Search Results (4,742)

Objectives: The aim of this study was to evaluate the stability of azacitidine (AZA) under clinical storage conditions (room temperature vs. refrigeration) to identify practical protocols that minimize waste and improve cost-effectiveness. Methods: AZA solutions (1 mg/mL) were stored at 23 ± 2 °C or 4 °C. Stability was assessed using a validated high-performance liquid chromatography (HPLC) method. Chromatographic separation was achieved on a Hypersil ODS C₁₈ column (250 mm × 4.6 mm, 5 μm) using an isocratic mobile phase of 50 mM potassium phosphate buffer (pH 7.0)-acetonitrile (98:2, v/v) at a flow rate of 1.0 mL/min, with UV detection at 245 nm and a 20 μL injection volume. The method demonstrated specificity for AZA and its main degradation product (DP), with LOD and LOQ of 12.56 μg/mL and 62.8 μg/mL, respectively. Linearity (R² = 0.9928), precision (RSD% < 5 for mid/high levels), and accuracy (mean recovery 96%) were established. Results: Azacitidine degraded rapidly at room temperature, with >85% loss within 24 h. In contrast, refrigeration at 4 °C significantly delayed degradation, with only ~26% loss observed over the same 24 h period. Chromatographic analysis confirmed the formation of a primary degradation product (tentatively identified as the open-ring hydrolytic species N-(formylamidino)-N′-β-D-ribofuranosylurea based on its chromatographic behavior and literature data), consistent with the known hydrolytic pathway. The applied HPLC-UV method offered an optimal balance of specificity and practicality for monitoring this main degradation trend under clinical storage conditions, distinguishing it from more complex techniques used primarily for structural elucidation. Conclusions: The pronounced instability of reconstituted AZA underscores the critical importance of strict adherence to immediate-use protocols. Refrigeration provides only a limited stability window. Based on our kinetic data, maintaining the reconstituted solution within an acceptable degradation limit (e.g., ≤10% loss) at 4 °C would require administration within a very short timeframe, supporting current handling guidelines to ensure therapeutic efficacy and minimize economic waste. Full article

Exploring green organic solvents is a global demand. Most of the currently used solvents pose some concerns regarding environmental sustainability and occupational health risks. In this work, propylene glycol was employed for the first time as a green solvent for mobile phase preparation in the reversed phase chromatographic separation of a mixture of two antioxidants, glutathione and ascorbic acid. The slight viscosity of propylene glycol was manipulated by using water as a co-fluidizing agent to facilitate pumping. Method optimization was performed using factorial design experimental Expert 13^® Software (Minneapolis, MN, USA) to achieve the maximum resolution and the minimum run time. The reported method was properly validated according to the International Conference on Harmonization criteria at the linearity range of 1–500 µg/mL, with acceptable accuracy and precision for both drugs. The method was effectively applied for the quantification of both drugs in their commercial pharmaceutical formulation. The proposed method was assessed for environmental and operator safety by means of global tools like AGREE and MoGAPI and has proved high degrees of greenness. Propylene glycol has several benign properties, such as low volatility, less toxicity, compatibility with UV detectors and very low flammability, that will soon assemble it as a promising alternative for the conventionally used solvents. Full article

Background/Objectives: Conventional next-generation sequencing (NGS) workflows often require more than two weeks to complete, delaying treatment decisions and limiting access to precision oncology in community settings. This report aimed to demonstrate the feasibility of performing rapid, comprehensive cell-free DNA (cfDNA)-based genomic profiling by introducing a fully automated NGS workflow in a community hospital environment. Case Presentation: A postoperative patient with pancreatic ductal adenocarcinoma and liver metastasis underwent cfDNA-based liquid biopsy using plasma collected in PAXgene^® Blood ccfDNA Tubes. Gene analysis was performed using the Oncomine Precision Assay GX5 on the Ion Torrent Genexus™ System (Thermo Fisher Scientific). Three pathogenic hotspot mutations—KRAS G12R, TP53 M246I/M246K, and GNA11—and one copy number gain in PIK3CA were identified, whereas no variants were detected in a healthy volunteer control. The total turnaround time from plasma separation to report generation was approximately 27 h, requiring only 40 min of total hands-on time. Discussion: This rapid, automated workflow enabled comprehensive cfDNA analysis within a clinically practical timeframe, overcoming key limitations of conventional multi-step NGS workflows that typically require external sample shipment and specialized personnel. The results confirm the technical feasibility of conducting high-quality molecular testing in a regional hospital setting. Conclusions: This report demonstrates that fully automated cfDNA-based NGS can achieve clinically meaningful genomic profiling within 27 h in a community hospital. This advancement addresses the time and cost barriers of traditional NGS analysis and represents a significant step toward promoting precision medicine in community healthcare. Full article

This study aimed to establish a fast and efficient method for the determination of N-nitrosamines (NAs) in meat products by integrating two sample preparation techniques—QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) and FaPEx (Fast Pesticide Extraction)—with liquid chromatography–tandem mass spectrometry (LC–MS/MS). Chromatographic separation was performed on a Poroshell 120 Phenyl Hexyl column using a gradient elution of acetonitrile and 0.01% formic acid at a flow rate of 0.3 mL/min and a column temperature of 25 °C. Under these conditions, nine NAs and one internal standard were completely separated within 11 min with selective reaction monitoring mode (SRM) for detection. Samples were first extracted with QuEChERS powder using acetonitrile containing 0.1% formic acid, followed by purification with a FaPEx-Chl cartridge. This combined approach demonstrated superior performance compared with traditional solvent extraction or QuEChERS extraction alone. The recoveries of the developed method ranged from 76% to 111% and 52% to 103% at spiking levels of 50 ng/g and 20 ng/g, respectively. The limits of detection (LOD) and quantification (LOQ) were 0.002–0.3 ng/g and 0.006–1.00 ng/g, respectively. The inter-day and intra-day precisions (RSD%) ranged from 2.7% to 17% and 2.9% to 17%, respectively. These results indicate that the proposed method is among the most time-efficient and effective analytical approaches currently available for the determination of NAs in meat products. Full article

Polyampholytes combine cationic and anionic groups in one macromolecular platform and are emerging as versatile components for energy storage and conversion. This review synthesizes how their charge balance, hydration, and architecture can be engineered to address ionic transport, interfacial stability, and safety across batteries, supercapacitors, solar cells, and fuel cells. We classify annealed, quenched, and zwitterionic systems, outline molecular design strategies that tune charge ratio, distribution, and crosslinking, and compare device roles as gel or solid electrolytes, eutectogels, ionogels, binders, separator coatings, and interlayers. Comparative tables summarize ionic conductivity, cation transference number, electrochemical window, mechanical robustness, and temperature tolerance. Across Li and Zn batteries, polyampholytes promote ion dissociation, homogenize interfacial fields, suppress dendrites, and stabilize interphases. In supercapacitors, antifreeze hydrogels and poly(ionic liquid) networks maintain conductivity and elasticity under strain and at subzero temperature. In solar cells, zwitterionic interlayers improve work function alignment and charge extraction, while ordered networks in fuel cell membranes enable selective ion transport with reduced crossover. Design rules emerge that couple charge neutrality with controlled hydration and dynamic crosslinking to balance conductivity and mechanics. Key gaps include brittleness, ion pairing with multivalent salts, and scale-up. Opportunities include soft segment copolymerization, ionic liquid and DES plasticization, side-chain engineering, and operando studies to guide translation. Full article

Designing ionic liquids (ILs) where a single functional group orchestrates a suite of enhanced properties remains a key challenge in materials science. Here, we introduce 1-butyl-3-methylimidazolium mandelate, [Bmim][Man], a novel IL where the hydroxyl group on the mandelate anion simultaneously enhances hydrogen bonding, thermal stability, antimicrobial activity, and extraction selectivity. The structure-property relationships of [Bmim][Man] were investigated through measurements of density, viscosity, and conductivity and were compared with analogous ILs. The presence of the hydroxyl group on the mandelate anion resulted in the highest density and viscosity among the series, attributed to strong hydrogen bonding and efficient ion packing. Notably, [Bmim][Man] exhibited a high molar conductivity that decouples from its high viscosity, suggesting an unusual degree of ion dissociation facilitated by the hydroxyl group. Thermogravimetric analysis revealed superior thermal stability. Furthermore, the investigated ionic liquid demonstrated a low critical aggregation concentration (CAC = 0.01982 mol·dm⁻³) in water, indicating a strong propensity for self-aggregation. [Bmim][Man] showed synergistic, enhanced antibacterial activity against E. coli and P. aeruginosa. Finally, the functional utility of this designed liquid was demonstrated in separation science, where [Bmim][Man]-based aqueous biphasic systems showed selective extraction capabilities for transition metals, a process driven by the same hydrogen-bonding and coordination interactions that define its bulk properties. These findings establish [Bmim][Man] as a promising multifunctional material where the mandelate anion concurrently dictates liquid microstructure, thermal resilience, antimicrobial performance, and application in extraction. Full article

Utilization of natural processes can reduce the complexity and production cost of any device by limiting the necessary steps in the production scheme, especially when it comes to fibers with periodic changes in refractive index. One such process is the nematic–isotropic phase separation of liquid crystal-based composite confined in 1D space. In this paper, we analyze the behavior of polymer-stabilized liquid crystal-based self-assembled periodic structures in an external electric field. We performed a detailed analysis regarding the reorientation of liquid crystal molecules under two orthogonal directions of the external electric field applied to the examined sample. It was demonstrated that the period of the polymerized structure remains constant until full reorientation, as the electric field induces the formation of new periodic defects in LC orientation. Consequently, the structure’s effective birefringence changes quite drastically, and this observed change depends on the direction of the electric field vector. The obtained results seem promising when it comes to application of the proposed periodic structures as voltage or electric field sensors operating as long-period fiber gratings or fiber Bragg gratings for the visible or near-infrared spectral regions. Full article

GC-rich sequences affect DNA replication, recombination and repair, as well as RNA transcription in vivo. Such sequences may also impede site-directed mutagenesis in vitro. P3a site-directed mutagenesis is a highly efficient method, but it has not been tested with plasmids possessing GC-rich sequences. Here we report that it is very efficient with a BRPF3 expression vector but unsuccessful with that for KAT2B. Because two GC-rich regions located within the synthetic CAG promoter and the KAT2B coding region may form guanine (G)-quadruplexes and hinder plasmid denaturation during PCR, we developed P3b site-specific mutagenesis, achieving an average efficiency of 97.5% in engineering ten KAT2B mutants. Importantly, deletion mutagenesis revealed that either of the two GC-rich regions are sufficient for rendering the plasmid incompatible with P3a mutagenesis. Consistent with this, only P3b mutagenesis worked efficiently with several widely used sgRNA/Cas9 expression vectors, which contain the CAG promoter, and with an expression vector for CDK13, which possesses an intrinsically disordered domain encoded by a GC-rich DNA fragment. Thus, this study highlights serious challenges posed by GC-rich sequences to site-directed mutagenesis and provides an effective remedy to address such challenges. The findings support that G-quadruplex formation is one mechanism whereby such sequences impede regular PCR-based mutagenesis methods. Full article

Encapsulated formulations have emerged as a promising tool for increasing nutrient absorption in the food supplement and cosmetic industries. Although the theoretical amplification factors for improving the bioavailability of encapsulated formulations are very high for poorly soluble active compounds, it has long been known that encapsulation can also enhance the absorption of water-soluble ingredients. These findings have led to the development of new technologies for encapsulating nutrients for use in the food industry. However, accurate quantification of nutrients in encapsulated formulations in the food supplement industry remains a challenge. This study presents the development and validation of novel analytical procedures for determining artemisinin in various food supplement formulations. Three formulations were prepared using different emulsifying procedures for artemisinin encapsulation. High-performance liquid chromatography with UV/Vis detection (HPLC-UV/Vis) was used for analysis. Separation was performed using a Waters ACQUITY Premier BEH C18 column. Specialized sample preparation procedures were designed to efficiently disrupt encapsulation and extract artemisinin for precise quantification. Three different sample preparation procedures were required to accurately determine the artemisinin content in the tested formulations. All methods were validated. The precision, linearity expressed as R², LOD, and LOQ of the chromatographic method were 0.39%, 0.9995, 18 µg/mL, and 26 µg/mL, respectively. Recoveries of the sample preparation methods were above 94%. The developed procedures enable accurate determination of artemisinin in encapsulated formulations, ensuring product quality and safety. These findings suggest that, for quality control of encapsulated food products, specialized analytical procedures for individual formulations may need to be developed and validated. Full article

Methenamine, a urinary antiseptic with antimicrobial properties, decomposes into toxic formaldehyde under acidic conditions. Its use is prohibited in dairy cattle in Korea to prevent harmful residues in milk. This study was designed to develop and validate a sensitive and reliable LC–MS/MS method for determining methenamine in raw milk and bovine muscle in compliance with the Positive List System (PLS) regulations. Samples were extracted with acetonitrile (ACN)–methanol (MeOH) (7:3, v/v) containing ammonia water, followed by defatting with n-hexane and purification with primary secondary amine (PSA). Chromatographic separation was performed on a hydrophilic interaction liquid chromatography (HILIC) column, and quantification was conducted using matrix-matched calibration to minimize matrix effects. The method showed excellent linearity (R² > 0.999), low limits of quantification (LOQ) (0.49 μg/kg for raw milk; 0.64 μg/kg for bovine muscle), and acceptable recoveries (78.1–102.8%) with precision (CV ≤ 8.75%), meeting Codex CAC/GL 71-2009 criteria. Stability studies demonstrated that methenamine remained stable in stock solutions, working standards and processed extracts under the storage and handling conditions used. Application to incurred samples resulted in the detection of methenamine in 2 of 32 raw milk samples (0.65 and 1.14 μg/kg) but in none of the 25 bovine muscle samples, with all detected levels below the Korean PLS limit. These findings confirm that the developed method is accurate, sensitive, and applicable for routine surveillance of methenamine residues to ensure consumer safety. Full article

Coal gangue, a major industrial solid waste from coal mining and processing, requires efficient alumina and silica separation for high-value utilization. This study focused on mineral reaction mechanisms and characteristics of coal gangue during calcination and alkaline leaching. Results showed calcination at 900–1200 °C altered its phase composition, affecting silica separation efficiency, with the optimal calcination range being 960–1120 °C. Poorly crystallized mullite and Al₂O₃ in calcined gangue were insoluble under low-alkaline and low-temperature conditions. On the contrary, amorphous silica is soluble and forms a sodium silicate solution in the proper alkaline conditions. This characteristic facilitates the efficient separation of alumina and silica. It was determined that the suitable conditions for silica removal from coal gangue are as follows: 1080 °C calcination for 90 min, leaching at 75 °C with 200 g/L NaOH (solid–liquid ratio of 1:4) for 4 h. Under these selected conditions, the silica leaching efficiency was 77.31%, the alumina leaching efficiency was 12.21%, the Na₂O content in the leached residue was 1.94%, and the mass ratio of alumina to silica (A/S) in the leached residue increased from 0.88 to 3.42. A potential desilication mechanism was also analyzed. Full article

Background/Objectives: Pregnancy represents a unique physiological state marked by extensive metabolic adaptations, particularly in lipid pathways essential for maternal adjustments, fetal development, and postpartum recovery. This study aimed to explore these changes through untargeted lipidomic profiling. Methods: This observational, comparative, non-interventional clinical study included 107 women, of which 65 were in the third trimester of pregnancy (mean age 27.9 ± 5 years) and 42 were in the early postpartum period (≤7 days, mean age 28.9 ± 5.9 years). Inclusion criteria were singleton, term pregnancies (37–41 weeks) with neonates weighing > 2500 g and no associated pregnancy-related pathologies; exclusion criteria included multiple gestation, use of lipid-altering medications, maternal age > 40 years, or diagnosed pregnancy complications. Plasma samples were analyzed using High-Performance Liquid Chromatography–Quadrupole Time-Of-Flight–Electrospray Ionization (positive mode)–Mass Spectrometry, data were processed with MetaboAnalyst 6.0 using multivariate and univariate analyses (Partial Least Squares–Discriminant Analysis, Volcano Plot, Random Forest, Receiver Operating Characteristic analysis), with statistical significance set at p < 0.05. Results: Multivariate analysis demonstrated a clear separation between groups with high predictive accuracy as reflected by strong classification metrics (Accuracy = 0.90, R² = 0.75, Q² = 0.68). Several discriminative lipids were consistently identified across statistical models, including 2-Methoxyestrone (AUC = 0.861), Eicosanedioic acid (AUC = 0.854), and Pregnenolone sulfate (AUC = 0.843). These biomarkers were further categorized into five major lipid classes: steroid hormones, long-chain fatty acids, lysophospholipids, ceramides/sphingolipids, and glycerolipids. Conclusions: Untargeted lipidomic profiling revealed distinct metabolic signatures that differentiate late pregnancy from early post-partum states. The identification of robust lipid biomarkers with high discriminative performance highlights their potential utility in maternal health monitoring, obstetric risk assessment, and postpartum recovery surveillance. Full article

Background: Platelet-rich fibrin (PRF) is an autologous biomaterial utilized as an adjunct in dental implant surgeries owing to its significant biocompatibility, supra-physiological concentration of growth factors, and ability to speed either soft or hard tissue regeneration. Methods: Today, PRF is available in both solid and liquid forms with an average resorption period of roughly 2 weeks. While various research endeavors have attempted to utilize Solid-PRF as a barrier membrane in guided bone regeneration (GBR) and various other applications, its two-week resorption period has limited its use as a solo “barrier” membrane owing to its faster-than-ideal resorption properties. Results: Recent studies have demonstrated that by heating and denaturing Liquid-PRF/albumin, the resorption properties of the heated albumin gel could be extended from 2 weeks to 4–6 months by utilizing the Bio-Heat technology. This emerging technology was given the working name ‘extended-PRF’ or e-PRF, with many clinical indications being proposed for further study. Numerous clinicians have now utilized extended-PRF (e-PRF) membranes as a substitute for collagen barrier membranes in various clinical applications, such as guided tissue/bone regeneration, recession coverage, and lateral window sinus lifts. Conclusions: This two-part case series paper aims to first illustrate the evolution of techniques developed taking advantage of this new technology in clinical practice for alveolar ridge preservation. This includes four different methods of fabrication of e-PRF along with its application in clinical practice. This article discusses the clinical outcomes, including the advantages/disadvantages of utilizing each of the four separate techniques to prepare and utilize e-PRF membranes for ridge preservation. Full article

Although intensified flotation cells have been introduced as fast-kinetic and plug-flow-type flotation machines, there is limited empirical verification and information about their fluid flow patterns and dispersion regimes. The present communication paper investigates this for an Imhoflot^TM G-06 cell operated in an open-circuit mode using an impulse method to measure and model the residence time of a liquid–gas system. For experimental measurements, a concentrated KCl solution was employed, and water conductivity was monitored for 20 min. By fitting several relevant models, such as large and small tanks in series (LSTS), Weller, N-Mixer, and perfect mixer, to the experimental data, it was revealed that the N-Mixer represented the dispersion pattern the best (N = 1.3–1.6). Further, the obtained practical mean retention time (MRT) of 4.11 ± 0.16 min was somewhat aligned with the theoretical value, i.e., 5.0 min per pass, indicating a back-calculated gas hold-up magnitude of 18%–22% in the separator. These results provide an in-depth perception of scale-up procedures and requirements for cell modification. Full article

Extracellular vesicles (EVs) secreted by Fusarium oxysporum play an important role in the process of its infestation of the host, but the in vitro research system for EVs of F. oxysporum (Fo-EVs) has not yet been improved, and the mechanism of its action remains unclear. In this study, particle size distribution, particle concentration, number of particles per unit of protein, number of particles per unit of mycelial biomass, and concentration of contaminated proteins were used as indicators to evaluate the yield and purity of Fo-EVs. The optimal method for Fo-EV preparation and extraction was screened by comparing liquid culture, solid culture, and solid culture with enzymatic cell wall hydrolysis. The optimal system for Fo-EVs separation and purification was screened by a pairwise combination of three primary methods (Ultracentrifugation (UC), Ultrafiltration (UF), and Polyethylene glycol precipitation method (PEG)) and two secondary methods (Size-exclusion chromatography (SEC) and Aqueous two-phase system (ATPS)), respectively. The protein composition was identified via mass spectrometry technology, followed by GO annotation and GO enrichment analysis using whole-genome proteins as the background. Based on these steps, a Fo-EV protein library was constructed to reveal Fo-EV’s most active biological functions. The results showed that solid culture combined with the UC-SEC method could effectively enrich Fo-EVs with a typical cup-shaped membrane structure. The obtained Fo-EVs had an average particle size of 253.50 nm, a main peak value of 200.60 nm, a particle concentration of 2.04 × 10¹⁰ particles/mL, and a particle number per unit protein of 1.09 × 10⁸ particles/μg, which were significantly superior to those of other combined methods. Through proteomic analysis, 1931 proteins enriched in Fo-EVs were identified, among which 350 contained signal peptides and 375 had transmembrane domains. GO enrichment analysis revealed that these proteins were mainly involved in cell wall synthesis, vesicle transport, and pathogenicity-related metabolic pathways. Additionally, 9 potential fungal EV markers, including Hsp70, Rho GTPase family, and SNARE proteins, were screened. This study constructed an isolation system and a marker database for Fo-EVs, providing a methodological and theoretical basis for in-depth analysis of the biological functions of Fo-EVs. Full article