High Glucose Concentrations Research Articles

Biomineralized ceria nanoparticles target the heart to improve diabetic cardiac remodeling by regulating mitochondrial oxidative stress and decreasing excessive mitophagy

Abstract Background An altered state of cardiomyocytes due to high glucose concentrations results in cell hypertrophy, leading to diabetic cardiomyopathy (DCM).The molecular regulatory process of mitochondrial dyshomeostasis and mitophagy, oxidative stress, and reactive oxygen species (ROS) production in diabetes is still not fully understood. Mitochondrial dyshomeostasis and mitophagy could be critical factors in the heart remodeling that occurs in diabetes. Purpose Our transcriptome sequencing of heart tissue revealed upregulation of the gene encoding the transferrin receptor (TfR) in the diabetic rat (Figure 1A). We then validated in vitro that high glucose increased TfR protein expression in the HL-1 cell line (Figure 1B). Herein, we developed a novel therapeutic ceria nanoparticle via biomineralization using transferrin (Tf) protein as a temple and assessed its targeted therapy for DM-induced cardiac remodeling via Tf-TfR mediated endocytosis. Method We synthesized Tf protein-based ceria NPs (CeO2@Tf) via biomineralization. Then the physicochemical and biological properties of NPs were characterized. Cellular uptake, cytotoxicity test, intracellularmitochondrial membrane potential (MMP), and ROS were verified with mouse cardiomyocytes HL-1 cells line. Afterward, the male C57BL/6N mice were randomly divided into control, DM, DM+Metformin (Met), and DM+NPs groups with eight weeks of treatment. Heart structure and function were measured by echocardiographic. The heart tissues were stained with hematoxylin, eosin (H&E), and Masson's trichrome stain to observe morphological changes and fibrosis. Western blot was used to determine mitochondrial homeostasis regulation-related proteins and mitophagy pathway. Results TEM and X-ray photoelectron spectroscopy characterized the properties successfully. MTT assay shows that HL-1 cell viabilities were not influenced by CeO2@Tf at 200 µM.High-glucose treatment led to increased ROS and decreased intracellular MMP in HL-1 cells. DM mice cardiac transcriptome sequencing results showed upregulation of transferrin receptor gene expression. Echocardiographic showed increased LAD and abnormal diastolic and systolic functions. All these abnormal changes were improved in the DM+ Met and DM+NPs groups. Our investigations revealed proteins linked to the PINK1-Parkin signaling pathway (Figure 2A-B) and found that the level of mitophagy in the diabetic group was overly high, which was improved after the application of CeO2@Tf. We observed the mitochondrial autophagosomes through transmission electron microscopy (Figure 2C). The autophagosomes in the diabetic group increased significantly and improved after treatment with CeO2@Tf. Conclusions These consistent biochemical and histopathological results suggest that the biomimetic mineralization of CeO2@Tf could inhibit oxidative stress and improve mitochondrial function in vitro induced by high-glucose and in vivo caused by DM, alleviating cardiac remodeling in DM mice.

Abstract 176: The Pathological Role of Aldose Reductase in Isolated Cardiomyocytes Undergoing Hypoxia / Reoxygenation After Prior Exposure to High Glucose Concentrations

Hyperglycemia increases the vulnerability of the cardiovascular system to ischemia/reperfusion (IR) injury. There are currently no effective therapeutic strategies. We hypothesized that hyperglycemia would increase glucose flux through the polyol pathway. Aldose reductase (AR) catalyzes the first step of this pathway by converting glucose to sorbitol. This reaction generates more oxidative stress due to the depletion of NADPH and osmotic stress caused by sorbitol accumulation, making the heart more susceptible to IR injury. This study aimed to better understand the injury mechanism in cardiomyocytes after high glucose exposure and IR injury. Confluent cardiomyocytes were exposed for 24 hours to 25, 50, or 100 mM glucose or 25 mM glucose with 25 or 75 mM sucrose concentrations added. The non-metabolizable sugar sucrose was used as an osmolarity control. Epalrestat, an AR inhibitor, was added at 0, 1, or 10 μM per well (n = 18 per group) during glucose or sucrose exposure. Cells were then subjected to 24 hours of hypoxic conditions (glucose-, sucrose- and serum-free media; 0.01% O 2 , 37 °C; hypoxic chamber) or normoxic conditions (complete media; 21% O 2 , 37 °C), followed by a 2-hour reoxygenation period in normoxic conditions. Cell injury was quantified by measurement of lactate dehydrogenase (LDH) release. Data were normalized to 25 mM glucose after control/normoxic conditions without AR inhibitor. A three-way ANOVA was conducted to compare groups, followed by the Holm-Šidák test. Increasing glucose concentrations lead to increased cell damage after hypoxia/reoxygenation. Adding the AR inhibitor during the glucose exposure period reduced LDH release after hypoxia/reoxygenation in a dose-dependent manner. The addition of sucrose did not increase cell injury. The AR inhibitor did not affect cells exposed to sucrose. The polyol pathway may play a dominant role in increased cell injury by hyperglycemia after hypoxia/reoxygenation.

Dose-dependent anti-hyperglycemic & anti-dyslipidemic potential of aqueous leaves extract of Typha elephantina in-vivo and in-vitro

Diabetes mellitus is among the fundamental causes of illness and millions of deaths around the globe are directly attributed to it each year. Current antidiabetic medications often lack sustained glycemic control and carry significant risks of side effects. As a result, the use of plant-based treatments has gained popularity. In this experimental study, we evaluated the aqueous extracts (LQE) of Typha elephantina (also known as Elephant grass) leaves collected from freshwater marshes, for their potential anti-hyperglycemic and anti-hyperlipidemic antioxidant effects in healthy streptozotocin caused diabetic-mice. We employed glucose adsorption tests at different glucose levels and glucose diffusion tests to assess the in-vitro antidiabetic action of plant extract. For the in-vivo trail, we measured fasting blood glucose (FBG), glucose tolerance (GTT), as well as long-term anti-diabetic, anti-hyperlipidemic, and antioxidant activities. Our results from the glucose diffusion test indicated that the extract was highly effective at both low glucose concentrations (5 mmol L) and high glucose concentrations (100 mmol L). However, the glucose-diffusion ability reached its peaked at an excessively high dosage of the aqueous extract, suggesting a dose-related effect. Similarly, we observed that high doses of TEL.AQ extracts (400 mg/kg body weight) significantly reduced blood glucose levels in healthy mice during the glucose tolerance test (GTT) at 3 h and fasting blood glucose studies (FBG) at 6 h. Furthermore, the high-dose TEL.AQ extract effectively reduced liver-related serum markers and blood-glucose concentration (BGC) in severely chronic diabetic rats. The extract dosage also influenced lipid profile, conjugate and unconjugated bilirubin levels, cholesterol, triglycerides, HDL, and total bilirubin levels. Additionally, after administering a high extract dose, we observed considerable improvement in the liver homogenate markers CAT, POD, and SOD. In contrast, the extract at a low dosage(100 mg/kg),showed minimal, while a moderate dose (200 mg/kg), yielded promising results.

Colorimetric detection of glucose with smartphone-coupled µPADs: harnessing machine learning algorithms in variable lighting environments

Recent advances in microfluidic paper-based analytical devices (µPADs) have shown immense potential for point-of-care testing (POCT) in resource-constrained settings, especially for monitoring glucose levels in patients with diabetes. However, the practical adoption of smartphone-coupled µPADs has been limited due to the variability in ambient lighting conditions, which affects the colorimetric detection. In this study, we have developed a machine learning-based algorithm together with a flash/no-flash technic, adaptable to various µPADs and capable of accurately monitoring plasma glucose levels across a wide range (25 µM to 30 mM), addressing issues related to environmental illumination variations, and shooting angles.The algorithm is implemented in an Android application, Gluco estimator. It utilizes images captured by smartphones and incorporates intelligent feature engineering. The XYZ color space is adopted for device-independent analysis. KI and TMB color indicators are used to enhance resolution for low and high glucose concentrations, respectively. The EBC classifier, combined with a handcrafted feature set, exhibits outstanding performance, achieving an accuracy of 95% and 91% for TMB and KI, respectively. Additionally, multiple linear regression (MLR) yields high reliability among regression models, with R2 values of 0.95 at 0.25 µM to 3 mM and 0.97 at 3–30 mM. This work represents a significant stride toward realizing a portable, high-resolution, and reliable smartphone-coupled µPAD in uncontrolled environments.

Effect of shortening the dry period on colostrum and milk quality, blood parameters and some reproductive parameters in high-producing Holstein cows of different body condition score.

This study aimed to evaluate the effect of shortening the dry period in high-yielding cows of different body condition scores (BCS). We report colostrum and milk quality, some serum metabolites, BCS changes, and some reproductive parameters with measurements being made over the first two months of lactation. Cows were grouped based on the length of the dry period (normal: about 50 d and short: about 28 d) and BCS (moderate: 2.75 to 3.5 and high ≥ 3.5). Short dry period decreased colostrum volume and, in combination with high BCS only, caused a decrease in milk production. Short dry period moderate BCS cows had the highest serum insulin concentration on day 14 after calving and highest glucose concentration on day 28, but neither differed significantly when measured over the whole period. By contrast, short dry period cows had significantly lower concentrations of non-esterified fatty acids and beta-hydroxybutyrate measured over the whole period. Post-partum loss of BCS was less in short and especially so in the short, moderate BCS group. Following a synchronization protocol at 35 d postpartum. The cows with a short dry period and moderate BCS had lower open days, days to first postpartum estrus and services per conception. It was concluded that short dry periods and moderate BCS had a positive influence on serum metabolites, BCS changes and reproductive parameters.

Periodontitis therapy with iron alginate and glucose oxidase based on the Fenton reaction

Periodontitis is a frequent chronic inflammatory disease brought on by bacterial infection in the biofilm that forms on the teeth. It can cause the gingival recession and tooth loss, gravely jeopardizing the oral health of people. Current therapies still suffer from issues like bleeding, pain, injured gums, limited treatment effectiveness, a lengthy treatment cycle, and bacterial resistance when using a combination of surgical intervention and antibiotic therapy. Herein, a simple one-step cross-linking process was used to create glucose oxidase (GOD) loaded upon iron alginate (FeAlg/GOD), which has a strong anti-biofilm activity, good biocompatibility, and was successfully used to treat periodontitis caused by biofilms. The results demonstrated that in the presence of a high glucose concentration in the gingival crevicular fluid, FeAlg/GOD could catalyze the conversion of glucose into gluconic acid and hydrogen peroxide. The produced hydrogen peroxide could convert Fe3+ to Fe2+ when it got to the deep periodontal pockets. Following that, the Fenton reaction was activated, producing large amounts of hydroxyl radical (OH). In conclusion, oral plaque biofilms were removed by the FeAlg/GOD. Additionally, successful in vivo treatment was accomplished in rat's experimental periodontitis models, as evidenced by a markedly reduced inflammatory response, decreased bone loss, and made healthier gum tissue. Overall, our biocompatible and antibacterial FeAlg/GOD study presents a novel strategy for effectively speeding periodontal tissue repair and eliminating oral pathogens to tackle the grave threat of biofilm-associated illnesses.

The concentration of glucose in the media influences the susceptibility of stallion spermatozoa to ferroptosis.

Although common in many commercial extenders, supraphysiological concentrations of glucose in the media may be detrimental to stallion spermatozoa. In this study, we present evidence that these elevated glucose levels may predispose spermatozoa to ferroptosis. Stallion spermatozoa depend on oxidative phosphorylation as their major source of ATP; however, the metabolism of these cells is complex and a great degree of metabolic plasticity exists. The composition of the media in which the spermatozoa are extended, or exposed to in the mare's reproductive tract, exerts a profound effect on sperm function and may even accelerate cell demise. Recent research indicates that high concentrations of glucose in the media, although common in commercial extenders, may be detrimental. To determine if supraphysiological glucose concentration may induce or predispose to ferroptosis (a caspase-independent form of programmed cell death, triggered by oxidative stress), stallion spermatozoa were incubated under different concentrations of glucose, 67 mM (HG) or 1 mM plus 10 mM pyruvate (LG-HP), in the presence or absence of known inductors of ferroptosis. Furthermore, we developed a single-cell flow metabolic assay to identify different metabolic phenotypes in spermatozoa. Storage and incubation of spermatozoa under high glucose concentrations led to an increase in the percentage of necrotic spermatozoa (P < 0.0001). Moreover, ferroptosis was induced more intensely in sperm in media with high glucose concentrations (P < 0.0001). Finally, we observed that induction of ferroptosis modified two proteins (oxoglutarate dehydrogenase and superoxide dismutase 2) in spermatozoa incubated in media containing 67 mM glucose but not in media containing 1 mM glucose and 10 mM pyruvate. The composition of the media, especially the concentration of glucose, exerts a major impact on the functionality and life span of the spermatozoa. The results reported here may pave the way for improvements in existing semen extenders.

Betaine regulates steroidogenesis, endoplasmic reticulum stress response and Nrf2/HO-1 antioxidant pathways in mouse Leydig cells under hyperglycaemia condition

We studied the effects of betaine on steroidogenesis, endoplasmic reticulum stress and Nrf2 antioxidant pathways of mice Leydig cells under hyperglycaemia conditions. Leydig cells were grown in low and high glucose concentrations (5 mM and 30 mM) in the presence of 5 mM of betaine for 24 h. Gene expression was determined using a real-time PCR method. The protein levels were determined by Western blot analysis. The testosterone production was evaluated by the ELISA method. Cellular contents of reduced and oxidised glutathione were measured by colorimetric method. Hyperglycaemia caused impaired steroidogenesis and ERS in Leydig cells associated with the down-regulation of 3β-HSD, StAR, P450scc, LH receptor and increased expression of GRP78, CHOP, ATF6 and IRE1. Betaine could improve cell viability, attenuate the ERS, and restore testosterone production in Leydig cells under hyperglycaemia conditions. Betaine can protect Leydig cells against the adverse effects of hyperglycaemia by regulating steroidogenesis, antioxidants, and ERS.

Krüppel-like factor 4 promotes autophagy in macrophages under high glucose concentration by inhibiting the AKT/mTOR signaling pathway

Background: Diabetic atherosclerosis (AS) is the main cause of disability and death in diabetes. In the progression of AS, autophagic activity plays an important role. Krüppel-like factor 4 (KLF4) is a member of the zinc finger protein transcription factor family and is believed to play a protective role in the pathogenesis of atherosclerosis. This study aimed to explore the role of KLF4 in diabetic atherosclerosis and the autophagic mechanism. Methods: A diabetic mouse model was established and the expression level of KLF4 protein in the aorta of the mice was detected after a high-fat diet. The effects of KLF4 on cholesterol content, apoptosis, autophagy-related proteins, and the AKT/mTOR signaling pathway of THP-1 macrophages were also evaluated. Results: The expression level of KLF4 protein in the aorta of diabetic mice was decreased. Meanwhile, overexpression of KLF4 in THP-1 macrophages significantly decreased cholesterol accumulation, increased beclin-1 expression, decreased P62 expression, enhanced LC3 fluorescence intensity decreased cell apoptosis and p-mTOR and p-AKT expression were decreased under the condition of high glucose. After the reduction of KLF4 expression, the result is reversed. Conclusion: KLF4 induces autophagy by inhibiting the AKT/mTOR pathway and alleviates cholesterol deposition in THP-1 macrophages under high glucose concentration.

OYSTER MUSHROOM (PLEUROTUS OSTREATUS) ETHANOLIC EXTRACT PROTECTS YEAST CELLS FROM HYPER- AND HYPO-GLYCEMIC STRESS

Glucose is the primary source of carbon and energy. Cells possess a sophisticated mechanism for sensing glucose and responding to it appropriately. However, hyperglycemia represents the predominant risk factor for the development of diabetes. On the other hand, nutrient starvation, including glucose, may lead to the development of malnutrition. Noteworthy, glucose restriction has been associated with significantly delayed aging. Thus, controlled glucose intake is vital for healthy living. The presented study shows the effect of the oyster mushroom Pleurotus ostreatus ethanolic extract on fission yeast Schizosaccharomyces pombe undergoing glucose-derived stress. Fruiting bodies of P. ostreatus were used for ethanolic extraction and its anti-hyper- and hypo-glycemic stress activity was investigated. Compared were cells treated with ethanolic extract of P. ostreatus (1 or 2%) with untreated control. Different glucose concentrations in growth media were used to analyze its impact on growth intensity, generation time (gt), metabolic activity, ROS generation, MDA content, and cell death and apoptosis of S. pombe cells. Glucose deprivation or its over-supplementation resulted in growth retardation and prolonged cell doubling time (gt) which was significantly adjusted by the oyster mushroom extract. Interestingly, oyster mushroom extract reduced excessive ROS and formed MDA that were produced as a consequence of glycemic stress. Unexpectedly, the extract was able to increase the metabolic activity of cells supplemented with high glucose content and thereby protected them from death. Taken together, our study suggests that growth media supplementation with the ethanolic extract of P. ostreatus protects S. pombe cells from glucose-mediated stress through alleviation of oxidative stress and enhancement of metabolic activity.

Evaluating the potential of semi-continuous itaconic acid fermentation by Aspergillus terreus: operational profile and experiences

Itaconic acid is an important bio-based chemical. The present study aims to evaluate the applicability of semi-continuous fermentation technique for itaconic acid production by Aspergillus terreus. The fermentation is planned to be connected with bipolar membrane electrodialysis unit for acid recovery. This process allows the reuse of residual glucose from the effluent. Our particular attention was focused on the effect of glucose concentration. Two different glucose supplementation strategies were tested: constant glucose concentration in the refilling medium and adjusted glucose concentration in order to maintain a continuously high – 120 g/L – glucose concentration in the fermentor. The itaconic acid titre, yield and productivity for the 24 h time periods between draining/refilling interventions were investigated. The constantly high glucose concentration in the fermentor resulted in doubled biomass formation. The average itaconic acid titre was 32.9 ± 2.7 g/L. The producing strain formed numerous spores during semi-continuous fermentation that germinated continuously. Yield and volumetric productivity showed a periodic pattern during the procedure.

Global and Spatial Metabolomics of Individual Cells Using a Tapered Pneumatically Assisted nano-DESI Probe.

Single-cell metabolomics has the potential to reveal unique insights into intracellular mechanisms and biological processes. However, the detection of metabolites from individual cells is challenging due to their versatile chemical properties and concentrations. Here, we demonstrate a tapered probe for pneumatically assisted nanospray desorption electrospray ionization (PA nano-DESI) mass spectrometry that enables both chemical imaging of larger cells and global metabolomics of smaller 15 μm cells. Additionally, by depositing cells in predefined arrays, we show successful metabolomics from three individual INS-1 cells per minute, which enabled the acquisition of data from 479 individual cells. Several cells were used to optimize analytical conditions, and 93 or 97 cells were used to monitor metabolome alterations in INS-1 cells after exposure to a low or high glucose concentration, respectively. Our analytical approach offers insights into cellular heterogeneity and provides valuable information about cellular processes and responses in individual cells.

GIP_HUMAN [22-51] Peptide Encoded by the Glucose-Dependent Insulinotropic Polypeptide (GIP) Gene Suppresses Insulin Expression and Secretion in INS-1E Cells and Rat Pancreatic Islets.

GIP_HUMAN [22-51] is a recently discovered peptide that shares the same precursor molecule with glucose-dependent insulinotropic polypeptide (GIP). In vivo, chronic infusion of GIP_HUMAN [22-51] in ApoE-/- mice enhanced the development of aortic atherosclerotic lesions and upregulated inflammatory and proatherogenic proteins. In the present study, we evaluate the effects of GIP_HUMAN [22-51] on insulin mRNA expression and secretion in insulin-producing INS-1E cells and isolated rat pancreatic islets. Furthermore, we characterize the influence of GIP_HUMAN [22-51] on cell proliferation and death and on Nf-kB nuclear translocation. Rat insulin-producing INS-1E cells and pancreatic islets, isolated from male Wistar rats, were used in this study. Gene expression was evaluated using real-time PCR. Cell proliferation was studied using a BrdU incorporation assay. Cell death was quantified by evaluating histone-complexed DNA fragments. Insulin secretion was determined using an ELISA test. Nf-kB nuclear translocation was detected using immunofluorescence. GIP_HUMAN [22-51] suppressed insulin (Ins1 and Ins2) in INS-1E cells and pancreatic islets. Moreover, GIP_HUMAN [22-51] promoted the translocation of NF-κB from cytoplasm to the nucleus. In the presence of a pharmacological inhibitor of NF-κB, GIP_HUMAN [22-51] was unable to suppress Ins2 mRNA expression. Moreover, GIP_HUMAN [22-51] downregulated insulin secretion at low (2.8 mmol/L) but not high (16.7 mmol/L) glucose concentration. By contrast, GIP_HUMAN [22-51] failed to affect cell proliferation and apoptosis. We conclude that GIP_HUMAN [22-51] suppresses insulin expression and secretion in pancreatic β cells without affecting β cell proliferation or apoptosis. Notably, the effects of GIP_HUMAN [22-51] on insulin secretion are glucose-dependent.

Alta producción de ácido cítrico en fermentación en estado sólido por Aspergillus brasiliensis sobre espuma de poliuretano

A statistical approach based on fractional and complete factorial designs to increase the citric acid yield was used. Polyurethane foam (PUF) was used as inert support for solid-state fermentation (SSF) due to its high-water retention capacity (up to 60-fold of its weight), resulting in the recovery of concentrated citric acid by squeezing it. The maximum yields for citric acid (0.62 Cmol/Cmol glucose) and polyols (0.11 Cmol/Cmol glucose) are obtained by growing Aspergillus brasiliensis in a well-defined culture medium (at 30°C for 120 h). Citric acid, polyols, and glucose consumption are negatively affected by a high glucose concentration (250 g/L). In contrast, enrichment of the culture medium with yeast extract only enhances citric acid production yield (0.54 Cmol/Cmol glucose). This study demonstrates that the citric acid production yield is negatively affected by phosphate (&gt; 4.54 g/L) and magnesium (&gt; 0.26 g/L). The PUF for SSF is an inexpensive and useful support for the commercial production of citric acid. The utilization of low-cost inert supports through SSF can contribute to achieving industrially viable and sustainable citric acid production. As far as we know, this work reports the highest citric acid production yield using SSF on an inert support such as PUF soaked with a defined medium.

Isoscutellarein from Bixa orellana Leaves Uncompetitively Inhibits Recombinant Human Aldose Reductase and Prevents Sorbitol Accumulation and Lens Opacity

Introduction: Enhanced aldose reductase activity results in increased accumulation of sorbitol. Therefore, inhibition of aldose reductase is an effective strategy to prevent or delay certain diabetic complications. Method: Various extracts of the leaves of B. orellana were tested for their inhibitory activity on the aldose reductase. Ethyl acetate extract that showed maximum inhibition was further fractionated and the inhibitor was identified as isoscutellarein by spectroscopic methods. IC50 of recombinant human aldose reductase by isoscutellarein was found to be 14 μM and the mode of inhibition was uncompetitive with a decrease in both Km and Vmax. Isoscutellarein was bound to the active site of aldose reductase (3RX3), namely to Ala-299, Leu-300, Leu-301, His-110 and Tyr-48. Result: Docking results exhibited a binding energy of -9.15 kJ/mol. Conclusion: The incubation of red blood cells with high glucose concentrations mimicking hyperglycemic conditions promoted sorbitol accumulation, which was effectively inhibited by isoscutellarein. Further, xylose-induced opacity of the lens was effectively inhibited by isoscutellarein.

Quality evaluation of therapeutic noodle from water yam, millet and wheat composite flour and its effect on lipid profile and blood glucose concentration using albino rats

This study investigated the development and sensory qualities of therapeutic noodle from wheat, pearl millet (Pennisetum glaucum) and water yam composite flour and the effect of feeding the product on lipid profile and blood glucose concentration using alloxan induced albino rats. The therapeutic noodle was produced using different ratios of flour blends of wheat, millet and water yam. Sensory analysis was carried out based on color, taste, aroma, slipperiness and overall acceptability on the basis of a 9-point hedonic scale. The therapeutic noodle samples competed well with the commercial noodle (control) sample as they were all rated above average (&gt;5.0) with noodle sample (WMF5) having a very high score of 8.40 in the overall acceptability. Alloxan induced male albino rats were fed the formulated therapeutic noodle for 21 days and assessed for their resultant body weight, blood glucose concentration and lipid profiles. The total cholesterol levels of all diabetic albino rats fed the formulated therapeutic noodle were reduced to within the normal range of below 200mg/dl ideal for adults after the 21days. The therapeutic noodles also helped to significantly (p &lt; 0.05) reduce the high blood glucose concentration of the diabetic rats to below 200mg/dl while diabetic rats fed control diet (100:0) had the highest concentration of blood glucose (485mg/dl). Due to the high anti-diabetic properties of the therapeutic noodles, it can be recommended as a functional food source for the treatment of overweight, obesity, and diabetes.

Blockage of DCLK1 in cardiomyocytes suppresses myocardial inflammation and alleviates diabetic cardiomyopathy in streptozotocin-induced diabetic mice

Diabetic cardiomyopathy (DCM) is a pathophysiological condition triggered by diabetes mellitus and can lead to heart failure. Doublecortin-like kinase protein 1 (DCLK1) is a multifunctional protein kinase involved in the regulation of cell proliferation, differentiation, survival, and migration. Current studies on DCLK1 mainly focus on cancer development; however, its role in non-tumor diseases such as DCM is yet to be deciphered. Our analysis revealed that DCLK1 was upregulated in cardiomyocytes of streptozotocin (STZ)-induced type 1 diabetic mouse, suggesting a correlation between DCLK1 and DCM progression. It was further demonstrated that either cardiomyocyte-specific DCLK1 knockout or pharmacological DCLK1 inhibitor DCLK1-IN-1 significantly alleviated cardiac hypertrophy and fibrosis in STZ-induced diabetic mice. RNA-seq analysis of heart tissues revealed that DCLK1 regulated the nuclear factor kappa B (NF-κB)-mediated inflammatory response in DCM. In vitro, DCLK1 activated NF-κB and the inflammatory response by inducing the IKKβ phosphorylation in high-concentration glucose (HG)-challenged cardiomyocytes. DCLK1-IN-1 also prevented HG-induced IKKβ/NF-κB activation and inflammatory injuries in cardiomyocytes. In conclusion, this study highlights the novel role of cardiomyocyte DCLK1 in regulating IKKβ/NF-κB, which aggravates inflammation to promote the pathogenesis of DCM. DCLK1 may serve as a new target for DCM treatment.

Effect of co-mingling non-littermates during lactation and feed familiarity at weaning on the performance, skin lesions and health of piglet

This study investigated how co-mingling of non-littermates before weaning and feed familiarity at weaning in a 2 × 2 factorial design can minimize the adverse effects related to conventional weaning. Before weaning, piglets were either conventionally reared or co-mingled by grouping piglets from 3 litters during 10 days before weaning. At weaning, piglets from 3 litters were distributed over 3 pens in the nursery. Conventionally reared piglets from 3 litters were mixed, whereas the co-mingled piglets were housed with those they were co-mingled with before weaning. At weaning, piglets were either or not provided with 0.538 kg creep feed per piglet in creep feeders, next to the weaner diet. Creep feed was the same provided the last 4 days before weaning. Hence, the treatment groups were conventional with (n = 9) or without creep feed (n = 9), and co-mingling with (n = 9) or without creep feed (n = 9). Co-mingling non-littermate piglets did not affect weaning weight, daily weight gain and creep feed intake during d-10-d0 before weaning. Co-mingling resulted in higher skin lesion scores of head and ears in piglets at d1 before weaning (P < 0.05), but in lower skin lesion scores at the snout, shoulders and flanks at d2 post-weaning compared to conventionally reared piglets (P < 0.05). Co-mingling did not affect salivary cortisol at d1 post-weaning . Hematocrit, hemoglobin, mean corpuscular volume and mean corpuscular hemoglobin were higher in the co-mingled piglets at d2 post-weaning (P < 0.05). Co-mingling resulted in higher glucose concentrations in blood (P < 0.05) and in a tendency for lower NEFA concentration in serum at d2 post-weaning (P < 0.10). Feed familiarity at weaning increased average daily feed intake and weight gain during the first 5 days post-weaning (P < 0.05). During d5–14 post-weaning average daily feed intake tended to be lower (P < 01.0) and daily gain was lower (P < 0.05) in creep feed fed piglets compared to no creep feed fed piglets. Nevertheless, feed familiarity did not affect overall piglet performance during the first 2 weeks after weaning. Feed familiarity did not affect blood hematology and biochemical blood parameters, except the level of basophils at d2 post-weaning. Neither housing nor feeding strategy affected piglet performance between d14–42 post-weaning and ear and tail damage at d42 post-weaning . In conclusion, pre-weaning co-mingling of piglets and feed familiarity at weaning each contributed to a better adaptation of the weaning process but did not affect long-term piglet performance.

ВЛИЯНИЕ РАЗЛИЧНЫХ КОНЦЕНТРАЦИЙ ГЛЮКОЗЫ В ПИТАТЕЛЬНОЙ СРЕДЕ НА МОРФОГЕНЕЗ РАСТЕНИЙ ВИНОГРАДА IN VITRO

To study the organic nutrition of grape plants in in vitro culture, we conducted research on the effect of glucose introduced into the nutrient medium on the morphogenesis of test-tube grape plants. The experiments were carried out on two model varieties (Sibirkovyi and Krasnostop Karpi) selected by us from the laboratory collection of in vitro healthy grape plants. These varieties, in addition to differences in genotypes, most noticeably differ in the features of development in in vitro culture, which we noted earlier when they were cultivated and deposited in the collection. The Sibirkovyi variety is one of the fastest growing, and Krasnostop Karpi is one of the slowest developing varieties in in vitro culture. In the experiment, glucose concentrations from 0.0 (sucrose control 10.0 g/l) to 40.0 g/l were used. The varietal response of the model varieties in the experiment with different concentrations of glucose in the nutrient medium was different. The variety Sibirkovyi was already one and a half times ahead of the variety Krasnostop Karpi in development in the first terms of accounting. At the same time, for the Sibirkovyi variety, the optimal range of concentrations for the development and preservation of microplants according to most indicators was the range from 5 to 20 g/l, and for the Krasnostop Karpi variety, the optimal range was narrower – 0-20 g/l. At high glucose concentrations of 30,0-40,0 g/l, the plants of both varieties experienced stress, which was manifested in the intensive release of phenols into the nutrient medium (this was most clearly manifested in the Sibirkovyi variety), and often the plants did not reach the top of the cultivation vessel during ontogenesis. besides, they showed earlier aging processes. At the same time, at high concentrations of glucose, a slowdown in growth processes and its ripening of shoots in individual microplants was noted.

Fabrications of the Flexible Non-Enzymatic Glucose Sensors Using Au-CuO-rGO and Au-CuO-rGO-MWCNTs Nanocomposites as Carriers.

A flexible, non-enzymatic glucose sensor was developed and tested on a polyethylene terephthalate (PET) substrate. The sensor's design involved printing Ag (silver) as the electrode and utilizing mixtures of either gold-copper oxide-modified reduced graphene oxide (Au-CuO-rGO) or gold-copper oxide-modified reduced graphene oxide-multi-walled carbon nanotubes (Au-CuO-rGO-MWCNTs) as the carrier materials. A one-pot synthesis method was employed to create a nanocomposite material, consisting of Au-CuO-rGO mixtures, which was then printed onto pre-prepared flexible electrodes. The impact of different weight ratios of MWCNTs (0~75 wt%) as a substitute for rGO was also investigated on the sensing characteristics of Au-CuO-rGO-MWCNTs glucose sensors. The fabricated electrodes underwent various material analyses, and their sensing properties for glucose in a glucose solution were measured using linear sweep voltammetry (LSV). The LSV measurement results showed that increasing the proportion of MWCNTs improved the sensor's sensitivity for detecting low concentrations of glucose. However, it also led to a significant decrease in the upper detection limit for high-glucose concentrations. Remarkably, the research findings revealed that the electrode containing 60 wt% MWCNTs demonstrated excellent sensitivity and stability in detecting low concentrations of glucose. At the lowest concentration of 0.1 μM glucose, the nanocomposites with 75 wt% MWCNTs showed the highest oxidation peak current, approximately 5.9 μA. On the other hand, the electrode without addition of MWCNTs displayed the highest detection limit (approximately 1 mM) and an oxidation peak current of about 8.1 μA at 1 mM of glucose concentration.