Search Results (560)

Hedgehog (Hh) signaling regulates cell survival and microenvironmental responses in various cancers, but its role in metabolic adaptation to the hypoxic bone marrow microenvironment in myeloid malignancies remains insufficiently defined. K562, KU812, and U937 cells were cultured under normoxic or hypoxic conditions and treated with the Smoothened (Smo) antagonist Cyclopamine or the agonist SAG (Smoothened Agonist). Cell proliferation, apoptosis- and autophagy-related markers, glycolysis-associated proteins, and metabolic parameters were assessed by viability assays, Western blotting, immunofluorescence, and biochemical assays. Selected findings were further evaluated in CRISPR/Cas9-mediated Smo knockout cells. Smo inhibition reduced cell proliferation, increased PARP cleavage, and decreased BNIP3 expression under both oxygen conditions. It also downregulated glucose transporter 1 (GLUT1), hexokinase 2 (HK2), lactate dehydrogenase (LDH), monocarboxylate transporter 1 (MCT1), carbonic anhydrases IX and XII (CAIX and CAXII), and was associated with reduced glucose consumption, lactate production, and ATP levels. These changes were associated with modulation of the AMPK–mTOR axis and with reduced phosphorylation of mTOR downstream effectors. These findings support a role for Hh signaling in hypoxia-driven metabolic adaptation in myeloid malignancy cell models and suggest a functional link between Smo activity, glycolytic remodeling, and AMPK-mTOR-related signaling. Full article

Ethanol consumption can modify both drug exposure and drug response. However, the clinical relevance of these interactions depends strongly on the timing and pattern of alcohol intake, the affected pharmacological pathway, the dosage form and organ reserve. This review summarizes current evidence on ethanol–drug interactions, particularly human crossover studies, phenotyping studies, cohort analyses and appropriate case reports. It distinguishes acute ethanol–drug co-exposure, chronic alcohol exposure, drug use during early abstinence after chronic drinking, and pharmacotherapy in alcohol-associated liver disease. Key mechanisms include ADH- and ALDH-dependent ethanol oxidation, acetaldehyde formation, NADH/NAD⁺ redox shift, CYP2E1 induction, carboxylesterase 1 (CES1) modulation, altered intestinal and hepatic first-pass handling, dose dumping from susceptible modified-release products, changes in protein binding in alcohol-associated liver disease, and ALDH inhibition with acetaldehyde accumulation in disulfiram-like reactions. At the molecular level, ethanol may promote acetaldehyde adduct formation with proteins and DNA, CYP2E1-driven reactive oxygen species generation, redox stress, intestinal barrier injury, and CES1-dependent transesterification of selected ester drugs. Acute ethanol intake mainly increases pharmacodynamic toxicity and causes short-term pharmacokinetic disturbances, including enhanced central nervous system depression, delayed gastric emptying, impaired glucose and lactate handling and altered hemodynamic responses. In contrast, chronic exposure, early abstinence and alcohol-associated liver disease are more often associated with hepatic enzyme and transporter remodeling, altered protein binding, reduced hepatic or renal reserve, and greater susceptibility to drug-related organ injury. The highest-risk scenarios involve older adults, polypharmacy, alcohol-associated liver disease, dehydration or acute illness, early abstinence, and the concurrent use of central nervous system depressants, glucose-lowering drugs, NSAIDs, antihypertensives, renally eliminated drugs or warfarin. Hence, ethanol exposure should be treated as a dynamic, context-dependent modifier factor that can acutely exacerbate pharmacodynamic toxicity, alter selected pharmacokinetic pathways and lower organ tolerance to drug-related injury. Full article

Background/Objectives: Glucose transporter type 1 deficiency syndrome (GLUT1DS) is a rare neurometabolic disorder with an expanding clinical spectrum, including mild and non-classical presentations. We report a boy with severe transient neonatal hyperkalemia, bilateral congenital cataracts, and later subtle neurological and neurocognitive symptoms, in whom genomic testing supported the diagnosis of mild GLUT1DS. Methods: This single-patient case report describes clinical follow-up from birth to nine years of age, including neurological, metabolic, neuropsychological, imaging, and genetic investigations. Whole-exome sequencing using next-generation sequencing technology was performed. Results: The patient required intensive care immediately after birth because of severe transient hyperkalemia of unclear etiology. Bilateral congenital cataracts were surgically corrected during infancy. Later, he developed two brief seizure episodes, reduced exercise tolerance, episodic fatigue, attentional difficulties, motor restlessness, and mild graphomotor impairment. Neuropsychological assessment showed overall average intellectual functioning, below-average verbal abilities, low-average non-verbal abilities, and attention-deficit/hyperactivity disorder. Repeated metabolic investigations, electroencephalography, and brain magnetic resonance imaging were unrevealing. Whole-exome sequencing identified an apparently de novo heterozygous SLC2A1 variant, NM_006516.4.1333_1335del, p.(Phe445del), supporting the diagnosis of mild GLUT1DS. Because of the mild phenotype and preserved everyday functioning, ketogenic diet therapy was not initiated. Conclusions: This case highlights the diagnostic challenges of mild GLUT1DS and the value of genomic testing in children with unexplained neurological or neurocognitive symptoms despite normal routine investigations. Although neonatal hyperkalemia and GLUT1DS coexisted in this patient, current evidence is insufficient to establish a causal relationship. Full article

The prevalence of diabetes has increased considerably in recent decades, representing a global health problem. Although several pharmacological approaches for the treatment of diabetes are available, it is pertinent to explore more effective alternatives with reduced adverse effects. In this work, the hypoglycemic activity of rhamnolipids was studied using the chicken embryo (in ovo) model. The results obtained demonstrated that rhamnolipids produced by Pseudomonas aeruginosa #112 and commercial rhamnolipids (RL-90) significantly reduced blood glucose levels of chicken embryos on embryonic day 11 (from 135 ± 11 mg/dL to 94–107 mg/dL). These values were similar to those achieved with human insulin (104 ± 20 mg/dL) and the rapid-acting human insulin analog FIASP^® (95 ± 18 mg/dL). It was also verified that rhamnolipids did not have a negative effect on chicken embryo development at the concentrations tested. Regarding the molecular mechanisms involved in the decrease in blood glucose levels, for both insulins, a reduction in the expression of genes encoding the glucose transporter 2 (glut2) and the gluconeogenic enzymes phosphoenolpyruvate carboxykinase 2 and fructose-1,6-biphosphatase 1 was observed. However, in the case of rhamnolipids, only a reduction in the expression of glut2 was observed. According to the results obtained, rhamnolipids are potential candidates for further studies on the development of new alternative treatments for diabetes symptoms. Full article

Gastroenteropancreatic neuroendocrine tumors (GEP-NETs) are characterized by substantial biological heterogeneity and complex regulation of apoptosis, metabolism, and angiogenesis. The aim of this study was to evaluate the concentrations of selected proteins: FGF2, HGF, Fas/CD95, CASP9, ALDH1A1, and GLUT1 in tumor and margin samples and assess correlations with clinical and demographic parameters. A total of 59 samples from patients with GEP-NETs were analyzed using multiplex immunoassay and ELISA methods. Significant differences in protein expression between tumor and margin tissues were observed. Fas/CD95 levels were lower in tumor samples, whereas HGF concentration was higher. Elevated HGF, FGF2 and Fas/CD95 levels were associated with advanced tumor stage. HGF and GLUT1 concentrations varied depending on nodal status, while FGF2, Fas/CD95, and CASP9 levels were increased in metastatic cases. Additionally, differences related to tumor localization and the influence of smoking and alcohol consumption were identified. Dysregulation of apoptotic, metabolic, and angiogenic pathways plays a crucial role in GEP-NETs progression and highlights the importance of the tumor microenvironment. GEP-NET exhibit biological heterogeneity and complex progression driven by multiple interacting molecular pathways. The factors analyzed may have potential significance as biomarkers of disease progression; however, their exact role requires further investigation in larger, prospective cohorts. Full article

Age-related hyperphosphatemia is increasingly recognized as a contributing factor in sarcopenia. This work studies the metabolic effects of elevated phosphate on muscle. C2C12 cells were differentiated in the absence or presence of 10 mM β-glycerophosphate (BGP), an exogenous phosphate donor. In addition, quadriceps muscles from four experimental groups of male C57BL/6J mice were analyzed: young (5 months) and old (24 months) fed with standard diet; old mice fed with hypophosphatemic diet or supplemented with the phosphate binder Velphoro^®, for the last three months of life. Mice were stratified according to sarcopenia degree based on muscle mass, strength and physical performance. Protein levels were determined by immunoblotting and mRNA expression by RT-qPCR. ATP levels were measured by luminescence and L-lactate production, citrate synthase and cytochrome c oxidase activities by colorimetric assays. Mitochondrial content, membrane potential and reactive oxygen species (ROS) were determined by fluorescence assay. BGP-treated cells showed increased glucose transporter 1 (GLUT1) and decreased NADH Dehydrogenase (CI-NDUFB8) protein expression, elevated hexokinase II (HK2), phosphoglycerate kinase 1 (PGK1) and lactate dehydrogenase A (LDHA) mRNA levels, reduced ATP levels, increased lactate production, and decreased mitochondrial enzyme activities. Moreover, BGP increased ROS, diminished mitochondrial membrane potential, and altered fusion–fission dynamics and mitophagy. In aged quadriceps, oxidative phosphorylation (OXPHOS) subunits and superoxide dismutase 2 (SOD2) expression were reduced. The hypophosphatemic diet improved all parameters, whereas Velphoro^® selectively increased Mitochondrial cytochrome C oxidase subunit 1 (CIV-MTCO1) expression. Several altered mitochondrial markers are associated with sarcopenia degree. Altogether, hyperphosphatemia induces metabolic changes that scale with the sarcopenic degree. Our findings show a relevant association between hyperphosphatemia and mitochondrial dysfunction, and they support the potential benefit of phosphate reduction as a strategy to prevent or mitigate sarcopenia. Full article

We aimed to investigate the potential antidiabetic effects of an ethanol extract derived from the fruit of Malus floribunda (MF) on insulin resistance, oxidative stress, inflammation, and apoptosis associated with diabetes. In our study, diabetes was induced through the administration of a 10% fructose solution and 40 mg/kg Streptozotocin (STZ). Once diabetes had been induced, metformin (Met) 300 mg/kg and the MF extract (250 mg/kg and 500 mg/kg) were administered orally once daily for 30 days. At the end of the experiment, markers of insulin resistance, oxidative stress, inflammation and apoptosis were evaluated in the serum, muscle and liver tissues of the different groups. The MF extract significantly improved the levels of HOMA-IR, insulin receptor (InR), insulin receptor substrate 1 (IRS-1) and glucose transporter 4 (GLUT4)—key components of peripheral insulin resistance associated with type 2 diabetes. Fructose/streptozotocin induced oxidative stress, inflammation, and apoptosis were mitigated by increasing Nuclear factor erythroid 2-related factor 2 (NRF2) expression, which restored antioxidant levels (Superoxide dismutase (SOD) and Glutathione (GSH)), significantly improved cytokine levels (Tumor necrosis factor alpha (TNF-α) and Interleukin-1 beta (IL-1β)), and downregulated apoptotic proteins (caspase-3 and caspase-9). We demonstrated the antidiabetic effect of MF extract using a fructose/streptozotocin-induced type 2 diabetes model. MF extract shows promise for future use in herbal medicine. Full article

Background: Glucose transporter 1 (GLUT1) is frequently upregulated in solid tumors and may reflect metabolic adaptation of malignant tissues. However, evidence regarding GLUT1 protein levels in salivary gland tumors remains limited. Methods: In this pilot study, GLUT1 protein concentrations were quantified in tissue homogenate supernatants from salivary gland tumors (n = 9) and non-malignant salivary gland tissue obtained from surgical margins (controls; n = 4) using a commercial ELISA kit (BlueGene Biotech; E01G0020) according to the manufacturer’s instructions. Supernatants were stored at −80 °C until analysis. Group comparisons were performed using a non-parametric Mann–Whitney U test. Results: GLUT1 levels showed substantial inter-individual variability. The tumor group exhibited higher values than controls [median (IQR): 15.53 (12.44–26.38) vs. 10.14 (7.40–13.26); mean ± SD: 19.26 ± 11.49 vs. 10.33 ± 4.39 (ng/mL)], although the between-group difference did not reach statistical significance (Mann–Whitney U = 27, two-sided p = 0.199). Conclusions: These preliminary data suggest heterogeneity of GLUT1 levels in salivary gland tumor tissue homogenates and numerically higher concentrations compared with non-malignant margin tissue. These findings should be interpreted as preliminary and hypothesis-generating. Larger, clinically annotated cohorts with orthogonal validation are required before any diagnostic, prognostic, or clinical relevance of GLUT1 can be considered. Full article

Epidemiological and biological evidence indicate a close connection between Alzheimer’s disease (AD) and type-2 diabetes mellitus. Glucose transporter 1 (GLUT1), encoded by the SLC2A1 gene, has a major role in glucose metabolism, the dysregulation of which has been implicated in both diseases. We conducted a case-control association study in a sample of 439 non-diabetic patients with late-onset AD and 304 cognitively healthy, non-diabetic elderly controls to determine the potential risk for developing AD associated with SLC2A1 rs841847 polymorphism. The rs841847 C/C genotype occurrence was higher in the AD group (AD: 60.4%, controls: 50.7%), while the minor T allele-containing genotypes were more frequent among controls (AD: 39.6%, controls: 49.3%). A multivariate logistic regression model adjusted for age, sex, and apolipoprotein E (APOE) ε4 status (ε4 allele carriers versus non-carriers) demonstrated that carriers of the T allele had a significantly reduced risk for AD compared to C/C homozygotes (OR = 0.672; 95% CI: 0.493–0.916; p = 0.012). Although the rs841847 polymorphism has been linked to type-2 diabetes mellitus, the present study investigated this gene variant in AD for the first time. Our findings indicate a moderate protective effect for the rs841847 T allele on the susceptibility to AD. We demonstrated the rs841847 polymorphism as a candidate single nucleotide polymorphism for further examination as a predisposing genetic factor for AD. Full article

Fermentation is widely used to enhance the bioactivity of herbal phytochemicals through microbial bioconversion. Rehmannia glutinosa contains catalpol, an iridoid glycoside with metabolic and immunomodulatory potential; however, its efficacy in the unfermented form is limited. This study investigated whether fermentation enhances catalpol production and improves metabolic and immune-regulating functions via glucagon-like peptide-1 receptor (GLP-1R) signaling. Rehmannia glutinosa extract was fermented under optimized conditions, and catalpol and iridoid precursor levels were quantified to assess bioconversion efficiency. Biological effects were evaluated in intestinal epithelial cells, macrophages, and an Artemia model, focusing on glucose transport, GLP-1 secretion, dipeptidyl peptidase-4 (DPP-4) expression, mucosal defense, and GLP-1R/protein kinase A/cAMP response element-binding protein (PKA/CREB) signaling. Fermentation significantly increased catalpol content while reducing iridoid precursors. The fermented extract suppressed intestinal glucose absorption by downregulating sodium–glucose cotransporter 1 (SGLT1) and glucose transporter 2 (GLUT2). It also enhanced GLP-1 secretion and reduced DPP-4 expression, leading to activation of GLP-1R/PKA/CREB signaling. This activation increased mucin 2 (MUC2) expression and promoted anti-inflammatory. Full article

Background/Objectives: Exploiting the metabolic properties of postbiotics is a novel strategy for managing metabolic disorders, including diabetes. Inactivated microorganisms, a major class of postbiotics, improve glycemic control in preclinical and clinical studies. Here, we examined whether heat-killed (HK) Mycobacterium aurum (M. aurum) exerts prophylactic or therapeutic anti-hyperglycemic effects in diabetic mice. Methods: Diabetes was induced in male BALB/c mice by streptozotocin (STZ; 150 mg/kg) injection. HK M. aurum (1 mg) was given orally (three prophylactic doses before STZ) or intradermally (six weekly therapeutic doses after STZ). We assessed glycemic parameters, serum C-peptide/insulin (ELISA), and tissue protein expression (Western blot). Results: Neither route altered body weight or glucose homeostasis in non-diabetic mice. In STZ-diabetic mice, oral prophylactic treatment significantly attenuated hyperglycemia (39–60% reduction weeks 5–8 post-STZ) and showed a trend toward improved serum C-peptide, but did not affect dysregulated expression of skeletal muscle (SM), hepatic, pancreatic and renal proteins involved in glucose transport (GLUT2, GLUT4, and SGLT2), glycolysis (α-LDH), mitochondrial uncoupling (UCP2 and UCP3), and antioxidant defense (CAT). Therapeutic intradermal administration significantly decreased blood glucose (~30% at week 5, ~40% at week 6) and modestly enhanced insulin secretion. Hepatic UCP2 and α-LDH and SM UCP3 protein levels were normalized toward non-diabetic levels, whereas hepatic GLUT2 and SM GLUT4 remained largely unchanged. These correlative findings suggest effects independent of insulin-dependent glucose transport, but do not demonstrate direct functional improvement in mitochondrial or redox status. Conclusions: HK M. aurum exerts partial anti-hyperglycemic effects in STZ-induced diabetic mice, but the associated protein changes require functional validation before its role as a postbiotic in β-cell dysfunction can be established. Full article

Diabetes Mellitus is a chronic metabolic disorder characterized by elevated blood glucose due to defects in insulin secretion or action, or both, leading to serious short- and long-term complications if not effectively managed. However, there is limited qualitative evidence exploring how youths diagnosed with Type 1 Diabetes Mellitus (T1DM) experience disease onset, management, complications, emotional adaptation, and education within the South African public healthcare system. The study aims to investigate the lived experiences of youths living with T1DM in a selected public hospital in Limpopo province, South Africa. The objectives were to explore and describe the lived experiences of youths living with T1DM. A qualitative, explorative, descriptive, and contextual design was used to gain a thorough understanding of the experiences of youths living with T1DM. A non-probability sampling technique was used to select 12 participants using a pre-determined criterion. Data were collected through individual semi-structured interviews using an interview guide. The data were analyzed using Colaizzi’s method, where themes and sub-themes were developed with the inclusion of an independent coder. Measures to ensure trustworthiness and ethical considerations were adhered to throughout the study. The findings revealed that, despite the participants sharing the same diagnosis, they experience multiple interrelated barriers that significantly hindered effective self-care management, such as limited access to diabetic diet, glucometers and supplies, treatment and informational-related barriers, school-related challenges, transportation constraints and inadequate social support. Furthermore, the findings highlighted gaps in early recognition of symptoms, standardized diabetes education, psychosocial support, and continuity of care. The study recommends the need for holistic, patient-centred, and contextualized interventions that do not only address medical management but the socioeconomic, educational, and psychological needs of youths. Full article

The molecular mechanism underlying male reproductive toxicity associated with Perfluorooctanoic acid (PFOA), a persistent environmental endocrine disruptor (EDC), has not yet been fully elucidated. Six-week-old male C57BL/6 mice were treated with PFOA by oral gavage at 0, 1.25, 5, 10, and 20 mg/kg/day for 35 days to explore its toxic effects on the male reproductive system and the underlying mechanisms. Analyses of semen quality, testicular histopathology, and blood–testis barrier (BTB) integrity revealed that PFOA caused dose-dependent structural and functional damage to the BTB, leading to markedly reduced semen quality. Based on transcriptomic sequencing and differential gene enrichment analysis, the glycolytic pathway was identified as a key regulatory target for PFOA-induced damage to the reproductive system. Further validation revealed that PFOA exposure inhibited glycolysis-related enzymes (Hexokinase 1 (HK1), Glucose Transporter 1 (GLUT1), and Lactate Dehydrogenase A (LDHA)), reduced lactate production and ATP synthesis, lowered Pan-Kla and H3K18la levels, and diminished H3K18la enrichment at the Hk1, Glut1, and Ldha promoters, whereas exogenous sodium lactate reversed these changes. This study is the first to identify the “glycolysis–lactate–H3K18la” chain as a key regulator in PFOA-induced BTB damage and spermatogenesis impairment, offering a new theoretical foundation for understanding EDC-induced male reproductive toxicity. Full article

Ertugliflozin is a sodium-glucose co-transporter-2 inhibitor that has demonstrated promise as a treatment for hyperinsulinaemia in horses. Despite the frequent use of ertugliflozin in equine clinical practice, the pharmacokinetics of this drug in horses has not been established. The aim of the present study was to determine the pharmacokinetics of one supratherapeutic dose (0.25 mg/kg) of ertugliflozin in eight horses. Horses were defined as being healthy by physical examination, haematological, blood biochemical and oral sugar test (OST) results. Plasma concentrations of ertugliflozin were quantified using high-performance liquid chromatography–tandem mass spectrometry 0, 0.25, 0.5, 0.75, 1, 2, 3, 4, 6, 8, 10, 14, 18, 24, 30, 36, 48, 60, 72, 96, and 120 h after drug administration enterally. Non-compartmental analysis led to determination of key pharmacokinetic variables, including mean ± SD time to maximum concentration (T_max) of 0.91 ± 0.13 h, maximum measured concentration (C_max) of 267.52 ± 25.37 ng/mL, terminal elimination half-life (T_1/2) of 17.65 ± 3.15 h and apparent oral clearance (CL/F) of 106.95 ± 27.53 mL/h/kg. No clinical signs of adverse effects or blood biochemical abnormalities occurred after drug administration. The results of this study suggest that a single supratherapeutic dose of ertugliflozin in healthy horses is safe. The pharmacokinetics of enterally administered ertugliflozin in horses are similar to pharmacokinetics of the drug in humans and the long T_1/2 makes ertugliflozin suitable for once daily dosing in horses. It is proposed that a starting dose for ertugliflozin in horses be in the range 0.05–0.1 mg/kg. Further pharmacokinetic studies are required to optimise the dose regimen for treating horses with hyperinsulinaemia. Full article

Metabolic dysfunction, a hallmark of diet-induced obesity (DIO), is increasingly attributed to alterations in intestinal nutrient and electrolyte transport. Yet the mechanisms that drive obesity-associated functional alterations of intestinal transporters remain incompletely understood. In this context, the effects of a high-fat diet (HFD) induced obesity on sodium-dependent glucose co-transporter 1 (SGLT1), Na⁺/H⁺ exchanger 3 (NHE3), and Cl⁻/HCO₃⁻ exchangers (DRA/PAT1), the primary glucose, sodium, and chloride absorptive pathways in mice small intestinal villus cells, were investigated. SGLT1 activity significantly increased in intact villus cells and brush border membrane vesicles (BBMV) from HFD-fed mice. Kinetic analysis demonstrated reduced Km without a change in Vmax, indicating enhanced transporter affinity. Notably, SGLT1 mRNA and protein expression, including BBM localization, were unchanged. Basolateral Na⁺/K⁺-ATPase activity was decreased, excluding enhanced Na⁺ gradient generation as the mechanism for SGLT1 stimulation. In contrast, DRA/PAT1 activity was significantly increased in HFD-fed mice, and kinetic studies revealed elevated Vmax without a change in Km, indicating increased transport capacity. DRA/PAT1 mRNA, total protein, and BBM expression were all significantly elevated. NHE3 activity and expression remained unchanged. These findings demonstrate that DIO enhances intestinal glucose absorption by increasing SGLT1 affinity and chloride absorption by upregulating DRA/PAT1 transcription. These transporter-specific alterations may amplify nutrient absorption and contribute to metabolic dysregulation in obesity. Full article