BACKGROUND: Recurrent COVID-19 remains a major public health challenge, yet the host factors driving susceptibility are poorly understood. Although genetic determinants of COVID-19 severity have been explored, their role in infection recurrence particularly in genetically diverse populations such as Malaysia has received limited attention. This study represents the first Malaysian assessment of the combined impact of ACE (I/D), TNF-alpha (-308G>A), and IL-6 (-572G/C) polymorphisms on susceptibility to recurrent COVID-19. METHODS: A total of 120 participants were recruited, comprising 60 individuals with documented recurrent COVID-19 (RT-PCR positive >90 days apart) and 60 matched SARS-CoV-2-negative controls. Genotyping was performed using PCR-based approaches. Genotype and allele frequencies were compared using Chi-square Tests, and odds ratios (ORs) were calculated to estimate relative risks. RESULTS: A significant association was observed for the ACE (I/D) polymorphism at both genotype (x(2)=25.94, df=2, P<0.001) and allele levels (x(2)=17.01, df=1, P<0.001). The D allele was more frequent in recurrent cases (50.8%) than controls (25%), with an OR of 0.50 (95% CI: 0.37-0.68). For TNF-alpha (-308G>A), no significant genotype association was found (x(2)=4.92, P=0.086), though the A allele was more common in recurrent cases (31.7%) vs. controls (19.2%), indicating a modest association (x(2)=4.95, p=0.037; OR=0.62, 95% CI: 0.41-0.93). The IL-6 (-572G/C) variant showed strong associations at both genotype (x2=18.31, P<0.001) and allele levels (x(2)=15.67, P<0.001), with the C allele more frequent in recurrent cases (45.8%) than controls (21.7%; OR=0.59, 95% CI: 0.43-0.82). CONCLUSIONS: ACE and IL-6 polymorphisms appear to be key genetic contributors to recurrent COVID-19 risk, with a suggestive role for TNF-alpha. This study provides novel evidence that pro-inflammatory genetic variants may predispose individuals to repeated SARS-CoV-2 infections, emphasising the importance of host genetic profiling in understanding COVID-19 recurrence.
Various noble metals have garnered attention in medicine due to their broad-spectrum biological activities, such as silver (Ag), the most focused material. Many studies have focused on developing eco-friendly and sustainable silver nanoparticles (AgNPs) nanomaterials due to their potential in wound healing. The current studies outline the protocol for the sustainable synthesis of bioactive AgNPs using Curcuma caesia (black turmeric) extract as a reducing and stabilising agent. Phytoextract-mediated conversion of precursor salt into AgNPs done at 80 degrees C for 30 30-minute incubation, results in phytoconjugated crystalline phyto-AgNPs with a particle size150f0.05 nm. In vitro assay studies reveal that the resulting phyto-AgNPs nanoparticles exhibit significant antioxidant, anti-inflammatory, and antidiabetic activity with IC50 values of 8.97 & micro;g/ mL, 12 & micro;g/mL and 5.24 & micro;g/mL, suggesting their potential in pharmaceutical and therapeutic interventions. Conclusively the study outcomes, demonstrate the standardization of sustainable and cost-effective approach for the AgNPs synthesis having better bio-stability, bio-functionality with minimal toxicity for biomedical application.
BACKGROUND: Fuchs' endothelial corneal dystrophy (FECD) is a primary disease of the corneal endothelium, characterized by structural modifications of the Descemet's membrane (DM), including focal excrescences known as guttae, which ultimately lead to edema. In this study, we evaluated the therapeutic potential of microvesicle-enriched plasma fraction for treating FECD and tested their effects on corneal thickening, curvature, and other FECD-like damages in rabbits. METHODS: A total of eight rabbits (16 corneas) were divided into three distinct groups: the negative control group, the positive control group, grouping eyes that underwent FECD induction with deep intrastromal injection of collagenase II, and the treated group, consisting of eyes subjected to FECD induction followed, after seven days, by a daily treatment with microvesicle-enriched plasma fraction for an additional 7 days. RESULTS: Pachymetric and keratometric measurements of the rabbit eyes showed that the deep intrastromal injection of collagenase II induced a significant increase in corneal thickness, while treatment with microvesicle-enriched plasma fraction restored the corneal thickness reaching control group stage. This finding was supported the histopathological examination of corneal sections showing that microvesicle-enriched plasma fraction reduced some FECD-like histological damage by eliminating cysts and reducing the thickening of the DM. CONCLUSIONS: Together, these data offer valuable insights into the role of microvesicle-enriched plasma fraction in corneal regeneration and the repair of histological damage in the rabbit model of FECD, emphasizing their potential as a novel biotechnological application of plasma-derived microvesicles for degenerative corneal diseases, such as FECD.
BACKGROUND: Diabetic nephropathy (DKD) is one of the most severe microvascular complications of diabetes. Resveratrol (RSV), a natural polyphenol, exhibits antioxidant, anti-inflammatory, and cytoprotective effects, but its mechanism in DKD remains incompletely understood. METHODS: An in vitro high glucose (HG)-induced MPC-5 podocyte injury model was established. Cell viability, membrane integrity, apoptosis rate, and podocyte-specific protein expression were assessed using CCK-8, LDH release assays, flow cytometry, and Western blot. NLRP3 inflammasome-related proteins and inflammatory cytokines were evaluated via Western blot, immunofluorescence, and ELISA, with NLRP3-specific inhibitor MCC950 and activator nigericin used for validation. A DKD mouse model was constructed to assess RSV's effects on blood glucose, body weight, renal function, and renal pathology. NLRP3 inflammasome activation in renal tissues was examined by immunofluorescence, ELISA, and Western blot. RESULTS: In vitro, HG significantly reduced MPC-5 cell viability, increased LDH release and apoptosis, and downregulated podocyte-specific proteins; RSV dose-dependently reversed these effects. HG upregulated NLRP3, ASC, IL-1(3, and IL-18 expression, promoted caspase-1 activation and GSDMD cleavage-effects inhibited by MCC950. RSV suppressed NLRP3 inflammasome activation, with nigericin partially reversing RSV's protection. In vivo, RSV lowered blood glucose and body weight, improved renal function, attenuated glomerular injury, restored podocyte-specific proteins, and reduced renal fibrosis in DKD mice. RSV also inhibited NLRP3 inflammasome activation in renal tissues. CONCLUSIONS: RSV effectively attenuates DKD-induced renal injury by inhibiting NLRP3 inflammasome-mediated pyroptosis. This discovery reveals the potential of RSV as a novel therapeutic strategy, offering fresh perspectives for the prevention and treatment of DKD.
BACKGROUND: The pathogenesis of Hypertensive intracerebral hemorrhage (HICH) critically involves endothelial cell dysfunction, but the specific signaling pathways involved require further investigation. The role of miR-223, a key regulator in cardiovascular disease, in HICHassociated EC injury is not well understood. METHODS: An in-vitro HICH model was established by inducing injury in hCMEC/D3 cells with thrombin, and the expression of miR-223 was measured. Bioinformatic prediction and dual-luciferase reporter assays were used to verify the targeting relationship between miR-223 and Insulin-like Growth Factor 1 Receptor (IGF1R). Subsequently, cells were transfected with miR-223 mimics or inhibitor, as well as si-IGF1R, and treated with PI3K/AKT pathway agonist (740Y-P) and inhibitor (LY294002) to assess the effects and molecular mechanisms of miR-223 on endothelial cell viability, proliferation, migration, invasion, angiogenesis, NO/ET-1 balance, blood-brain barrier (BBB) integrity, inflammatory response, oxidative stress, and apoptosis. Finally, an in vivo HICH rat model was constructed, and the effects of intracerebral injection of miR-223 inhibitor were evaluated. RESULTS: Thrombin treatment significantly upregulated miR-223 expression in hCMEC/D3 cells, resulting in impaired endothelial cell viability and proliferation, enhanced migration and invasion, inhibited angiogenesis, reduced NO release, increased ET-1 levels, increased BBB permeability, downregulated tight junction proteins, elevated inflammatory cytokine and ROS release, and aggravated apoptosis. miR-223 mimics further exacerbated these injuries, whereas miR-223 inhibitor significantly reversed these adverse effects. Downregulation of miR-223 targeted IGF1R, thereby activating the PI3K/AKT/ mammalian target of rapamycin (mTOR)/p70 Ribosomal protein S6 Kinase (p70S6K) signaling axis and ameliorating endothelial cell dysfunction. In-vivo experiments further demonstrated that intracerebral injection of miR-223 inhibitor significantly alleviated neurological deficits, brain edema, brain tissue pathological damage, and apoptosis in HICH rats. CONCLUSIONS: This study confirms that downregulation of miR-223 ameliorates endothelial cell injury and brain damage induced by HICH via targeting IGF1R and activating the PI3K/AKT/mTOR/p70S6K axis.
BACKGROUND: The isolation of DNA or RNA is a crucial step that must be carried out before many biochemical and diagnostic procedures can be performed. To date, several methods have been developed for the purification of biological entities. The most common magnetic materials/systems used in biological separations involve the use of nanoparticles with an iron oxide core. However, the use of such systems for the separation of nucleic acids from biological fluids does not seem to be free of problems. It required laborious, multi-step and lengthy procedures, skilled chemists and expensive reactants. Such considerations explain the growing demand for alternative techniques. A patented process developed by some of us, could be used for the production of reliable magnetic adsorbents. The process uses commercially available zeolites as raw materials and the final product is a metal-ceramic nanocomposite. The same metal-ceramic nanocomposites, which have magnetic properties, have already been successfully used for a wide range of applications. The aim of this study was to compare RNA extraction from blood samples from pediatric patients with RSV bronchiolitis extracted with the manual commercial Promega ReliaPrep silica column kit and a manual protocol using magnetic nanocomposites derived from zeolite. METHODS: Thirty pediatrics bronchiolitis patients and 10 healthy pediatrics control. Each sample was extracted in duplicate, with both automated extraction and nanocomposites extraction. These extracts were compared in terms of yield and quality, measured by traditional UV spectroscopy, and amplifiability, the latter measured by evaluating the results obtained from type 1 interferon signature with real-time PCR. RESULTS: The zeolite-derived nanocomposite method for RNA extraction yielded a higher RNA quantity compared to the commercial magnetic silica bead kit (Promega), although with lower purity and amplification efficiency. Nevertheless, both extraction methods consistently confirmed the interferon signature data. CONCLUSIONS: Although the zeolite-based method produced higher RNA yields, residual contaminants likely compromised RNA purity and reduced amplification efficiency compared to the commercial kit. Nonetheless, the final IFN score was comparable between methods. Ongoing optimizations aim to improve nanocomposite purity and amplification efficiency, highlighting their potential as low-cost alternatives to commercial magnetic systems for diagnostic and research applications.
BACKGROUND: Cervical cancer (CC) is caused by human papillomavirus (HPV) infection and has serious impacts on women's health. While some HPV vaccines are already available, there is still a need for deeper understanding of the pathogenesis of CC and better ways to diagnose and treat it. METHODS: qRT-PCR was used to determine the level of miR-141-5p expression in CC. A CCK-8 assay, colony formation experiment, and Transwell assay were used to determine the influence of miR-141-5p on biological characteristics. A dual luciferase assay was employed to examine the correlation between miR-141-5p and MRVI1. The expression of MRVI1 and proteins linked to epithelial-mesenchymal transition (EMT) were identified by western blot. Knockdown ofMRVI1 was performed to investigate its revertive effect on miR-141-5p down-regulation. A subcutaneous tumor-graft model was constructed to explore the impacts of down-regulating miR-141-5p on tumors in vivo, and the positivity of Ki-67 and MRVI1 in CC tissues was detected by immunohistochemistry. RESULTS: The level of miR-141-5p was markedly increased in CC. miR-141-5p knockdown reduced the viability, migration, and invasion of CC cells and hindered their EMT process. The opposite result was obtained when miR-141-5p was over-expressed. MRVI1 was screened as a target gene of miR-141-5p. MiR-141-5p negatively regulated MRVI1 expression, and silencing MRVI1 reversed the impact of miR-141-5p down-regulation on CC cells. Knockdown of miR-141-5p upregulatedMRVI1 expression in tumor tissues and inhibited CC formation in vivo. CONCLUSIONS: miR-141-5p was markedly increased in CC and regulated CC malignant progression by targeting and negatively regulating MRVI1. Additionally, knockdown of miR-141-5p inhibited CC progression in vivo.
BACKGROUND: Vitamin D influences cellular functions through vitamin D receptor (VDR)-mediated signaling pathways. Beyond its established role in calcium and phosphorus homeostasis, it plays a vital role in immune responses, cell proliferation, and cardiovascular health. Numerous studies have associated vitamin D deficiency with various clinical conditions; however, a systems biology approach to pinpoint key genes involved in VDR signaling and related clinical phenotypes remains unexplored. Biological networks are complex with several hubs and functional modules that spread among intertwined layers, therefore identifying crucial genes can often be challenging but offer deep insight of disease pathogenesis and theragnostic. METHODS: A network theory approach was employed to identify crucial genes in terms of modularity. This process involved constructing a protein-protein interaction (PPI) network of genes associated with the VDR signaling pathway, followed by modular analysis and topological scoring using metrics such as degree distribution, clustering coefficient, and neighborhood connectivity. Using this approach, 5 (BMP2, TNFSF11, IRS1, SLC2A1, and PDGFRB) Motif Hubs (MHs) were identified as key regulators, which were deeply embedded from top to bottom throughout the network, influenced the network down to the motif level. RESULTS: The identified five key regulators played a central role in the highly clustered module 2 and were essential for the stability of the entire network, indicating quick accessibility to the molecules and rapid information processing. Functional analysis of the top five modules displayed their strong correlation with enzyme binding, signaling receptor binding, and ligand activity. Similarly, disease association analysis also predicted their relationship with neoplasms, nervous system disorders, and immune system disorders. CONCLUSIONS: The current findings warrant experimental biochemical validation of the observed key regulators, which may offer potential therapeutic targets for treating cardiovascular diseases, cancer, diabetes, and neurological disorders associated with vitamin D deficiency.
BACKGROUND: Human activities in diverse sectors contribute to the surge of antimicrobial resistance. This study assessed wastewater and soil from selected environments in Lagos, Nigeria, for the presence of antibiotic-resistant bacteria (ARB), as well as their resistance genes. Lagos is a city with densely populated urban and slum settlements characterized by poor drainage systems into which waste effluents are emptied. This highlights Lagos as an ideal site for the study, as it could be an epicenter for the breeding and transmission of ARB. METHODS: A total of 12 samples, comprising four soil and eight water samples, were collected and analyzed from selected environments, including an abattoir, a tertiary hospital, a pharmaceutical company, and farms. The physicochemical parameters of the samples were characterized using standard protocols. Bacterial strains were isolated through culture, and antimicrobial resistance profiles were determined using the Kirby-Bauer method. Resistance genes were detected through specific primer-dependent real-time quantitative PCR, from the extracted genome. RESULTS: The physicochemical analyses of the wastewaters revealed higher biochemical oxygen demand (BOD: 326.7 +/- 4.51-821.0 +/- 2.0 mg/L), dissolved oxygen (DO: 1.05 +/- 0.07-4.27 +/- 0.14 mg/L), and total dissolved solids (TDS: 0.14 +/- 0.0-5.18 +/- 0.012 g/L) than the recommended limits. Soil samples contained total organic carbon, organic matter (OM), and phosphate, ranging from 0.14 (+/- 0.031)-9.77 (+/- 0.091), 0.24 (+/- 0.050)-16.90 (+/- 0.164), and 2.22 (+/- 0.046)-5.28 (+/- 0.308) mg/kg, respectively. Klebsiella pneumoniae (21.5%) predominated among the isolated bacteria, which also included Enterobacter spp., Providencia stuartii, Salmonella spp., Pseudomonas spp., and Vibrio parahaemolyticus. Enterobacter aerogenes isolated from abattoir effluents exhibited resistance to 7 antibiotics with a Multi-Antibiotic Resistance Index of 0.58. Thirty-four (52.3%) bacterial strains were resistant to pefloxacin. blaTEM resistance gene was detected in all (100%) samples with mean Ct values ranging from 15.60 +/- 0.12 to 27.93 +/- 0.06, while blaCMY and qnrS were detected in 58% and 83% of the samples respectively. CONCLUSIONS: This study reveals that effluents from the assessed environments are hotspots for the dissemination of antibiotic resistance within the ecosystem.
BACKGROUND: Leukocyte telomere shortening can occur with physiological age-related changes in the body as well as due to several damaging factors. We investigated the relationships between early markers of kidney damage and the leukocyte relative telomere length (RTL) in individuals with type 2 diabetes (T2D). METHODS: This study included 86 persons with T2D which were divided into two groups according to the estimated glomerular filtration rate (eGFR): group I (N.=21), with an eGFR <60 mL/min/m2, and group II (N.=65), with an eGFR >= 60 mL/min/m2. To determine the RTL standardized method proposed by Cawthon was used. RESULTS: The assessment of RTL depending on the level of the eGFR revealed a significant difference between the groups of patients (1.10 [0.97-1.18] vs. 1.18 [1.06-1.41] in group I and group II, respectively, P=0.031). A positive significant correlation was established between RTL and the eGFR only in a reduced eGFR group (r=0.44; P=0.042). In multivariate logistic analysis RTL, ACR in daily urine, and T2D duration were independent predictors of eGFR decrease (<60 mL/min/m2). The AUROC of the model was 0.79 (95% CI 0.684-0.897; P<0.001) with a sensitivity of 71.4% and a specificity of 73.8%. CONCLUSIONS: RTL shortening, a longer duration of diabetes, and albuminuria are early prognostic factors independently associated with decline of renal function in patients with DKD. (Cite this article as: Yerokhovych V, Karpenko O, Ilkiv Y, Gurianov V, Krasnienkov D, Sulaieva O, et al. Relationship between leukocyte telomere length and early markers of kidney damage in people with type 2 diabetes. Minerva Biotechnol Biomol Res 2025;37:115-22. DOI: 10.23736/S2724-542X.25.03266-3)
BACKGROUND: Gastric cancer (GC) is commonly found among gastrointestinal tract cancers, and it often exhibits a poor prognosis. MiRNA-767-5p, a microRNA (miRNA), has been related to the progression and prognosis of numerous human cancers. Currently, the level of miR-767-5p expression and its function have not been thoroughly investigated in GC. METHODS: We examined miR-767-5p and target gene hypoxia-inducible factor-3a (HIF3A) mRNA expression in GC and normal tissues, and also in GES-1 cells, N87 cells, and GAS cells using qRT-PCR. We used MTT, plate cloning, and EdU staining assays to measure the variations in the biological properties of GC cells. The Epithelial-mesenchymal transition (EMT) process and the effects of miR-767-5p and HIF3A on the PI3K/AKT signaling pathway in GC cells were analyzed through Western blotting. RESULTS: MiR-767-5p exhibited a high level of expression in GC. Inhibitors of miR-767-5p hindered the proliferation, clonal formation capability, and EMT process in GC cells. In addition, there was a negative correlation between miR-767-5p and HIF3A mRNA expression. The silencing of HIF3A boosted proliferation and colony formation abilities in GC cells, and partially offset the suppression inflicted by miR-767-5p inhibitors on these cells. Silencing miR-767-5p impeded the PI3K/AKT pathway, whereas silencing HIF3A activated this pathway. The use of PI3K/AKT pathway agonists amplified the effects of HIF3A silencing, which could further enhance GC cell proliferation and EMT, as opposed to the effects of PI3K/AKT pathway inhibitors. CONCLUSIONS: MiR-767-5p negatively regulates HIF3A and is involved in the proliferation and EMT process via the PI3K/AKT signaling pathway in GC.
BACKGROUND: Human endogenous retroviruses (HERVs) make up approximately 8% of the human genome and play a role in modulating inflammatory and immune responses. Burns are complex injuries that induce both local and systemic effects, primarily due to prolonged inflammatory responses. This study investigates the regulation of HERV expression after burn injury by analyzing the transcript levels of the pol genes of HERV-H, HERV-K and HERV-W in whole blood from burn patients and healthy controls. METHODS: The transcript levels of the HERV-H, HERV-K and HERV-W pol genes were quantified using quantitative real-time PCR (qRT-PCR) in whole blood samples collected from burn patients both on admission to hospital and on discharge from hospital. RESULTS: The pol genes of HERV-H, HERV-K and HERV-W were transcriptionally active in both healthy subjects and burn patients. However, their expression was significantly lower in patients with severe burns on admission than in healthy controls or in the same patients on discharge suggesting a dynamic expression pattern linked to injury and recovery phases. CONCLUSIONS: The observed reduction in HERV pol gene expression in burn patients may be related to the decreased IFN-gamma levels reported in literature suggesting a potential link between immune response dysregulation and endogenous retrovirus modulation in burn injury. These findings highlight HERVs' potential role as biomarkers and modulators of immune status following severe burn injury.
BACKGROUND: Traumatic brain injury (TBI) can cause pathological disruption of the blood-brain barrier, leading to neurological damage. Recent studies suggest that microRNA-208b-3p (miR-208b-3p) in serum exosomes may play a role in TBI, but its precise mechanism remains unclear. This study aims to investigate the effect of serum exosomal miR-208b-3p on brain microvascular endothelial cells (BMVECs) and its regulatory mechanism involving Nemo-like kinase (NLK). METHODS: Serum exosomes were extracted from patients with TBI and co-cultured with BMVECs following scratch injury. The expression levels of tight junction proteins, inflammatory factors, and NLK in BMVECs were measured using Western blotting and quantitative real-time PCR (QRT-PCR). The levels of miR-208b-3p in exosomes were also assessed. Bioinformatics analysis and a dual luciferase assay were used to confirm the targeting relationship between miR-208b-3p and NLK. RESULTS: Serum exosomes from TBI patients were found to be rich in miR-208b-3p. miR-208b-3p was shown to regulate apoptosis in BMVECs and directly target NLK, leading to its reduced expression following scratch injury. Downregulation of NLK was associated with increased endothelial cell damage and impaired nerve repair. CONCLUSIONS: Serum exosomal miR-208b-3p exacerbates traumatic injury by downregulating NLK expression in BMVECs, thereby aggravating blood-brain barrier disruption and impeding neural repair. These findings provide a potential therapeutic target for the treatment of traumatic brain injuries.
BACKGROUND: Osteoarthritis (OA) is a leading cause of disability worldwide, and its impact is exacerbated by the long-term effects of COVID-19, which can lead to structural and functional disorders in various organ systems, including hemostasis. This retrospective observational study aimed to evaluate changes in the hemostasis system's procoagulant, anticoagulant, and fibrinolytic links in patients with OA following recovery from COVID-19. METHODS: The study involved patients aged 45 to 68 with knee OA, divided into three groups: Control (healthy donors), OA (stage II or III knee OA patients), and OA+COVID-19 (OA patients who had COVID-19). The concentrations of & rcy;r & ocy;thrombin, protein C, thrombomodulin (TM), plasminogen, tissue plasminogen activator (tPA), and plasminogen activator inhibitor-1 (PAI-1) in blood plasma were measured by enzyme-linked immunosorbent assay. The activity of prothrombin, antithrombin III (AT III), plasminogen, alpha-2-antiplasmin (a2-AP) and tPA was assessed using chromogenic substrates. Soluble fibrin monomer complexes were evaluated with o-phenanthroline test, and the activity of a1-antitrypsin (a1-AT) and a2-macroglobulin (a2-MG) was measured spectrophotometrically. RESULTS: In analyzing hemostasis system indicators in patients with OA and those who had experienced COVID-19, a decrease in prothrombin content and activity was established, alongside an increase in SFMC concentration. There was inhibition of the protein C, AT III, and TM. The fibrinolysis system was impaired, as evidenced by decreases in the concentration of plasminogen, its tissue activator, the PAI-1 inhibitor, and a2-AP activity. Abnormal levels of the proteinase inhibitors a1-AT and a2-MG were also detected. CONCLUSIONS: The findings indicate that the thrombotic risk in patients with OA remains high even months after the acute COVID-19 phase. This is attributed to suppressed anticoagulant activity and reduced fibrinolytic potential in plasma.
