Search Results (120)

Plasminogen activator inhibitor-1 (PAI-1), encoded by SERPINE1, is the principal physiological inhibitor of tissue-type and urokinase-type plasminogen activators and a central regulator of fibrinolysis. Beyond its canonical hemostatic role, PAI-1 has emerged as a pleiotropic mediator of tissue remodeling, fibrosis, metabolic dysfunction, cancer progression, cellular senescence, and age-associated immune dysregulation. A central argument of this review is that PAI-1 should be understood not only as a downstream biomarker of aging-associated pathology, but also as an active effector linking senescence-associated secretory phenotype (SASP) signaling, chronic low-grade inflammation, impaired immune surveillance, fibrotic extracellular matrix remodeling, and a prothrombotic state. In this framework, PAI-1 may function as an immune-aging checkpoint: a molecular node through which senescent, stromal, malignant, and inflammatory cells reinforce immune evasion and tissue dysfunction. Structure-guided drug discovery has enabled the development of small-molecule PAI-1 inhibitors, including TM5275, TM5441, TM5509, and TM5614. Among these, TM5614 is an orally available investigational compound that has progressed to clinical evaluation. Preclinical studies support anti-thrombotic, anti-fibrotic, anti-inflammatory, anti-senescent, and tumor-microenvironment-modulating effects of PAI-1 inhibition, while early clinical studies have evaluated TM5614 in chronic myeloid leukemia, immune-checkpoint-refractory malignant melanoma, non-small-cell lung cancer, and COVID-19-associated pneumonia. This review summarizes the biology of PAI-1, expands the discussion of immunoaging, reviews representative preclinical and clinical data, compares available PAI-1 inhibitors, and discusses the translational opportunities and safety considerations for TM5614 and related compounds. Full article

Given its highly aggressive nature and poor clinical outcome, pancreatic ductal adenocarcinoma (PDAC) requires physiologically relevant in vitro models that more accurately reflect tumor biology and drug response. In this study, adhesive and non-adhesive hydrogel microenvironments were comparatively evaluated for pancreatic cancer spheroid modeling using PANC-1 and MIA PaCa-2 cells. Gelatin methacryloyl (GelMA) hydrogels were synthesized, photocrosslinked, and optimized in terms of stability, swelling, degradation, and cytocompatibility, while 3% agarose was used as a non-adhesive counterpart. Although the optimized GelMA formulation showed adequate structural stability and no cytotoxicity, it did not support spheroid formation. In contrast, agarose enabled the formation of compact, viable, and proliferative spheroids in both cell lines. Agarose-derived spheroids exhibited time-dependent growth, positive Ki-67 staining, and increased HIF-1α expression under 3D conditions, indicating the establishment of hypoxia-associated tumor-like microenvironments. Gemcitabine treatment induced a time-dependent reduction in spheroid viability, while viable cell populations persisted throughout exposure, reflecting the heterogeneous therapeutic response typical of 3D tumor models. Overall, these findings provide a comparative, microenvironment-based assessment of pancreatic cancer spheroid modeling, indicating that hydrogel-dependent differences in adhesivity and structural dynamics are important determinants of spheroid assembly, hypoxia-associated molecular adaptation, and chemotherapeutic response. Overall, these findings provide a comparative, microenvironment-based assessment of pancreatic cancer spheroid modeling, indicating that hydrogel-dependent differences in adhesivity and structural dynamics are important determinants of spheroid assembly, hypoxia-associated molecular adaptation, and chemotherapeutic response. Overall, these findings provide a comparative, microenvironment-based assessment of pancreatic cancer spheroid modeling, indicating that hydrogel-dependent differences in adhesivity and structural dynamics are important determinants of spheroid assembly, hypoxia-associated molecular adaptation, and chemotherapeutic response. Full article

The human genome produces a large repertoire of non-coding RNAs (ncRNAs) with important regulatory roles in development, physiology, and most of diseases. Among these, long non-coding RNAs (lncRNAs) have emerged as key modulators of gene expression, chromatin organization, and cellular homeostasis, despite displaying remarkably low primary-sequence conservation across species. This apparent evolutionary paradox questions the limitations of predicting biological function based on conservation, particularly across different biological domains. Here, we examine current evidence on lncRNA evolution, with a focus on their roles in metabolic regulation compared with neurobiological processes. We hypothesize that lncRNAs involved in ancient and conserved pathways such as metabolism may be under stronger evolutionary constraint than those associated with higher-order, species-specific traits, although available data support a more nuanced interpretation. Functional importance often correlates poorly with linear sequence conservation and instead appears to depend on higher-level features, including RNA secondary or tertiary structure, genomic context, regulatory architecture, and interactions with conserved molecular partners. We propose a systematic comparative framework to empirically assess conservation among metabolism- and neuro-associated lncRNAs using phylogenetic, syntenic, structural, and expression-based metrics. Finally, we discuss the therapeutic implications of lncRNA biology, highlighting how a deeper understanding of their evolutionary and mechanistic properties may inform the development of more precise and effective RNA-targeting strategies. Together, these insights underscore the non-coding transcriptome as a critical frontier for both fundamental biology and precision medicine. Full article

Coinfection between hepatitis C virus (HCV) and human T-lymphotropic virus 1/2 (HTLV-1/2) remains poorly investigated in the Northern Region of Brazil despite its clinically important condition. The objective of this study was to determine the prevalence and describe the epidemiological and behavioral risk factors for HCV/HTLV-1/2 coinfection in Belém, Pará. This observational, descriptive, and cross-sectional study analyzed 192 samples from patients previously diagnosed with HCV: 127 participants recruited between May 2023 and June 2025 and 65 samples previously stored in the Virology Laboratory of UFPA. Data were collected through a structured survey. Serological screening for HTLV-1/2 was performed by enzyme-linked immunosorbent assay (ELISA) and confirmed by INNO-LIA and molecular biology (qPCR). HCV/HTLV-1/2 coinfection was observed in 4 individuals (2.1%), of whom 1.6% had HCV/HTLV-1 coinfection and 0.5% HCV/HTLV-2. There was no statistically significant association when comparing the sociodemographic, clinical characteristics, or risk factors of HCV monoinfected and HCV/HTLV-1/2 coinfected individuals. Although the results show a low prevalence of HTLV-1/2 and HCV coinfection in Belém do Pará, they still reinforce the importance of including HTLV in testing protocols for patients with hepatitis C in the North region of Brazil. Full article

Understanding species-dependent siRNA length generation provides both fundamental and applied insights. At the basic level, it highlights an underappreciated dimension of RNAi diversity in insects, emphasizing that antiviral immunity cannot be fully understood from Drosophila melanogaster alone. At the applied level, these findings have direct implications for the design of dsRNA-based pest management strategies, where tailoring siRNA production to the target insect order could enhance both efficacy and specificity. Previously studies demonstrated that Dicer-2-generated siRNAs exhibit distinct, species-dependent length distributions: dipterans (D. melanogaster) and coleopterans (Tribolium castaneum) mainly produce 21-nt siRNAs; and hymenopterans (Bombus terrestris) and orthopterans (Locusta migratoria) generate siRNAs enriched at 22 nt; whereas lepidopterans such as Spodoptera exigua and Trichoplusia ni predominantly produce 20-nt siRNAs. The central biological question motivating this study was whether structural divergence in Dicer-2 may explain these lineage-specific differences in siRNA length profiles and antiviral RNAi efficiency. To address this, we interpreted observed structural variations in the context of the “molecular ruler” hypothesis and integrated them with previous experimental data on siRNA length variation across insect taxa. Future studies that combine high-resolution structural biology, comparative genomics, and functional assays will be essential to experimentally test whether the structural correlations proposed here determine Dicer-2 cleavage length in vivo and in vitro, and to leverage this knowledge for both agricultural and biomedical applications. Full article

Metapneumoviruses comprise a genus of negative-sense RNA viruses that cause significant respiratory disease across human and avian hosts. Human metapneumovirus (hMPV) is a globally prevalent pathogen associated with acute lower respiratory tract infections in infants, older adults, and immunocompromised individuals. Avian metapneumovirus (aMPV) imposes substantial economic losses on the poultry industry through respiratory disease, reproductive impairment, and high mortality in the presence of secondary infections. Despite their distinctive host ranges, hMPV and aMPV share a conserved genomic architecture and encode homologous structural and non-structural proteins that mediate viral entry, replication, assembly, and evasion of host innate immunity. Comparative analysis highlights that both have deeply conserved polymerase and nucleocapsid functions, and yet have a wide range of diversity in the attachment glycoprotein (G) and small hydrophobic protein (SH), reflecting divergent evolutionary pressures in human versus avian hosts that have led to such distinctive differences. The recent emergence and detection of aMPV/A and aMPV/B across the previously aMPV-free United States beginning in late 2023, combined with rising cases globally of hMPV post-SARS-CoV-2 pandemic, underscore the continued challenges of metapneumovirus surveillance and control in humans and animals. This review aims to highlight the current knowledge on the history, molecular virology, pathogenesis, epidemiology, diagnostics, and control strategies for aMPV while drawing mechanistic parallels to hMPV. By contextualizing shared biology and structure alongside host-specific adaptations, we aim to identify key gaps that shape vaccine design, antiviral development, and future research priorities aimed at mitigating the health and economic burden posed by metapneumoviruses found in both birds and humans. Full article

Therapeutic proteins face a critical pharmacokinetic challenge: rapid clearance from circulation limits their clinical efficacy. Albumin-binding domains (ABDs) offer an elegant solution by enabling therapeutic proteins to “hitchhike” on serum albumin’s favorable 19-day half-life through FcRn-mediated recycling. Clinical validation through approved therapeutics like ozoralizumab demonstrates the success of this approach, with preclinical studies showing fusion to an ABD extended half-life to 18 days. This review provides an analysis of ABD-fusion protein design, integrating structural biology, computational prediction, and rational engineering principles. We catalog the major classes of albumin-binding modalities, including bacterial three-helix bundle domains, engineered peptides, antibody-derived binders, and alternative scaffolds, comparing their binding properties, size contributions, cross-species reactivity, and production cost. Critical examination of linker architectures reveals that flexible glycine-serine linkers (particularly the widely successful (GGGGS)₃ motif) provide optimal balance between domain independence and molecular economy, though linker choice profoundly influences not only spatial separation but also binding affinity, folding, stability, and pharmacokinetics. We evaluate the utility and limitations of the structure prediction tools for ABD-fusion design. We establish practical guidelines for integrating computational screening with experimental validation. This review provides protein engineers and synthetic biologists with a comprehensive framework for rational design of albumin-binding therapeutics, emphasizing the synergistic integration of structural insight, computational prediction, and systematic experimental validation to accelerate development of next-generation long-acting biotherapeutics. Full article

Direct methods provide a model-free approach to solving the crystallographic phase problem and deliver unbiased atomic structures. However, conventional iterative projection algorithms such as Hybrid Input–Output (HIO) face two critical challenges: discontinuous density modification at the protein-solvent boundary and inaccurate molecular envelope reconstruction that fails to account for trapped solvent, particularly in crystals with solvent content approaching the lower limits of direct phasing applicability. We introduced four continuous iterative projection algorithms, including our improved continuous version, which implements smooth density modification at protein-solvent interfaces. To address envelope inaccuracy, we developed a two-step refined reconstruction scheme using sequential large-radius and small-radius Gaussian filters to identify trapped solvent molecules within surface cavities and internal channels. This scheme enhances the performance of both continuous and classical algorithms, including HIO, the difference map, and our improved versions. Benchmarking on 28 protein structures (solvent contents 55–78%, resolutions 1.46–3.2 Å, reported R-factor less than 0.22) showed that the refined envelope scheme increased average success rates of continuous algorithms by 45.7% and classical algorithms by 60.5%. The performance of continuous algorithms and improved classical algorithms proved comparable to the well-established HIO algorithm, forming a top-tier group that exceeded other classical algorithms. Integrating a genetic algorithm co-evolution strategy further enhanced average success rates by approximately 2.5-fold and accelerated convergence through population-wide information sharing. Although the success rate correlates with solvent content, our strategy improved success probability at any given solvent level, extending the practical boundaries of direct methods. The high success rate enabled averaging of multiple independent solutions, which reduced mean phase error by approximately 6.83° and yielded atomic models with backbone root-mean-square deviation (RMSD) typically below 0.5 Å relative to structures reported in the Protein Data Bank (PDB). This work introduces novel algorithms, a refined envelope reconstruction methodology, and an effective optimization strategy with genetic algorithm evolution. The complete framework enhances the capability and reliability of direct methods for phasing protein crystals with limited solvent content and provides a toolkit for addressing challenging cases in structural biology. Full article

Background: Multiple sclerosis (MS) is a neuroinflammatory disease characterized by autoimmune-driven inflammation in the central nervous system that damages axons and destroys myelin. It is difficult to diagnose multiple sclerosis due to its complexity, and different people may react differently to different treatments. While the exact cause of multiple sclerosis (MS) and the reasons for its increasing prevalence remain unclear, it is widely believed that a combination of genetic predisposition and environmental influences plays a significant role. Methods: Finding biomarkers for complicated diseases like multiple sclerosis (MS) is made more promising by the emergence of network and system biology technologies. Currently, using tools like Network Analyst to apply network-based gene expression profiling provides a novel approach to finding potential medication targets followed by molecular docking and MD Simulations. Results: There were 1200 genes found to be differentially expressed, with CD44 showing the highest degree score of 15, followed by CDC42 and SNAP25 genes, each with a degree score of 14. To explore the regulatory kinases involved in the protein–protein interaction network, we utilized the X2K online tool. The present study examines the binding interactions and the dynamic stability of four ligands (Obeticholic acid, Chlordiazepoxide, Dextromethorphan, and Hyaluronic acid) in the Hyaluronan binding site of the human CD44 receptor using molecular docking and molecular dynamics (MD) simulations. Docking studies demonstrated a significant docking score for Obeticholic acid (−6.3 kcal/mol), underscoring its medicinal potential. MD simulations conducted over a 100 ns period corroborated these results, revealing negligible structural aberrations (RMSD 1.3 Å) and consistent residue flexibility (RMSF 0.7 Å). Comparative examinations of RMSD, RMSF, Rg, and β-factor indicated that Obeticholic acid exhibited enhanced stability and compactness, establishing it as the most promising choice. Conclusions: This integrated method underscores the significance of dynamic validations for dependable drug design aimed at CD44 receptor-mediated pathways. Future experimental techniques are anticipated to further hone these findings, which further advance our understanding of putative biomarkers in multiple sclerosis (MS). Full article

This systematic review investigates the impact of chronic stress on treatment outcomes among cancer patients with divergent survival rates, focusing on breast, prostate, pancreatic, and ovarian cancers. The analysis explores how chronic stress influences molecular pathways and tumor progression while comparing cancers with five-year survival rates above and below 50%. A comprehensive literature search was conducted in PubMed and Scopus for studies published between 2014 and 2025 using combinations of keywords related to “chronic stress,” “psychological stress,” “psychotherapy,” and selected cancer types. All studies met the inclusion criteria according to the PRISMA 2020 guidelines. Evidence suggests that chronic stress is associated with the activation of neuroendocrine and immune mechanisms, including β-adrenergic and glucocorticoid signaling. These multifactorial processes are associated with disease progression and survival, particularly in pancreatic and ovarian cancers; however, these links remain primarily associative rather than causative. Conversely, psychotherapeutic interventions alleviate stress-related biological responses, improve quality of life, and may indirectly enhance therapeutic efficacy. By structuring the evidence around cancers with higher versus lower five-year survival, our review provides a survival informed synthesis of cancer type specific stress biology and stress-mitigating interventions, highlighting potentially targetable pathways and clear evidence gaps for future trials. The findings underscore the need to integrate psychological care into oncological practice to improve overall outcomes. Full article

Heme released during blood digestion represents a major oxidative challenge for hematophagous insects, promoting the generation of reactive oxygen species (ROS) and redox imbalance. Although Aedes aegypti has evolved specialized mechanisms to mitigate heme toxicity, how these responses vary across developmental stages remains poorly understood. Here, we applied quantitative proteomics to compare the effects of heme exposure in larvae and adult females. In larvae, heme treatment predominantly led to downregulation of metabolic and antioxidant proteins, consistent with a shift toward energy conservation and growth regulation. Nonetheless, selective upregulation of proteins associated with mitochondrial MnSOD activity, lipid remodeling, and cytoskeletal organization indicates the engagement of complementary protective mechanisms. In contrast, adults exhibited a coordinated bioenergetic response, characterized by enrichment of mitochondrial pathways, redox-related proteins, and molecular chaperones, reflecting enhanced resilience to oxidative stress. Enrichment of cuticle-associated proteins in both stages further suggests heme-induced structural remodeling. Together, these findings demonstrate that A. aegypti employs divergent, stage-specific proteomic strategies to cope with heme toxicity: larvae suppress metabolic activity while maintaining structural and redox homeostasis, whereas adults reinforce mitochondrial function and proteostatic defenses. These insights advance our understanding of mosquito redox biology and highlight stage-specific vulnerabilities that may be exploited for innovative vector control strategies. Full article

The livestock industry experiences significant economic losses as a result of viral infections. Building on recent advances in biotechnological research, recombinant viral vector vaccines have emerged as promising platforms for next-generation vaccines. These vaccines can overcome many limitations of conventional vaccines, as they provide stronger protective immune profiles, stability, and improved safety profiles for various diseases. Bovine herpesviruses serve as viral vector platforms utilized due to their large genome capacity, potential for multigenic antigen delivery, and significant immune stimulation. In this review, we explored the structural characteristics and genomic organization of bovine alphaherpesviruses (BoHV-1, BoHV-4, and BoHV-5), covered BoHV-5 biology and attenuation strategies as part of the comparative platform analysis, and summarised the latest advancements in molecular tools used for viral genome editing. We further highlight the development of vaccines against bovine and zoonotic pathogens, discuss applications of BoHV-based vectors, and deliberate on future directions to improve vaccine efficacy. It also discussed the current state of research in the field, considered prospects, and outlined strategies for impending research. BoHV vectors are promising candidates as next-generation vaccine platforms in veterinary medicine and will play an important role in integrated disease control in livestock. Full article

The silver carp (Hypophthalmichthys molitrix) is a filter-feeding fish species, characterized by significant morphological transformations in its filter-feeding apparatus, particularly the gill rakers, which are closely associated with dietary changes throughout its development. Despite the importance of these morphological innovations, the molecular mechanisms driving these changes remain largely unexplored. To investigate this, we employed an integrative approach combining scanning electron microscopy (SEM) and comparative transcriptomics to examine the gill rakers at five critical developmental stages (6, 15, 30, 45, and 60 days post-hatching, dph). SEM analysis revealed a structural evolution from sparse, bump-like protrusions to a dense, interlocking mesh. Simultaneously, transcriptomic analysis identified 10,184 differentially expressed genes (DEGs), which showed significant enrichment in pathways such as Focal Adhesion, ECM-Receptor Interaction, and the PI3K-Akt Signaling Pathway. Gene Set Enrichment Analysis (GSEA) indicated a coordinated upregulation of collagen and integrin gene families during the early developmental transition (6 vs. 15 dph), highlighting their crucial role in the formation of the sieve structure. This study reveals the molecular mechanisms of gill raker development in silver carp, providing initial insights into genetic regulation of morphology for ecological adaptation. The findings connect developmental biology, evolutionary biology, and ecology. Full article

Time-qualified dietary interventions, including time-restricted eating (TRE), intermittent fasting (IF), and periodic fasting-mimicking diets (FMDs), have emerged as strategies to improve metabolic health. While preclinical studies consistently demonstrate robust effects on energy metabolism, cardiometabolic function, and longevity, translation to humans remains heterogeneous. In free-living settings, most metabolic improvements observed with TRE and IF appear primarily driven by spontaneous caloric restriction rather than meal timing per se, and isocaloric randomized controlled trials generally show no additional benefits compared to standard calorie restriction. Evidence supporting circadian-specific advantages, particularly for early TRE, is promising but inconsistent and often context-dependent. Important uncertainties also persist regarding long-term efficacy, lean mass preservation, safety in specific populations, and the physiological impact of extended fasting windows. Despite these controversies, time-qualified diets represent a paradigm shift in nutritional science by integrating chronobiology with dietary patterns. Future directions include tailoring eating windows to individual chronotypes, combining fasting regimens with high-quality dietary patterns and structured physical activity, and clarifying the molecular mechanisms that may mediate calorie-independent benefits. Large, long-term, mechanistically informed human trials are essential to determine whether aligning eating behaviors with circadian biology can produce durable clinical improvements. Such work will ultimately shape the role of personalized chrononutrition in preventive and therapeutic nutrition. Full article

Blueberry (Vaccinium spp.) is a high-value crop due to its growing global demand, recognized nutraceutical properties, and strong linkage with emerging technologies in precision agriculture and postharvest management. To characterize the scientific evolution and intellectual structure, we conducted a bibliometric analysis of 474 documents indexed in Scopus between 1987 and 2025. A systematic search strategy based on taxonomic, agronomic, and technological descriptors was applied, followed by data cleaning and analysis with Bibliometrix and VOSviewer. Performance indicators and science-mapping techniques were used to examine temporal growth, geographical distribution, institutional and author leadership, and thematic structure. Scientific output shows a sustained upward trend with a maximum of 42 articles in 2024, confirming the consolidation of blueberry as a model crop for interdisciplinary research. Research articles represent over 75% of the total (359/474), evidencing an application-oriented and experimentally grounded field. Agricultural and Biological Sciences dominate (382 documents), followed by Engineering (70) and Biochemistry, Genetics, and Molecular Biology (66), reflecting increasing integration of crop management, technological innovation, and food science. Thematic mapping identified five main clusters: physiology and health, plant protection, agronomic management and digitalization, processing and stability of phenolic compounds, and analytical characterization. The analysis reveals gaps in the integration of physiology, food science, and metabolomics, as well as in the biological validation of biomarkers and the study of peripheral Vaccinium species. Overall, the field exhibits a consolidated and sustainability-oriented interdisciplinarity, highlighting opportunities to advance toward more comparable analytical protocols, digital traceability, and artificial-intelligence-assisted decision support along the blueberry value chain. Full article