Non-diabetic Controls Research Articles

Dietary resistant starch enhances immune health of the kidney in diabetes via promoting microbially-derived metabolites and dampening neutrophil recruitment

BackgroundDietary-resistant starch is emerging as a potential therapeutic tool to limit the negative effects of diabetes on the kidneys. However, its metabolic and immunomodulatory effects have not yet been fully elucidated.MethodsSix-week-old db/db mice were fed a diet containing 12.5% resistant starch or a control diet matched for equivalent regular starch for 10 weeks. db/m mice receiving the control diet were utilised as non-diabetic controls. Freshly collected kidneys were digested for flow cytometry analysis of immune cell populations. Kidney injury was determined by measuring albuminuria, histology, and immunohistochemistry. Portal vein plasma was collected for targeted analysis of microbially-derived metabolites. Intestinal histology and tight junction protein expression were assessed.ResultsResistant starch limited the development of albuminuria in db/db mice. Diabetic db/db mice displayed a decline in portal vein plasma levels of acetate, propionate, and butyrate, which was increased with resistant starch supplementation. Diabetic db/db mice receiving resistant starch had a microbially-derived metabolite profile similar to that of non-diabetic db/m mice. The intestinal permeability markers lipopolysaccharide and lipopolysaccharide binding protein were increased in db/db mice consuming the control diet, which was not seen in db/db mice receiving resistant starch supplementation. Diabetes was associated with an increase in the kidney neutrophil population, neutrophil activation, number of C5aR1+ neutrophils, and urinary complement C5a excretion, all of which were reduced with resistant starch. These pro-inflammatory changes appear independent of fibrotic changes in the kidney.ConclusionsResistant starch supplementation in diabetes promotes beneficial circulating microbially-derived metabolites and improves intestinal permeability, accompanied by a modulation in the inflammatory profile of the kidney including neutrophil infiltration, complement activation, and albuminuria. These findings indicate that resistant starch can regulate immune and inflammatory responses in the kidney and support the therapeutic potential of resistant starch supplementation in diabetes on kidney health.

An exploratory study to understand the relationship between diabetes and various pulpal conditions: An Indian perspective.

Diabetes mellitus (DM) affects an estimated 100 million Indians, ranking second globally in diabetic prevalence. Despite this, the correlation between DM and specific pulpal diagnoses remains underexplored. This study compares pulpal conditions in DM patients and nondiabetic controls. The study was started after taking ethical approval. Two thousand and five hundred and sixty teeth were examined over 4 months at the institute, evenly distributed between diabetic and nondiabetic cases, we assessed diagnoses - normal pulp, reversible pulpitis (RP), symptomatic irreversible pulpitis, asymptomatic irreversible pulpitis, pulp necrosis (PN), missing teeth (MT), and root canal-treated teeth (RCT). Statistical analysis used the Chi-square test. PN and MT prevalence in DM patients significantly exceeded the control group. Conversely, the control group showed higher RP prevalence in the older subgroup. PN prevalence was higher in diabetics, suggesting reduced pulp sensitivity in individuals over 60 years. This diminished sensitivity might lead to delayed dental treatment, increasing PN prevalence.

Diabetic Wound Keratinocytes Induce Macrophage JMJD3-Mediated Nlrp3 Expression via IL-1R Signaling.

Macrophage (Mφ) plasticity is critical for normal wound repair; however, in type 2 diabetic wounds, Mφs persist in a low-grade inflammatory state that prevents the resolution of wound inflammation. Increased NLRP3 inflammasome activity has been shown in diabetic wound Mφs; however, the molecular mechanisms regulating NLRP3 expression and activity are unclear. Here, we identified that diabetic wound keratinocytes induce Nlrp3 gene expression in wound Mφs through IL-1 receptor-mediated signaling, resulting in enhanced inflammasome activation in the presence of PAMPs and DAMPs. We found that IL-1 alpha is increased in human and murine wound diabetic keratinocytes compared to non-diabetic controls and directly induces Mφ Nlrp3 expression through IL-1 receptor signaling. Mechanistically, we report that the histone demethylase, JMJD3, is increased in wound Mφs late post-injury and is induced by IL-1 alpha from diabetic wound keratinocytes, resulting in Nlrp3 transcriptional activation through an H3K27me3-mediated mechanism. Using genetically engineered mice deficient in JMJD3 in myeloid cells (Jmjd3fl/fllyz2cre+), we demonstrate that JMJD3 controls Mφ-mediated Nlrp3 expression during diabetic wound healing. Thus, our data suggest a role for keratinocyte-mediated IL-1 alpha/IL-1R signaling in driving enhanced NLRP3 inflammasome activity in wound Mφs. These data also highlight the importance of cell crosstalk in wound tissues and identify JMJD3 and the ILR signaling cascade as important upstream therapeutic targets for Mφ NLRP3 inflammasome hyperactivity in nonhealing diabetic wounds.

Dysregulation of long non-coding RNA gene expression pathways in monocytes of type 2 diabetes patients with cardiovascular disease

BackgroundMonocytes play a central role in the pathophysiology of cardiovascular complications in type 2 diabetes (T2D) patients through different mechanisms. We investigated diabetes-induced changes in lncRNA genes from T2D patients with cardiovascular disease (CVD), long-duration diabetes, and poor glycemic control.MethodsWe performed paired-end RNA sequencing of monocytes from 37 non-diabetes controls and 120 patients with T2D, of whom 86 had either macro or microvascular disease or both. Monocytes were sorted from peripheral blood using flow cytometry; their RNA was purified and sequenced. Alignments and gene counts were obtained with STAR to reference GRCh38 using Gencode (v41) annotations followed by batch correction with CombatSeq. Differential expression analysis was performed with EdgeR and pathway analysis with IPA software focusing on differentially expressed genes (DEGs) with a p-value < 0.05. Additionally, differential co-expression analysis was done with csdR to identify lncRNAs highly associated with diabetes-related expression networks with network centrality scores computed with Igraph and network visualization with Cytoscape.ResultsComparing T2D vs. non-T2D, we found two significantly upregulated lncRNAs (ENSG00000287255, FDR = 0.017 and ENSG00000289424, FDR = 0.048) and one significantly downregulated lncRNA (ENSG00000276603, FDR = 0.017). Pathway analysis on DEGs revealed networks affecting cellular movement, growth, and development. Co-expression analysis revealed ENSG00000225822 (UBXN7-AS1) as the highest-scoring diabetes network-associated lncRNA. Analysis within T2D patients and CVD revealed one lncRNA upregulated in monocytes from patients with microvascular disease without clinically documented macrovascular disease. (ENSG00000261654, FDR = 0.046). Pathway analysis revealed DEGs involved in networks affecting metabolic and cardiovascular pathologies. Co-expression analysis identified lncRNAs strongly associated with diabetes networks, including ENSG0000028654, ENSG00000261326 (LINC01355), ENSG00000260135 (MMP2-AS1), ENSG00000262097, and ENSG00000241560 (ZBTB20-AS1) when we combined the results from all patients with CVD. Similarly, we identified from co-expression analysis of diabetes patients with a duration ≥ 10 years vs. <10 years two lncRNAs: ENSG00000269019 (HOMER3-AS10) and ENSG00000212719 (LINC02693). The comparison of patients with good vs. poor glycemic control also identified two lncRNAs: ENSG00000245164 (LINC00861) and ENSG00000286313.ConclusionWe identified dysregulated diabetes-related genes and pathways in monocytes of diabetes patients with cardiovascular complications, including lncRNA genes of unknown function strongly associated with networks of known diabetes genes.

Clinical profile and risk factors for type-2 diabetes – A cross-sectional study

Objectives Type 2 diabetes is a chronic metabolic disorder characterised by elevated blood glucose levels due to insulin resistance or insufficient insulin production. Understanding the prevalence, characteristics, and markers of this disease is essential for effective prevention, management and treatment. The study aims to explore the association between type 2 diabetes and its prevalence, characteristics and markers. Material and Methods Adults with type 2 diabetes and matched healthy controls were enrolled in the study. Statistical calculations were used to establish the sample size. Information on demographics, physical examinations and lab tests were collected. We assessed plasma glucose, glycated haemoglobin (HbA1c), serum lipids and serum DPP4. Results The mean age (SD) of the population under research was 55.7 (6.12) years for the study subjects and 55.7 (6.11) years for the controls. A positive family history of diabetes was present in 34 (or 33%) of the diabetes patients compared to 11 (or 11%) of the non-diabetic patients (p 0.001). The mean HbA1c in the diabetic group was substantially greater than that of the non-diabetic controls (5.47 1.89%), as expected (7.23 2.69%), p 0.001. It’s interesting to note that total cholesterol was markedly higher in the diabetes participants (5.59 2.24 mmol/L) than in the non-diabetic controls (6.48 1.54 mmol/L), p = 0.001. Conclusion Due to common risk factors and underlying mechanisms, type 2 diabetes and hypertension may be related, as suggested by the high prevalence of hypertension in the diabetic group. Type 2 diabetes was substantially related to elevated HbA1c and fasting plasma glucose levels, indicating poor glycaemic control. The higher mean serum Dipeptidyl peptidase-4 (DPP4) level in the group with type 2 diabetes shows a link between high DPP4 levels and the disease, which may have an effect on incretin hormone activity, insulin resistance and therapeutic options.

Exploring auditory temporal resolution and dichotic listening skills among individuals with type 2 diabetes mellitus

The study aimed to explore the auditory temporal resolution and dichotic listening skills in patients with type 2 diabetes mellitus (T2DM) and identify associated health-related factors. Using a cross-sectional design, 87 adults with T2DM and 48 non-diabetic controls, all with normal hearing, participated. The two central auditory processing (CAP) skills were assessed through the Gaps-In-Noise (GIN) and Dichotic-Digits Listening (DDL) tests. T2DM participants underwent blood tests to measure various health-related factors. In the GIN test, the shortest gap threshold (GapTh) obtained across both ears was significantly higher in the diabetic group (9.1 ± 2.4 ms) compared to the non-diabetic group (7.5 ± 1.5 ms), and the score of correctly identified gaps (GapSc) in the diabetic group (45±11 %) was significantly lower than GapSc in the non-diabetic group (52±9 %), p < 0.001. In the DDL test, the free-recall score (73.8 ± 18.5 %) across both ears and the right-ear advantage (-1.3 ± 20.6 %) in the diabetic group were significantly lower than the free-recall score (85.8 ± 11.9 %) and right-ear advantage (6.9 ± 11.9 %) in the non-diabetic group, p < 0.005. Furthermore, the duration of diabetes, eGFR level, retinopathy, carotid plaque, fasting blood glucose level, and HDL-C (good cholesterol) level were factors significantly associated with performances in the GIN and/or DDL tests for T2DM participants. In conclusion, individuals with T2DM are at risk of reduced auditory processing skills in temporal resolution and dichotic listening, impacting their speech understanding. Six health-related factors were identified as significantly associated with CAP skills in T2DM patients.

Association of Circulating Markers of Microbial Translocation and Hepatic Inflammation with Liver Injury in Patients with Type 2 Diabetes.

Virtually the entire spectrum of liver disease is observed in association with type 2 diabetes mellitus (T2DM); indeed, T2DM is now the most common cause of liver disease in the U.S. We conducted a pilot study to investigate the relevance of increased microbial translocation and systemic inflammation in the development of liver injury in patients with T2DM. Patients with T2DM (n = 17) and non-diabetic controls (NDC; n = 11) aged 25-80 yrs. participated in this study. Serum levels of endotoxin, calprotectin, soluble CD14 and CD163, and several inflammatory cytokines were measured. In addition to standard liver injury markers, ALT and AST, novel serum markers of liver injury, keratin 18 (K-18) M30 (apoptosis-associated caspase-cleaved keratin 18), and M65 (soluble keratin 18) were evaluated. Statistical analyses were performed using the Mann-Whitney test to assess differences between study groups. Pearson's correlation analysis was performed to determine the strength of association between two variables using GraphPad Prism 9.5.0 software. Patients with T2DM had significantly higher levels of sCD14 in comparison to NDC, suggesting an increase in gut permeability, microbial translocation, and monocyte/macrophage activation. Importantly, relevant to the ensuing inflammatory responses, the increase in sCD14 in patients with T2DM was accompanied by a significant increase in sCD163, a marker of hepatic Kupffer cell activation and inflammation. Further, a positive correlation was observed between sCD163 and endotoxin and sCD14 in T2DM patients but not in NDC. In association with these changes, keratin 18 (K-18)-based serum markers (M65 and M30) that reflect hepatocyte death were significantly higher in the T2DM group indicating ongoing liver injury. Notably, both M65 and M30 levels correlated with sCD14 and sCD163, suggesting that immune cell activation and hepatic inflammation may be linked to the development of liver injury in T2DM. These findings suggest that the pathogenic changes in the gut-liver axis, marked by increased microbial translocation, may be a major component in the etiology of hepatocyte inflammation and injury in patients with T2DM. However, larger longitudinal studies, including histological evidence, are needed to confirm these observations.

Association between continued metformin use during hospital admission and hospital-acquired complications.

The safety of continuing metformin during a hospital admission has not been robustly demonstrated. We evaluated the association of continuing metformin in hospital with the risk for a hospital-acquired complication (HAC). This is a retrospective observational study of patients admitted to a medical or surgical ward. We considered those with diabetes who continued metformin (DM/MET group), those who discontinued metformin upon admission (DM/MET-STOP), and those with diabetes not on metformin just prior to and during admission (DM/NoMET). We prepared propensity score-matched (PSM) control groups from admitted patients without diabetes. The likelihood of a HAC was determined using a Kaplan-Meier survival analysis. A Cox proportional hazards model was employed to calculate the hazard ratio, adjusted for covariates. Of the 4446 (14%) patients with diabetes, 3331 (10%) were prescribed metformin on admission, and it was continued in 2557 patients. HAC occurred in 5.5% of DM/MET group and 6.4% of the PSM control group. Continuation of metformin was associated with a lower likelihood of HAC, adjusted hazard ratio 0.85 (95% CI 0.69, 1.04), p = 0.117 compared to a PSM-matched control group without diabetes. The DM/NoMET and DM/MET-STOP groups had an increased risk for HAC, adjusted HR 1.77 (1.44, 2.18), p < 0.001 and 2.57 (2.10, 3.13), p < 0.001, as compared to their respective PSM control groups. An individualized assessment to continue metformin during hospital admission was associated with a reduced likelihood of HAC, with the caveat that there was limited matching to non-diabetes controls. This finding warrants further exploration.

Therapy Combining Glucagon-Like Peptide-1 Receptor Agonist with Sodium-Glucose Cotransporter 2 Inhibitor Suppresses Atherosclerosis in Diabetic ApoE-Deficient Mice.

Glucagon-like peptide-1 receptor agonists (GLP-1RAs) and sodium-glucose cotransporter 2 inhibitors (SGLT2i) have beneficial effects on cardiovascular disease in addition to their glucose-lowering effects. In this study, the effects of these drugs, when used individually or in combination, on cardiovascular atherosclerotic lesion development were compared in diabetic ApoE-deficient (ApoE KO) hyperlipidemic mice. ApoE-KO mice were treated with streptozotocin and nicotinamide, generating a type 2 diabetes model. The mice were randomly divided into four groups: vehicle-treated (untreated), liraglutide (LIRA), ipragliflozin (IPRA), and combination therapy (combo). These mice, as well as non-diabetic controls, were fed a high-fat diet. After 8 weeks of drug administration, the heart and aorta were removed and analyzed. Atherosclerotic lesions evaluated by oil red O (ORO) staining were significantly larger in the untreated group (13.4±0.8% of the total aortic area) than in the non-diabetic controls (4.4±0.5%, p<0.01), while being reduced in the combo group (6.0±1.0%, p<0.01) as compared with the untreated group. The ORO stain-positive area in the LIRA and IPRA groups tended to be reduced but their differences were not statistically significant. Transcript levels of Mcp1 and Sirt1 were significantly reduced and increased, respectively, in the combo compared with the untreated group, while no significant changes were observed in the monotherapy groups. The data suggest that combination therapy with liraglutide and ipragliflozin may be an efficient regimen for preventing the development of atherosclerosis in diabetic mice deficient in ApoE.

An ensemble-based machine learning model for predicting type 2 diabetes and its effect on bone health

BackgroundDiabetes is a chronic condition that can result in many long-term physiological, metabolic, and neurological complications. Therefore, early detection of diabetes would help to determine a proper diagnosis and treatment plan.MethodsIn this study, we employed machine learning (ML) based case-control study on a diabetic cohort size of 1000 participants form Qatar Biobank to predict diabetes using clinical and bone health indicators from Dual Energy X-ray Absorptiometry (DXA) machines. ML models were utilized to distinguish diabetes groups from non-diabetes controls. Recursive feature elimination (RFE) was leveraged to identify a subset of features to improve the performance of model. SHAP based analysis was used for the importance of features and support the explainability of the proposed model.ResultsEnsemble based models XGboost and RF achieved over 84% accuracy for detecting diabetes. After applying RFE, we selected only 20 features which improved the model accuracy to 87.2%. From a clinical standpoint, higher HDL-Cholesterol and Neutrophil levels were observed in the diabetic group, along with lower vitamin B12 and testosterone levels. Lower sodium levels were found in diabetics, potentially stemming from clinical factors including specific medications, hormonal imbalances, unmanaged diabetes. We believe Dapagliflozin prescriptions in Qatar were associated with decreased Gamma Glutamyltransferase and Aspartate Aminotransferase enzyme levels, confirming prior research. We observed that bone area, bone mineral content, and bone mineral density were slightly lower in the Diabetes group across almost all body parts, but the difference against the control group was not statistically significant except in T12, troch and trunk area. No significant negative impact of diabetes progression on bone health was observed over a period of 5-15 yrs in the cohort.ConclusionThis study recommends the inclusion of ML model which combines both DXA and clinical data for the early diagnosis of diabetes.

The effect of long-term magnesium treatment on changes in hepatic autophagic and apoptotic ability of type 2 diabetic female rats and their offspring

AimWe investigated the role of magnesium sulfate (MgSO4) and insulin administration on improved autophagy and apoptosis to prevent diabetes in high-fat diet (HFD)-diabetic parents and their offspring. MethodsA HFD and a low dose of streptozotocin (STZ) were used to induce diabetes. Animals were assigned to four groups: non-diabetic control (NDC), diabetic control (DC), the diabetic group received insulin (INS), and the diabetic group received magnesium (MG). The duration of the research was 6 months, and offspring were fed a normal diet for 4 months. Blood glucose was determined weekly in rats (parents and offspring). The gene expression of adenosine monophosphate-activated protein kinase (AMPKα1), Bcl-2-interacting myosin-like coiled-coil protein (Becline-1), P62, Microtubule-associated protein 1 light chain 3 (LC3-II), and caspase-9 of liver tissues were assessed using Real-time PCR. ResultThe analysis of maternal gene expression data showed that treatment with MG and INS considerably augmented the AMPKα1 and LC3-II genes expression and diminished P62 and caspase-9 expression. Also, a significant decrease in the expression of AMPKα1, LC3-II, P62, and Becline-1 was observed in the female offspring in the MG and INS groups. Significant alterations in levels of AMPKα1, Beclin-1, P62, and caspase-9 mRNA were not observed between groups in male offspring, but the expression of LC3-II considerably diminished in the groups compared with the DC group. DiscussionOur data demonstrated that MG supplementation strengthened the mother's autophagy and reduced the transmission of damage to the female offspring. In the male offspring, the transmission of damage was generally less, and MG improved the autophagy condition in the male offspring. Our findings support the presence of a link between autophagy induction and MG-modified biomaterials and determine a potential mechanism of MG-mediated diabetes and autophagy.

Data-based modeling of cerebral hemodynamics quantifies impairment of cerebral blood flow regulation in type-2 diabetes

We studied the regulation dynamics of cerebral blood velocity (CBv) at middle cerebral arteries (MCA) in response to spontaneous changes of arterial blood pressure (ABP), termed dynamic cerebral autoregulation (dCA), and end-tidal CO2 as proxy for blood CO2 tension, termed dynamic vasomotor reactivity (DVR), by analyzing time-series data collected at supine rest from 36 patients with Type-2 Diabetes Mellitus (T2DM) and 22 age/sex-matched non-diabetic controls without arterial hypertension. Our analysis employed a robust dynamic modeling methodology that utilizes Principal Dynamic Modes (PDM) to estimate subject-specific dynamic transformations of spontaneous changes in ABP and end-tidal CO2 (viewed as two “inputs”) into changes of CBv at MCA measured via Transcranial Doppler ultrasound (viewed as the “output”). The quantitative results of PDM analysis indicate significant alterations in T2DM of both DVR and dCA in terms of two specific PDM contributions that rise to significance (p < 0.05). Our results further suggest that the observed DVR and dCA alterations may be due to reduction of cholinergic activity (based on previously published results from cholinergic blockade data) that may disturb the sympatho-vagal balance in T2DM. Combination of these two model-based “physio-markers” differentiated T2DM patients from controls (p = 0.0007), indicating diabetes-related alteration of cerebrovascular regulation, with possible diagnostic implications.

Cross-sectional associations of self-perceived stress and hair cortisol with metabolic outcomes and microvascular complications in type 2 diabetes.

Increasing evidence supports chronic psychological stress as a risk factor for the development of type 2 diabetes. Much less is known, however, about the role of chronic stress in established diabetes. The aim of the current study was to comprehensively assess chronic stress in a sample of 73 patients with type 2 diabetes and 48 non-diabetic control participants, and to investigate associations with indicators of glycemic control (HbA1c), insulin resistance (HOMA-IR), β-cell functioning (C-peptide), illness duration, and the presence of microvascular complications. Chronic stress was measured using questionnaires [the Perceived Stress Scale (PSS), the Screening Scale of the Trier Inventory of Chronic Stress (SSCS), the Perceived Health Questionnaire (PHQ) as well as the Questionnaire on Stress in Patients with Diabetes-Revised (QSD-R)]; hair cortisol was used as a biological indicator. We found that patients with type 2 diabetes had higher levels of hair cortisol in comparison to the control group (F(1,112) = 5.3; p = 0.023). Within the diabetic group, higher hair cortisol was associated with a longer duration of the illness (r = 0.25, p = 0.04). General perceived stress did not show significant associations with metabolic outcomes in type 2 diabetes patients. In contrast, higher diabetes-related distress, as measured with the QSD-R, was associated with lower glycemic control (r = 0.28, p = 0.02), higher insulin resistance (r = 0.26, p = 0.03) and a longer duration of the illness (r = 0.30, p = 0.01). Our results corroborate the importance of chronic psychological stress in type 2 diabetes. It appears, however, that once type 2 diabetes has developed, diabetes-specific distress gains in importance over general subjective stress. On a biological level, increased cortisol production could be linked to the course of the illness.

Metabolic changes enhance necroptosis of type 2 diabetes mellitus mice infected with Mycobacterium tuberculosis.

Previously, we found that Mycobacterium tuberculosis (Mtb) infection in type 2 diabetes mellitus (T2DM) mice enhances inflammatory cytokine production which drives pathological immune responses and mortality. In the current study, using a T2DM Mtb infection mice model, we determined the mechanisms that make T2DM mice alveolar macrophages (AMs) more inflammatory upon Mtb infection. Among various cell death pathways, necroptosis is a major pathway involved in inflammatory cytokine production by T2DM mice AMs. Anti-TNFR1 antibody treatment of Mtb-infected AMs from T2DM mice significantly reduced expression of receptor interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like (MLKL) (necroptosis markers) and IL-6 production. Metabolic profile comparison of Mtb-infected AMs from T2DM mice and Mtb-infected AMs of nondiabetic control mice indicated that 2-ketohexanoic acid and deoxyadenosine monophosphate were significantly abundant, and acetylcholine and pyridoxine (Vitamin B6) were significantly less abundant in T2DM mice AMs infected with Mtb. 2-Ketohexanoic acid enhanced expression of TNFR1, RIPK3, MLKL and inflammatory cytokine production in the lungs of Mtb-infected nondiabetic mice. In contrast, pyridoxine inhibited RIPK3, MLKL and enhanced expression of Caspase 3 (apoptosis marker) in the lungs of Mtb-infected T2DM mice. Our findings demonstrate that metabolic changes in Mtb-infected T2DM mice enhance TNFR1-mediated necroptosis of AMs, which leads to excess inflammation and lung pathology.

Reduced expression of central innate defense molecules in pancreatic biopsies from subjects with Type 1 diabetes

Aims/HypothesisDefensins play a crucial role in the innate immune system's first defense against microbial threats. However, little is known about the defensin system in the pancreas, especially in relation to Type 1 diabetes. We explore the expression of defensins in different disease stages of Type 1 diabetes and correlated obtained findings to the degree of inflammation, providing new insights into the disease and the innate immune system.Material and methodsPancreases from non-diabetic human organ donors of different age groups and donors with Type 1 diabetes with different disease duration were examined. Sections from head, body and tail of the pancreas were stained for eight different defensins and for immune cells; CD3+, CD45+, CD68+ and NES+ (granulocytes).ResultsIn non-diabetic adult controls the level of expression for defensins Beta-1,Alpha-1, Cathelicidin and REG3A correlated with the level of inflammation. In contrast, individuals with Type 1 diabetes exhibit a reduction or absence of several central defensins regardless of the level of inflammation in their pancreas. The expression of Cathelicidin is present in neutrophils and macrophages but not in T-cells in subjects with Type 1 diabetes.ConclusionsObtained findings suggest a pancreatic dysfunction in the innate immune system and the bridging to the adaptive system in Type 1 diabetes. Further studies on the role of the local innate immune system in Type 1 diabetes is needed.

Skeletal muscle myosin heavy chain expression and 3D capillary network changes in streptozotocin-induced diabetic female mice.

It is not well-understood how type 1 diabetes (T1DM) affects skeletal muscle histological phenotype, particularly capillarisation. This study aimed to analyze skeletal muscle myosin heavy chain (MyHC) fibre type changes and 3D capillary network characteristics in experimental T1DM mice. Female C57BL/6J-OlaHsd mice were categorized into streptozotocin (STZ)-induced diabetic (n = 12) and age-matched non-diabetic controls (n =12). The muscle fibre phenotype of the soleus, gluteus maximus, and gastrocnemius muscles were characterized based on the expression of MyHC isoforms, while capillaries of the gluteus maximus were assessed with immunofluorescence staining, confocal laser microscopy and 3D image analysis. STZ-induced diabetic mice exhibited elevated glucose levels, reduced body weight, and prolonged thermal latency, verifying the T1DM phenotype. In both T1DM and non-diabetic mice, the gluteus maximus and gastrocnemius muscles predominantly expressed fast-twitch type 2b fibers, with no significant differences noted. However, the soleus muscle in non-diabetic mice had a greater proportion of type 2a fibers and comparable type 1 fiber densities (26.2 ± 14.6% vs 21.9 ± 13.5%) relative to diabetic mice. T1DM mice showed reduced fiber diameters (P = 0.026), and the 3D capillary network analysis indicated a higher capillary length per muscle volume in the gluteus maximus of diabetic mice compared to controls (P < 0.05). Overall, T1DM induced significant changes in the skeletal muscle, including shifts in MyHC fibre types, decreased fibre diameters, and increased relative capillarisation, possibly due to muscle fibre atrophy. Our findings emphasize the superior detail provided by the 3D analytical method for characterizing skeletal muscle capillary architecture, highlighting caution in interpreting 2D data for capillary changes in T1DM.

Urine Exosomal MicroRNA Profiling in the Family Investigation of Nephropathy and Diabetes (FIND)

About 1 in 3 adults with either type 1 or type 2 diabetes (T2D) has some degree of renal injury or diabetic nephropathy (DN). There is a need to develop more sensitive and specific means of detecting and gauging DN progression so that therapies can be started as early as possible. We hypothesized that UE microRNAs (miRs), small, non-coding 21-25 nucleotide species, a relatively stable readout of renal metabolic health, could be utilized in this fashion. From the NIH, NIDDK Central Repository, we obtained 299 urine samples (300 ml) from African American subjects previously enrolled in the FIND study. The FIND study was initially designed to elucidate genetic determinants of DN in related patient populations using both T2D and non-diabetic control (CTRL), male (M) and female (F) subjects. Our samples included: 20 CTRL-M, 46 CTRL-F, 43 T2D-M, and 90 T2D-F). Other deidentified demographic and clinical data was obtained from the NIH. We measured urine albumin and creatinine in the samples and calculated ratios (mg albumin/g creatinine). Patients were categorized as normal (N, &lt; 30, 120 subjects), microalbuminuric (MIC, 30-300, 33 subjects), or macroalbuminuric (MAC, &gt;300, 46 subjects). Urine exosomes (UE) were isolated from each urine using the Total Exosome Isolation (from urine) kit (Thermofisher), which allows for precipitation of water-insoluble exosomes via centrifugation at medium speed (10,000g) for 1 hour. Small RNA was isolated followed by cDNA synthesis using commercially-available kits (Qiagen). Previously, we found miR-451a to be more highly expressed in UE from human subjects with chronic kidney disease, than in control subjects. We also found this miR to be a potential predictive biomarker for albuminuria in diabetic rats. Therefore, first we used conventional qRT-PCR to evaluate UE levels of miR-451a, as well as, the anti-sense product, miR-451b. MiR-127 was used as the internal standard, as it has been reported not to change with T2D. In MAC subjects, we found that the UE median miR-451a level increased with a fold change (FC) of 3.5 (relative to N). Similarly, miR-451b showed a 3.6-fold increase. FC in the MIC groups were 2.9 and 2.5 for miR-451a and b, respectively (relative to N). Next we conducted an unbiased screen of commonly excreted miRs in these subjects using a miR Array (miRCURY LNA miRNA Focus PCR panel, Qiagen) configured with 88 targets highly expressed in UE. We screened 19 subjects representing a range of conditions, i.e., albuminuric and normal, diabetic and non-diabetic, male and female. We found positive signals in 5 or more subjects for 16 miRs. Of those, 12, i.e., let-7b-5p, miR-16-5p, miR-23b-3p, miR-26a-5p, miR-27a-3p, miR-30a-5p, miR-30b-5p, miR-30c-5p, miR30e-5p, miR-141-3p, miR-203a, and miR-204-5p were expressed at mean levels 2-fold higher in MAC versus N subjects. Furthermore, 13 of these miRs were also over 2-fold higher when subjects were divided as T2D versus CTRL, i.e., the above list plus miR-200c-3p and miR-30d-5p, but not let-7b-5p in this case. When subjects were not divided by groups, miR-598-5p had the highest positive correlation with urinary albumin excretion, R = 0.53 and p = 0.035 (for slope ≠ 0). Using miRNet (McGill University), a freely available miR analysis software tool, we mined for common targets of miR-451(a &amp; b) and miR-598. One putative target of both miR-451b and miR-598 was vascular endothelial growth factor A (VEGFA), which has been shown to play a role in DN. In sum, these miRs represent candidate targets to undergo additional scrutiny. We conclude UE miRs may hold clinical value in the non-invasive assessment of DN severity and predicting progression. NIH/NINDS U01-DK103225 (Feldman, PI); Ecelbarger Pilot. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

IL-17A-mediated JMJD3 regulation of Cxcl1 gene expression in diabetic keratinocytes increases neutrophil recruitment and impairs wound repair.

Abstract Diabetic keratinocytes display an overexpression of chemokines that contribute to prolonged inflammation and non-healing. Using sequencing, we identified that Th17 cells and an IL-17A signature predominate in human diabetic wound tissue compared to non-diabetic controls. The effects of increased IL-17A in diabetic skin on keratinocyte function is unknown. We subjected isolated murine keratinocytes to a 5hr rIL-17A stimulation and performed bulk RNA-seq. We observed increased neutrophil chemoattract genes Cxcl1, Cxcl3, Cxcl5 compared to their controls. We analyzed epigenetic enzymes known to increase gene expression and identified that JMJD3, an activating demethylase, was increased both in human and murine diabetic keratinocytes. IL-17A stimulation increased Jmjd3 expression in murine keratinocytes, and keratinocytes from mice with a keratinocyte-specific IL-17RA deficiency were unable to increase Jmjd3 with rIL-17A. Treatment with a JMJD3-specific inhibitor decreased chemokine genes in IL-17A-stimulated keratinocytes. Keratinocytes from mice with a keratinocyte-specific JMJD3-deficiency displayed decreased chemokines compared to their littermate controls with rIL-17A, suggesting that JMJD3 is relevant to IL-17A-mediated regulation of chemokines. We then analyzed immune cells recruited to the wounds of normal and diabetic mice via flow cytometry and found that Ly6G+ neutrophils were significantly increased in diabetic mice on day 5, a timepoint at which IL-17A is elevated.

Impaired coronary flow reserve in patients with poor type 2 diabetes control: Preliminary results from prospective microvascular dysfunction registry.

Type 2 diabetes (DM) is a common comorbidity associated with cardiovascular disease, especially when poor glucose control is present. Extracardiac microcirculatory complications prevalence is well documented, however coronary microcirculatory dysfunction (CMD) seem to be underreported in this group. The present study analyzed coronary physiology measurements (coronary flow reserve [CFR], index of microcirculatory resistance [IMR], resistance reserve ratio [RRR]) in 47 diabetic patients (21 subjects with poor glycemia control defined as fasting glucose levels > 7.2 mmol/L and 26 with normal fasting glucose), and compared to 54 non-diabetic controls, who had undergone coronary angiography due to symptoms of chronic coronary syndrome. The median age of patients was 65.5 [59.0; 73.0] years old, 74% male, similar in terms of cardiovascular risk factors and prior myocardial infarction. Insulin was used by 19% of diabetic patients with poor glucose control and by 15% of those with DM and low fasting glucose. Prevalence of CMD was 38% in poor glycemia control patients, 27% in DM-patients with proper glucose control and 31% of non-diabetics. Median CFR values were the lowest in poor DM control patients compared to both, normal fasting glucose (1.75 [1.37; 2.32] vs. 2.30 [1.75; 2.85], p = 0.026) and to non-diabetics (1.75 [1.37; 2.32] vs. 2.15 [1.50; 2.95], p = 0.045). Levels of IMR, RRR and MRR did not differ significantly between compared groups (p > 0.05 for all comparisons). Poor glycemia control in type 2 DM might be associated with a higher prevalence of CMD driven by decreased coronary flow reserve, however, further research in larger groups of patients should be performed to confirm this observation.

Brain-Derived Neurotrophic Factor is Associated with Self-Reported Quality of Sleep in Type 2 Diabetes Patients in Ghana.

Sleep disturbances are common in patients with type 2 diabetes (T2DM), and this exacerbates disease severity and results in poor quality of life. Brain-derived neurotrophic factor (BDNF) has been reported to mediate the association between T2DM and poor sleep health. The burden of self-reported poor sleep quality and duration in T2DM and their association with serum BDNF levels were investigated. In this case-control design, the Pittsburgh Sleep Quality Instrument was used to assess self-reported sleep quality and duration in 100 patients with T2DM and 80 nondiabetic controls. Sociodemographic data and medical history were collected from case notes and/or using a structured questionnaire. Fasting venous blood samples (5 mL) were collected to measure plasma lipid profile and serum BDNF levels. patients with T2DM had low levels of BDNF, poor sleep quality (61.9% vs 27.5%, p<0.001), and shorter sleep duration (6.1±2.2 vs 6.9±1.1 h, p=0.003). T2DM status was associated with doubling the odds of poor sleep quality [OR (95%CI)=2.06 (1.07-6.43), p=0.039] and 1.6 times the odds of short sleep duration [1.63 (1.03-3.79), p=0.028]. Multivariable logistic regression analysis revealed no association between serum BDNF levels and sleep status. However, there was a negative biological interaction between T2DM and BDNF levels on poor sleep quality, resulting in 0.28 relative excess risk due to the interaction and a 12% attributable proportion due to the interaction. In this study population, patients with T2DM had a high burden of self-reported poor quality of sleep and shorter sleep duration compared to the nondiabetic controls. T2DM interacts negatively with serum BDNF levels to affect sleep quality.