Search Results (14,981)

The potential health hazards caused by microwave exposure have attracted increasing attention. Microwave radiation has been reported to induce oxidative stress in neural tissues, which is considered one of the primary mechanisms underlying its adverse effects on central nervous system function. The hippocampus is sensitive to microwave radiation, whereas underlying cellular and molecular mechanisms remain incompletely understood. In this study, microwave-exposed mice exhibited significantly impaired performance in the Go/No-go, Y-maze, and novel object recognition tests at 6 h and 7 days post-exposure, indicating deficits in hippocampus-dependent working memory. Single-cell RNA sequencing of hippocampal tissues from control and microwave-exposed mice yielded 94,088 high-quality cells across eight major cell types. Astrocyte sub-clustering identified five transcriptionally distinct subpopulations, with Astrocyte_S100a6 and Astrocyte_Son proportions increased and Astrocyte_Serpinf1 decreased in the radiation group. Analysis of astrocyte transcriptional state transitions showed microwave-exposed astrocytes were preferentially distributed toward terminal reactive states with depletion at early homeostatic nodes. Cell–cell communication analysis revealed increased total interactions and interaction strength following radiation. Astrocyte outgoing signaling was increased for pathways associated with vascular remodeling, phagocytic regulation, and neuroinflammation, while pathways related to trophic support were decreased. Incoming signaling showed increased activity in pathways linked to phagocytic recruitment and inflammatory mediation. Taken together, these findings indicate that microwave exposure is associated with hippocampus-dependent working memory deficits accompanied by transcriptional remodeling of astrocyte subpopulation composition, directional astrocyte state transitions toward reactive phenotypes, and broad alterations in astrocyte-centered intercellular communication, providing a cellular and molecular framework for understanding astrocyte involvement in microwave radiation-associated hippocampal dysfunction. Full article

The increasing resistance of agricultural pests and disease-vectoring arthropods to synthetic pesticides underscores the urgent need for novel and sustainable biocidal agents. This study evaluates, for the first time, the insect antifeedant and ixodicidal activities of essential oils derived from ten Amazonian Piper species and their major constituents. Antifeedant effects were assessed against Spodoptera littoralis, Myzus persicae, and Rhopalosiphum padi, whereas ixodicidal activity was tested on Hyalomma lusitanicum. Additionally, the effects of these oils on the plant-parasitic nematode Meloidogyne javanica were investigated. Essential oils from Piper mituense (51.6% apiol) and P. sancti-felicis (76.1% apiol) exhibited the highest bioactivity, achieving more than 75% feeding inhibition across all insect species and 100% tick mortality. P. mituense consistently demonstrated greater potency, suggesting possible synergistic interactions among its minor constituents. Principal component analysis linked apiol-rich chemotypes with broad-spectrum activity. In contrast, oils rich in oxygenated caryophyllene derivatives, particularly those from P. casapiense, showed strong selective antifeedant effects against R. padi. Pure apiol displayed activity across all assays, whereas no nematicidal effects were observed. Molecular docking analyses supported these findings, indicating that apiol can interact with acetylcholinesterase in addition to its known effect on cytochrome P450 targets. Overall, these results identify complementary Piper chemotypes with promising potential as dual-purpose biopesticides for integrated pest management strategies. Full article

Background/Objectives: White–Sutton syndrome (WHSUS; OMIM 616364) is a rare neurodevelopmental disorder caused by pathogenic variants in the POGZ gene and characterized by developmental delay, intellectual disability, speech impairment, autism spectrum features, and dysmorphic traits. Although most reported cases are sporadic, inherited forms are exceptionally rare. We describe a familial case of WHSUS involving an affected mother and two children carrying a heterozygous POGZ nonsense variant, highlighting marked intra-familial phenotypic variability and expanding the clinical spectrum of the disorder. Methods: Clinical evaluation included multidisciplinary assessments. Genetic testing was performed using clinical exome sequencing (CES) with a virtual neurodevelopmental disorder (NDD) gene panel, followed by Sanger confirmation and segregation analysis in family members. The POGZ transcript reference NM_015100.3 was used for variant nomenclature and verified with the Mutalyzer tool. CNV detection from NGS data was performed using the Alissa CNV caller (Agilent) and visualized via IGV; the Xp11.22 microduplication was confirmed by chromosomal microarray (aCGH) and parental segregation analyses. Results: CES identified the heterozygous pathogenic POGZ variant c.1522C>T (p.Arg508*) in the female proband (III₆), an infant presenting with global developmental delay, hypotonia, speech impairment, gait abnormalities, and characteristic dysmorphic features. Segregation analysis demonstrated maternal inheritance and confirmed the presence of the variant in her affected brother (III₄), who also carries a de novo 1.79 kb microduplication at Xp11.22, while the maternal grandparents tested negative, indicating a de novo origin in the mother. The mother exhibited an attenuated phenotype, including mild neuropsychiatric and gastrointestinal manifestations. The variant is predicted to undergo nonsense-mediated decay (NMD), consistent with a moderate clinical presentation; however, experimental validation was not performed. Conclusions: This report documents a rare familial occurrence of WHSUS with highly variable expressivity. Our findings broaden the phenotypic and molecular characterization of POGZ-related disorders and emphasize the importance of comprehensive segregation studies and early genomic diagnosis. While experimental data link POGZ deficiency to DNA repair defects, no longitudinal clinical studies have demonstrated increased cancer risk in WHSUS; therefore, formal malignancy screening guidelines cannot be established at present, and this issue deserves future study in larger cohorts or registries. Full article

Recently, there has been great interest in AI-based approaches for de novo design of novel drug candidates. However, the generation of useful lead drug candidate compounds requires more than predicting engagement with the desired protein target. Candidate molecules must also be anchored in the real world of medicinal chemistry for their synthesis and modification as well as satisfying multiple drug development-related criteria. Here, we present Nevermore, an AI target-conditioned, database-grounded workflow for prioritizing candidate ligands from large compound libraries. Nevermore uses a geometry-aware protein–ligand affinity oracle to score target-specific binding and perform sparse integer edits in count-based Morgan fingerprint space. Nevermore then retrieves the most structurally similar molecules from public chemical databases. This design enables multi-objective search over predicted affinity and absorption, distribution, metabolism, excretion, and toxicity (ADMET) proxies while keeping all candidates anchored to valid database compounds. We evaluated Nevermore’s performance across three biologically distinct targets: Menin, a protein-interaction target relevant to leukemia; SARS-CoV-2 M^pro, a viral cysteine protease relevant to antiviral discovery; and epidermal growth factor receptor (EGFR), a kinase-superfamily oncology target with extensive experimentally tested compounds. Nevermore retrieved candidate sets with favorable predicted affinity–property trade-offs. These results support database-grounded fingerprint steering as a practical computational strategy for lead prioritization and for generating testable molecular hypotheses, although the prioritized candidates remain predictions, requiring follow-up experimental validation. Full article

The rapid expansion of molecular point-of-care (POC) diagnostics into decentralized settings, including emergency departments, retail pharmacies, and home environments, has shifted the burden of diagnostic performance from laboratory professionals to heterogeneous, often non-expert users. While traditional evaluation frameworks focus on analytical and clinical validity, they often overlook the impact of human-system interactions on real-world reliability. This review introduces the concept of Operational Validity: the ability of a diagnostic system to preserve its intended performance when operated by intended users within the constraints of real-world workflows and environments. To establish a rigorous foundation for this concept, this study provides a critical comparative analysis contrasting Operational Validity against traditional clinical evaluation dimensions (analytical validity, clinical validity, and clinical utility) and post-market metrics. While existing literature outlines isolated usability principles, the significance of this study lies in its synthesis of these fragmented concepts into a formalized, lifecycle-based “Operational Validity” framework that explicitly maps the causal mechanisms connecting initial user interaction directly to downstream clinical outcomes. By synthesizing international standards (IEC 62366-1) alongside the newly finalized May 2026 U.S. Food and Drug Administration (FDA) guidance on the Content of Human Factors Information in Medical Device Marketing Submissions, we examine how human factors engineering (HFE) and usability engineering serve as the methodological foundation for operational validity. We analyze the specific complexities of molecular workflows, identify key parameters of use-related failure modes in pre-analytical and interpretation stages, and detail the mandatory role of iterative formative and final summative usability testing in mitigating these risks. Finally, we propose a lifecycle-based approach to HFE that integrates design, simulated-use validation, and post-market surveillance. Establishing operational validity is essential to ensure that the high analytical sensitivity of molecular POC platforms translates into consistent clinical utility across the full spectrum of decentralized care. Full article

Objective: Our objectives were to describe molecular profiling in a real-life cohort of patients with radioiodine-resistant (RAI-R) differentiated or poorly differentiated thyroid cancer (DTC or PDTC) treated with lenvatinib and to focus on factors potentially influencing the quality of tissue samples for molecular analysis, including the impact of storage time, defined as the interval between tissue collection and molecular testing. Design: We retrospectively included all lenvatinib-treated RAI-R DTC or PDTC patients tested with DNA- and/or RNA-based next-generation sequencing (NGS) in our center, also analyzing the results of fluorescence in situ hybridization (FISH) for RET fusions if the sample did not satisfy quality criteria for RNA-based NGS analysis. We investigated differences in terms of histotype, biopsy site, or storage time between adequate and inadequate samples for RNA-based NGS. Results: At least one gene alteration was detected in 50% of the cohort (18 out of 36 patients); RAS and BRAF were the most frequent mutations, while gene fusions accounted for 5.6% of cases. Tissue samples were more frequently adequate for DNA-based NGS compared to RNA-NGS analysis (93.9% vs. 58.3%, p < 0.001). The median storage time was significantly longer in the case of inadequate samples for RNA-based NGS compared with adequate specimens (41.5 vs. 9.5 months, p = 0.016); samples archived for ≥3 years led more frequently to an inadequate result. Conclusions: Advanced RAI-R TC candidates for systemic therapy often harbor gene alterations. An adequate result was less frequently achieved in cases of RNA-based NGS than in DNA-based NGS, especially if the interval between tissue collection and molecular analysis was longer; nevertheless, the limited cohort size precludes definitive conclusions. Full article

Background: Microsatellite instability (MSI) and mismatch repair (MMR) deficiency are key predictive biomarkers for immune checkpoint inhibitors (ICIs) in metastatic colorectal cancer (mCRC). In real-world practice, however, diagnostic pathways often involve heterogeneous testing modalities, which may lead to discordant or inconclusive results. Methods: We conducted a retrospective study of patients with mCRC who underwent at least one MSI/MMR assessment between 2015 and 2025. Diagnostic modalities included IHC, tissue-based and liquid-based MSI testing. A predefined decision algorithm classified results as conclusive or inconclusive; discordant cases underwent adjudication that integrated a pathology review, molecular features, and technical considerations. Patients were ultimately assigned to definitive MSS or definitive MSI groups. Clinical characteristics, treatment patterns, and outcomes—particularly in relation to immunotherapy—were evaluated. Results: Among 727 evaluable patients, the MSI/MMR status was conclusive in 695 (95.6%) and inconclusive in 32 (4.4%). Inconclusive cases resulted from isolated MMR protein loss, heterogeneous or equivocal staining, inter-tumoral discordance, or discrepancies between tissue- and liquid-based assays. After adjudication, 54 patients (7.4%) were classified as definitive MSI and 673 (92.6%) as definitive MSS. Definitive MSI tumors were associated with female sex, right-sided primaries, high-grade histology, nodal involvement, and BRAF V600E mutations. Among the definitive MSI patients, 31 (57.4%) received immunotherapy, achieving a complete response rate of 48.4% and an overall response rate of 71.0%. Median PFS and OS were not reached in the definitive MSI group, whereas definitive MSS patients treated with ICIs experienced significantly poorer outcomes. Conclusive and adjudicated MSI groups demonstrated comparable responses to immunotherapy. Conclusions: In real-world practice, a meaningful proportion (4%) of mCRC patients experience inconclusive MSI/MMR assessment, with important clinical implications. Both technical and biological factors contribute to diagnostic uncertainty. Integrating orthogonal testing modalities and applying structured adjudication improves classification accuracy and ensures appropriate access to immunotherapy. Full article

Since January 2023, the Russian Federation has implemented expanded neonatal screening for 36 hereditary disorders, which has changed the diagnostic algorithm for hyperphenylalaninemia/phenylketonuria (HPA/PKU) by introducing tandem mass spectrometry (MS/MS) on the second day of life, followed by confirmatory biochemical and molecular testing in newborns at risk. We analyzed 1247 newborns aged 5–15 days with elevated phenylalanine levels (≥120 µmol/L) and a phenylalanine to tyrosine ratio of at least 1 detected during the first stage of screening using MS/MS. At the reference center, newborns underwent repeat biochemical testing and stepwise molecular analysis of HPA-associated genes. Two pathogenic variants in HPA-associated genes were identified in 538 newborns, including 534 newborns with biallelic pathogenic variants in PAH and 4 with BH4-deficient forms (PTS, QDPR). The incidence of molecularly confirmed HPA was 1:4518 newborns (95% CI: 1:4152–1:4925). The PAH variant spectrum was dominated by p.Arg408Trp (c.1222C>T) (33.4%). Genotype-based analysis indicated that 73 newborns (13.7%) were likely responsive to cofactor therapy, whereas 222 (41.6%) were potentially responsive. These findings define the molecular epidemiology of HPA in Russia and support early genetic stratification for diagnosis and treatment. Full article

H9N2 avian influenza viruses inherently carry cross-species transmission potential, making continuous surveillance critical for pandemic prevention. This study focused on monitoring the 2024 H9N2 epidemic in Jiangsu Province’s external environment, analyzing its molecular evolution and receptor binding properties, assessing cross-species transmission and pandemic risks, and investigating serological antibody levels across different human populations. Environmental samples were collected from live poultry markets, farms, slaughterhouses, and bird habitats across Jiangsu, screened via quantitative PCR (qPCR), with positive samples used for virus isolation and whole-genome sequencing. Receptor binding properties were tested by hemagglutination assay, and H9N2 antibody levels were measured in 370 occupationally exposed individuals and 240 non-exposed individuals using hemagglutination inhibition (HI) assays. Among the 5779 collected samples, 6.89% tested H9N2-positive, and 12 strains belonging to the Eurasian lineage Y280-like clade G57 genotype were successfully isolated. All strains carried the HA-Q226L mutation, with 11 showing preferential binding to human α-2,6 receptors and one strain possessing dual receptor binding capability. Internal genes harbored mammalian adaptation mutations, and M2 proteins contained mutations conferring complete resistance to amantadine-class antiviral drugs. Serological tests revealed antibody positive rates of 4.05% in exposed populations and 2.5% in non-exposed populations, with no statistically significant difference between groups. These findings confirm that Jiangsu’s circulating H9N2 viruses have acquired human receptor preference and mammalian adaptation, posing silent infection and pandemic risks. Enhanced surveillance and the development of candidate vaccine stockpiles are strongly recommended. Full article

The subcellular localization of long noncoding RNAs (lncRNAs) is a central determinant of their function, yet its molecular determinants remain incompletely defined, and most existing predictors rely on the primary sequence. Because RNA-binding proteins (RBPs) are the proximal effectors of RNA compartmentalization, this study tested whether the composition of RBPs bound to a lncRNA is predictive of its nuclear or cytoplasmic localization. Enhanced crosslinking and immunoprecipitation (eCLIP) occupancy for 139 RBPs in K562 cells was integrated with the cytoplasmic–nuclear relative concentration indices (CN-RCIs) derived from matched subcellular fractionation, and localization was modeled under chromosome-grouped cross-validation with nested regularization. RBP-occupancy composition predicted localization beyond the transcript size and total binding amount (incremental cross-validated coefficient of determination, delta-R-squared = 0.17; receiver-operating-characteristic area under the curve, AUC = 0.73, a moderate-strength association; Freedman–Lane permutation, p = 0.005). This increment persisted (delta-R-squared = 0.12; p = 0.005) against an expanded baseline that additionally absorbed the transcript abundance, intron content and exon number, indicating predictive information that is not reducible to these transcript features, and the classifier was well calibrated (Brier score = 0.10; expected calibration error = 0.02). The signed coefficient profile separated RBP function systematically: factors acting in nuclear processes (splicing, 3′-end processing, and nuclear-matrix association) carried negative, nuclear-direction weights, whereas factors acting in cytoplasmic processes (translation and messenger RNA stability) carried positive, cytoplasmic-direction weights (Mann–Whitney p = 0.013). The profile generalized across cell lines: a K562-trained model predicted HepG2 localization (transfer AUC = 0.71 using 76 shared RBPs), and HepG2 reproduced the association independently (AUC = 0.77). The association is correlational and of moderate strength; it is presented as an interpretable, RBP-occupancy-based complement to sequence-based predictors of lncRNA localization. Full article

The genus Tamarix L. includes several species widely used in traditional medicine for their therapeutic properties. This study aims to evaluate the bioactive potential of Tamarix gallica extracts from Western Algeria using an integrated in vitro and in silico approach. GC–MS analysis with BSTFA derivatization was performed to characterize the chemical profile of the methanolic fraction. In addition, total phenolic, flavonoid, and tannin contents were determined in methanolic extracts of leaves and stems. The biological activities were assessed using antioxidant (DPPH, ABTS, β-carotene, FRAP, O-phenanthroline, and cupric reducing assays), antimicrobial, antidiabetic, and anti-Alzheimer in vitro assays. Molecular docking was conducted to evaluate the inhibitory potential of selected flavonoids against α-amylase, acetylcholinesterase, and butyrylcholinesterase. Results revealed a rich metabolite profile dominated by long-chain aliphatic alcohols (including hentriacontan-12-ol), phytosterols (β-sitosterol), fatty acids, phenolic derivatives, and sugar alcohols. The extracts exhibited strong antioxidant activity (IC₅₀ = 1.34 ± 0.43 and 12.32 ± 0.36 μg·mL⁻¹), significant antimicrobial effects against the tested pathogens, and notable antidiabetic and anticholinesterase activities (IC₅₀ = 78.65 ± 1.43 and 98.37 ± 1.07 μg·mL⁻¹). Molecular docking analysis supported these findings, showing strong binding affinities of quercetin and rhamnetin toward the target enzymes. Overall, T. gallica exhibits promising multifunctional bioactivities with potential pharmaceutical relevance. Full article

Objectives: To compare the adjunctive efficacy of a MicroRepair^® mouthwash containing an antibacterial complex (ABX), composed of cetylpyridinium chloride, magnolol, and honokiol, with 0.12% chlorhexidine (CHX) in the management of generalized plaque-induced gingivitis, assessing clinical periodontal parameters, salivary activated matrix metalloproteinase-8 (aMMP-8) levels, and patient-reported outcomes over 6 months. Methods: A randomized, controlled, parallel-group clinical trial included 40 systemically healthy adults with generalized gingivitis and was reported in accordance with CONSORT 2025 guidelines. Following professional oral hygiene according to the Guided Biofilm Therapy (GBT) protocol, participants were randomly allocated to ABX or 0.12% CHX, used twice daily for 14 days. Clinical parameters, including Full-Mouth Bleeding Score (FMBS, primary outcome), Full-Mouth Plaque Score (FMPS), Probing Pocket Depth (PPD), Clinical Attachment Level (CAL), Gingival Recession (REC), and Modified Lobene Stain Index (MLSI), were recorded at baseline, 2 weeks, 1, 3, and 6 months. Salivary aMMP-8 levels were assessed at baseline and 2 weeks. Heavy smokers were excluded, and smoking status was evaluated as a potential covariate. Non-parametric tests were applied (p < 0.05). Results: Both groups showed significant reductions in FMBS and FMPS over time (p < 0.05), with no intergroup differences for the primary outcome at any follow-up at the patient level. Patient-level analyses did not reveal consistent differences across secondary parameters. At the tooth level, lower FMPS values were observed in the trial group at 2 weeks and 1 month (p < 0.05), with earlier PPD reduction. CAL, and REC remained stable. Salivary aMMP-8 levels decreased significantly in both groups without intergroup differences. Patient-reported outcomes were comparable. Smoking status was balanced between groups and was not significantly associated with treatment allocation or the main clinical outcomes. Conclusions: No significant differences were observed between ABX and CHX for the main clinical and molecular outcomes, supporting its potential use as an adjunct in gingivitis management. Full article

PVP-stabilized silver nanoparticles (Ag NPs) were functionalized with sildenafil (Sil), leading to spherical NPs (Ag@Sil NPs) with a size of about 30 nm as observed through transmission electron microscopy and dynamic light scattering. Fourier-transformed IR spectroscopy confirmed the covering of the particles with Sil. The Ag@Sil NPs were incorporated into a 0.1 wt% ointment and tested for the treatment of acute anal fissures in a preliminary medical study involving 50 patients. Typical symptoms such as pain, bleeding, itching, and mass sensation were improved in the intervention group with no adverse effects. Molecular docking showed strong interactions with docking scores slightly above −10 kcal/mol between sildenafil and two different model complexes [Ag–Sil]⁺ for the Ag-bound sildenafil with either piperazine-N- or pyrazole-N-bound Ag⁺ ions and the muscarinic M2 and the nicotinic acetylcholine α3β4 receptor, which are both involved in anal sphincter regulation. All three showed superior binding compared with nitroglycerin and L-arginine. The residue analysis revealed a higher number of relevant interactions for the sildenafil and the two Ag⁺ complexes, compared to nitroglycerin and L-arginine, fully in line with the differences in the docking scores. Full article

Cross-presentation of exogenous antigens by dendritic cells (DCs) relies on the cytosolic pathway, enabling proteasomal processing and subsequent loading of antigenic peptides onto major histocompatibility complex class I (MHC-I) molecules. Although this pathway is central to CD8⁺ T-cell activation, the molecular mechanisms that regulate intracellular antigen processing and redistribution during cross-presentation remain incompletely defined. In this study, we investigated the contribution of the large-pore channel Pannexin-1 (Panx1) to antigen handling during cross-presentation. Using confocal microscopy and quantitative image analysis in granulocyte–macrophage colony-stimulating factor/interleukin-4 (GM-CSF/IL-4)-derived inflammatory bone marrow-derived dendritic cell (BMDC)-like cellsexposed to ovalbumin (OVA)–Alexa Fluor 488, we observed time-dependent changes in intracellular antigen distribution that were altered upon pharmacological inhibition of Panx1 with the blocking peptide 10Panx1. In parallel, functional assays revealed that Panx1 inhibition significantly reduced SIINFEKL peptide-dependentactivation of B3Z CD8⁺ T-cell hybridomas following pulsing with full-length OVA. Similar effects were observed in the cross-presentation-competent MUTU1940 dendritic cell line. Importantly, Panx1 inhibition did not significantly affect dendritic-cell viability or LPS-induced activation under the experimental conditions tested. In contrast, pharmacological inhibition or genetic deficiency of P2X7 receptor (P2X7) did not produce comparable reductions in cross-presentation, and combined inhibition did not result in additive effects under the experimental conditions tested. Together, these findings provide functional evidence supporting a role for Panx1 in regulating intracellular antigen redistribution associated with cross-presentation. While not establishing direct genetic causality, our data identify Panx1 as a modulatory component influencing antigen-processing events that culminate in CD8⁺ T-cell activation, thereby expanding the current framework of intracellular antigen-processing mechanisms involved in dendritic-cell-mediated cross-presentation. Full article

Appendiceal mucinous neoplasms (AMNs) are a rare but clinically significant category of gastrointestinal tumors, ranging from low-grade appendiceal mucinous neoplasm (LAMN), the main precursor of pseudomyxoma peritonei (PMP), to high-grade appendiceal mucinous neoplasm (HAMN), poorly differentiated and signet-ring-cell adenocarcinomas, and goblet cell adenocarcinoma. Although current WHO and PSOGI classifications provide well established diagnostic criteria, controversies persist regarding the biological behavior and prognostic significance of the most aggressive subtypes and the relationship between HAMN and mucinous adenocarcinoma. While appendectomy is sufficient for localized LAMN, cytoreductive surgery with hyperthermic intraperitoneal chemotherapy (CRS/HIPEC) is the treatment of choice for peritoneal dissemination This review integrates the histopathological and molecular classification of AMN and PMP with the evolution of intraperitoneal chemotherapy. Key findings indicate that KRAS and GNAS mutations are central drivers of mucin overproduction and peritoneal spread, that tumor grade and mucin cellularity remain the strongest prognostic determinants, and that the evidence supporting HIPEC and PIPAC derives largely from observational rather than randomized data. As a novel insight, we highlight the emerging role of patient-derived organoids as translational models for functional drug testing. Progress will depend on integrating molecular characterization, critical appraisal of intraperitoneal therapies, and organoid-based testing to advance individualized treatment for peritoneal surface malignancies. Full article