Butyrylcholinesterase (BChE) plays a key role in preserving appropriate cholinergic neurotransmission that is essentially altered in the brains of advanced Alzheimer's disease (AD), hence a therapeutic target. This study employed a machine learning (ML) bioactivity predictive model to explore the chemical space of potential BChE inhibitors. A cheminformatics pipeline was explored to create a ML model for BChE inhibition using structural insights complemented with a comprehensive variance importance plot (VIP) and correlation matrix analysis. Specifically, a compiled library of 2179 secondary metabolites (SMs) from 50 Nigerian medicinal plants with reported cholinergic activity was investigated using the ML model. After which, molecular modelling was used to further screen the active SMs (827). The final predicted models demonstrated significant robustness, with a correlation coefficient of 0.8981. Molecular docking investigation of the 827 SMs identified the top five candidates based on their scores: three triterpenoids (adipedatol, lupenone and β-amyrin), one steroid (9 (11)-dehydroergosterol benzoate) and one flavonoid (tiliroside). These leads exhibited favourable ADMET properties and promising safety profiles. Among these five, lupenone (-48.78 kcal/mol), β-amyrin (-49.19 kcal/mol) and adipedatol (-48.87 kcal/mol), from Peltophorum pterocarpum, Bryophyllum pinnatum and Alchornea laxiflora, respectively, were the most promising leads with significant binding free energy compared to decamethonium (-17.64 kcal/mol) and more favourable van der Waals, electrostatics and nonpolar solvation energetics. Furthermore, the binding of these leads resulted in optimised interaction profiles that preserved the structural integrity of the BChE and aligned well with desirable drug-like characteristics. These findings position the leads as promising candidates for therapeutic applications targeting BChE for AD management, subject to further in vitro and in vivo validation investigations.
Antibiotic resistance has emerged as a major global health challenge, particularly in biofilm-forming pathogens that exhibit enhanced tolerance to antimicrobial therapies. K. pneumoniae, a multidrug-resistant pathogen, is a leading cause of hospital-acquired infections, including pneumonia, septicemia, and device-associated infections. Its robust biofilm-forming capacity facilitates immune evasion, restricts antibiotic penetration, and contributes to recurrent infections. Therefore, the identification of agents capable of targeting both planktonic bacterial growth and biofilm architecture is of considerable therapeutic importance. In the present study, the antibacterial, antibiofilm, and antivirulence potential of 6-Aminoflavone was investigated against K. pneumoniae. Antibacterial activity was evaluated using growth inhibition assays, time-kill kinetics, and clonogenic survival analysis. Mechanistic investigations revealed significant intracellular ROS accumulation, increased oxidative stress susceptibility, and induction of apoptosis-like cell death, suggesting ROS-mediated antibacterial activity. In addition to suppressing bacterial proliferation, 6-Aminoflavone exhibited promising antibiofilm efficacy, inhibiting biofilm formation by 86.23% and eradicating 84.77% of established biofilms. These effects were associated with a substantial reduction in cell surface hydrophobicity (∼45.39%), and EPS levels were reduced to ∼2.33% as compared to untreated controls, indicating severe destabilization of the biofilm matrix. Furthermore, eDNA, a key structural component of the biofilm scaffold, exhibited a maximum ∼2.13-fold reduction following treatment. Confocal microscopy confirmed marked disruption and collapse of biofilm architecture. Additionally, quorum sensing (QS)-regulated virulence factors, including urease (∼51.92%), protease (∼55.18%), and lipase (∼44.90%), were significantly attenuated. These findings demonstrate that 6-Aminoflavone exerts antimicrobial activity by inducing oxidative stress-mediated apoptosis-like bacterial death, disrupting biofilm structural components, and suppressing QS-regulated virulence in K. pneumoniae.
Plasmodium falciparum is the most virulent human malaria parasite and is responsible for numerous deaths annually. The increasing resistance of P. falciparum to antimalarial drugs necessitates the development of improved diagnostic tools for timely malaria detection. Malaria biomarkers such as PfHSP70 and PfLDH are highly valuable for malaria detection because they are essential for parasite survival and are consistently expressed during infection. PfHSP70 is associated with the parasite's stress adaptation and proteostasis mechanisms under febrile and drug-induced conditions, while PfLDH plays a central role in glycolytic metabolism and redox balance. Their functional importance, parasite specificity and elevated expression during active infection make these proteins reliable molecular indicators for the sensitive and specific detection of P. falciparum, thereby supporting their potential application in rapid diagnostic and biosensing platforms for malaria surveillance and disease management. In this study, an M13 phage-displayed single-chain variable fragment (scFv) antibody library was used to screen for antibodies against recombinant PfHSP70 and PfLDH. Four rounds of biopanning were conducted to enrich high-affinity binders, followed by ELISA, UV-visible spectroscopy and microscale thermophoresis (MST) to evaluate specificity and binding affinity. Functional interactions were assessed in Escherichia coli expressing PfHSP70 or PfLDH. Recombinant PfHSP70 and PfLDH were successfully expressed and purified from E. coli, with approximately 50% of selected colonies demonstrating significant binding to both targets, confirming the enrichment of antigen-specific phages. Specific phages were validated using ELISA and transmission electron microscopy (TEM). Selected scFvs exhibited strong target binding, with MST-determined dissociation constants (Kd) of 7.47 μM for PfHSP70 and 3.64 μM for PfLDH. Exposure to scFv-displaying phages impaired the survival of PfHSP70- and PfLDH-expressing E. coli and induced spectral shifts consistent with protein-antibody interactions. This study validates phage display as a robust platform for isolating high-affinity scFvs against P. falciparum targets. These binders have the potential to develop into low-cost diagnostic tools, addressing the urgent need for novel diagnostic interventions. Nevertheless, further studies are required to confirm binding specificity and assess translational applicability.
Pomelo (Citrus maxima) peel essential oil is a volatile-terpenoid-rich plant extract that may exhibit antioxidant-associated effects in biological systems. However, the relationship among its chemical composition, conventional antioxidant capacity, and cellular effects under oxidative stress remains insufficiently defined. This study integrated gas chromatography-mass spectrometry (GC-MS) profiling with chemical antioxidant assays and an SH-SY5Y cell-based oxidative-stress model. Under the selected analytical and data-processing conditions, limonene was the only compound confidently identified and integrated by GC-MS. In chemical assays, pomelo peel essential oil showed moderate DPPH radical-scavenging and ferric-reducing activities compared with the hydrophilic reference antioxidants Trolox and ascorbic acid. Cellular effects were evaluated separately in SH-SY5Y cells exposed to H2O2-induced oxidative stress. Pomelo peel essential oil was tested at 25, 50, and 100 μg/mL using cotreatment and pretreatment protocols. In the cotreatment model, cells were exposed simultaneously to the essential oil and H2O2, whereas in the pretreatment model, cells were incubated with the essential oil before H2O2 challenge. Pomelo peel essential oil attenuated intracellular ROS accumulation and lipid peroxidation and improved cell viability, with clearer effects at 50 and 100 μg/mL. Pretreatment generally produced stronger cytoprotective effects than cotreatment. These findings indicate that conventional chemical antioxidant assays alone may not fully predict cellular antioxidant-associated effects, particularly for lipophilic essential oils. Pomelo peel essential oil therefore showed cytoprotective activity in an SH-SY5Y oxidative-stress model, although the contributing constituents and underlying molecular mechanisms require further investigation.
Repetitive traumatic brain injury (RTBI) can cause long-term complications, including persistent neuroinflammation, which can extend beyond the central nervous system, impacting various peripheral organs as liver. This study aimed to explore the neuroprotective and hepatoprotective effects of arbutin treatment in a rat model of mild RTBI (mRTBI), focusing on nerve growth factor (NGF)/tropomyosin receptor kinase A (TrkA) signaling pathway along with the crosstalk between brain injury and hepatic dysfunction. Animals were randomly assigned into three groups: one served as a normal control (NC) group, while the other two groups were exposed to one blow for 5 days and either left for one week after the fifth blow (mRTBI) or received arbutin intraperitoneally (100 mg/kg/day for 7 days, mRTBI + ARB). Biochemical and histopathological changes were monitored in the brain cortex and the liver. This study revealed that arbutin treatment offered neuroprotection and preserved most of the neuronal structures. Arbutin demonstrated a significant increase in the cortical NGF and TrkA contents, along with a marked upregulation in cortical phosphoinositol-3 kinase (PI3K) and protein kinase B (AKt) mRNA levels compared to the mRTBI group. Furthermore, arbutin decreased cortical and serum inflammatory markers, reflecting its anti-inflammatory power. Peripherally, arbutin treatment resulted in a substantial decrease in hepatic inflammatory markers, Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) and caspase-3. These effects preserved hepatocellular histoarchitecture and reduced liver injury markers. Collectively, arbutin effectively modulated the NGF/TrkA signaling pathway, diminished inflammation, and alleviated the detrimental effects of mRTBI on both the brain and liver.
Venoms from Bothrops species and Daboia russelii are among the leading causes of venom-induced coagulopathy and snakebite-related deaths. In this study, we comparatively evaluated the biochemical composition and hemostatic effects of venoms from seven Bothrops species and D. russelii using protein profiling and functional coagulation assays. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed broadly similar molecular weight distributions between the two genera, consistent with the presence of snake venom metalloproteinases, serine proteases, and phospholipases A2, but with notable interspecific variability among Bothrops venoms. Enzymatic assays demonstrated heterogeneous metalloproteinase, serine protease, and phospholipase A2 activities across Bothrops species, whereas D. russelii venom showed strong phospholipase A2 activity and limited fibrinogenolytic activity. Functional coagulation analyses revealed marked differences between the genera. Bothrops venoms induced rapid clot initiation, with alterations in clot formation observed in a species-dependent manner, accompanied by pronounced fibrinogen degradation, consistent with consumption coagulopathy driven by fibrinogenolysis and clot destabilization. In contrast, D. russelii venom exhibited a strong procoagulant profile, significantly reducing prothrombin time and activated partial thromboplastin time, with minimal direct fibrinogen degradation. Previous studies have shown that D. russelii venom also promotes rapid clot formation in rotational thromboelastometry assays. Together, these findings demonstrate that Bothrops spp. and D. russelii venoms converge functionally in disrupting hemostasis but do so through distinct biochemical strategies, providing experimental support for the divergent coagulation phenotypes observed clinically.
The physiological age-related decline in skeletal muscle mass, power, and function is challenging for humans. Skeletal muscle has been recently recognized as a secretory organ, with human myogenic progenitor cells (hMPCs) releasing extracellular vesicles (EVs). Here, we investigate the role of hMPC-derived EVs as mediators in skeletal muscle aging. This heterologous approach enables the analysis of age-related variations in EV burden and their impact on human muscle stem cell function. Therefore, we isolated EVs from hMPCs obtained from vastus lateralis muscle biopsies of young and elderly subjects. Then, we characterized EVs for specific marker, size, and concentration and analyzed their miRNA expression and proteomic profiles to delineate the bioactive cargo that influences recipient cell signaling. Next, we tested the ability of EVs to modulate on hMPCs. Specifically, we treated elderly hMPCs with young EVs and vice versa to analyze viability and differentiation. Our results demonstrate that EVs released by young hMPCs carry regenerative signals that mitigate the functional decline of aged muscle stem cells. Conversely, the EVs derived from elderly hMPCs compromise the regenerative capacity of their younger counterparts. Therefore, these results suggest that hMPCs release EVs and that their cargo is modulated by donor age. Moreover, the EVs significantly modulated hMPCs’ viability and differentiation in cell culture.
Despite the urgent need for new therapies to treat multidrug-resistant virulent Plasmodium falciparum, many promising antimalarial candidates fail to progress to clinical use because their pharmacokinetic (PK) profile is poorly characterized. This study predicted the PK profile of a new compound, 3-chloro-4-(4-chlorophenoxy)aniline (ANI), in silico using the pkCSM online platform. In vitro cytotoxicity in Vero E6 cells and antiplasmodial activity against chloroquine-sensitive (P. falciparum 3D7) and chloroquine-resistant (P. falciparum W2) strains were screened using the MTT colorimetric and the [3H]-hypoxanthine incorporation assays, respectively. In vivo antimalarial efficacy was evaluated in Plasmodium berghei ANKA and piperaquine-resistant (PQR) P. berghei parasites in mouse models employing early and established infection tests. In silico predictions show that ANI is a substrate for CYP 3A4 and CYP 2D6 and is unlikely to inhibit the hERG potassium ion channel. ANI exhibited low cytotoxicity, with a CC50 value of 7.90 × 102 ± 86.40 μM, and potent antiplasmodial activity, with IC50 values of 1.72 ± 0.09 μM and 1.84 ± 0.21 μM against the 3D7 and W2 strains, respectively. The selectivity index of greater than 400 indicates that ANI has a broad safety margin. In vivo, ANI demonstrated antimalarial efficacy with ED50 values of 3.07 mg/kg/day against P. berghei ANKA and 2.81 mg/kg/day against PQR P. berghei parasites. Notably, mice treated with ANI in the established infection model survived for up to 30 days without observable adverse effects. These findings highlight ANI as a potential antimalarial compound and support further drug evaluation and development.
This study investigates the phytochemical composition and biological activities of Heliotropium indicum and Caesalpinia bonduc, two medicinal plants widely used in traditional medicine in Benin. In ethnomedicinal practice, H. indicum is commonly prepared as decoctions or infusions of the aerial parts and administered orally or topically to treat inflammatory conditions, wounds, fever, and infections, whereas C. bonduc roots are typically used in decoction for the management of urinary tract infections, pain, diabetes, and reproductive disorders. The present research aims to generate preliminary scientific data supporting these traditional applications. Aqueous and hydroethanolic (70% ethanol) extracts were prepared from the aerial parts of H. indicum and the roots of C. bonduc. Preliminary phytochemical screening was performed using standard qualitative colorimetric and precipitation reactions. The antimicrobial activities were evaluated against multiresistant uropathogenic strains using the agar disk diffusion method and broth microdilution for determination of minimum inhibitory concentrations (MICs). Antibiofilm activity was assessed using a crystal violet colorimetric assay. Analgesic and diuretic activities were evaluated in vivo in Wistar rats using hydroethanolic extracts. Acute oral toxicity was assessed following a limit test at 2000 mg/kg in rats. Phytochemical analysis revealed the presence of alkaloids, flavonoids, glycosides, saponins, and tannins in both plant species. Among the tested samples, only the hydroethanolic extract of C. bonduc exhibited antibacterial activity against the tested uropathogenic strains, with inhibition zones ranging from 14.6 to 16.6 mm and MIC values between 6 and 6.5 mg/mL. The aqueous extract of C. bonduc and both extracts of H. indicum did not exhibit detectable antibacterial activity under the experimental conditions. However, all extracts demonstrated antibiofilm activity, inhibiting biofilm formation in Escherichia coli and Staphylococcus spp. strains by 40.52%-71.93%. Hydroethanolic extracts of both plants showed significant analgesic activity (p < 0.05) in the acetic acid-induced writhing test and exhibited measurable diuretic effects in rats. In the acute toxicity study, no mortality or clinical signs of toxicity were observed at 2000 mg/kg during the 14-day observation period. Hematological and biochemical parameters remained within normal ranges, and histopathological examination of the liver and kidneys revealed no treatment-related alterations. These findings provide preliminary experimental support for some of the traditional uses of H. indicum and C. bonduc and suggest that hydroethanolic extracts of these plants warrant further investigation for the isolation and characterization of bioactive compounds.
Inflammation-induced dysregulation of microRNAs, particularly miR-138-5p, compromises trophoblast cell function and may contribute to pregnancy-related complications. While Elsholtzia species have been reported to possess therapeutic properties, the anti-inflammatory effects of Elsholtzia kachinensis Prain (EKP) in trophoblast cells have not been explored. In this study, a 70% ethanolic extract of EKP was fractionated into hexane (HEX), dichloromethane (DCM), ethyl acetate (ETAC), and water (WT) fractions. Each fraction was subjected to phytochemical profiling, antioxidant assessment, and bioactive compound identification by GC-MS and high-performance liquid chromatography (HPLC). Among them, the ETAC fraction (EKP-ETAC) showed the highest antioxidant activity, reflected by the lowest IC50 values in ABTS and DPPH assays, and contained the greatest levels of phenolic and flavonoid compounds. Nontoxic concentrations were determined by an MTT assay. Pretreatment with EKP-ETAC alleviated lipopolysaccharide (LPS)-induced impairment of proliferation, migration, and invasion in HTR-8/SVneo trophoblast cells, while reducing apoptosis, intracellular ROS generation, and inflammatory cytokine expression. Mechanistic analysis revealed that EKP-ETAC suppressed LPS-induced upregulation of miR-138-5p and restored the expression of FOXC1. The dual luciferase reporter assay confirmed that FOXC1 is a direct target of miR-138-5p, mediating the effects of EKP-ETAC. LC-MS and HPLC analyses identified rosmarinic acid as the predominant bioactive component. These findings suggest that rosmarinic acid-enriched EKP-ETAC exerts protective effects on trophoblast cells by mitigating oxidative stress and inflammation through the miR-138-5p/FOXC1 axis.
Metabolic dysfunction-associated steatotic liver disease (MASLD) is defined as the presence of excess triglyceride storage in the liver in the presence of at least one cardiometabolic risk factor. This term highlights the connection between fatty liver and metabolic dysfunction. Dietary factors, such as excessive consumption of saturated fats and sugar, contribute significantly to the accumulation of lipids in organs not specialized for fat storage, such as the liver. Hepatic lipid accumulation initiates dynamic changes in mitochondrial function and promotes the development and progression of MASLD. High-fat, high-fructose diet models have provided crucial insights into how nutritional factors induce mitochondrial dysfunction, which is characterized by impaired fatty acid oxidation, excessive generation of reactive oxygen species, and damage to mitochondrial DNA. Lifestyle modifications, including dietary adjustments such as calorie restriction and weight loss, are essential for the early prevention and long-term treatment of MASLD. However, in recent years, several pharmacological options have emerged for the treatment of MASLD, primarily for the management of its comorbidities. This review explores the mechanisms of mitochondrial dysfunction promoted by the accumulation of hepatic lipids, analyzes the evidence of mitochondrial alterations in the liver of dietary models, and summarizes some of the main therapeutic interventions for MASLD and their effects on mitochondrial function.
Background:The escalating rise of multi-drug-resistant (MDR) bacterial strains significantly threatens global health, creating a "silent pandemic" prompted by natural selection, gene mutation, and horizontal gene transfer. This crisis is worsened by the deficit in the development of new treatments, necessitating the innovative discovery of new potent antibacterial agents. Objective:This review examines animal venom, a complex mixture of an evolutionary array of bioactive molecules, as an important emergent source of broad-spectrum antimicrobial peptides (AMPs), creating potential drug templates for next-generation therapeutics. Results:We highlight numerous identified AMPs from various venomous taxa, including scorpions, snakes, spiders, frogs, bees, and wasps, characterized by their bactericidal activity against both Gram-positive and Gram-negative bacteria. They exhibit diverse mechanisms of action, characterized by rapid membrane disruption models, biofilm inhibition, bacterial enzyme dysregulation, immunomodulatory effects, and the control of intracellular targets. These bioresources serve as a structural base for the development of analogs with enhanced potency, higher selectivity, and less systemic toxicity. We also discuss repurposing strategies applied to the native AMPs, the potential application of nanoparticle technologies and the usage of computational methods. Conclusion:These advanced approaches accelerate the examination of large databases to optimize structure-function characteristics, providing a roadmap for the development of future potential antimicrobial treatments derived from the rich reservoir of animal venom bioactive molecules.
This investigation examined the long-term impact of stimulant use, assessing systematic dysfunction of the nervous system and how these changes were reflected in the gene expression of two addiction-related genes, SLC6A3 (dopamine transporter) and CARTPT (cocaine- and amphetamine-regulated transcript). Study subjects included 100 men over the age of 18 who have used stimulants (crystal and Captagon) as well as a group of 30 men of the same age who were healthy. Blood samples were taken from each subject, and plasma levels of dopamine, serotonin, norepinephrine, and total protein were measured using enzyme-linked immunosorbent assay (ELISA). Gene expression was analyzed in peripheral blood mononuclear cells (PBMCs) using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Relative expression of SLC6A3 and CARTPT was calculated using the 2-ΔΔCt method with GAPDH serving as an internal control. Stimulant users had significantly higher concentrations of plasma dopamine and norepinephrine compared to non-users, while also having much lower levels of both plasma serotonin and total protein compared to controls (p < 0.0001). qRT-PCR showed that the expression of SLC6A3 and CARTPT was significantly increased in the stimulant group (5-7-fold increase). These findings, which reflect both biochemical and gene expression level changes, indicate increased dopaminergic activity, decreased serotonergic activity, and changes in the neuropeptide pathways related to stress and reward due to chronic use of stimulants. Overall, the combination of the biochemical and gene expression data indicates that chronic crystal and Captagon users demonstrate significant systemic dysfunction. The significant changes in the levels of monoamines and the activation of the SLC6A3 and CARTPT genes indicate that these two genes could serve as peripheral biomarkers in assessing stimulant use disorder. These data give a mechanistic basis for developing new diagnostic and treatment methods based on dopaminergic and neuropeptide signaling pathways involved in the use of stimulants.
Pothos scandens, belonging to the Araceae family, locally known as "batilata," is traditionally used for treating various chronic diseases in folk medicine, such as epilepsy, seizures, convulsions, wounds, snakebites, skin issues, asthma, muscle problems, and diarrhea. This research examined the anxiolytic, antidepressant, analgesic, and anti-inflammatory potentials of the acetone extract of P. scandens whole plant (APS) on in vivo mice and rat models. The elevated plus maze (EPM) and hole board tests (HBTs) were carried out to assess the anxiolytic effects, while antidepressant activity with forced swimming test (FST) and tail suspension test (TST) and the acetic acid-induced writhing and formalin-induced licking tests on Swiss albino mice were used to evaluate the analgesic activity, and the carrageenan-induced rat paw edema model was applied for anti-inflammatory effect. APS at 400 mg/kg exhibited significant anxiolytic (p < 0.001) and antidepressant (p < 0.01) effects. This study also demonstrated that APS has dose-dependent and significant analgesic effects for acetic acid-induced writhing (p < 0.05) and formalin-induced licking tests (p < 0.01), respectively, as well as anti-inflammatory (p < 0.001) activities compared to the standard diclofenac sodium. Furthermore, 11 compounds were detected in the extract through GC-MS. These phytochemicals exhibited significant binding affinities, excellent pharmacokinetics, and a safe toxicological profile, as reported by in silico molecular docking and ADMET analysis. The outcomes of this study confirmed that the presence of bioactive compounds in APS offers it considerable potential as a multimodal therapeutic substance with neuropharmacological, analgesic, and anti-inflammatory properties.
Objective:Colon adenocarcinoma (COAD) remains a leading cause of cancer-related mortality, necessitating reliable prognostic biomarkers. This study explored the potential prognostic role of C-C motif chemokine ligand 3 (CCL3) and its association with immune-related signatures in COAD. Methods:Multiple publicly accessible databases, including TCGA, GEPIA2, UALCAN, TIMER, and Kaplan-Meier (KM) plotter, were used to examine the expression and prognostic profiles of CCL3 in various cancers. The association between CCL3 expression and clinical stage, immune infiltration, and immune gene markers was investigated using TIMER, GEPIA2, and TISIDB databases. Furthermore, we conducted functional enrichment analyses (GO and KEGG) of genes coexpressed with CCL3, assessed methylation status using OncoBD and MEXPRESS, and developed a gene-miRNA interaction network using miRWalk to elucidate their potential roles in COAD. Results:The expression of CCL3 was significantly upregulated in COAD and most other malignancies. Furthermore, high CCL3 expression was significantly associated with poor overall survival (OS) and relapse-free survival (RFS), with a univariate HR = 1.52 (95% CI: 1.12-2.06, p < 0.01). Multivariate Cox regression confirmed CCL3 as an independent prognostic factor after adjusting for the TNM stage, age, gender, and grade (adjusted HR = 1.42, 95% CI: 1.05-1.92, p = 0.023). CCL3 showed strong positive correlations with immune cell infiltration, particularly macrophages (Cor = 0.72, p < 0.001) and neutrophils (Cor = 0.74, p < 0.001), suggesting its role in shaping an immunosuppressive tumor immune microenvironment (TIME). Additionally, preliminary correlations indicate that high CCL3 may be associated with chemotherapy resistance. Conclusion:This study suggests that CCL3 could serve as a potential prognostic marker for COAD and other cancers. Because CCL3 expression was strongly associated with immune modulation within the tumor microenvironment, this pointed out that CCL3 could serve as a promising therapeutic target with significant implications for immunotherapy and chemotherapy response.
Oxidative stress and neuroinflammation are key components in neurodegenerative diseases, where early intervention using natural treatments may offer neuroprotective effects. This study shows that isoliquiritigenin (ISL), hesperidin (HES), and curcumin (CUR) can mitigate lipopolysaccharide (LPS)-induced neuroinflammation and oxidative stress in vitro and in vivo. The compounds were initially tested for cytotoxicity and found to reduce nitric oxide (NO) production, especially CUR and ISL. They were able to restore antioxidant enzyme activities both in vivo and in vitro. All treatments reduced inducible nitric oxide synthase (iNOS) expression compared to the untreated LPS control group. Behavioral assessments indicated that LPS impaired spatial and nonspatial memory, but treatments improved cognitive performance. Biomarker analyses revealed that ISL, HES, and CUR reduced the interleukin (IL)-1β and nuclear factor erythroid 2-related factor 2 (Nrf2) ratio in the hippocampus. Moreover, they decreased the level of caspase-3 demonstrated by western blotting and tumor necrosis factor-α (TNF-α) level. Thereby they inhibited LPS elicited apoptosis. Likewise, their anti-inflammatory effects were illustrated in the histopathological examination. Furthermore, they decreased the expression of amyloid-β. The study reinforces the potential of these natural compounds as protective and therapeutic cost-effective alternatives for managing neuroinflammation and neurodegeneration.
Background:While dyslipidemia is associated with chronic kidney disease (CKD), conventional observational studies cannot establish causality, and previous Mendelian randomization (MR) findings on low-density lipoprotein cholesterol (LDL-C) and CKD remain inconsistent. Objective:To further investigate the causal relationship between LDL-C and CKD using MR and evaluate its independence. Methods:We employed a two-sample and multivariable MR (MVMR) framework. Initially, two-sample and reverse MR analyses were performed for four lipid traits on CKD using genetic data from the IEU Open GWAS and the UK Biobank separately, with a Bonferroni-corrected significance threshold of p < 0.0125. Subsequently, MVMR was conducted to assess the independent effect of LDL-C after adjusting for other lipid traits (p < 0.05). Finally, an MVMR model incorporating LDL-C, smoking, alcohol consumption, and body mass index (BMI) was fitted to test the independence from these lifestyle confounders, followed by a sensitivity analysis with linkage disequilibrium-based confounder filtering to verify robustness. Results:Two-sample MR showed a significant causal effect of LDL-C on increased CKD risk (IEU source: OR = 1.13, 95% CI: 1.03-1.23, p < 0.01; consistent results from UK Biobank), with no evidence of reverse causation. MVMR confirmed the independence of this association: LDL-C remained significantly associated with CKD after adjusting for other lipid traits (OR = 1.21, 95% CI: 1.08-1.35, p = 0.0007) and after further adjustment for BMI, smoking, and alcohol (OR = 1.14, 95% CI: 1.04-1.25, p = 0.068). The effect direction remained consistent in the stringent sensitivity analysis. Conclusion:LDL-C may be an independent risk factor for CKD, the independence of which warrants further validation.
Background and Objective:Renal clear cell carcinoma (RCCC) stands out as a prevalent and aggressive subtype of kidney cancer characterized by a challenging prognosis. The need to enhance patient outcomes in RCCC underscores the significance of identifying prognostic biomarkers and therapeutic targets. MicroRNAs (miRNAs) and the signaling pathways orchestrating RCCC pathogenesis emerge as promising candidates for such endeavors. Methods:This study utilized publicly available gene expression data to compare miRNA profiles in nine RCCC and 11 normal kidney tissues. Rigorous bioinformatics analyses were employed to identify differentially expressed miRNAs and their associated gene targets. Prognostic significance was assessed, and a protein-protein interaction network was constructed to highlight pivotal RCCC hub genes. The expression and prognostic value of key hub genes and miRNAs were further validated in independent cohorts, including the GEO dataset GSE76351 and the TCGA-KIRC cohort via the Kaplan-Meier plotter. Expression of RUNX2 was confirmed using real-time PCR in five cancer and five normal renal tissues. Results:Fifteen DEMs were identified alongside 74 hub genes. The downregulation of miR-26a-1-3p, miR-144-3p, and miR-144-5p was associated with a poorer prognosis in RCCC. The overexpression of CDK1 and RUNX2 was validated in an independent GEO dataset and correlated with decreased patient survival in the TCGA-KIRC cohort. Furthermore, a statistically significant but modest inverse association was observed between miR-26a-1-3p and RUNX2 expression, indicating a possible miRNA-mRNA relationship. Significant enrichment was observed in pathways related to PI3K-Akt, MAPK, apoptosis, and cell cycle. The overexpression of RUNX2 was confirmed in our patient samples (p value< 0.05). Conclusion:This multistep validation study confirms that specific miRNAs and hub genes, particularly the miR-26a-1-3p/RUNX2 axis, are potential prognostic indicators in RCCC. A comprehensive understanding of these biomarkers and their enriched signaling pathways provides deeper insight into the molecular underpinnings of RCCC, uncovering potential therapeutic opportunities.
Bromelain is one of the protease enzymes found in all parts of pineapple (Ananas comosus (L.) Merr.), including the crown and peel. This enzyme has been widely used in various fields of life, including the food industry, health, pharmaceuticals, and cosmetics. However, pineapple processing often focuses on the flesh of the fruit, leaving behind substantial agricultural waste, such as crown and peel waste. The waste is often collected and stored before being used, causing the bromelain enzyme to decrease or even dissipate. Therefore, this study aims to determine the effect of time and the condition of storage of pineapple crown and peel waste on total protein content and protease activity. The extracted bromelain was precipitated with ethanol and then dried, and total protein content and protease activity were determined. The results showed that pineapple crown and peel waste can be stored for 7 days at 29 ± 1°C and humidity of 70 ± 2% and 20 days at 4 ± 0.5°C and humidity of 40 ± 2%, respectively. The total protein content and protease activity were 169.94 ± 2.68 μg/mL and 46.35 ± 0.69 IU/mg for crown bromelain, while those for peel bromelain were 229.75 ± 15.61 μg/mL and 29.10 ± 1.98 IU/mg, respectively. In conclusion, pineapple crown waste has the potential to be developed as a source of bromelain.
