were performed on 88 white male Wistar rats weighing 170-210 g. The animals were divided into three groups: 1 – intact (n=10); 2 – control (n=40); 3 – experimental (n=38) with a model of diabetes mellitus, which was reproduced by intraperitoneal administration of streptozotocin company “Sigma” (USA), diluted in 0.1 M citrate buffer with pH 4.5, at a rate of 60 mg/kg body weight. An equivalent dose of 0.1 M citrate buffer solution with a pH of 4.5 was intraperitoneally administered to the control group of animals. Pulmonary tissue collection for electron microscopic examination was performed under thiopental anesthesia 14, 28, 42, 70 days after streptozotocin administration. Pieces of lung tissue were fixed in 2.5 % glutaraldehyde solution, followed by fixation in 1 % osmium tetroxide solution. After dehydration, the material was poured into epon-araldite. Sections obtained on an ultramicrotome “Tesla BS-490” were studied in an electron microscope “PEM-125K”. It was found that in the early stages of diabetes mellitus (14-28 days) there is a violation of the rheological properties of blood, as evidenced by erythrocyte aggregates, excessive accumulation of neutrophils, their adhesion and aggregation in the hemocapillaries of the alveolar wall. With the extension of the experiment (42-70 days) there is a progressive violation of the ultrastructural organization of hemocapillaries of the alveolar wall and pronounced changes in the rheological properties of blood. Erythrocyte sludges and leuco-platelet aggregates are determined in the lumen of microvessels. Increased permeability of hemocapillaries of the alveolar wall leads to the emigration of neutrophilic granulocytes into the interstitium and the lumen of the alveoli with the development of interstitial and intraalveolar edema. Thus, streptozotocin-induced diabetes is accompanied by the development of acute lung damage in the pathogenesis of which the leading role belongs to neutrophilic granulocytes. The nature and severity of changes in the lungs depends on the duration of exposure to hyperglycemia.

Ethanolic Extract of Centella asiatica Treatment in the Early Stage of Hyperglycemia Condition Inhibits Glomerular Injury and Vascular Remodeling in Diabetic Rat Model.

Background Diabetes mellitus (DM) is marked by oxidative stress, inflammation, and vascular dysfunction that caused diabetic nephropathy that resulted in end-stage renal disease (ESRD). Vascular dysfunction is characterized by an imbalance in vasoconstrictor and vasodilator agents which underlies the mechanism of vascular injury in DM. Additionally, diminished podocytes correlate with the severity of kidney injury. Podocyturia often precedes proteinuria in several kidney diseases, including diabetic kidney disease. Centella asiatica (CeA) is known as an anti-inflammatory and antioxidant and has neuroprotective effects. This research aimed to investigate the potential effect of CeA to inhibit glomerular injury and vascular remodeling in DM. Methods The DM rat model was induced through intraperitoneal injection of streptozotocin 60 mg/kg body weight (BW), and then rats were divided into 1-month DM (DM1, n = 5), 2-month DM (DM2, n = 5), early DM concurrent with CeA treatment for 2 months (DMC2, n = 5), and 1-month DM treated with CeA for 1-month (DM1C1, n = 5). The CeA (400 mg/kg BW) was given daily via oral gavage. The control group (Control, n = 5) was maintained for 2 months. Finally, rats were euthanized and kidneys were harvested to assess vascular remodeling using Sirius Red staining and the mRNA expression of superoxide dismutase, podocytes marker, ACE2, eNOS, and ppET-1 using RT-PCR. Results The DM groups demonstrated significant elevation of glucose level, glomerulosclerosis, and proteinuria. A significant reduction of SOD1 and SOD3 promotes the downregulation of nephrin and upregulation of TRPC6 mRNA expressions in rat glomerular kidney. Besides, this condition enhanced ppET-1 and inhibited eNOS and ACE2 mRNA expressions that lead to the development of vascular remodeling marked by an increase of wall thickness, and lumen wall area ratio (LWAR). Treatment of CeA, especially the DMC2 group, attenuated glomerular injury and showed the reversal of induced conditions. Conclusions Centella asiatica treatment at the early stage of diabetes mellitus ameliorates glomerulosclerosis and vascular injury via increasing antioxidant enzymes.

Diabetic retinal neurodegeneration as a form of diabetic retinopathy.

To review the evidence supporting diabetic retinal neurodegeneration (DRN) as a form of diabetic retinopathy. Review of literature. DRN is recognized to be a part of retinopathy in patients with diabetes mellitus (DM), in addition to the well-established diabetic retinal vasculopathy (DRV). DRN has been noted in the early stages of DM, before the onset of clinically evident diabetic retinopathy. The occurrence of DRN has been confirmed in animal models of DM, histopathological examination of donor's eyes from diabetic individuals and assessment of neural structure and function in humans. DRN involves alterations in retinal ganglion cells, photoreceptors, amacrine cells and bipolar cells, and is thought to be driven by glutamate, oxidative stress and dysregulation of neuroprotective factors in the retina. Potential therapeutic options for DRN are under evaluation. Literature is divided on the temporal relation between DRN and DRV, with evidence of both precedence and simultaneous occurrence. The relationship between DRN and multi-system neuropathy in DM is yet to be evaluated critically.

Chronic Hyperglycemia Drives Functional Impairment of Lymphocytes in Diabetic INSC94Y Transgenic Pigs.

People with diabetes mellitus have an increased risk for infections, however, there is still a critical gap in precise knowledge about altered immune mechanisms in this disease. Since diabetic INSC94Y transgenic pigs exhibit elevated blood glucose and a stable diabetic phenotype soon after birth, they provide a favorable model to explore functional alterations of immune cells in an early stage of diabetes mellitus in vivo. Hence, we investigated peripheral blood mononuclear cells (PBMC) of these diabetic pigs compared to non-diabetic wild-type littermates. We found a 5-fold decreased proliferative response of T cells in INSC94Y tg pigs to polyclonal T cell mitogen phytohemagglutinin (PHA). Using label-free LC-MS/MS, a total of 3,487 proteins were quantified, and distinct changes in protein abundances in CD4+ T cells of early-stage diabetic pigs were detectable. Additionally, we found significant increases in mitochondrial oxygen consumption rate (OCR) and higher basal glycolytic activity in PBMC of diabetic INSC94Y tg pigs, indicating an altered metabolic immune cell phenotype. Thus, our study provides new insights into molecular mechanisms of dysregulated immune cells triggered by permanent hyperglycemia.

Evaluation of miR-181b and miR-126-5p expression levels in T2DM patients compared to healthy individuals: Relationship with NF-κB gene expression

BackgroundType 2 diabetes mellitus (T2DM) is a progressive metabolic disorder whose prevalence is rising very fast across the world. Diagnosis of this disease in early stages (pre-diabetic stage) plays an important role in reducing mortality associated with this disorder. miRNAs, as key players in the pathogenesis of T2DM, have been investigated in several studies. Furthermore, their expression profile changes in the early stages of diabetes mellitus in body fluids such as serum, peripheral blood, and peripheral blood mononuclear cell (PBMC) have been studied. Due to their high stability and the presence of non-invasive sensitive methods for their measurement, such as real-time PCR, they can be used for early diagnosis of T2DM as a biomarker. In this experimental study, the expression levels of miR-181b, miR-126-5p, and NF-κB were measured in patients with T2DM, pre-diabetic subjects, and healthy controls in a Yazd population. Material and methodNinety asymptomatic subjects including 30 T2DM, 30 pre-diabetic, and 30 healthy subjects (diagnosis based on WHO criteria) were included in this study. Real-time PCR was used to measure the expression levels of miR-181b and miR-126-5p. Moreover, the NF-κB expression level was also measured to determine its relationship with these two microRNAs. ResultIn this study, the expression level of miR-181b and miR-126-p decreased gradually in pre-diabetic as well as T2DM subjects compared to healthy controls. Furthermore, our study showed a significant negative correlation between these two miRNAs and NF-κB for the first time. ConclusionThese results introduce these anti-inflammatory miRNAs as powerful tools for early diagnosis of T2DM.

Thickness of Intraretinal Layers in Patients with Type 2 Diabetes Mellitus Depending on a Concomitant Diabetic Neuropathy: Results of a Cross-Sectional Study Using Deviation Maps for OCT Data Analysis

Optical coherence tomography (OCT) supports the detection of thickness changes in intraretinal layers at an early stage of diabetes mellitus. However, the analysis of OCT data in cross-sectional studies is complex and time-consuming. We introduce an enhanced deviation map-based analysis (MA) and demonstrate its effectiveness in detecting early changes in intraretinal layer thickness in adults with type 2 diabetes mellitus (T2DM) compared to common early treatment diabetic retinopathy study (ETDRS) grid-based analysis (GA). To this end, we obtained OCT scans of unilateral eyes from 33 T2DM patients without diabetic retinopathy and 40 healthy controls. The patients were categorized according to concomitant diabetic peripheral neuropathy (DN). The results of MA and GA demonstrated statistically significant differences in retinal thickness between patients and controls. Thinning was most pronounced in total retinal thickness and the thickness of the inner retinal layers in areas of the inner macular ring, selectively extending into areas of the outer macular ring and foveal center. Patients with clinically proven DN showed the strongest thinning of the inner retinal layers. MA showed additional areas of thinning whereas GA tended to underestimate thickness changes, especially in areas with localized thinning. We conclude that MA enables a precise analysis of retinal thickness data and contributes to the understanding of localized changes in intraretinal layers in adults with T2DM.

PERBANDINGAN RETINAL GANGLION CELL THICKNESS DAN PERIMETRI PASIEN DIABETES MELITUS TIPE 2

&lt;p&gt;&lt;em&gt;Diabetic retinopathy (RD) is a microvascular complication that often occurs in diabetes mellitus (DM). Retinal ganglion cells (RGC) are reduced in the early stages of DM, even before microvascular abnormalities are seen on the retina. &lt;strong&gt;Methods&lt;/strong&gt;: Sample of 35 eyes of non RD RD type 2 patients and 35 mild non proliferative diabetic retinopathy (NPDR) eyes After examination of ophthalmological status, perimetry and optical coherence tomography (OCT) were examined. &lt;strong&gt;Results:&lt;/strong&gt; The mean MD on non-RD type 2 DM -2.74 ± 3.5 mild NPDR -5.61 ± 4.5 with p value 0.414. The average non-RD type 2 PSD DM was -3.35 ± 3.3, mild NPDR was 4.16 ± 1.4 with a p value of 0.206. The mean RGC thickness of patients with non-RD type 2 DM was 83.8 ± 7.4 µm, mild NPDR 82.7 ± 8.1 μm. There was a decrease in the value of RGC thickness, MD and PSD in patients with type 2 DM, but it was not statistically significant . &lt;strong&gt;Conclusion&lt;/strong&gt;: No significant difference was found in RGC thickness in DM patients between non-RD and mild NPDR. No significant difference was found between MD and PSD in DM patients between non-RD and mild NPDR. No significant relationship was found between RGC thickness and perimetry in DM patients between non-RD and mild NPDR. &lt;/em&gt;&lt;/p&gt;

Proteome profile of neutrophils from a transgenic diabetic pig model shows distinct changes

INSC94Y transgenic pigs develop a stable diabetic phenotype early after birth and therefore allow studying the influence of hyperglycemia on primary immune cells in an early stage of diabetes mellitus in vivo. Since immune response is altered in diabetes mellitus, with deviant neutrophil function discussed as one of the possible causes in humans and mouse models, we investigated these immune cells in INSC94Y transgenic pigs and wild type controls at protein level. A total of 2371 proteins were quantified by label-free LC-MS/MS. Subsequent differential proteome analysis of transgenic animals and controls revealed clear differences in protein abundances, indicating a deviant behavior of granulocytes in the diabetic state. Interestingly, abundance of myosin regulatory light chain 9 (MLC-2C) was increased 5-fold in cells of diabetic pigs. MLC-2C directly affects cell contractility by regulating myosin ATPase activity, can act as transcription factor and was also associated with inflammation. It might contribute to impaired neutrophil cell adhesion, migration and phagocytosis.Our study provides novel insights into proteome changes in neutrophils from a large animal model for permanent neonatal diabetes mellitus and points to dysregulation of neutrophil function even in an early stage of this disease.Data are available via ProteomeXchange with identifier PXD017274. SignificanceOur studies provide novel basic information about the neutrophil proteome of pigs and contribute to a better understanding of molecular mechanisms involved in altered immune cell function in an early stage diabetes. We demonstrate proteins that are dysregulated in neutrophils from a transgenic diabetic pig and have not been described in this context so far. The data presented here are highly relevant for veterinary medicine and have translational quality for diabetes in humans.

The effect of high glucose on the biological characteristics of nucleus pulposus-derived mesenchymal stem cells.

Diabetes mellitus (DM) is a dependent risk factor in the progression of intervertebral disc degeneration (IVDD). High glucose supply has negative effects on nucleus pulpous (NP) cell and mesenchymal stem cell (MSC) biology. However, the effect of hyperglycaemia on the biological characterization of nucleus pulpous-derived mesenchymal stem cell (NPMSC) has not been investigated previously. Therefore, further exploration of the effects of DM-associated hyperglycaemia on NPMSC biology is important to better understand and develop endogenous repair strategies of DM patient-associated IVDD. Therefore, the cell biological characteristics were compared between NPMSC cultured in media with low glucose concentration (LG-NPMSC) and high glucose concentration (HG-NPMSC). The results demonstrated that HG-NPMSC showed significantly decreased cell proliferation, colony formation ability, migration and wound-healing capability compared with those of LG-NPMSC. HG-NPMSC also showed significantly decreased expressions of stemness genes and mRNA and protein expressions of silent information regulator protein 1 (SIRT1), SIRT6, hypoxia inducible factor-1α (HIF-1α) and glucose transporter 1 (GLUT-1), whereas increased cell apoptosis, cell senescence and caspase-3 expression. These results suggest that high glucose may decrease proliferation and stemness maintenance ability and increase apoptosis and senescence of NPMSC. SIGNIFICANCE OF THE STUDY: We found that high glucose concentration significantly decreased cell proliferation, colony formation ability, migration and wound-healing capability of nucleus pulposus-derived mesenchymal stem cells. Moreover, high glucose cultured nucleus pulposus-derived mesenchymal stem cells showed significantly decreased expression of stemness genes, related mRNA and protein, whereas increased cell apoptosis, cell senescence and expression of caspase-3. The present study indicated that better control of high concentration glucose in the early stage of diabetes mellitus should be recommended to prevent or limit intervertebral disc degeneration.