High Glucose Concentrations Research Articles

High glucose potentiates Zika virus induced-astroglial dysfunctions.

Zika virus (ZIKV) is a neurotropic flavivirus that induces congenital Zika syndrome and neurodevelopmental disorders. Given that ZIKV can infect and replicate in neural cells, neurological complications in adult brain are also observed. Glial cells may emerge to delay and/or prevent the development of ZIKV-induced neurodegeneration. These cells actively participate in metabolic, inflammatory and redox processes, and consequently, in the pathophysiology of neurodegenerative diseases, including diabetic encephalopathy. In this sense, changes in glucose metabolism can support the inflammatory activity of astroglial cells; however, the effects of increased glucose concentration during ZIKV infection have not yet been explored in astroglial cells. Here, we evaluated functional parameters of astroglial cells exposed to ZIKV upon normal and high glucose concentrations, focusing on inflammatory profile, oxidative stress, and expression of critical genes for astroglial functions. High glucose potentiated the pro-inflammatory and oxidative effects of ZIKV, as well as potentiated the downregulation of signaling pathways, such as Nrf-2 (nuclear factor erythroid derived 2 like 2), sirtuin 1 (SIRT1), peroxisome proliferator activated receptor gamma coactivator 1-alpha (PGC-1α), and poly (ADP-ribose) polymerase (PARP). In summary, our results suggest that high glucose can favor the activation of inflammatory signaling while impairing cytoprotective pathways in astroglial cells exposed to ZIKV and reinforce the hypothesis that this virus is highly neurotrophic, with significant impact in glial cells.

Molecular dynamics and solvation structures of the β-glucosidase from Humicola insolens (BGHI) in aqueous solutions containing glucose

The β-glucosidase enzyme is a glycosyl hydrolase that breaks down the β-1,4 linkage of cellobiose. It is inhibited by glucose at high concentrations due to competitive inhibition. However, at lower glucose concentrations, the glucose-tolerant β-glucosidase from Humicola insolens (BGHI) undergoes stimulation. Proteins, in aqueous sugar solutions, tend to be preferentially hydrated, which generally promotes their stabilization. Thus, solvation phenomena may contribute to both glucose tolerance and stimulation processes. We have performed atomistic classical Molecular Dynamics (MD) simulations of BGHI at different glucose concentrations to mimic the conditions found in the catalytic experiments. A detailed examination of the solvent environment through the calculation of minimum distance distribution functions (MDDFs) and Kirkwood-Buff (KB) integrals was performed. The enzyme is preferentially hydrated in the presence of glucose at all concentrations. Nevertheless, the hydration does not prevent the glucose from directly interacting with the BGHI surface or from entering the active site. Based on the obtained results, we hypothesize that preferential hydration is beneficial for enzyme activity. At the same time, product inhibition has little effect at lower concentrations of glucose, and at higher glucose concentrations, competition for the active site becomes predominant and the enzyme is primarily inhibited.

Increased glucose concentration modifies TGF-β1 and NFκB signaling pathways in aniridia limbal fibroblasts, in vitro

To determine the impact of increased glucose concentration on gene expression of primary healthy human limbal fibroblasts (LFCs) and congenital aniridia human limbal fibroblasts (AN-LFCs), in vitro.LFCs (n = 8) and AN-LFCs (n = 8) were isolated and cultured in serum containing DMEM, including either normal glucose (17.5 mM) or increased glucose (70 mM) concentration for 48h or 72h, respectively. mRNA and protein expression of transforming growth factor beta 1 (TGF-β1), alpha-smooth muscle actin (ACTA)2A1, SMAD 2/3, hypoxia markers such as nuclear factor kappa B (NFκB), inducible nitric oxide synthase (iNOS), hypoxia-inducible factor 1-alpha (HIF-1ɑ), oxidative stress markers such as nuclear factor erythroid 2-related factor 2 (Nrf2) and Catalase (CAT) were analyzed using qPCR and Western blot.In 70 mM glucose concentration medium for 48 h, TGF-β1 mRNA expression was significantly lower (p = 0.001, p < 0.001), Nrf2 (p = 0.001, p = 0.001) and CAT (p = 0.001, p = 0.001) mRNA expression was significantly higher in LFCs and AN-LFCs, than using 17.5 mM glucose concentration medium. In addition, in 70 mM glucose concentration medium for 48 h, SMAD 2, SMAD 3, NFκB, HIF-1ɑ mRNA expression was significantly lower in AN-LFCs, than in 17.5 mM glucose concentration medium (p = 0.003, p = 0.002, p = 0.008, p = 0.020). At this time-point in 70 mM glucose concentration medium, at protein level, TGF-β1, SMAD2/3 and NFκB were significantly lower in AN-LFCs, than in 17.5 mM glucose concentration medium (p = 0.041, p = 0.002, p = 0.012).In 70 mM glucose concentration medium for 72h, TGF-β1 was significantly higher (p < 0.001, p < 0.001) and Nrf2 (p = 0.001, p = 0.001) and CAT (p < 0.001, p < 0.001) mRNA were significantly lower in LFCs and AN-LFCs, than in 17.5 mM glucose concentration medium. At this time-point, in 70 mM glucose concentration medium, NFκB mRNA was significantly higher (p < 0.001) in LFCs, than in 17.5 mM glucose concentration DMEM medium. In 70 mM glucose concentration medium for 72 h, TGF-β1 and NFκB protein were significantly lower in AN-LFCs, than in 17.5 mM glucose concentration medium (p < 0.001, p < 0.001).Our study confirmed that high glucose concentration has an impact on TGF-β1 and NFκB signaling both in AN-LFCs and LFCs. These findings highlight that prolonged exposure to high glucose levels may contribute to cellular stress and dysfunction in LFCs and AN-LFCs.

Investigation on the Effect of Sacoglottis gabonensis on Blood Glucose, Body Weight and Behavioural Changes of Mice Exposed to Non-nutritive Sweeteners during Gestation Period

Aim: Aim of this research was to investigate the role of Sacoglotts gabonensis on blood sugar, body weight and behavioural changes of mice exposed to artificial sweeteners. Study Design: A total of 16 female mice (mean weight 18.6±2.32g) divided into four groups were used in this study. Methodology: Group A received no sweeteners or S. gabonensis, B received 50mg of sweeteners only, C received 50mg of sweeteners and 250mg/kg/bw of S. gabonensis, D received 250mg/kg/bw of S. gabonensis only for 8weeks. Results: show that group A and D had significant (p&lt;0.05) decrease in blood glucose concentration with 7.27±3.53 mmol/L and 8.22±1.87 mmol/L respectively compared to group B and C with 9.12±1.03 mmol/L and 8.22±1.87 mmol/L. There was a significant (p&lt;0.05) increase in body weight in groups A 32.60±7.34g and B 31.38±4.53g compared to groups C, 22.45±15.49g and D, 24.00±14.71g. Gestation length was between 19 and 20 days for the different groups and behavioural changes such as climbing, running, nesting and restlessness were observed in all the groups. Conclusion: The administration of S. gabonensis did not alter gestation length and behavioural changes during gravid period in experimental animals but altered the blood glucose concentrations However, administration of S. gabonensis is recommended for individuals who are at risk of developing high blood glucose concentration and increased body weight as it has shown modulatory effect on blood glucose and body weight gain.

Oxophilic Sn to promote glucose oxidation to formic acid in Ni nanoparticles.

The electrochemical glucose oxidation reaction (GOR) presents an opportunity to produce hydrogen and high-value chemical products. Herein, we investigate the effect of Sn in Ni nanoparticles for the GOR to formic acid (FA). Electrochemical results show that the maximum activity is related to the amount of Ni, as Ni sites are responsible for catalyzing GOR via the NiOOH/Ni(OH)2 pair. However, the GOR kinetics increases with the amount of Sn, associated with an enhancement of the OH- supply to the catalyst surface for Ni(OH)2 reoxidation to NiOOH. NiSn nanoparticles supported on carbon nanotubes (NiSn/CNT) exhibit excellent current densities and direct GOR via C-C cleavage mechanism, obtaining FA with a Faradaic efficiency (FE) of 93% at 1.45 V vs. reversible hydrogen electrode. GOR selectivity is further studied by varying the applied potential, glucose concentration, reaction time, and temperature. FE toward FA production decreases due to formic overoxidation to carbonates at low glucose concentrations and high applied potentials, while acetic and lactic acids are obtained with high selectivity at high glucose concentrations and 55 °C. Density functional theory calculations show that the SnO2 facilitates the adsorption of glucose on the surface of Ni and promotes the formation of the catalytic active Ni3+ species.

Abstract 4140181: Mechanisms of SGLT-2 Inhibitor Empagliflozin in Attenuating Intramitochondrial Stress and Restoring Mitochondrial Function in Hyperglycemic Cardiomyocyte

Systemic hyperglycemia causes tissue damage and triggers cardiovascular disease (CVD). Sodium-glucose cotransporter-2 inhibitors (SGLT-2i) are a novel class of glucose-lowering agents that have shown unexpected benefits in clinical trials for the treatment of CVDs. We aim to investigate the underlying mechanisms of how SGLT-2 inhibitors alleviate CVDs associated with elevated glucose stress. iPSC-derived cardiomyocytes (iPSC-CM) were incubated with high glucose concentrations for 72 hours. Mitochondrial function in these cardiomyocytes was assessed by flow cytometry with JC-1 staining and ATP luminescence assay. Intracellular reactive oxygen species (ROS) and intramitochondrial calcium stress were measured using CellROX, MitoSOX, and Rhod-2 AM staining. Patch clamp was employed to determine ion current changes in the cardiomyocytes. Mitochondrial oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were determined using the Seahorse XFe96 analyzer. In addition, qPCR, Western blot, and DM mouse heart histological analysis were performed to assess the regulation of associated molecules. The results indicated that exposure to a high glucose environment caused cardiomyocyte injury and impaired mitochondrial biosynthesis. Empagliflozin exhibited a beneficial effect on mitochondrial function by reducing ROS production and calcium deposition. It also mitigated the reduction in respiratory OCR of cardiomyocytes induced by high glucose incubation. Furthermore, molecular analysis revealed that Empagliflozin attenuated the dysregulation of mitochondrial calcium channels and biosynthesis by reducing associated gene expression, including Bcl2 , Mfn1 , Mx2 , Oas1 , Ant3 , Mcu , Micu1 , Vdac1 , Ryr2 , and Cypd-ppid . Histological analysis of DM mouse hearts demonstrated that reduced MFN2 and ZBP1 were target molecules for hyperglycemia-induced reduction of calcium channel currents in cardiomyocytes and could be restored by Empagliflozin treatment. This study concludes that high glucose stress diminishes mitochondrial calcium channel regulators MFN2 and ZBP1 in cardiomyocyte, which reduces calcium channel currents and leads to sensitization of cardiomyocyte to arrhythmogenesis, resulting in VT/VF. It provides experimental evidence for the clinical efficacy of Empagliflozin in ameliorating CVDs and managing diabetes-related CVDs.

Obese male zucker rats show basilar dendritic retraction in the medial prefrontal cortex

Obesity, a prevalent disorder, predisposes individuals to metabolic syndrome, type 2 diabetes mellitus, and high blood pressure. Obesity has been investigated in various organisms that display genetic, high-fat, and high-carbohydrate diet (HFCD)-induced obesity. Recent studies have found that both male and female Zucker rats, which are genetically obese, exhibit alterations in dendritic arborization of neurons in certain structures of the central nervous system. Therefore, the present study aimed to analyze dendritic arborization and dendritic spine density of pyramidal neurons in the medial prefrontal cortex (mPFC) of obese adult male Zucker rats using the Golgi-Cox method and Sholl analysis. Obese male Zucker rats exhibit increased body weight and high concentrations of glucose, triglycerides, and cholesterol. Analysis of mPFC pyramidal neurons in these rats revealed basilar dendritic retraction at a medium distance from the soma, in addition to a reduction in total basilar dendritic length, without any changes in dendritic spine density. These findings are consistent with previous reports, indicating that changes in dendritic retraction may occur as a result of the leptin receptor mutation itself, in addition to the reduction in dendritic arborization observed in other regions of the central nervous system in rats. Furthermore, we suggest the possibility that biological processes modulated by the mPFC, such as foraging and social behavior, are also affected.

Study on the Mechanism of Alpinia officinarum Hance in the Improvement of Insulin Resistance through Network Pharmacology, Molecular Docking and in vitro Experimental Verification.

Research has elucidated that the pathophysiological underpinnings of non-alcoholic fatty liver disease and type 2 diabetes mellitus are intrinsically linked to insulin resistance (IR). However, there are currently no pharmacotherapies specifically approved for combating IR. Although Alpinia officinarum Hance (A. officinarum) can ameliorate diabetes, the detailed molecular mechanism through which it influences IR has not been fully clarified. To predict the active components of A. officinarum and determine the mechanism by which A. officinarum affects IR. The active compounds and molecular mechanism underlying the improvement of IR by A. officinarum were predicted via network pharmacology and molecular docking. To further substantiate these predictions, an in vitro model of IR was induced in HepG2 cells using high glucose concentrations. Cytotoxicity and oxidative stress levels were evaluated using Cell Counting Kit-8, reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) assay kits. The putative molecular mechanisms were corroborated through Western blot and RT-PCR analyses. Fourteen principal active components in A. officinarum, 133 potential anti-IR gene targets, and the top five targets with degree values were ALB, AKT1, TNF, IL6, and VEGFA. A. officinarum was posited to exert its pharmacological effects on IR through mechanisms involving lipid and atherosclerosis, the AGE-RAGE signaling pathway in diabetic complications, the PI3K-AKT signaling pathway, fluid shear stress, and atherosclerosis. Intriguingly, network pharmacology analysis highlighted (4E)-7-(4-hydroxy-3-methoxyphenyl)-1-phenylhept-4-en-3- one (A14) as the most active compound. Molecular docking studies further confirmed that A14 has a strong binding affinity for the main targets of PI3K, AKT, and Nrf2. The experiments demonstrated that A14 significantly diminished the ROS and MDA levels while augmenting the SOD activity. Moreover, A14 was found to elevate the protein expression of PI3K, AKT, Nrf2, and HO-1, and increase the mRNA levels of these targets as well as NQO1. A. officinarum could play a therapeutic role in IR through multiple components, targets, and pathways. The most active component of A. officinarum responsible for combating IR is A14, which has the ability to regulate oxidative stress in IR-HepG2 cells by activating the PI3K/AKT/Nrf2 pathway. These findings suggest a potential pharmacological intervention strategy for the treatment of IR.

Periostin Induces Epithelial-Mesenchymal Transition via p38-MAPK Pathway in Human Renal Tubular Cells by High Glucose.

Periostin mediates inflammation and fibrosis by regulating extracellular matrix adhesion, migration, and differentiation in multiple organ diseases. Studies have shown periostin mainly located in the dilated mesangium, tubulointerstitial and fibrotic regions of the diabetic kidney disease, which was negatively correlated with renal function. However, the underlying mechanism remains poorly explored. The expression of periostin in HK-2 cells was investigated under high glucose and high concentration of TGF-β1. The signaling pathway of periostin involved in epithelial-mesenchymal transdifferentiation of HK-2 cells was also validated. The expression of periostin were investigated by RT-PCR, western blot analysis and immunofluorescence assays with different concentrations of glucose and TGF-β1. The expression of E-Cad, α-SMA and p38 proteins were also detected. The effects of periostin, E-Cad, and α-SMA in high glucose were investigated by p38 inhibitors. To demonstrate the interaction among periostin, p38 and EMT markers, periostin under high glucose and high TGF-β1 was knocked down, resulting p38 and phosphorylated p38 was evaluated. The combined of high glucose (HG, 22 mmol/L) and high TGF-β1 (10 ng/mL) upregulated the expression of periostin obviously, stimulating the expression of α-SMA and p38 while inhibiting the expression of E-Cad. p38 inhibitors reduced the expression of periostin and α-SMA while promoted E-Cad protein expression in HK-2 cells under HG conditions. Additionally, p38-MAPK signal pathway was involved in epithelial-mesenchymal transition of human renal tubules in high glucose environment. Significant, knockdown periostin expression effectively inhibited the expression of p38 and phosphorylated p38 under the combination of HG and high TGF-β1, verifying the interaction of periostin with the p38-MAPK signaling pathway. Periostin, a downstream factor of TGF-β1, is positively regulated by TGF-β1 under HG condition, affecting the epithelial-interstitial differentiation of HK-2 cells via p38-MAPK signaling pathway. Therefore, periostin may serve as a biomarker of renal fibrosis in diabetic kidney disease.

Targeting the histone methyltransferase SETD7 rescues diabetes-induced impairment of angiogenic response by transcriptional repression of semaphorin 3G

Abstract Background Peripheral artery disease (PAD) is highly prevalent in patients with diabetes (DM) and associates with a high rate of limb amputation and poor prognosis. Surgical and catheter-based revascularization have failed to improve outcome in DM patients with PAD. Hence, a need exists to develop new treatment strategies able to promote blood vessel growth in this setting. Mono-methylation of histone 3 at lysine 4 (H3K4me1) - a specific epigenetic signature induced by the histone methyltransferase SETD7 - favours an open chromatin thus enabling gene transcription. Purpose To investigate whether SETD7-dependent epigenetic changes modulate angiogenic response in diabetes. Methodology Primary human aortic endothelial cells (HAECs) were exposed to normal glucose (NG, 5 mM) or high glucose (HG, 25 mM) concentrations for 48 hours. Unbiased gene expression profiling was performed by RNA sequencing (RNA-seq) followed by Ingenuity Pathway Analysis (IPA). In vitro assays, namely cell migration and tube formation were employed to study angiogenic properties in HAECs. SETD7 and H3K4me1 levels were investigated by Western blot and Chromatin immunoprecipitation (ChIP). Pharmacological blockade of SETD7 was achieved by using the highly selective inhibitor (R)-PFI-2. Mice with streptozotocin-induced diabetes were orally treated with (R)-PFI-2 or vehicle and underwent hindlimb ischemia by femoral artery ligation for 14 days. Blood flow recovery was analysed at 30 minutes, 7 and 14 days by laser Doppler imaging. Our experimental findings were also translated in gastrocnemius muscle samples from patients with and without diabetes. Results RNA-seq in HG-treated HAECs revealed a profound upregulation of the methyltransferase SETD7, an enzyme involved in mono-methylation of lysine 4 at histone 3 (H3K4me1). SETD7 upregulation in HG-treated HAECs was associated with increased H3K4me1 levels as well as with impaired endothelial cell migration and tube formation. Both SETD7 gene silencing and pharmacological inhibition by (R)PFI-2 rescued hyperglycemia-induced impairment of HAECs migration and tube formation, while SETD7 overexpression blunted the angiogenic response. RNA-seq and ChIP assays showed that SETD7-dependent H3K4me1 regulates the transcription of the angiogenesis inhibitor semaphorin-3G (SEMA-3G). Moreover, SEMA-3G overexpression blunted migration and tube formation in SETD7-depleted HAECs. In diabetic mice with hindlimb ischemia, treatment with (R)-PFI-2 improved limb vascularization and perfusion as compared to vehicle. Finally, SETD7/SEMA3G axis was upregulated in muscle specimens from T2D patients as compared to controls. Conclusion Targeting SETD7 represents a novel epigenetic-based therapy to boost neovascularization in diabetic patients with PAD.

Unveiling the impact of carbon sources on phosphorus release from sediment: Investigation of microbial interactions and metabolic pathways for anaerobic phosphorus recovery

The aim of this study was recovery of phosphorus (P) from marine sediment, and our results revealed the influence of P release from the sediment stimulated with different types and concentrations of carbon sources. During the 15-day anaerobic operation, the sediments stimulated with 1 g/L propionic acid and glucose exhibited more prominent effects compared to other trials, with 5.98 mg/L and 6.44 mg/L of P released, respectively, with a total solid content of 4 %. Notably, the excessive addition of carbon sources was shown to can partially inhibit P release. As microbial activity intensified, P was utilized for microbial synthesis, resulting in a decreased P in the supernatant. For example, in glucose-fed systems with concentrations of 5 g/L and 10 g/L, the P concentration decreased from 5 mg/L on Day 3 to approximately 3 mg/L on Day 15. The sequencing results indicated distinct evolutions within different carbon source-fed systems over the 15-day operations. Feeding high concentrations of glucose resulted in rapid enrichment of fermentative bacteria under anaerobic conditions, while sulfate-reducing bacteria promoted P release in volatile fatty acids-fed systems. Metabolic analysis revealed that carbon sources not only influence gene expression in different systems, but also impact the metabolic pathways involved in nutrient cycling, which can be interrelated. For example, a significant positive correlation was observed between the abundance of P and sulfur cycling functional genes (phoD, cysD).

Regulation of hepatic transporters OATP1A2 and OATP1B1 by the action of nitric oxide (II)

Nitric oxide II (NO) is a signaling molecule that has a wide range of physiological effects, including the regulation of gastrointestinal processes. The liver actively expresses the clinically significant transporters OATP1A2 and OATP1B1, which are involved in the influx of biologically active and medicinal substances. That is why it seems relevant to determine the pathways of regulation of hepatic transporters under the influence of NO. Aim. To study the effect of NO on the relative amount and expression of the transporters OATP1A2 and OATP1B1 in vitro in HepG2 cells. Materials and methods. The study was performed on a culture of HepG2 cells, which were cultured in 6-well plates at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) with a high glucose content (4500 mg/l) containing L-glutamine (4 mM), 10% fetal bovine serum, 100 U/ml penicillin and 100 mg/ml streptomycin (all components from Sigma-Aldrich, Germany). S-nitrosoglutathione (Sigma-Aldrich, Germany) was added to the culture medium at concentrations of 1, 10, 50, 100 and 500 µM, incubated for 24 and 72 hours. Water for injection (solvent) was added to control cells in an equivalent volume S-nitrosoglutathione). The relative amounts of OATP1A2 and OATP1B1 proteins were assessed by Western blot, and the expression of SLCO1A2 and SLCO1B1 by real-time PCR. The results of the study. In the course of this study, it was shown that the addition of S-nitrosoglutathione in the concentration range of 10-500 μM and exposure duration of 24 and 72 hours causes an increase in the intracellular level of nitric oxide metabolites, which indicates the adequacy of the use of this NO donor. At the same time, under the influence of NO, there was an increase in the relative amount of the studied transporters - OATP1A2 at an exposure period of 24 hours and S-nitrosoglutathione concentrations of 50 and 100 μM, OATP1B1-24 and 72 hours, at concentrations of 10-500 μM, a similar trend was noted for the expression genes SLCO1A2 and SLCO1B1. Conclusion. The NO donor - S-nitrosoglutathione causes an increase in the relative amount of OATP family transporters - OATP1A2 and OATP1B1, due to increased expression of the SLCO1A2 and SLCO1B1 genes, in vitro in HepG2 cells.

Liver-expressed antimicrobial peptide 2 is a hepatokine regulated by ghrelin, nutrients, and body weight.

Liver-expressed antimicrobial peptide 2 (LEAP2) is a peptide that counteracts the hunger hormone ghrelin-induced functions. Recently, we showed that vertical sleeve gastrectomy (VSG) did not alter the serum LEAP2 concentration in individuals with obesity. Here, we investigated the effects of VSG in both chow diet (CD)-fed and high-fat diet (HFD)-fed mice. In CD-fed mice, VSG increased plasma LEAP2 levels and hepatic Leap2 mRNA levels while decreasing body weight, blood glucose levels, and ghrelin levels. Intraperitoneal (ip) administration of ghrelin reversed these changes. These effects were found in both male and female mice. In contrast, VSG or weight loss in HFD-induced obese mice decreased LEAP2 levels. After fasting, the plasma LEAP2 concentration was in the following order: hepatic vein > abdominal aorta > portal vein. A high glucose concentration robustly increased the plasma LEAP2 concentration in the hepatic vein and abdominal aorta but not in the portal vein. In addition, corn oil or palmitate increased LEAP2 expression and secretion. The increase in LEAP2 levels after the meal tolerance test was delayed in the human subjects with diabetes. Our data suggest that various factors (metabolic, hormonal, and nutritional) regulate LEAP2, and the liver is the predominant site for the production and secretion of LEAP2. Furthermore, the interaction between ghrelin and LEAP2 is involved in the pathogenesis of obesity and diabetes.

Identification of Candida albicans in the Urine of Type 2 Diabetes Mellitus Patiens at Puskesmas Jaten 2, Karanganyar Regency

Background: Candida albicans is an opportunistic fungus that thrives in environments with high glucose concentrations, which makes patients with Diabetes Mellitus (DM) especially vulnerable to infection. Elevated blood glucose levels in DM can lead to glycosuria, providing a rich nutrient source for fungal growth. This research is critical because there is a lack of specific studies focusing on fungal infections in diabetic patients in Indonesia, despite the increasing prevalence of DM. This research provides a new perspective by highlighting a local diabetic population that has not been extensively studied before. Objective: The objective of this study is to identify the presence of Candida albicans in the urine of Type 2 Diabetes Mellitus patients at Puskesmas Jaten 2 in Karanganyar Regency. Materials and Methods: This descriptive study included 20 urine samples from Type 2 Diabetes Mellitus patients. The inclusion criteria were fasting blood glucose levels of 200 mg/dl or higher, and all patients were enrolled in the chronic disease management program (Prolanis) at Puskesmas Jaten 2. The samples were collected and examined macroscopically in February 2024 to detect the presence of Candida albicans. Results: The results showed that Candida albicans was found in 6 out of 20 urine samples (30%), while the fungus was not detected in 14 samples (70%). The majority of positive cases occurred in patients aged 50 and older, suggesting an age-related susceptibility to fungal infection in this population. Patients with higher glucose levels also showed a greater tendency for Candida growth. Conclusions: Based on the results of this study, it can be concluded that Candida albicans is present in the urine of Type 2 Diabetes Mellitus patients at Puskesmas Jaten 2, with a notable prevalence of 30%. This finding highlights an increased susceptibility to fungal infections in older patients and those with elevated glucose levels, suggesting the need for careful monitoring and potential intervention in this population. Further studies with larger sample sizes are suggested to confirm these results and explore additional preventive measures.

High glucose-induced senescence contributes to tubular epithelial cell damage in diabetic nephropathy

Dysfunctional renal tubular epithelial cells, induced by high glucose, are commonly observed in the kidney tissues of diabetic nephropathy (DN) patients. The epithelial-mesenchymal transition (EMT) of these cells often leads to renal interstitial fibrosis and kidney damage in DN. High glucose also triggers mitochondrial damage and apoptosis, contributing further to the dysfunction of renal tubular epithelial cells. Cellular senescence, a recognized characteristic of DN, is primarily caused by high glucose. However, it remains unclear whether high glucose-induced cellular senescence in DN exacerbates the functional impairment of tubular epithelial cells. In this study, we examined the relationship between EMT and cellular senescence in kidney tissues from streptozotocin (STZ)-induced DN and HK-2 cells treated with high glucose (HG). We also investigated the impact of HG concentrations on tubular epithelial cells, specifically mitochondrial dysfunction, cellular senescence and apoptosis. These damages were primarily associated with the secretion of cytokines (such as IL-6, and TNF-α), production of reactive oxygen species (ROS), and an increase of intracellular Ca2+. Notably, resveratrol, an anti-aging agent, could effectively attenuate the occurrence of EMT, mitochondrial dysfunction, and apoptosis induced by HG. Mechanistically, anti-aging treatment leads to a reduction in cytokine secretion, ROS production, and intracellular Ca2+ levels.

Pattern recognition of drying lysozyme–glucose droplets using machine learning classifiers

Out-of-equilibrium processes, such as sessile droplet drying, often result in distinctive macroscopic residual patterns in systems containing molecules, proteins, and colloids. Protein–glucose mixtures are particularly effective models for studying the behavior of complex fluids containing biomolecules. This study investigates the drying patterns of lysozyme droplets with varying initial glucose concentrations. Without glucose, the crack patterns are chaotic and dispersed throughout the droplet. Interestingly, cracks predominantly form around the droplet edges at intermediate glucose concentrations, while the deposits become uniform and crack-free at high glucose concentrations. To understand and classify the unique patterns related to the initial compositional changes, we developed an automated pattern recognition pipeline. We used two methods for analyzing images captured throughout the drying process. The first method involved extracting statistical textural parameters from the images as quantitative features for machine learning classifiers. The second method utilized a neural network-based classifier to directly classify the images, achieving an accuracy of 97%. The results demonstrate the effectiveness of using images from the entire drying process, not just the final images, for pattern classification. This approach may be useful in gaining a fundamental understanding of unique crack pattern that emerge when glucose is added to a protein solution.

Double (W1/O/W2) emulsions prepared with natural vegetable oils and varied inner glucose concentration: Microstructure, rheology and stability

The objective was to study the microstructure, rheology and stability of double (W1/O/W2) emulsions prepared with Tween 80 as hydrophilic emulsifier and coconut, palm or sunflower oil. The effects of glucose concentration in inner aqueous phase and crystallization of lipid phase were analyzed. Aggregation of oil globules by partial coalescence was observed in emulsions prepared with coconut or palm oil because of the crystallization of the lipid phase. The increase of glucose concentration increased the amount of inner water by osmotic swelling and produced a higher partial coalescence degree due to a higher collision frequency between oil globules. Consequently, higher elastic and viscous modules were observed at higher glucose concentration. Fat crystals in lipid phase allowed a partial retention of inner water in the absence of glucose, but they also favored the release of inner water due to partial coalescence. Creaming stability was enhanced by the presence of glucose, because of the formation of a three-dimensional network of aggregated oil globules in emulsions with coconut or palm oil, and a higher packing and global density of oil globules in systems with sunflower oil. The obtained results contribute to the understanding of partial coalescence in W1/O/W2 emulsions, including its effects on inner water release and creaming stability. This work could be relevant for the design and stabilization of vegetable, lipid-reduced food emulsions prepared with natural oils.

Folic acid ameliorates high glucose–induced neurotoxicity in human forebrain organoids: Insights from proteomics

AbstractPregestational diabetes (PGDM) has been associated with an elevated risk of congenital abnormalities, particularly those affecting the nervous system. The efficacy of folic acid (FA) supplementation in reducing the incidence of neurodevelopmental damage caused by PGDM has been well documented. However, the exact mechanism is unclear. Here, a human forebrain organoid model, which replicates the three‐dimensional structure of the early fetal neural tissue, was employed to study the neuroprotective effects of FA in PGDM. In this study, the forebrain organoids were cultured at high glucose (HG) concentrations from Days 20 to 40 with or without FA. Immunostaining revealed that the supplementation of FA significantly decreased HG‐induced neuron apoptosis. The proteomics examination suggested HG caused an increase in glial fibrillary acidic protein expression, a marker of astrocytes, leading to the upregulation of metallothionein expression and perturbation of mineral absorption, whereas FA reversed this effect. Proteomics analysis further showed that FA reduced HG‐induced cell migration. Moreover, Western blot analysis verified that FA mitigated HG‐induced apoptosis and cell migration via AMPK/FOXO pathway. Overall, current findings indicate that FA, as a functional food ingredient, has a protective effect on HG‐induced abnormal fetal neurodevelopment.

VCAM-1 targeted nanocarriers of shRNA-Smad3 mitigate endothelial-to-mesenchymal transition triggered by high glucose concentrations and osteogenic factors in valvular endothelial cells

Endothelial to mesenchymal transition (EndMT) of valvular endothelial cells (VEC) is a key process in the development and progression of calcific aortic valve disease (CAVD). High expression of the Smad3 transcription factor is crucial in the transition process. We hypothesize that silencing Smad3 could hinder EndMT and provide a novel treatment for CAVD. We aimed at developing nanoparticles encapsulating short-hairpin (sh)RNA sequences specific for Smad3 targeted to the aortic valve. We synthesized VCAM-1-targeted lipopolyplexes encapsulating shRNA-Smad3 plasmid (V-LPP/shSmad3) and investigated their potential to reduce the EndMT of human VEC. VEC incubation in a medium containing high glucose concentrations and osteogenic factors (HGOM) triggers EndMT and increased expression of Smad3. Exposed to lipopolyplexes, VEC took up efficiently the V-LPP/shSmad3. The latter reduced the EndMT process in VEC exposed to HGOM by downregulating the expression of αSMA and S100A4 mesenchymal markers and increasing the expression of the CD31 endothelial marker. In vivo, V-LPP/shSmad3 accumulated in the aortic root and aorta of a murine model of atherosclerosis complicated with diabetes, without affecting the liver and kidney function. The results suggest that targeting activated VEC with lipopolyplexes to silence Smad3 could be an effective, novel treatment for CAVD mediated by the EndMT process.

Development of a Competitive Nutrient-Based T-Cell Immunotherapy Designed to Block the Adaptive Warburg Effect in Acute Myeloid Leukemia.

Background: T-cell-based adoptive cell therapies have emerged at the forefront of cancer immunotherapies; however, failed long-term survival and inevitable exhaustion of transplanted T lymphocytes in vivo limits clinical efficacy. Leukemia blasts possess enhanced glycolysis (Warburg effect), exploiting their microenvironment to deprive nutrients (e.g., glucose) from T cells, leading to T-cell dysfunction and leukemia progression. Methods: Thus, we explored whether genetic reprogramming of T-cell metabolism could improve their survival and empower T cells with a competitive glucose-uptake advantage against blasts and inhibit their uncontrolled proliferation. Results: Here, we discovered that high-glucose concentration reduced the T-cell expression of glucose transporter GLUT1 (SLC2A1) and TFAM (mitochondrion transcription factor A), an essential transcriptional regulator of mitochondrial biogenesis, leading to their impaired expansion ex vivo. To overcome the glucose-induced genetic deficiency in metabolism, we engineered T cells with lentiviral overexpression of SLC2A1 and/or TFAM transgene. Multi-omics analyses revealed that metabolic reprogramming promoted T-cell proliferation by increasing IL-2 release and reducing exhaustion. Moreover, the engineered T cells competitively deprived glucose from allogenic blasts and lessened leukemia burden in vitro. Conclusions: Our findings propose a novel T-cell immunotherapy that utilizes a dual strategy of starving blasts and cytotoxicity for preventing uncontrolled leukemia proliferation.