Search Results (1,700)

Median overall survival remains a central endpoint in oncology, but it can obscure a clinically meaningful long tail of patients with advanced solid tumors who survive well beyond the median. One biological context in which this pattern may be relevant is tumor microenvironment (TME) acidosis. Driven by aerobic glycolysis, hypoxia, impaired perfusion, and proton-export programs, acidic TME is increasingly implicated in invasion, therapeutic resistance, and immune suppression. This narrative review examines TME acidosis as the primary biological framework and considers long-tail survival as a clinical lens through which its implications may be interpreted. We summarize the biological basis and heterogeneity of acidic TME, review current approaches to clinical and translational assessment of tumor acidity, including acidoCEST magnetic resonance imaging (MRI) and positron emission tomography (PET)-based approaches, and discuss the potential and limitations of alkalization-oriented interventions such as buffering and diet-based strategies. Particular attention is given to the distinction between direct measurements of tumor acidity and clinically feasible but indirect markers such as urinary pH, which should not be interpreted as a direct surrogate for local tumor extracellular pH. From a science-based medicine perspective, long-tail survival is treated here as a hypothesis-generating clinical signal rather than proof of causality. Overall, alkalization-oriented interventions appear biologically plausible and clinically testable, but current clinical evidence remains limited and context-dependent. Future progress will require mechanistically informed biomarkers, careful safety evaluation, and trial designs capable of detecting delayed separation of survival curves and tail-oriented patterns of benefit. Full article

Vaccination remains the core strategy for the prevention and control of Newcastle disease (ND). The inherent thermosensitivity of traditional Newcastle disease virus (NDV) vaccines imposes major limitations on their transportation, storage, and field application. To address these challenges, a novel liquid, thermostable, live ND vaccine was developed in the present study. Firstly, Tris/HCl buffer at near-neutral pH was identified as the optimal basic buffer system. On this basis, further screening and formulation optimization of vaccine stabilizers were conducted, and NDV strains with excellent thermal stability were used to verify the stability-conferring properties of the developed stabilizer. The results showed that the formulation composed of 0.5% gelatin, 4% trehalose, 0.1% L-glutamic acid, and 0.5% thiourea was confirmed as the optimal stabilizer for ND liquid vaccines. This formulation maintained the stable storage of the tested NDV for 12 months at 4 °C and exhibited promising stability for 30 days at 25 °C, marking a significant advancement toward development thermostable NDV vaccines that are independent of a continuous cold chain. More importantly, the liquid vaccine stored at 4 °C for 12 months still induced high levels of NDV-specific antibodies in specific pathogen-free chicks and provided 100% protective efficacy against challenge with virulent NDV. In conclusion, the liquid vaccine stabilizer developed in this study not only significantly enhanced the thermostability of the vaccine but also effectively maintained its immunogenicity, thereby providing an important theoretical basis for the research and development of liquid ND vaccines. Full article

Pectate lyase (PeL) is a key cell wall-degrading enzyme in the infection process of plant-parasitic nematodes, with a large gene family exhibiting functional redundancy. The dominant PeL isoform during the initial infection time course remains unclear. In this study, 21 Ddpel genes were identified in Ditylenchus destructor Thorne, 1945, 7 of which were differentially expressed during the initial infection time course of this nematode. The purified proteins of these seven DdPeLs showed pathogenicity toward both sweet potato and tobacco, and their optimal enzymatic pH varied significantly. Prior to host infection, D. destructor preferentially expresses Ddpel genes encoding pectate lyase with higher activity at pH 5.8. However, within 5 days post-inoculation with nematodes, the expression of genes encoding acidic DdPeL enzymes (enzymes with optimal activity in acidic pH) was upregulated, while genes encoding alkaline DdPeL enzymes (optimal activity in alkaline pH) were concurrently downregulated. Through site-directed mutagenesis, we demonstrated that the loss of enzymatic activity in DdPeLs abolished their ability to induce plant cell death. Furthermore, when acidic or alkaline DdPeLs were pre-treated with dialysis in their respective optimal pH buffers prior to infiltration, their pathogenicity was significantly enhanced. Together, these findings demonstrate that enzymatic activity, governed by protein structure and local pH, is a key determinant of pathogenicity. Previous studies have reported that phytopathogens can secrete organic acids during the initial infection phase, leading to localized acidification of the host microenvironment. We therefore hypothesize that, during the initial infection time course, nematodes may actively acidify the host microenvironment to specifically enhance the enzymatic activity of acidic DdPeLs, thereby promoting cell wall degradation and facilitating infection establishment. Full article

The persistence of cadmium (Cd) immobilization in acidic paddy soils is exacerbated by acidification and fluctuating redox conditions that promote Cd re-mobilization. While biochar is a promising amendment, its long-term efficacy in Cd immobilization relative to conventional lime and the underlying mechanisms remain incompletely resolved. This study tested the hypothesis that biochar’s superior effect lies in its durable enhancement of soil pH buffering capacity (pHBC), not merely in increasing initial pH. Using six acidic paddy soils amended with three biochars (corn straw, peanut straw, and seeded sunflower plate) and pH-matched lime [Ca(OH)₂] controls, we quantified pHBC changes, resistance to simulated acidification, and Cd dynamics during a flooding-drying cycle. Results showed that biochar amendments increased pHBC by 24.7–110%, significantly more than lime. Under acid stress, biochar-treated soils maintained higher pH and released 40–85% less soluble and extractable Cd than lime controls at equivalent pH range. Correlation and regression analyses established that the biochar-induced change in pHBC (ΔpHBC) was the strongest predictor of reduced Cd availability, exerting twice the influence of native soil pHBC. During the redox cycle, enhanced pHBC directly attenuated soil re-acidification upon drainage, minimizing Cd re-mobilization. Thus, the durable enhancement of soil pHBC is the central mechanism for biochar’s sustained Cd immobilization, advocating a strategic shift from transient pH adjustment to building inherent soil buffering resilience for long-term remediation security. Full article

This study presents a novel, integrated membrane–resin hybrid platform for the high-efficiency purification of N-acetylneuraminic acid (sialic acid, NANA) from complex microbial fermentation broths. By synergistically combining four sequential stages—ceramic microfiltration (50 nm), ultrafiltration (3 kDa), nanofiltration (150 Da), and dual-resin purification (macroporous adsorption + cation-exchange)—the process achieves stepwise removal of cells, proteins, pigments, monovalent salts, and divalent metal ions without using organic solvents or high-salt buffers. Critically, each stage demonstrates high target recovery: 76.2% (CM), 67.3% (UF), and 77.5% (NF), with near-quantitative retention (>95%) during resin treatment due to NANA’s low hydrophobicity and electrostatic repulsion at pH 6.8. Following optimised acidification crystallisation (acetic acid dosage = 3 × concentrate volume; sialic acid concentrate concentration = 333.49 g/L), the final product reaches 97.9% purity with a crystalline yield of 78.6%. This scalable, green purification strategy eliminates major bottlenecks in downstream processing and enables industrial-scale production of pharmaceutical-grade sialic acid, with broad applicability to other high-value acidic biomolecules. Full article

The translation of nucleic acid amplification into practical point-of-care and biosensor-integrated diagnostics is still significantly impeded by the necessity for rapid sample preparation. For this reason, a broad comparison of seven commercially available kits for DNA/RNA extraction containing their temperature-related adjustments was performed. Extracts isolated from SARS-CoV-2-positive nasopharyngeal swabs, viral stocks, as well as laboratory-prepared suspensions of clinically relevant Gram-positive and Gram-negative bacteria were evaluated by recombinase polymerase amplification (RPA) and real-time PCR. In addition, the impact of transport media for SARS-CoV-2 samples was investigated. Extraction performance varied markedly according to the kit, pathogen, sample background. For SARS-CoV-2, rapid extraction was more effective for samples collected in viral transport medium than in inactivation buffer. Across bacterial targets, performance was species dependent, highlighting substantial differences in compatibility between simplified extraction workflows and downstream amplification. Among the rapid methods tested, a simplified QuickExtract protocol (95 °C, 5 min) provided the most consistent overall results, although it did not uniformly match the reference silica-based method for all targets. In conclusion, these results demonstrate that rapid nucleic acid extraction must be thoroughly evaluated as an essential element of the entire sample-to-answer workflow, rather than being chosen as a standalone preprocessing step for point-of-care molecular diagnostics. Full article

Background: Post-episiotomy wound healing remains largely managed through supportive care, despite growing evidence that local biochemical conditions critically influence tissue regeneration. Lactic acid is of particular interest in this context because it is both an endogenous metabolic intermediate and a physiologic component of the vaginal microenvironment, where it contributes to acidic pH maintenance, microbial homeostasis, and mucosal protection. Beyond these local effects, lactate has emerged as a signaling metabolite involved in angiogenesis, immune regulation, and extracellular matrix remodeling, making it a relevant candidate for regenerative wound care. Methods: This narrative translational review integrates evidence from molecular biology, biomaterials science, and clinical obstetrics to examine the therapeutic potential of lactic acid-loaded hydrogels for post-episiotomy tissue repair. Literature from PubMed, Scopus, and Web of Science was analyzed to evaluate physicochemical design parameters, lactate-mediated signaling pathways, and available clinical outcomes. Results: Lactic acid may function both as a microenvironmental regulator and as a metabolic signal capable of stabilizing hypoxia-inducible factor-1α signaling, enhancing vascular endothelial growth factor expression, modulating macrophage polarization, and influencing fibroblast-mediated extracellular matrix synthesis. Hydrogel matrices provide tunable platforms for controlled lactate release, pH buffering, and mucosal compatibility. Clinical studies suggest improved epithelialization, reduced infection risk, and lower pain scores following topical lactic acid formulations in episiotomy repair. In parallel, platelet-rich plasma provides autologous growth factor enrichment that may complement regenerative signaling pathways. Conclusions: Integrating microenvironment stabilization through lactic acid-based hydrogels with biologically active regenerative strategies represents a promising direction for post-episiotomy wound healing. Further controlled trials and standardized biomaterial characterization are required to define optimal therapeutic protocols and confirm long-term clinical benefit. Full article

Background/Objectives: Mass spectrometry-based multi-attribute methods (MAM) have the potential to transform therapeutic protein analysis by enabling comprehensive monitoring of multiple quality attributes in a single assay. However, the widespread adoption of MAM is hindered by significant challenges, most notably artificial oxidation during sample preparation and analysis. This report summarizes long-term operational observations and several case studies that substantiate this concern. Methods: A tryptic digest, high-resolution LC-MS MAM workflow was applied to an Fc-fusion protein and multiple antibody-based therapeutics, with a frozen reference standard analyzed in each run for system suitability and longitudinal trending. Oxidation excursions were investigated by comparing laboratories, consumables, LC-MS configurations, and other method parameters. Results: In a seven-year trending record, apparent total methionine oxidation in the Fc-fusion protein reference standard showed an abrupt, sustained increase (up to ~5-fold); the shift was traced to a specific bag of microcentrifuge-tubes used during buffer exchange and resolved after those tubes were discontinued. In an antibody–drug conjugate, observed methionine oxidation was strongly influenced by the sample preparation procedure. In other antibodies, variability of observed methionine oxidation was attributed to on-column oxidation, which produced a broad and noisy peak that interferes with automated peak integration. EDTA flushing reduced this feature, implicating exposure to metal ions. Conclusions: While advances continue to address many MAM challenges, artificial oxidation remains unpredictable and constitutes a major obstacle to robust implementation in regulated QC environments. Enhanced control strategies and further research are urgently needed to ensure reliable therapeutic protein analysis. Such control strategies include consumable qualification and change control, system suitability/trending using a reference standard, metal management across LC flow path/column lifecycle, reduction of trifluoracetic acid (TFA) exposure, data analysis to safeguard excessive on-column oxidation, etc. Full article

Background: Chromophobe renal cell carcinoma (ChRCC) is characterized by the accumulation of abnormal mitochondria, a high rate of mitochondrial DNA (mtDNA) mutations, and altered oxidative metabolism. There are no existing circulating biomarkers to distinguish metastatic ChRCC from clear cell renal cell carcinoma (ccRCC). Methods: High-throughput plasma proteomic profiling using the SomaScan platform was performed in 18 ChRCC (including 16 metastatic ChRCC) and 197 metastatic ccRCC patients. Data were harmonized to generate a unified 7K-protein matrix. Results: Differential expression analysis was performed using limma (version 3.62.2). Of 7272 quantified human plasma proteins, 209 were differentially expressed between ChRCC and ccRCC. Upregulated proteins in ChRCC included essential β-oxidation enzymes such as ECH1 (enoyl-CoA hydratase 1) and ECI1 (enoyl-CoA delta-isomerase 1), suggesting increased long-chain fatty acid degradation. Creatine and energy-buffering pathways were also represented, with increased CKMT1A (Creatine Kinase, Mitochondrial 1A) in ChRCC. KIM-1 (Kidney Injury Molecule-1) and leptin were lower in ChRCC, consistent with the known upregulation of these proteins in ccRCC. Pathway enrichment analyses revealed an overrepresentation of mitochondrial protein degradation, fatty acid β-oxidation, and respiratory electron transport in ChRCC, suggesting that ChRCC sheds a unique mitochondrial signature into the peripheral circulation. A bootstrap-based LASSO logistic regression restricted to upregulated mitochondrial proteins in ChRCC vs. ccRCC consistently selected ECI1 and CKMT1A. The LASSO model achieved an AUROC of 0.964. Conclusions: Compared to ccRCC, the plasma proteome of metastatic ChRCC is dominated by mitochondrial metabolic enzymes, revealing a systemic metabolic phenotype strikingly aligned with the known histologic accumulation of abnormal mitochondria in ChRCC cells. Full article

Background/Objectives: Sea urchin gonads (“roe”) are a valuable seafood product and a chemically complex matrix whose composition varies with physiology and environment. We present a biphasic extraction and ¹H NMR workflow to build a reusable reference inventory of polar metabolites and apolar lipid features in Paracentrotus lividus. Methods: Gonads from 37 adults (23 males, 14 females) collected at two sites (Alicante and Jávea–Dénia, Spain; October 2024) were lyophilized, extracted with methanol/chloroform/water, and analyzed by 400 MHz ¹H NMR in buffered aqueous solution (polar) and CDCl₃ (apolar). Polar metabolite identification combined 1D patterns with database matching and ¹H–¹³C HSQC confirmation on representative samples, yielding 71 annotated resonances corresponding to 37 metabolites spanning amino acids, osmolytes/quaternary amines, carbohydrates/aminosugars, and nucleoside/purine-related compounds. Results: Polar fingerprints enabled supervised modelling: PLS-LDA separated sexes with low cross-validated error, and SPA/COSS ranking highlighted glycine, alanine, creatine and osmolyte-associated signals as key discriminants; pathway mapping supported the enrichment of amino-acid and one-carbon/purine networks. Apolar spectra were annotated at the motif level and used for lipid-index estimation, indicating substantial unsaturation but low docosahexaenoic acid (DHA) and modest sex effects. Conclusions: The curated peak lists and reporting framework facilitate reproducible NMR annotation and future comparative studies of P. lividus gonads. Full article

Poor aqueous solubility remains a major limitation for the oral delivery of many active pharmaceutical ingredients (APIs). Deep eutectic solvents (DESs) exhibit remarkable drug-solubilization capacity, yet rapid precipitation upon aqueous dilution can compromise their ability to sustain supersaturation. This study investigates polymer-embedded DES (PEDES) systems as liquid supersaturating drug delivery platforms in which hydration and polymer chemistry jointly govern thermodynamic solubilization and kinetic stabilization. A choline chloride/DL-malic acid DES was prepared with 5% or 15% (w/w) water and combined with polyvinylpyrrolidone (PVP) or polyacrylic acid (PAA). Griseofulvin (GRF) was used as a precipitation-prone model drug. Structural characterization (ATR-FTIR, ¹H-NMR), equilibrium solubility measurements, storage stability studies, and non-sink dissolution testing were conducted to elucidate formulation behavior. The DES systems enhanced GRF solubility by up to ~59-fold relative to phosphate buffer (PBS, pH 6.8). Polymer incorporation produced hydration- and concentration-dependent effects. These results suggest the presence of competitive or cooperative interaction regimes. At 5% water, PEDES formulations failed to prevent recrystallization and showed limited supersaturation maintenance. In contrast, PEDES systems containing 15% water exhibited improved stability, with the formulation containing 4% PAA sustaining elevated drug concentrations for 120 min under non-sink conditions. Low-frequency solution-state ¹H-NMR confirmed stronger GRF–PAA interactions relative to PVP, supporting the role of polymer–drug association in supersaturation stabilization. These findings demonstrate that PEDES performance emerges from a hydration-dependent balance between solvent structuring and drug–polymer interactions, highlighting hydration and polymer functionality as key parameters for the rational design of liquid supersaturating systems. Full article

To promote the high-value utilization of fly ash (FA) and address the prolonged setting time and limited strength associated with conventional single-alkali activation, this study proposes a synergistic dual-alkali activation strategy using Ca(OH)₂ and Na₂SiO₃ in combination with microwave-assisted curing for low-calcium fly ash. Samples containing varying amounts of Ca(OH)₂ were systematically characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), compressive strength testing, and pore structure analysis. The results show that Ca(OH)₂ facilitates the formation of calcium aluminosilicate hydrate (C-A-S-H) gel, while Na₂SiO₃ sustains the alkaline environment and enhances the dissolution of SiO₂ and Al₂O₃ from FA. The dual-alkali synergistic system, when coupled with microwave treatment, markedly refines the pore structure, increases the degree of polymerization, and improves compressive strength from 0.5 MPa to 1.7 MPa with increasing Ca(OH)₂ content. In addition, the prepared fly ash-based geopolymer (FABG) demonstrates pronounced pH-buffering capacity in acidic environments and exhibits antibacterial activity, primarily attributable to its sustained release of alkalinity. This work highlights that integrating dual-alkali activation with microwave curing can simultaneously enhance microstructural development, chemical stability, and functional performance in low-calcium FA systems, thereby offering a viable route for the development of sustainable and multifunctional green building materials derived from industrial solid waste. Full article

Ultra-processed foods (UPFs) represent a distinct dietary paradigm characterized by structurally simplified food matrices and chronic exposure to multiple additives, including emulsifiers, artificial sweeteners, and preservatives. Rather than acting in isolation, these compounds operate within a multi-additive environment that reshapes the gut ecosystem through convergent mechanisms. Emerging evidence suggests that additive-rich ultra-processed dietary environments may disrupt the gut ecosystem through three interconnected layers: (1) structural impairment of the intestinal barrier, including mucus erosion and tight-junction destabilization; (2) microbial metabolic shifts marked by short-chain fatty acid depletion, altered bile acid signaling, and enrichment of lipopolysaccharide-producing taxa; and (3) immune and inflammatory reprogramming promoting low-grade systemic inflammation. These processes collectively reduce ecosystem resilience—the capacity of the gut microbiota to resist and recover from perturbation. Vulnerability to additive-driven dysbiosis varies across the life course. During infancy, incomplete ecosystem stabilization may increase susceptibility to long-term ecological imprinting, whereas in older age, reduced microbial diversity and immune remodeling may impair recovery capacity following dietary stressors. In contrast, fiber-rich, minimally processed dietary patterns appear to enhance microbial resilience by reinforcing functional redundancy, metabolic buffering, and barrier integrity. Although much mechanistic evidence has been derived from experimental models, accumulating human data support the biological plausibility of additive-associated microbiota alterations. By integrating multi-additive exposure, ecosystem disruption, life-course modulation, and resilience within a unified framework, this review provides a mechanistically coherent model linking ultra-processed dietary environments to microbiota-mediated chronic disease risk. Here, we formalize this integrative perspective as the Three-Layer Ecosystem Disruption (TLED) Model. Full article

Moist chilling is widely used to overcome seed dormancy in zoysiagrass (Zoysia japonica Steud.), but the coordinated physiological and molecular basis remains unclear. Here, freshly matured seeds were subjected to moist chilling at 4 °C in darkness for 0 (Control), 1 (CS1), 2 (CS2), 3 (CS3), or 4 weeks (CS4) and then transferred to germination conditions (30/20 °C, day/night). Prolonged moist chilling progressively improved dormancy release: final germination percentage increased from 40.5% (Control) to 73.5% (CS4), accompanied by a higher germination index and earlier, faster cumulative germination dynamics. Moist chilling also enhanced early seedling vigor, with stronger treatment differentiation in root elongation than in shoot growth. Physiologically, abscisic acid (ABA) content declined while gibberellic acid (GA) content increased, resulting in an elevated GA/ABA ratio with prolonged chilling. Metabolic activation was evidenced by increased α-amylase activity, greater soluble sugar and soluble protein accumulation, and stimulated oxygen uptake. In addition, CAT, SOD, and POD activities were enhanced under prolonged moist chilling, whereas H₂O₂ levels remained relatively stable, suggesting that redox adjustment during dormancy release was characterized by strengthened antioxidant buffering rather than pronounced oxidative accumulation. qRT-PCR supported a mechanistic transition from dormancy maintenance to germination execution, showing moist chilling-associated regulation of ABA/GA metabolism and signaling genes (e.g., NCED, CYP707A, ABI3/ABI5, and GA20ox) and downstream metabolic modules (e.g., GAMYB, AMY, ISA, INV, and HXK1), together with concurrent modulation of respiration- and ROS-related markers (e.g., AOX1a, RBOH, and CAT). Correlation analysis linked germination performance most strongly with α-amylase activity, oxygen uptake, and the GA/ABA ratio. Collectively, our data support a working model in which moist chilling rebalances the ABA–GA gate and activates downstream metabolic and redox adjustment modules to promote dormancy release and improve germination performance in zoysiagrass, providing practical markers for optimizing seed establishment through moist chilling treatment. Full article

Immobilized microorganism technology offers a promising approach for remediating heavy metal-contaminated soils. This study developed a novel bio-mineral composite (B-AM) by coupling acid-modified maifanite (AM) with Bacillus mucilaginosus to enhance lead (Pb) immobilization. Comparative experiments demonstrated that B-AM outperformed conventional amendments, including oyster shell, pristine maifanite, AM and B. mucilaginosus in Pb immobilization. The B-AM treatment optimized soil pH, improved soil fertility with increases in available potassium (1.06-fold) and available phosphorus (1.28-fold). Additionally, B-AM transformed Pb into more stable fractions, reducing labile Pb fractions by 52.52% while increasing the residual fraction by 88.36%. These improvements resulted in an 83.24% reduction in Pb accumulation and a 63.95% increase in the fresh root weight of radish. Mechanistic insights revealed that the enhanced remediation performance stems from both the individual contributions of AM (adsorption capacity) and B. mucilaginosus (biosorption and biomineralization) and their synergistic interaction. Specifically, AM acts as a carrier and pH buffer, promoting microbial proliferation and reducing Pb remobilization from cell lysis. The resulting sustained microbial activity further leads to the formation of stable Pb minerals. Collectively, our results establish a theoretical and practical basis for using B-AM to remediate Pb-contaminated soils. Full article