The emergence of carbapenem-resistant Klebsiella pneumoniae (CR-Kp) has become a major global public health concern. Infections caused by CR-Kp strains that also possess hypervirulence traits present significant challenges for clinical management and are associated with increased morbidity and mortality. This study aimed to investigate the prevalence of β-lactamase genes and hypervirulence-associated markers among clinical CR-Kp isolates collected in Cyprus. A total of 96 K. pneumoniae isolates were investigated. Species-level identification and antimicrobial susceptibility testing were performed using the VITEK-2 system. Carbapenem resistance was assessed using the Modified Hodge test (MHT). Hypermucoviscous phenotype was determined by string test. The presence of carbapenemase genes (blaOXA-48, blaNDM, blaIMP, and blaVIM) and hypervirulence-associated genes (iucA, peg-344 and iroB) were investigated by conventional polymerase chain reaction. Among isolates, 75 (78.1
Immunotherapy through adoptive cell therapy (ACT) has become an effective cancer treatment method, using genetically modified immune cells or immune cells grown outside the body. The ACT methods use chimeric antigen receptor T (CAR-T) cells, with proven results for blood cancers, while tumor-infiltrating lymphocytes (TILs) and T-cell receptor (TCR)–engineered T cells serve as effective methods to fight against solid tumors and intracellular antigens. The current medical field uses natural killer (NK) cell-based therapies because of their natural ability to destroy cells, their lower incidence of graft-versus-host disease, and their capability to generate readily available therapeutic products through allogeneic medical procedures. The clinical utilization of ACTs has been facing multiple obstacles, stemming from antigen diversity, immune system resistance, treatment-related adverse effects, and difficulties in production. The researchers are advancing multiple solutions to resolve current challenges through developing multi-targeted receptor designs and gene-editing technologies, enhanced cell persistence strategies, and scalable manufacturing platforms. The next-generation ACT platforms will achieve improved therapeutic results through a mix of combination therapies, biomarker-driven patient selection, and advanced manufacturing technologies. Emerging areas of biological research demonstrate how adoptive cell therapies can develop into key components of precision cancer immunotherapy. In this review, we discussed the principles, benefits, and challenges of ACT, with a focus on potential solutions to overcome these obstacles.
The eukaryotic signal recognition particle (SRP) consists of six proteins and one SRP RNA. This ribonucleoprotein complex assembles inside the nucleus. Nucleocytoplasmic transport is an essential process for the biogenesis of signal recognition particles (SRPs) as well as for the survival of a cell. There are studies on cells that indicate the import receptor is responsible for import of SRP proteins into nucleus, but there is a lack of evidence that SRP proteins directly bind with import receptors. Coding sequences of SRP 14 and importin α were amplified from synthesized cDNA and genomic DNA, respectively, of Plasmodium falciparum cultivated in vitro culture. The amplified products were cloned and expressed in E. coli, followed by purification. A binding study was conducted on glutathione-agarose as well as in a 96-well plate format at different concentrations of SRP 14 with immobilized importin α. This is the first report of direct binding between importin α and a eukaryotic signal recognition particle 14 (SRP 14). A cost-effective 96-well plate-based assay has also been developed to study the binding of cargoes of importin α.
Mealybugs (Hemiptera: Pseudococcidae) are economically important invasive pests that cause substantial losses in agricultural and horticultural crops worldwide. However, information on their diversity, genetic variation, and population structure in India remains limited. This study investigated the diversity, phylogenetic relationships, and population genetic structure of economically important mealybugs across different agroclimatic regions of India using mitochondrial cytochrome c oxidase subunit I (mtCOI) sequences. A total of 134 mealybug specimens were analysed using an integrative taxonomic approach combining morphological identification and mtCOI-based DNA barcoding. Phylogenetic relationships, haplotype composition, population differentiation, and demographic patterns were assessed using phylogenetic reconstruction, haplotype networks, analysis of molecular variance, neutrality tests, and mismatch distribution analyses. Fourteen mealybug species belonging to eight genera were identified, with phylogenetic analyses resolving distinct species-specific clades and supporting the utility of mtCOI for species identification. Phenacoccus solenopsis and Planococcus minor exhibited multiple haplotypes and pronounced geographic population structuring. Haplotype networks revealed both widely distributed and geographically restricted haplotypes, indicating historical dispersal, geographic isolation, and localized diversification. Demographic analyses suggested contrasting population histories among geographic regions, including evidence of recent expansion in some populations. The study provides a comprehensive molecular assessment of mealybug diversity and population structure in India. The findings reveal substantial geographic differentiation and complex evolutionary patterns among economically important mealybugs and provide a molecular basis for accurate species identification, invasion monitoring, biosecurity surveillance, and region-specific integrated pest management. These results improve understanding of mealybug dispersal and diversification and support effective management of invasive pest populations in India.
Dimethyl Phthalate (DMP) is a phthalate ester, widely present in nail polishes, hairsprays, coatings, textiles, and certain insect repellents, contributing to its pervasive environmental and occupational presence. DMPs are now considered priority emerging contaminants due to limited regulation and potential long-term adverse effects on human health. Despite inhalation being a major route of exposure, data on the pulmonary toxicity of phthalates, particularly DMP, remain limited. To investigate the cellular responses associated with DMP-induced pulmonary toxicity using the human lung adenocarcinoma-derived alveolar epithelial cell line A549 as an in vitro model. A549 cells were exposed to varying concentrations of DMP for different durations to investigate its cytotoxic effects. Cell morphology, viability, oxidative stress, mitochondrial superoxide generation and apoptosis were evaluated using phase-contrast microscopy, MTT assay, DCFH-DA, MitoSOX staining, and Annexin V/PI flow cytometry. Inflammatory and fibrotic responses were assessed by measuring cytokine and fibronectin expression using RT-qPCR, ELISA and ICC. Exposure to DMP resulted in significant concentration- and time-dependent cytotoxicity, accompanied by elevated intracellular and mitochondrial reactive oxygen species generation. DMP treatment also induced necrotic cell death and significantly increased the mRNA expression of the pro-inflammatory cytokines TNF-α, IL-6, IL-8, and enhanced the secretion of IL-6 and IL-8. Furthermore, DMP exposure enhanced fibronectin expression and accumulation, suggesting activation of remodeling-associated responses. These findings demonstrate that DMP elicits oxidative stress–driven epithelial damage, inflammatory responses, and fibronectin accumulation in human lung epithelial cells that overlap with pathways implicated in chronic respiratory diseases, including COPD.
Hypoxia modulates immune responses during intracellular infections, yet quantifiable cytokine thresholds for prognostication remain undefined in experimental models. The immunological basis of increased mortality when hypoxemia coexists with bacterial infection remains incompletely understood. In a randomized study, 175 male Sprague-Dawley rats were stratified into seven groups combining normoxia (21.5
Organ fibrosis is the ultimate common pathway resulting from dysregulated tissue repair caused by chronic inflammation, it accounts for approximately 45
Chemoresistance remains a major barrier to effective cancer therapy, limiting durable responses across multiple malignancies. Proteases, including matrix metalloproteinases, cathepsins, ubiquitin-specific proteases, proteasome components, and mitochondrial proteases, contribute to chemoresistance through diverse mechanisms, including modulation of drug transport, extracellular matrix remodeling, epithelial–mesenchymal transition, apoptosis, and pro-survival signaling. Collectively, these processes promote a tumor microenvironment that facilitates therapeutic escape. This review critically synthesizes current mechanistic evidence together with available clinical studies on protease-mediated chemoresistance, emphasizing the gap between strong biological rationale and limited clinical success. Broad-spectrum protease inhibition has shown limited clinical success owing to insufficient isoform selectivity, compensatory signaling, inadequate biomarker-guided patient stratification, and limited pharmacodynamic validation. Emerging strategies focus on context-specific protease targeting, biomarker guided patient selection, rational combination therapy, and advanced drug delivery platforms, including protease-activated prodrugs and nanoparticle-based systems. Integration of proteomic profiling with functional validation may facilitate identification of protease dependencies and support more precise therapeutic interventions. Collectively, these advances suggest that protease inhibition may evolve from empirical enzyme blockade toward mechanism-driven approaches for overcoming chemoresistance, although further clinical validation remains necessary.
Neurodegenerative disorders are characterized by progressive neuronal loss and remain a major global health concern, largely due to their complex and multifactorial nature. These conditions, including Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease, involve overlapping mechanisms such as oxidative stress, chronic inflammation, mitochondrial dysfunction, and impaired neurotransmission. In recent years, natural bioactive compounds have gained attention as potential therapeutic candidates. Among them, apigenin, a dietary flavonoid widely found in fruits and vegetables, exerts neuroprotective effects through multiple cellular pathways. This review highlights the potential role of apigenin in modulating key pathological processes in the central nervous system. Apigenin reduces oxidative stress by scavenging reactive oxygen species and activating endogenous antioxidant systems, particularly through Nrf2 signaling. It also exhibits anti-inflammatory effects by inhibiting pathways such as NF-κB and MAPK, thereby lowering pro-inflammatory cytokine production and regulating microglial activation. In addition, apigenin supports mitochondrial function and inhibits apoptosis by modulating Bcl-2 family proteins and caspase activity. Its role extends beyond maintaining blood–brain barrier integrity to regulating autophagy and mitophagy and enhancing neurotrophin signaling, especially through the BDNF/TrkB pathways. Moreover, apigenin influences neurotransmission by balancing excitatory and inhibitory signaling and modulating cholinergic and monoaminergic systems. Despite promising preclinical findings, its clinical application remains limited due to bioavailability challenges and insufficient human studies.
Klebsiella pneumoniae is an important opportunistic pathogen with a high capacity to acquire antimicrobial resistance determinants. However, data on the distribution and co-occurrence of extended-spectrum β-lactamase (ESBL), carbapenemase and plasmid-mediated colistin resistance genes in poultry-associated K. pneumoniae in Pakistan remain limited. This study investigated the prevalence, antimicrobial resistance profiles and molecular characteristics of K. pneumoniae recovered from commercial broiler farms, backyard poultry flocks and retail poultry markets. A total of 900 samples were collected from commercial broiler farms, backyard poultry flocks and retail poultry markets in Faisalabad Division, Pakistan, between September 2024 and April 2025. K. pneumoniae isolates were identified using conventional biochemical testing, API 20E, MALDI-TOF MS and rpoB gene amplification. Antimicrobial susceptibility testing was performed according to CLSI 2024 recommendations, and PCR was used to detect ESBL, carbapenemase, plasmid-mediated quinolone, aminoglycoside, sulfonamide, tetracycline, chloramphenicol and colistin resistance genes. K. pneumoniae was recovered from 303/900 samples (33.7
Periodontitis is a chronic inflammatory disease characterized by persistent periodontal tissue destruction and alveolar bone loss, which is closely associated with multiple systemic disorders. Emerging evidence indicates that ferroptosis and metabolic reprogramming are jointly involved in the progression of periodontitis; however, their causal linkage, hierarchical regulatory network, and unified core mechanism remain unclear. This review proposes a mitochondria-centered unifying axis that links metabolic reprogramming and ferroptosis in periodontitis. Specifically, periodontal pathogens trigger mitochondrial structural and functional collapse, which in turn drives comprehensive metabolic disturbances covering iron, lipid, amino acid, and glucose metabolism, thereby facilitating ferroptosis. Exacerbated ferroptosis further aggravates mitochondrial damage, forming a self-sustaining pathological vicious cycle that underlies refractory inflammation and sustained alveolar bone resorption. We systematically elaborate the molecular basis of this mitochondria-metabolism‐ferroptosis axis and outline a translational framework encompassing mitochondria-targeted therapeutics and advanced nano-delivery systems. This review provides a novel mechanistic basis for addressing therapeutic resistance and frequent recurrence in periodontitis, with profound theoretical value and translational promise for clinical application.
Antimicrobial resistance (AMR) poses a major global health threat, driven by bacterial defense mechanisms such as efflux pumps, enzyme-mediated drug inactivation, target site modification, and biofilm formation. Metal nanoparticles (NPs), including silver, gold, iron oxide, and zinc oxide, offer a potential multi-target strategy to dismantle these resistance pathways. Their antimicrobial activity stems from direct membrane disruption, induction of oxidative stress, and, crucially, the modulation of bacterial gene expression, including downregulation of efflux pump and biofilm-related genes and disruption of quorum-sensing networks. The efficacy and specificity of NPs are significantly enhanced through advanced functionalization strategies, such as organic ligand conjugation, antibody coupling, and green synthesis approaches using biological resources, enabling targeted drug delivery and reduced off-target toxicity. Despite promising translational applications in antimicrobial coatings, wound dressings, and adjuvant therapies, clinical adoption faces hurdles including potential cytotoxicity, unpredictable pharmacokinetics, and the need for standardized manufacturing. Future directions involve the development of environmentally responsive “smart” NPs, nanoparticle-mediated delivery of siRNA to suppress resistance genes, and the integration of multi-omics approaches to decipher NP-bacteria interactions at the molecular level. Collectively, engineered metal NPs represent a promising platform for next-generation antimicrobial strategies and may help address multidrug resistance through simultaneous molecular and physical effects.
Accurate species delimitation within electric rays of the genus Narcine has been hindered by overlapping morphological characters and limited molecular resolution in previous single-locus studies. This study aims to evaluate phylogenetic relationships and species boundaries within the Narcine species complex across the Western Atlantic using complete mitochondrial genomes. Seven complete mitogenomes were newly assembled from individuals representing distinct morphotypes sampled across geographically widespread Western Atlantic localities and analyzed together with publicly available reference sequences. Mitochondrial protein-coding genes (PCGs) were examined using concatenated nucleotide and amino acid datasets under partitioned maximum-likelihood frameworks. Both approaches recovered highly congruent topologies, consistently supporting a single, well-defined western Atlantic mitochondrial lineage with low internal divergence (0.04–2.13
Oral squamous cell carcinoma (OSCC) is a leading cause of cancer mortality, and reliable molecular markers are needed, particularly in resource-limited settings. This pilot study evaluated the expression of two microRNAs, hsa-miR-21-5p and hsa-miR-26a-5p, in OSCC tissue compared with normal oral tissue, as a hypothesis-generating step toward biomarker development. In this single-centre case–control pilot study, 16 histopathologically confirmed OSCC tissues and 16 histologically normal control tissues were analysed. Total RNA was extracted and reverse-transcribed, and hsa-miR-21-5p and hsa-miR-26a-5p were quantified by qRT-PCR using U6 snRNA as reference gene and the 2^−ΔΔCt method. Group differences were assessed using the Mann–Whitney U test with Hodges–Lehmann median differences, and receiver operating characteristic (ROC) analysis with bootstrap and leave-one-out internal validation. hsa-miR-26a-5p was consistently upregulated in OSCC (Hodges–Lehmann ΔCt difference 3.59, 95
Egregia menziesii (E. menziesii) is recognized for its anticancer and hypoglycemic properties. Chemically, it contains compounds with potential anti-inflammatory activity. Within the immune system, macrophages play an essential role in containing and eradicating pathogens through phenotypic polarization. The ethanol and water (1:1 v/v) (EtOH: H2O), extract of E. menziesii was obtained and fractionated. The chemical profiles of the extracts featured signals linked to sulfated polysaccharides and mannitol. Cell viability was evaluated at various concentrations with MTT assay and confirmed the safety (≥ 90
The combination of ethyl acetate fractions from Uncaria sp. and Selaginella doederleinii (EAF-BC) holds potential as an anticancer agent; however, its underlying molecular mechanism remains unclear. This study aimed to determine the anticancer potential of these combinations by evaluating the hallmarks of apoptosis using a Saccharomyces cerevisiae model. Yeast cells were incubated with EAF-BC at concentrations of 5, 10, and 100 ppm for 24 h. The analyzed parameters included petite colony frequency, mitochondrial activity, cell surface morphology, phosphatidylserine externalization, DNA fragmentation, and apoptosis-related gene regulation (FSH3, AIF1, and YCA1). The results showed that EAF-BC induced mitochondria-driven apoptosis in a dose-dependent manner. The 100-ppm concentration caused severe mitochondrial damage, yielding 85
Neuroinflammation involves the activation of glial cells, particularly astrocytes, and microglia, in response to pathological stimuli. Astrocyte activation is characterized by increased pro-inflammatory cytokines and oxidative stress. Given their role in neuroinflammation, targeting astrocytic mechanisms presents potential therapeutic opportunities. Ellagic acid (EA), a phenolic compound, exhibits antioxidant, anti-inflammatory, and neuroprotective effects. This study investigated the effect of EA on astrocytes from neonatal rats exposed to lipopolysaccharide (LPS), a potent proinflammatory agent widely used to induce neuroinflammation in vitro. Astrocytes were pretreated with EA (50, 100, and 200 µM) for 48 h and subsequently exposed to LPS (1 µg/mL) for 3h. Cell viability and proliferation, inflammatory and oxidative stress parameters were evaluated. LPS decreased astrocytic viability and increased cell proliferation and EA prevented these alterations. EA also attenuated the LPS-induced increase in reactive oxygen species and nitrite levels and the decrease in sulfhydryl content and antioxidant enzyme activities. LPS induced an increase in gene expression of interleukin 1β and tumor necrosis factor-alpha, and EA treatment attenuated these alterations. Conversely, EA increased interleukin 6 (IL-6) and glycogen synthase kinase-3 beta mRNA expression compared with LPS-only exposure. LPS increased IL-6 levels and decreased interleukin 10 (IL-10) levels in astrocytes, and EA at 100 µM prevented the reduction in IL-10 levels but did not significantly modify IL-6 levels compared with LPS group. EA modulates oxidative and inflammatory markers in LPS-stimulated astrocytes. Further studies are needed to clarify the underlying mechanisms and evaluate these effects in more complex models of neuroinflammation.
The nuclear factor erythroid 2-related factor 2 (NRF2) signaling pathway is a major regulator of cellular response to oxidative stress. While NRF2 protects the normal cells against oxidative stress, its abnormal signaling activity in the context of cancer helps tumor cells evade oxidative stress, survive, proliferate, and develop resistance to treatment. Increasing evidence has established the tight connection between NRF2 signaling and ferroptosis, a type of programmed cell death based on iron-dependent lipid oxidation. In this review, the role of NRF2 in regulating important pathways as glutathione metabolism, lipid peroxidation, and iron homeostasis that contribute to the process of ferroptosis is discussed. This review highlights the latest progress that has been made in therapies that are designed to target the NRF2–ferroptosis pathway in cancers, by providing an overview of the mechanism involved as well as relevant pre-clinical and current clinical studies. The latest advancements regarding the NRF2 modulators and ferroptosis inducers, both natural and synthetic, along with new studies regarding the ability of such compounds to overcome drug resistance and increase antitumor effectiveness are reviewed.In addition, it is addressed current studies in biomarker discovery, precision medicine, drug targeting in terms of future direction. Moreover, the existing limitations related to clinical application of NRF2 and ferroptosis modulation therapies are also described, including tumor heterogeneity, treatment selectivity, biomarker validation, and possible toxicity of such therapy to normal tissue.Overall, these developments provide the rationale for the clinical application of the NRF2–ferroptosis pathway as a therapeutic target in overcoming drug resistance. NRF2 signaling regulates ferroptosis susceptibility and therapeutic resistance in cancer. Targeting NRF2 can enhance ferroptosis induction in therapy-resistant cancer cells. Natural and synthetic NRF2 modulators offer promising strategies to overcome ferroptosis and drug resistance. Combining NRF2 modulation with ferroptosis inducers may improve antitumor efficacy while limiting off-target toxicity. NRF2-mediated ferroptosis is a vital target for precision oncology.
This study aimed to evaluate the oncolytic efficacy of a combination of vaccine-strain measles virus (MeV) and mumps virus (MuV) against colorectal cancer (CRC) cells in vitro and in a nude mouse xenograft model. HT-29 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) and infected with MeV, MuV, or a combination of MeV and MuV (MM; MeV: MuV = 1:1, v/v). Cell viability, synergistic effects, and apoptosis were assessed using MTT and flow cytometry assays, respectively, in vitro. A nude mouse xenograft model was established to evaluate the in vivo oncolytic efficacy of the MM against CRC xenograft tumors. The MM demonstrated synergistic effects at 48, 72, and 96 h (CI = 0.473, 0.692, and 0.633, respectively). The MM-treated group showed significantly lower cell viability (p < 0.05) and a markedly higher apoptotic rate (p < 0.01) compared with single-virus–infected groups in vitro. In the xenograft model, the MM-treated group exhibited significantly slower tumor growth (p ≤ 0.001), significantly prolonged survival (p < 0.05), reduced mortality, and increased proportions of innate immune cell populations in the spleen compared with single-virus–treated groups. The MM demonstrated synergistic and superior oncolytic efficacy against HT-29 colorectal cancer cells compared with single-virus therapies in both in vitro and in vivo xenograft models.
Circular RNAs (circRNAs) have emerged as pivotal regulators in cancer biology, attributed to their stability, abundance, and tissue-specific expression. Dysregulation of the Hippo pathway is associated with various cancers, with circRNAs influencing its activity. These circRNAs exhibit aberrant expression in multiple cancers, correlating with clinical features and prognosis. The interaction between circRNAs and the Hippo signaling pathway highlights their potential as diagnostic and prognostic biomarkers. By modulating Hippo pathway activity, circRNAs significantly impact tumorigenesis and cancer progression. Therapeutic strategies targeting circRNA-Hippo interactions have demonstrated promise in preclinical models. This review delves into the complex roles of circRNAs in Hippo pathway modulation and their implications for cancer progression. Future research should aim to elucidate the mechanisms of circRNA-Hippo interactions comprehensively and translate these insights into clinical applications, thereby advancing cancer diagnosis, prognosis, and treatment. Integrating circRNA profiling into clinical practice could revolutionize cancer management and enhance patient outcomes.
