Search Results (20,375)

Background: Transmitted drug resistance (TDR) remains a critical challenge in HIV-1 management, particularly in treatment-naïve populations. Baseline genotypic resistance testing is recommended to optimize antiretroviral therapy (ART), yet data from Saudi Arabia remain limited. This study aimed to characterize HIV-1 genetic diversity and baseline resistance-associated mutations among newly diagnosed, ART-naïve individuals. Methods: We conducted a multi-centre, retrospective cross-sectional study across three hospitals in Saudi Arabia between January 2023 and December 2024. Adult, treatment-naïve patients with confirmed HIV infection who underwent genotyping using Sanger sequencing were included. Mutations in reverse transcriptase (RT), protease (PI), and integrase (INSTI) genes were analyzed using the Stanford HIV Drug Resistance Database. Demographic, clinical, and virological data were collected, and comparative analyses between regions were performed. Results: A total of 614 patients were included, predominantly male (85.5%) and aged 25–44 years. Most patients presented with high viral loads (≥10,000 copies/mL, 89.7%), and 25.3% had CD4 counts <200 cells/mm³. HIV-1 subtype distribution was highly diverse, with subtype C (19.5%), CRF02_AG (15.6%), and subtype G (12.7%) predominating. Although mutations were frequently detected (RT: 85.2%, PI: 94.7%, INSTI: 36.4%), the majority were subtype-associated polymorphisms rather than major drug resistance mutations. Clinically significant mutations including M184V/I (1.17%), K103N (0.83%), and K65R (0.17%) were observed at low frequencies. No major INSTI resistance mutations were detected. Multi-class mutation patterns were common but largely driven by accessory variants. Conclusions: Despite the high prevalence of detected mutations, clinically significant TDR remained low, occurring in approximately 2.8% of patients. Most detected variants were polymorphic or accessory mutations, while susceptibility to INSTIs remained largely preserved. Continued baseline genotyping and molecular surveillance remain important to monitor emerging resistance patterns. Full article

Bacterioruberin (BR), a C₅₀ carotenoid produced by halophilic archaea, is emerging as a bioactive molecule with potential anticancer relevance, but its activity in solid tumour and multidrug-resistant (MDR) models remains poorly defined. This in vitro study evaluated the cytotoxic, antiproliferative and growth-inhibitory effects of a chemically characterized bacterioruberin-rich carotenoid extract (BRCE) from Haloferax (H.) mediterranei in A549 lung adenocarcinoma, BT-549 triple-negative breast cancer and WM115 melanoma cells, as well as in paired sensitive/multidrug resistant (MDR) lung cancer models. Metabolic activity and proliferation were assessed by thiazolyl blue tetrazolium bromide (MTT) and carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) assays, intracellular reactive oxygen species (ROS) by 2′,7′-dichlorodihydrofluorescein diacetate (H₂DCFDA) staining, and apoptosis-associated morphology by acridine orange/ethidium bromide (AO/EB) staining. Growth inhibition in A549/A549-CDR2 and NCI-H460/NCI-H460/R cells was analysed by sulforhodamine B (SRB) assay, while P-glycoprotein (P-gp) function and expression were examined using Rhodamine 123 (Rh123) accumulation and Western blotting. BRCE reduced metabolic activity and proliferation in a concentration- and time-dependent manner, increased intracellular ROS levels, and induced apoptosis-associated morphological changes in A549 cells. In MDR models, BRCE retained comparable growth-inhibitory activity in sensitive and resistant cells and partially attenuated P-gp-related drug efflux. These findings support further mechanistic investigation of BR in solid tumour and MDR cancer models. Full article

The emergence of multidrug-resistant (MDR) bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA), represents a major global health challenge by limiting the current therapeutic options. In this context, antimicrobial peptides (AMPs) have been widely studied for their potent antimicrobial properties. In this study, we evaluated the anti-MRSA effect of a novel AMP, LyeTx I mnΔKL, derived from a toxin of Lycosa erythrognatha. Its activity was evaluated in vitro by minimal inhibitory and bactericidal concentrations (MIC and MBC), antibiofilm effect, membrane interaction, cytotoxicity, synergistic interaction with vancomycin, and in vivo in an MRSA murine wound/abscess infection model. LyeTx I mnΔKL showed enhanced antimicrobial activity against clinical MRSA isolates compared to its prototype (LyeTx I mnΔK), with MIC₅₀ and MBC₅₀ of 2 and 8 µM and 16 and 32 µM, respectively. Furthermore, LyeTx I mnΔKL exhibited a rapid bactericidal effect and a pronounced ability to inhibit biofilm formation and disrupt mature biofilms. LyeTx I mnΔKL interacts with bacterial membranes, adopts an α-helical structure, and induces membrane disruption and leakage of intracellular material. In vivo, topical treatment with LyeTx I mnΔKL reduced MRSA burden compared with LyeTx I mnΔK and untreated controls (log₁₀ CFU/g of wound of 2.4, 4.4, and 6.5 for LyeTx I mnΔKL, LyeTx I mnΔK, and the saline group, respectively). However, increased cytotoxicity remains a significant limitation. Overall, LyeTx I mnΔKL is a promising anti-MRSA candidate for topical use with potent antibiofilm and in vivo activity. Full article

Tuberculosis (TB) continues to be a significant public health challenge in Europe, despite a sustained decline in disease incidence over recent decades. This narrative review briefly traces the historical development of TB diagnosis and focuses on the evolution of TB control protocols from the early twentieth century to the present across Europe, through a longitudinal comparative analysis of its geographical regions. A literature search was conducted using publications, guidelines, and surveillance reports from the World Health Organization (WHO), the European Centre for Disease Prevention and Control (ECDC), and national public health authorities. The analysis follows a geographical framework encompassing Eastern, Western, Northern, and Southern Europe, reflecting historical, socioeconomic, and healthcare system differences. This study presents the transition from traditional diagnostic approaches based on clinical assessment, chest radiography, and smear microscopy to modern molecular and immunological techniques, including Xpert MTB/RIF assays and interferon-gamma release assays (IGRAs). Similarly, treatment strategies have evolved from sanatorium-based supportive care to standardized, evidence-based short-course regimens employing first- and second-line anti-TB drugs. However, marked regional differences remain in the implementation of contemporary protocols. Western and Northern European countries have largely adopted advanced diagnostic technologies and comprehensive surveillance systems and are approaching TB elimination targets. In contrast, Eastern Europe continues to bear a disproportionate disease burden, driven by multidrug-resistant TB, HIV co-infection, and socioeconomic disparities. TB control protocols in Southern Europe are progressively converging with those of Western Europe through the adoption of modern diagnostic approaches, standardized treatment regimens, and WHO-endorsed guidelines. The findings of this study underscore the need for greater harmonization of TB control protocols across Europe through an initiative coordinated by the ECDC/WHO. Accelerating progress toward TB elimination in the European Region will depend on expanding access to modern diagnostic technologies, implementing targeted interventions in high-burden settings, and strengthening cross-border collaboration through coordinated public health policies. Full article

Disorders of the central nervous system (CNS) are intricate and often resistant conditions that create a significant global impact, affecting millions of individuals each year. The blood–brain barrier (BBB) acts as a protective mechanism for the brain against external substances, but it also prevents most therapeutic agents from entering the CNS, leading to inadequate drug absorption and reduced effectiveness after diagnosis. Nasal drug delivery has emerged as a viable approach to bypass the BBB, facilitating direct access to the brain through the olfactory and trigeminal nerve routes. Although considerable research focuses on innovative nasal formulations with proven clinical promise, a critical gap persists: a systematic framework that bridges laboratory breakthroughs with clinical implementation. This review addresses this unmet need by integrating recent basic research advances with practical clinical requirements. We summarize nasal transport pathways, targeted design strategies, formulation optimization, and device engineering. Crucially, we propose a structured clinical evaluation framework built upon five essential pillars: targeting precision, pharmacokinetic performance, multi-organ safety profiling, device–drug clinical compatibility, and anatomical translation from animal models to humans. By mapping current research capabilities against clinical readiness criteria, this framework identifies translational bottlenecks and provides actionable guidance to accelerate the bench-to-bedside transition of intranasal drug delivery systems for CNS disorders. Full article

Background: Acute sore throat (pharyngitis) accounts for 10–30% of ambulatory visits annually. Bacterial etiology, primarily Group A Streptococcus, is confirmed in only 20–34% of cases, and inappropriate antibiotic administration remains frequent, contributing to antimicrobial resistance. Community pharmacists play a key role in symptomatic management and stewardship, but real-world practices remain underexplored. Methods: A 16-item online questionnaire surveyed 629 Italian community pharmacists via professional networks, covering demographics, diagnostic tools, symptom assessment, treatment preferences, and follow-up. Responses were analyzed using descriptive statistics and interpreted within a multidisciplinary expert framework. Results: Italian community pharmacists (n = 629), predominantly mid-career and experienced individuals, prioritize targeted symptom questioning (flu-like signs (59.96%), fever (48.73%), pain intensity (49.64%)) and medical referral over formal diagnostic tools like the Centor criteria (unused in 53.66% of cases). A total of 46.74% report performing a Strep A test in fewer than 20% of patients. Topical sprays (83.33%) and lozenges (44.69%), especially containing flurbiprofen (94.19%), dominate recommendations, followed by systemic analgesics (45.79%). For mild cases, anti-inflammatory/analgesic sprays/lozenges (35.42%) and antiseptic lozenges (35.61%) dominate, while in severe cases, drugs with anti-inflammatory or analgesic effects are often preferred (72.47%). Most pharmacists request a follow-up (73.22%). Conclusions: Italian community pharmacists report a predominantly topical-first approach focused on symptomatic treatment and referral when appropriate. Diagnostic gaps and inconsistent follow-up represent actionable targets. The findings inform training outputs prioritizing simplified triage, expanded symptom checklists, spray/lozenge optimization, and 3–5-day call-backs. This work aims to promote pharmacists’ antimicrobial stewardship to reduce antibiotic use while enhancing patient-centred sore throat care in Italy’s pharmacy network. Full article

Anti-CD38 monoclonal antibodies have substantially improved outcomes in multiple myeloma (MM). Although daratumumab and isatuximab target the same antigen, accumulating evidence indicates that they differ in epitope recognition, biological activity, and immunomodulatory properties, suggesting these agents may not be therapeutically interchangeable. This review summarizes the molecular and immunological mechanisms underlying their distinct antitumor effects and their implications for treatment selection. Isatuximab binds near the catalytic site of CD38, resulting in potent enzymatic inhibition, enhanced antibody internalization, FOXM1 suppression, and reactive oxygen species-mediated cytotoxicity, which may preferentially target MM cells harboring 1q21 amplification. In contrast, daratumumab exerts prominent Fc-dependent immune effects, including trogocytosis-mediated downregulation of CD38 and VLA-4, suppression of cell adhesion-mediated drug resistance, and modulation of the immune microenvironment, potentially enhancing subsequent T-cell-redirecting therapies. We further discuss the relevance of these mechanistic differences to measurable residual disease, extramedullary disease, and sequencing with BCMA- and GPRC5D-directed immunotherapies. Finally, we propose a biology-guided treatment-selection model integrating genomic alterations, tumor biology, and immune remodeling to support precision medicine for patients with MM. Full article

Background: Drug resistance and treatment-associated toxicity remain major limitations of conventional chemotherapy for triple-negative breast cancer (TNBC). Thymoquinone (TQ), a bioactive phytochemical derived from Nigella sativa, has demonstrated anticancer properties and may enhance the therapeutic efficacy of docetaxel (DTX) through complementary molecular mechanisms. Objective: To investigate whether TQ potentiates the antitumor activity of DTX in MDA-MB-231 TNBC cells by affecting apoptosis, oxidative stress, wound closure, and PI3K/AKT pathway-related gene expression. Methods: MDA-MB-231 TNBC cells and HaCaT keratinocytes were treated with TQ, DTX, or their combination. Cell viability was determined using the MTT assay, and drug interactions were evaluated by the Chou–Talalay combination index (CI) method. Apoptosis, intracellular reactive oxygen species (ROS) production, ROS rescue experiments using N-acetyl-L-cysteine (NAC), caspase-9 expression, wound closure, and gene-expression changes were assessed using Annexin V/PI flow cytometry, DCFH-DA-based flow cytometric and fluorescence analyses, immunocytochemistry, wound-healing assay, and quantitative real-time PCR (qRT-PCR), respectively. Bioinformatic analyses were performed to identify signaling pathways associated with the observed molecular alterations. Results: The TQ + DTX combination demonstrated synergistic cytotoxicity and significantly increased apoptotic cell death compared with either monotherapy. Combination treatment markedly enhanced intracellular ROS accumulation, whereas NAC pretreatment significantly attenuated ROS generation and partially reversed the cytotoxic and pro-apoptotic effects, suggesting the involvement of ROS in the observed antitumor effects. Caspase-9 immunoreactivity was markedly increased following combination treatment, suggesting the involvement of the intrinsic apoptotic pathway. Furthermore, the combination significantly suppressed wound closure and downregulated BCL2, PIK3CA, and AKT1 while upregulating BAX, CASP9, and PTEN. Bioinformatic analyses identified apoptosis, p53, PI3K/AKT, mTOR, and MAPK signaling as the principal pathways potentially associated with the observed gene expression changes. Conclusions: TQ potentiates the antitumor activity of DTX, with the involvement of oxidative stress, apoptotic signaling, suppression of wound closure, and regulation of PI3K/AKT pathway-related gene expression in TNBC cells. These findings provide evidence supporting further preclinical investigation of the TQ + DTX combination as a promising therapeutic strategy for triple-negative breast cancer. Full article

Epilepsy is a multifactorial disorder, yet routine management still focuses on neuronal excitation and insufficient inhibition, with antiseizure medications (ASMs) as the primary therapeutic strategy. This approach fails in roughly one-third of patients who develop drug-resistant epilepsy (DRE). Converging evidence links DRE with neuroinflammation, oxidative stress (OS), and mitochondrial dysfunction—an interconnected distal pathophysiological triad that progressively lowers seizure thresholds yet remains peripheral to clinical epilepsy management. We map this triad mechanistically and show that ASMs modulate it beyond their anticonvulsant activity, while triad-targeting pharmacological, dietary, and botanical interventions independently reduce seizure susceptibility. Common precipitants are reinterpreted as acute activators of the distal triad, linking precipitant identification and patient agency to threshold elevation. Integrating these elements, we propose a threshold management framework for DRE, built on a revised reservoir model, and translate it into three structural priorities: mechanistic phenotyping to stratify patients by pathophysiological domain, dual-mechanism drug development, and trial designs suited to multicomponent, context-dependent interventions. Together, these proposals reframe epilepsy management from sequential pharmacological trials toward coordinated optimization of the full seizure threshold landscape. Full article

Background: The dissemination of extensively drug-resistant Pseudomonas aeruginosa strains across the human–food interface represents a critical threat to global public health, complicating empirical therapeutic choices and challenging traditional nosocomial restriction paradigms. Objective: This study investigated the prevalence, phenotypic resistome landscapes, and underlying integron-mediated mobilization mechanics of P. aeruginosa across the One Health continuum using an optimized real-time quantitative PCR (qPCR) platform. Methods: A total of 116 samples comprising human clinical specimens (n = 61) and retail food matrices (n = 55) were screened. Phenotypic resistance was profiled against 16 antipseudomonal agents, with polymyxin and colistin resistance validated via reference broth microdilution. A novel real-time SYBR Green qPCR assay backed by derivative melting curve analysis was developed to track Class 1 (intI1) and Class 2 (intI2) integrons. Results: Pseudomonas aeruginosa was recovered from 36.2% (n = 42/116) of total samples, exhibiting a homogeneous distribution across clinical (32.8%) and retail food matrices (40.0%, p = 0.445). Antibiogram characterization revealed a critical resistance burden: 100% of isolates fell into extreme epidemiological tiers, with 71.4% (n = 30) exhibiting an extensively drug-resistant (XDR) phenotype and 28.6% (n = 12) reaching absolute pandrug-resistant (PDR) status. Absolute phenotypic resistance (100%) was recorded across both cohorts for meropenem, ceftazidime, aztreonam, tobramycin, polymyxins, and novel combination therapeutics (ceftazidime–avibactam and ceftolozane–tazobactam). Pairwise tracking demonstrated an exceptionally strong positive correlation in resistance distribution patterns between human and food isolates (Spearman’s rho = 0.948, p < 0.001). Analytical validation of the qPCR assay demonstrated tight, reproducible specific derivative melting peaks (T_m) at 83.2 °C for intI1 and 83.8 °C for intI2. Genotypic screening revealed universal conservation of plasmid-borne intI1 (100%), whereas plasmidic intI2 was variably distributed (61.9%). Crucially, intI2 carriage served as a definitive marker for the transition to complete pandrug resistance, being detected in 100% (n = 12/12) of PDR isolates compared to 46.7% (n = 14/30) of XDR strains (p = 0.001), supported by a strong positive correlation with the PDR phenotype (rho = 0.496, p = 0.0008). Conclusions: These findings reveal an overlapping, extreme resistome across the food-clinical interface and identify intI2 as a genetic tipping point for absolute pandrug resistance. The validated real-time assay provides a highly sensitive, proactive molecular surveillance framework capable of mapping high-risk mobile genetic platforms before they manifest as catastrophic empirical treatment failures. Full article

Oligonucleotide therapeutics represent an expanding class of medicines that can regulate gene expression, RNA processing, protein translation, immune signaling, and biomolecular recognition through sequence-specific or structure-dependent mechanisms. Despite clinical progress, their application remains constrained by nuclease degradation, rapid clearance, inefficient tissue and cellular delivery, endosomal sequestration, off-target activity, immune recognition, and mechanism-specific requirements for target engagement. Chemical modification is central to oligonucleotide therapeutic development because it can mitigate some of these limitations while influencing target affinity, protein binding, pharmacokinetics, and intracellular activity. This review examines chemical modification strategies to improve the biological stability and functional performance of therapeutic oligonucleotides and is organized around major classes of chemical modification, including phosphate and backbone-linkage modifications, sugar and conformational modifications, backbone-replacement analogs, and conjugation-based approaches. Rather than presenting these chemistries as uniformly beneficial, this review emphasizes that the same modification can be enabling in one therapeutic mechanism and disruptive in another, so its value cannot be judged apart from the modality and molecular architecture in which it is placed. Clinically successful oligonucleotide designs are likely to rely on combinations of chemical features, including modified backbones, modified sugars, stereochemical control, terminal stabilization, and ligand- or formulation-based delivery strategies. Understanding how these features interact is essential to develop more predictable and mechanism-appropriate oligonucleotide therapeutics. Full article

Background: Gram-negative bacteria resistant to multiple drugs are a major cause of illness and death worldwide. Their remarkable capacity to develop resistance to antibiotics makes them a serious concern in medical practice. Methods: A simple, green, novel method is used to synthesize ZnO nanoparticles (ZnO NPs) using ethanolic extracts of Diplazium esculentum via precipitation. Results: ZnO NPs exhibit a hexagonal structure with a particle size of ~30 nm and a band gap of 3.24 eV. The defect sites formed in ZnO NPs were estimated using prominent peaks in the photoluminescence spectra. ZnO NPs displayed a more than 4-log reduction in multi-drug-resistant E. coli and K. pneumoniae clinical isolates at a 500 μg/mL concentration. Moreover, ZnO NPs significantly reduced the biofilm bacterial cell viability of clinical isolates of Gram-negative bacteria. Complete eradication of biofilms was achieved for drug-resistant E. coli clinical isolates using a combination of sub-MIC of gentamicin and 500 μg/mL ZnO NPs. Green-synthesized ZnO NPs did not induce oxidative stress in mice, as indicated by unchanged GST, GSH, and thiol levels across all the tested organs. ZnO NPs showed both antibacterial and antibiofilm efficacy against drug-resistant strains of E. coli, K. pneumoniae, and S. aureus and completely eradicated E. coli biofilm in combination with gentamicin. Conclusions: Our study focuses on the sustainable synthesis of biocompatible ZnO NPs for the treatment of infections caused by pathogens belonging to the high-priority ESKAPE group. Full article

Background: Tuberculosis remains a major public health challenge in Latin America, where social inequalities, fragmented health systems, and barriers to care continue to undermine disease control. Although effective treatment is available, treatment abandonment and loss to follow-up remain persistent obstacles to successful outcomes. Methods: A structured critical analysis of the contemporary literature on tuberculosis treatment abandonment and retention in care was conducted using a narrative evidence synthesis approach. Forty scientific documents published between 2018 and 2026 were included. Of these, 23 Latin American studies informed the results synthesis, while 13 international studies supported the comparative critical reflection. Results: Findings were organised into five domains: social vulnerability, clinical complexity, health system barriers, priority populations, and protective factors and innovation. The evidence indicates that treatment abandonment is not an isolated individual behaviour, but a phenomenon associated with the convergence of social, clinical, and institutional determinants across the care trajectory. Poverty, migration, drug resistance, incarceration, weak follow-up systems, and fragmented care were repeatedly associated with discontinuity, whereas social support, active follow-up, digital tools, and economic measures were linked to improved adherence and retention in care. Conclusions: From a biopsychosocial perspective, treatment abandonment should be understood as a systemic and preventable failure across the tuberculosis care continuum. The proposed Latin American Integrated Model for Retention in Tuberculosis Care (MILERTB) offers an anticipatory, equity-oriented, and person-centred framework to strengthen retention in care and guide future implementation research. Full article

In spite of at least six discrete classes of drugs available for cancer treatment, the quest for more biologic drugs continues. One type of biologic molecule that occurs naturally in the body is microRNA. MicroRNAs regulate post-transcriptional gene expression and can be under expressed in cancer (tumor suppressor microRNAs) or over expressed (oncogenic microRNAs). Strand-specific mimics of microRNAs have been developed and used successfully in vitro, in vivo, and in clinical trials, to control multiple aspects of cancer including metastasis, apoptosis and proliferation. Each microRNA is capable of binding a specific target mRNA or mRNAs, sometimes simultaneously interfering with multiple genes in a single pathway, or binding with a single nodal mRNA. Some microRNAs can facilitate chemotherapy that has stopped working, addressing the issue of drug resistance. Without chemical modification, microRNAs are too vulnerable to have lasting therapeutic value. Chemical modifications to microRNAs have provided nuclease resistance and greater stability and are the basis for microRNA mimics that can be used therapeutically. However, without a vehicle, microRNA mimics do not cross cell membranes. These nanoparticles can cause inflammatory reactions in patients. Additional modifications that enabled microRNA mimics to cross cell membranes include substituting uracil with 5-fluorouracil. Lessons from an siRNA therapeutic called Patisiran offer a roadmap for future success for microRNAs in cancer. This review provides a historical perspective of the continuing evolution of microRNA mimics for cancer treatment. Full article

Background/Objectives: Antimicrobial resistance in pediatric urinary tract infections (UTIs) is a growing concern. We evaluated resistance patterns of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa urinary isolates in a tertiary care Greek hospital from 2015 through 12 September 2025. Methods: We retrospectively analyzed pediatric urinary isolates collected from 2015 through September 2025. Antibiotic analysis was restricted to EUCAST UTI-relevant organism-drug pairs. Resistance was summarized for the prespecified antibiotics, and presumptive ESBL and carbapenem resistance phenotypes were assessed. Results: Among 776 isolates (567 E. coli, 104 K. pneumoniae, 105 P. aeruginosa), E. coli showed the lowest resistance among non-carbapenem options to nitrofurantoin (0.4%), amikacin (1.2%), and gentamicin (7.8%), and the highest to trimethoprim (98.8%), ampicillin (53.5%), and trimethoprim-sulfamethoxazole (31.5%). K. pneumoniae had the lowest resistance among non-carbapenem options to amikacin (8.6%) and gentamicin (15.5%), with high cephalosporin resistance (oral cefuroxime 40.9%, intravenous cefuroxime 39.7%). Presumptive ESBL and carbapenem resistance in K. pneumoniae were 42.3% and 20.2%, respectively. Corresponding prevalences in E. coli were 12.4% and 0.7%. P. aeruginosa retained low resistance to meropenem (1.9%) and piperacillin-tazobactam (2.1%). No age or sex association with the presumptive resistance phenotypes remained significant after false-discovery-rate adjustment. Conclusions: E. coli showed very high ampicillin resistance, low resistance to gentamicin and amikacin, and moderate resistance to guideline-relevant cephalosporins. P. aeruginosa retained low resistance to key antipseudomonal agents. K. pneumoniae showed the highest burden of cephalosporin resistance and of presumptive ESBL and carbapenem resistance phenotypes. These data support guideline-based empirical coverage more strongly for E. coli than for K. pneumoniae in this setting. Full article