Preparation Method Research Articles

A Fast, High-Sensitivity 96-Well Plate-Based MICROFASP Method for Processing Low Microgram Proteomics Sample within 1.5 h.

A rapid, sensitive, and high-throughput sample preparation method is of paramount significance for proteomics analysis. Here, we report a fast, high-sensitivity MICROFASP method that is capable of completing sample preparation within 1.5 h, enhancing the throughput by over 13 times compared to the previous reports. Protein digestion time was significantly cut from 17 h to 20 min in a limited volume. Simultaneous reduction and alkylation occurred within 30 min. The label-free quantitation intensities of proteins from the fast and conventional MICROFASP methods were highly correlated (r = 0.91), validating the reliability of the fast-MICROFASP method. When starting with 1 μg of K562 cell lysate, the fast-MICROFASP method identified over 6 times more protein groups and 19 times more peptides than did the iST method. A 96-well plate-based version was developed to process 8 brain tissue samples from APP/PS1 transgenic mice in parallel. Averagely, with only 1 μg of protein lysate, 2826 protein groups (n = 8, RSD = 0.7%) and 12,972 peptides (n = 8, RSD = 1.5%) were identified from each sample. Amyloid-beta protein was successfully identified as a highly expressed protein, which shows its potential for detecting diagnostic markers and proteome profiling with low-microgram samples. We anticipate the high-sensitivity 96-well plate-based fast-MICROFASP method will have wide application in high-throughput and rapid preparation of large cohorts of low-microgram samples (e.g., clinical biopsy) for comprehensive proteome profiling. Data are available via ProteomeXchange with the identifier PXD053720.

Nanoscale Probing of the Organic Binder in Artists' Paint Layers: Organic Phases and Chemical Heterogeneity.

Understanding paint structures at the nanoscopic level can address key questions related to artistic techniques, paint formulation, and long-term preservation of artworks. This involves examining spatial chemical complexity, the formation of molecular networks, and interactions between organic and inorganic constituents. Depending on the paint preparation methods, proteins and drying oils, the most common binders in traditional artistic practices, can be integrated to produce paints with diverse structures and nanoscale chemical intricacies. In this study, we utilize atomic force microscopy-based infrared spectroscopy (AFM-IR) to investigate the spatial chemical complexity and reaction pathways of organic species in artists' paints, including oil, tempera, and mixed-media tempera grassa. By analyzing these paints at the nanoscale, we established connections between their structural organization, chemistry, and formulation.

Mechanistic Insights into Amorphous Solid Dispersions: Bridging Theory and Practice in Drug Delivery.

Improving the bioavailability of poorly water-soluble drugs presents a significant challenge in pharmaceutical development. Amorphous solid dispersions (ASDs) have garnered substantial attention for their capability to augment the solubility and dissolution rate of poorly water-soluble drugs, thereby markedly enhancing their bioavailability. ASDs, characterized by a metastable equilibrium where the active pharmaceutical ingredient (API) is molecularly dispersed, offer enhanced absorption compared to crystalline forms. This review explores recent research advancements in ASD, emphasizing dissolution mechanisms, phase separation phenomena, and the importance of drug loading and congruency limits on ASD performance. Principal occurrences such as liquid-liquid phase separation (LLPS) and supersaturation are discussed, highlighting their impact on drug solubility, absorption and subsequent bioavailability. Additionally, it addresses the role of polymers in controlling supersaturation, stabilizing drug-rich nanodroplets, and inhibiting recrystallization. Recent advancements and emerging technologies offer new avenues for ASD characterization and production and demonstrate the potential of ASDs to enhance bioavailability and reduce variability, making possible for more effective and patient-friendly pharmaceutical formulations. Future research directions are proposed, focusing on advanced computational models for predicting ASD stability, use of novel polymeric carriers, and methods for successful preparations.

Fabrication and Modification of Hydrotalcite-BasedPhotocatalystsand TheirCompositesfor CO 2 Reduction: A Critical Review.

Layered double hydroxides(LDHs), whichresemble hydrotalcite, are a type ofmaterials withcationic layers and exchangeable interlayer anions. Theyhave drawn lots of curiosity as a high-temperature CO2adsorbentbecause of its quick desorption/sorption kinetics andrenewability.Due to its extensive divalent or trivalent cationic metals,high anion exchange property, memory effect, adjustable behavior, bio-friendliness, easy to prepareand relatively low cost, the LDHs-based materials are becoming increasingly popular for photocatalytic CO2reduction reaction (CO2RR). Fabricationand modificationaregood waysto move forward the advancement of LDHs-based catalysts, which will help chemistry and materials science makegreat progress.In this review we discussed structural characteristics and the methods for preparation and modification of LDHs-based photocatalysts.We also highlighted and discussed the major developments andapplications in photocatalytic CO2RR as well asthe photocatalytic mechanism. The goal of the present review is to give a broad summary of the various LDHs-basedphotocatalystsand the correspondingdesign strategies, which could motivate more excellent research works to explore this kind of CO2RR photocatalysts to further increase CO2conversion yield and selectivity.

Assessment of Long-Term Cultivated In Vitro Willow Clones according to Chromosomal Analysis.

Creation and long-term in vitro maintenance of valuable genotype collection is one of the modern approach to conservation of valuable gene pool of woody plants. However, during prolonged cultivation, genetic variability of cells and tissues may accumulate and lead to the loss of valuable characteristics of parental plants. It is therefore important to assess the genetic (including cytogenetic) stability of collection clones. The purpose of this work was to study the karyotype features (number, ploidy level and chromosome size) of various willow clones (S. dasyclados Wimm., S. viminalis L., S. purpurea L., S. caspica Pall., х S. palustrisHost.) under conditions of long-term in vitro storage. No such research has been conducted on willow so far. Nevertheless, there is evidence of significant influence of ploidy level on growth, productivity, and composition of wood for willow plants. Our study was based on the plants of five micropropagated willow clones rated as promising for plantation forestry. The plants used in the study were maintained in vitro for a long period (14 years) by rare subculturing (once in 5 months) on a hormone-free1/2 WPM nutrient medium under standard cultivation conditions (25± 2°C, 16 h light/8 h dark, 2.0 klx). Throughout the entire in vitro cultivation period, the plants showed proper growth and development, high regeneration activity, and no visible signs of somaclonal variation. Willow is one of the rather difficult objects for karyotype analysis. The authors improved the method of preparation and analysis of specimens. During the long-term cultivation, the clones showed cytogenetic stability, maintaining the ploidy (2n = 2х = 38 or 2n= 4х = 76) and the mixoploid nature of the original plants. Data obtained gives an update on the sizes of chromosomes of clones with different ploidy. The absolute chromosome length for diploid clones varies from 0.8 to 2.1 μm; tetraploid, from 0.9 to 2.5 μm. There were no statistically significant differences in the average chromosome length between diploid and tetraploid clones. All studied willow clones during long-term (for 14years) in vitro cultivation on a hormone-free nutrient media retain the karyotype of the respective species.

The role of aging and various surface preparation methods in the repair of nanohybrid composites

BackgroundRepairing composite resins is a less invasive alternative to complete restoration replacement. To achieve a successful bond between the existing and newly applied composite materials, various surface preparation methods, such as sandblasting and acid etching, have been explored. The aim of the study was to evaluate the effect of different surface treatments on the repair bond strength of a universal nanohybrid composite resin restorative material before and after thermal aging, by utilizing a micro-shear bond strength (µSBS) test.MethodsFor the micro-shear bond strength test, a total of 120 cylindrical (3mmX2mm) nanohybrid resin based composite specimens were prepared. The prepared specimens were divided into three groups (n = 40/per group) based on surface treatment methods: a non-aged group, 10,000 thermal cycle aging and 50,000 thermal cycle aging. The aged and non-aged specimens were further divided into four groups according to adhesive application modes and surface pretreatment methods: 1.universal adhesive/self-etch mode, 2.aluminum oxide sandblasting + universal adhesive/self-etch mode, 3.universal adhesive/etch-and-rinse mode, 4.aluminum oxide sandblasting + universal adhesive/etch-and-rinse mode. Subsequently, 0.8mmX2mm disc shape light cure resin based composite specimens were applied with a direct placement technique on the treated surfaces of all samples for repair. µSBS test was performed using a universal testing machine at a crosshead speed of 1 mm/min. Kruskal-Wallis and Mann-Whitney tests were performed for statistical analysis.ResultsThe µSBS values of the non-aged group were higher than those of the 10,000 and 50,000 thermal cycle groups, with no significant differences within the non-aged subgroup (p > 0.05). In the aged groups, significant differences were observed between adhesive application modes and surface treatments. Specifically, the etch-and-rinse mode showed higher bond strengths than the self-etch mode after 50,000 thermal cycles (p < 0.05). Sandblasting combined with universal adhesive (self-etch mode) improved bond strength, especially in the 10,000 thermal cycle group (p < 0.05).ConclusionAging reduced the bond strength of composite resin repairs, with the etch-and-rinse mode outperforming the self-etch mode in aged specimens. Sandblasting alone did not enhance bond strength. These findings highlight the importance of considering aging and adhesive strategies to optimize repair outcomes, with further research needed on long-term durability.

Gel-Based Sample Fractionation with SP3-Purification for Top-Down Proteomics.

Precise prefractionation of proteome samples is a potent method for realizing in-depth analysis in top-down proteomics. PEPPI-MS (Passively Eluting Proteins from Polyacrylamide gels as Intact species for MS), a gel-based sample fractionation method, enables high-resolution proteome fractionation based on molecular weight by highly efficient extraction of proteins from polyacrylamide gels after SDS-PAGE separation. Thereafter it is essential to effectively remove contaminants such as CBB and SDS from the PEPPI fraction prior to mass spectrometry. In this study, we developed a complete, robust, and simple sample preparation workflow named PEPPI-SP3 for top-down proteomics by combining PEPPI-MS with the magnetic bead-based protein purification approach used in SP3 (single-pot, solid-phase-enhanced sample preparation), now one of the standard sample preparation methods in bottom-up proteomics. In PEPPI-SP3, proteins extracted from the gel are collected on the surface of SP3 beads, washed with organic solvents, and recovered intact with 100 mM ammonium bicarbonate containing 0.05% (w/v) SDS. The recovered proteins are subjected to mass spectrometry after additional purification using an anion-exchange StageTip. Performance validation using human cell lysates showed a significant improvement in low-molecular-weight protein recovery with a lower coefficient of variation compared to conventional PEPPI workflows using organic solvent precipitation or ultrafiltration.

Facile and atom-economical synthesis of highly efficient chitosan-based flame retardants towards fire-retarding and antibacterial multifunctional coatings on cotton fabrics.

The development of bio-based flame retardants has garnered significant attention, however, significant challenges remain in achieving efficient flame retardancy and eco-friendly preparation methods. Herein, we propose a facile, atomic-efficient, and eco-friendly strategy for synthesizing a trinity chitosan-based flame retardant, phosphite-protonated chitosan (PCS). The chemical structure was systematically analyzed and the impact of varying degrees of protonation on the dissolution behavior and rheological properties were investigated. Benefiting from the promotion of dehydration and carbonization facilitated by phosphite groups, PCS exhibits high intrinsic flame retardancy with an LOI value of 80.7 %. Moreover, its favorable rheological and film-forming properties make it well-suited for easy application as a multifunctional coating in fabric finishing through blade coating processes. The finished cotton and polyester/cotton blended fabrics exhibit excellent flame retardancy, as evidenced by increased LOI values, successful passage of vertical burning tests, reductions of up to 65.0 % and 50.3 % in pHRR and THR values, respectively. Additionally, PCS imparts superior antibacterial properties to the fabrics, achieving a 99.99 % antibacterial rate against both E. coli and S. aureus. This study introduces a straightforward and atom-economical approach for preparing highly efficient chitosan-based flame retardants, along with the development of a transparent, green, and efficacious multifunctional coating system on textiles.

Unveiling the impact of preparation methods, matrix/carrier type selection and drug loading on the supersaturation performance of amorphous solid dispersions.

Amorphous solid dispersions (ASDs) are widely recognized for their potential to enhance the solubility of poorly water-soluble drugs, with factors such as molecular mobility, intermolecular interactions, and storage conditions playing critical roles in their performance. However, the influence of preparation methods on their performance remains underexplored, especially regarding their supersaturation performance. To address this gap, the present study systematically investigates ASDs of ibuprofen (IBU, used as a model drug) prepared using two widely utilized techniques (solvent evaporation, SE, and melt-quench cooling, M-QC). Three different matrices/carriers (Soluplus®, SOL, povidone, PVP, and copovidone, PVPVA) were employed to evaluate the combined influence of preparation method, matrix/carrier type, and drug loading on ASD performance. Supersaturation behavior during dissolution, particularly its dependence on the Sink Index (SI), was a key focus. All ASDs showed successful amorphization, but molecular near-order structures differed based on the preparation method. ATR-FTIR spectroscopy revealed stronger molecular interactions in M-QC ASDs (compared to SE). Dissolution studies under supersaturation conditions (SI = 0.1 and SI = 0.2) highlighted significant performance differences. M-QC ASDs consistently exhibited higher in vitro AUC(0→t) values under non-sink conditions compared to crystalline IBU. Conversely, SE ASDs showed improved supersaturation primarily under low SI conditions, especially with SOL at low drug loadings. The findings underscore the need for a systematic approach in developing ASDs, considering preparation method, matrix type, drug loading and dissolution study conditions collectively. These factors significantly influence dissolution behavior and supersaturation, emphasizing that they should not be independently studied but evaluated comprehensively to optimize ASD performance.

Structural Regulation and Performance Enhancement of Carbon-Based Supercapacitors: Insights into Electrode Material Engineering.

The development of carbon-based supercapacitors is pivotal for advancing high energy and power density applications. This review provides a comprehensive analysis of structural regulation and performance enhancement strategies in carbon-based supercapacitors, focusing on electrode material engineering. Key areas explored include pore structure optimization, heteroatom doping, intrinsic defect engineering, and surface/interface modifications. These strategies significantly enhance electrochemical performance through increasing surface area, improving conductivity, facilitating charge transfer, introducing additional pseudocapacitive reactions, and optimizing the density of states at the Fermi level, among other mechanisms. After introducing these fundamental concepts, the review details various preparation methods and their effects on supercapacitor performance, highlighting the interplay between material structure and electrochemical properties. Challenges in scaling advanced fabrication techniques and ensuring the long-term stability of functionalized materials are discussed. Additionally, future research directions are proposed, emphasizing the development of cost-effective, scalable methods and interdisciplinary approaches to design next-generation supercapacitors, thereby meeting the growing demand for efficient and sustainable energy storage solutions.

Cellulose-based delivery systems for bioactive ingredients: A review.

Considering the outstanding advantages including abundant resources, structure-performance designability, impressive mechanical strength, and 3D network structure-forming ability, cellulose is an ideal material for encapsulating bioactive ingredients. Due to its low solubility in water, large-scaled morphology and poor flexibility, cellulose is unsuitable for the construction of carriers. Consequently, the majority of cellulose is employed following physical or chemical modification. Cellulose and its derivatives are extensively employed in the food industry, including fat replacement, food packaging composites, food additives, 3D-printed food and delivery systems. Their benefits in food delivery systems are particularly pronounced. Therefore, the distinguishing features, preparation methods, recent developments and effectiveness of different cellulose-based delivery systems for bioactive ingredients are discussed. Cellulose-based delivery systems offer unique advantages in terms of environmental impact reduction, modification facilitation, stimuli-responsive release as well as tailored design, and their application has gained widespread recognition. However, they are facing challenges in the application process comprising modification methods for cellulose-based materials, new methods for commercial preparation on a wide scale, cellulose-based multifunctional conveyance systems and systematic evaluation using in vivo experiments. In conclusion, this review provides theoretical references for the development of novel delivery carriers as well as the efficient application and popularization of cellulose-based delivery systems.

The Efficacy and Safety of Prophylactic Treatment Methods for Cutibacterium Acnes Infection in the Shoulder: A Systematic Review of Level of Evidence I Randomized Controlled Trials.

Cutibacterium acnes (C. acnes), formerly Propionibacterium acnes (P. acnes), is the most common pathogen responsible for postoperative shoulder infections. Prevention of shoulder infection is crucial to improving patient outcomes, prolonging implant longevity, and reducing costs. The purpose of this study is to evaluate the effectiveness of various prophylactic treatments for C. acnes in the shoulder. A systematic review of 15 randomized controlled trials was conducted to evaluate the effectiveness of various skin preparation methods in the reduction of C. acnes for patients undergoing shoulder surgery. Three electronic databases were queried as follows: PubMed, Embase, and the Cochrane Library databases. Data extraction for this study was performed by three independent reviewers, and only level I studies were included. Outcomes were assessed based on the effectiveness of the method used; side effects were also analyzed. Fifteen studies were included in this review. Seven studies utilized a regimen of at-home treatment in the days leading up to culture sampling, while the other eight studies administered treatment just prior to sampling. Seven studies found their respective treatment methods to be statistically significant. No preparation methods applied immediately prior to sampling showed conclusive superiority. The application benzoyl peroxide in the days prior to sampling showed significantly lower rates of colonization when compared to control groups in five separate studies. There remains no consensus on the optimal skin preparation method immediately prior to surgery, however, acne cream applied in the days leading up to surgery has been shown to significantly reduce the bacterial load of C. acnes. While BPO appears to be the superior method of prophylactic treatment, it is also associated with higher rates of complications and skin reactions following treatment; thus, caution is warranted. Further studies should investigate whether this reduction in bacterial load translates to meaningfully lower rates of periprosthetic joint infections.

The Influence of Rice Husk Ash Incorporation on the Properties of Cement-Based Materials.

Rice husk ash is a kind of biomass material. Its main component is silicon dioxide, with a content of up to 80%. It has high pozzolanic activity and can react with hydroxide in cement. When treating rice husks, rice husk ash with high volcanic ash activity and a good microaggregate filling effect can be obtained by selecting a suitable incineration environment. These advantages make rice husk ash an ideal concrete admixture, replacing the traditional admixture such as fly ash and slag in concrete. This paper summarizes the preparation methods and physical and chemical properties of rice husk ash, as well as the physical and chemical properties of rice husk ash concrete, such as mechanical properties, temperature resistance, freezing resistance, permeability resistance and chemical erosion resistance. The results show that using 20% rice husk ash as a substitute material for cement improves the resistance strength, compressive strength, flexural strength, and permeability of concrete. In short, the incorporation of rice husk ash can effectively improve the performance of cement-based materials, which will be conducive to the green development of the building material industry and the implementation of the two-carbon strategy.

Cryo-electron tomography pipeline for plasma membranes

Cryo-electron tomography (cryoET) provides sub-nanometer protein structure within the dense cellular environment. Existing sample preparation methods are insufficient at accessing the plasma membrane and its associated proteins. Here, we present a correlative cryo-electron tomography pipeline optimally suited to image large ultra-thin areas of isolated basal and apical plasma membranes. The pipeline allows for angstrom-scale structure determination with subtomogram averaging and employs a genetically encodable rapid chemically-induced electron microscopy visible tag for marking specific proteins within the complex cellular environment. The pipeline provides efficient, distributable, low-cost sample preparation and enables targeted structural studies of identified proteins at the plasma membrane of mammalian cells.

Energized dispersive guided extraction (EDGE): a promising tool for analytical chemistry applications.

Analytical chemistry demands precise sample preparation methods to ensure accurate qualitative and quantitative determinations, especially those capable of clean-up and preconcentration of target analytes. Extraction plays a crucial role in enhancing the selectivity and sensitivity of analytical procedures. Thus, Energized Dispersive Guided Extraction (EDGE) has emerged as an innovative alternative to traditional methods, such as Soxhlet, maceration, and percolation, as well as modern techniques like Accelerated Solvent Extraction (ASE), Supercritical Fluid Extraction (SFE), and Microwave or Ultrasound Assisted Extraction (MAE and UAE). By combining dispersive solid-phase extraction (DSPE) with pressurized liquid extraction (PLE), this technique offers distinct advantages, including faster extraction times, reduced solvent consumption, and improved reproducibility. Its automated system also minimizes operator intervention, ensuring greater precision and optimizing workflows. However, there is a significant gap in studies exploring its various potential applications and perspectives. In this scenario, this is the first paper to broaden the range of applications of the EDGE system by exploring its use in food, pharmaceutical, and environmental areas. These sectors benefit from EDGE's ability to preserve analyte integrity, increase extraction yields, and promote environmentally sustainable practices. Moreover, EDGE's flexibility and efficiency make it a powerful tool for addressing emerging challenges, such as detecting contaminants in complex matrices and complying with stricter regulatory standards. Expanding EDGE's applications in these areas underscores its potential to transform analytical processes, driving advancements in food safety, pharmaceutical quality control, and environmental monitoring.

CRISPR knock-in of a chimeric antigen receptor into GAPDH 3'UTR locus generates potent B7H3-specific NK-92MI cells.

CAR-NK therapy is becoming a promising approach to treat solid tumors. However, the random insertion of the CAR gene and inflexible CAR expression caused by common preparation methods significantly impact its efficacy and safety. Here we successfully established a novel type of CAR-NK cells by integrating CAR sequences into the GAPDH 3'UTR locus of NK-92MI cells (CRISPR-CAR-NK), achieving site-specific integration of the CAR gene and allowing endogenous regulatory components to govern CAR expression. CRISPR-CAR-NK cells had comparable growth capacity but displayed superior anti-tumor activity compared with their lentiviral counterparts. They activated and degranulated more effectively when co-cultured with tumor cells, due to increased expression of activating receptors and decreased expression of inhibitory molecules. They also enhanced the production of Granzyme B and IFN-γ, and more effectively triggered the IFN-γ pathway. Moreover, CRISPR-CAR-NK cells demonstrated distinct properties from conventional CAR-NK concerning metabolic features and signal dependence. Notably, CRISPR-CAR-NK cells exhibited lower metabolic levels without compromising antitumor activity, and their function was less reliant on the PI3K-AKT pathway, implying that the CRISPR-CAR-NK cells have significant potential for enhanced synergy with AKT inhibitors and adaptation to nutrient stress within the tumor microenvironment. These findings provide a novel potential strategy for cancer immunotherapy and an experimental foundation and paradigm for optimizing CAR-NK cells utilizing CRISPR technology, highlighting the potential of CRISPR to advance immunotherapies.

Nanosuspension Innovations: Expanding Horizons in Drug Delivery Techniques.

Nanosuspensions (NS), with their submicron particle sizes and unique physicochemical properties, provide a versatile solution for enhancing the administration of medications that are not highly soluble in water or lipids. This review highlights recent advancements, future prospects, and challenges in NS-based drug delivery, particularly for oral, ocular, transdermal, pulmonary, and parenteral routes. The conversion of oral NS into powders, pellets, granules, tablets, and capsules, and their incorporation into film dosage forms to address stability concerns is thoroughly reviewed. This article summarizes key stabilizers, polymers, surfactants, and excipients used in NS formulations, along with ongoing clinical trials and recent patents. Furthermore, a comprehensive analysis of various methods for NS preparation is provided. This article also explores various in vitro and in vivo characterization techniques, as well as scale-down technologies and bottom-up methods for NS preparation. Selected examples of commercial NS drug products are discussed. Rapid advances in the field of NS could resolve issues related to permeability-limited absorption and hepatic first-pass metabolism, offering promise for medications based on proteins and peptides. The evolution of novel stabilizers is essential to overcome the current limitations in NS formulations, enhancing their stability, bioavailability, targeting ability, and safety profile, which ultimately accelerates their clinical application and commercialization.

The sensitivity of reconstructed carbon dioxide concentrations to stomatal preparation methods using a leaf gas exchange model

The sensitivity of reconstructed carbon dioxide concentrations to stomatal preparation methods using a leaf gas exchange model

The effect of the natural antioxidants extracted from Glycyrrhiza glabra (licorice) on the cryopreservation of chicken breast meat

Background: Chicken meat is highly nutritious, valued for its unique properties and versatility in culinary applications. Also, preparation methods and natural additives to its diet play an important role in estimating the nutritional value of its meat by increasing consumer acceptance of it from health, nutritional, and material aspects. Objective: This study was conducted to determine the effect of licorice as a natural antioxidant on cold-preserved chicken meat. Methods: Phenolic compounds were quantified using the colorimetric method with Folin's reagent, measuring absorbance at 765 nm, and results were expressed in terms of gallic acid equivalents per gram of dry weight. Gallic acid was used as a reference solution to prepare the standard curve with concentrations ranging between 20-120 μg/mL to determine the quantity of phenolic compounds. Determination of antioxidant activity was done using the DPPH method by taking 0.5 mL of the extract solution at concentrations ranging from 50-300 μg/mL and mixing with 3 mL of methanolic DPPH solution. The mixture was mixed for 1 min with a vortex mixer, and samples were measured by spectrophotometer at 517 nm. Moisture, ash, fat, protein, and pH were determined according to the methods mentioned by AOAC. Thiobarbituric acid-reactive substances (TBARS) were estimated by mixing 0.5 g of the sample with 2.5 mL of TBA solution. The mixture was heated in a boiling water bath for 10 min to orange color, chilled by adding 1 mL of chloroform and centrifuged at 5000 rpm for 25 min, and the absorbance was read at 532 nm wavelength. Total microorganisms, molds, and yeast were counted. Results: Protein content increased proportionally with higher concentrations of ethanolic licorice root extract. The gradual increase in protein content in samples is attributed to their antimicrobial effect, which leads to the preservation of meat from microbial growth and the action of proteolytic enzymes. The increase in protein during refrigerated storage is due to the decrease in moisture and increase in dry matter reflected in the increase in protein. The reduced acidity in chicken meat treated with licorice root ethanolic extract is attributed to its antimicrobial effects, which limit the growth of microorganisms responsible for metabolizing nitrogenous compounds. Licorice extracts, rich in phenolic compounds, reduce lipid oxidation. The observed decrease in thiobarbituric acid-reactive substances with increasing extract concentration is due to the antioxidant properties of these phenolics with the hydroxyl group attached to the aromatic ring capable of giving hydrogen atoms and inhibiting free radicals. This study found that the growth of microorganisms could be lowered by licorice extract containing phenolic compounds, flavonoids, tannins, and alkaloids, which showed a significant antimicrobial effect. Conclusion: This study examined how natural antioxidants, like licorice Glycyrrhiza glabra, affected the cryopreservation of chicken breast. Keywords: Licorice root extract, chicken breast, antioxidants, total phenol, Meat science

Preparation of Colored Polymer Microspheres.

Colored polymer microspheres have attracted significant attention in both academia and industry due to their unique optical properties and extensive application potential. However, achieving a uniform distribution of dyes within these microspheres remains a challenge, particularly when heavy concentrations of dye are used, as this can lead to aggregation or delamination, adversely affecting their application. Additionally, many dyes are prone to degradation or fading when exposed to light, heat, or chemicals, which compromises the long-term color stability of the microspheres. Consequently, the preparation of colored polymer microspheres with high stability continues to be a significant challenge. This review offers a comprehensive overview of the preparation techniques for colored polymer microspheres and their dyeing mechanisms, introducing the fundamental concepts of these microspheres and their applications in various fields, such as biomedicine, optical devices, and electronic display technologies. It further presents a detailed discussion of the different preparation methods, including physical adsorption, chemical bonding, and copolymerization. The advantages, limitations, and potential improvements of each method are explored, along with an analysis of the interactions between dyes and the polymer matrix, and how these interactions influence the properties of the microspheres, including their color uniformity, stability, and durability. Finally, the review discusses future perspectives on the development of colored polymer microspheres, highlighting the advancement of novel materials, innovations in preparation technology, and the exploration of potential new application areas.