Protective effects and molecular mechanisms of Litopenaeus vannamei treated with l-arginine/l-lysine against myofibrillar proteins oxidation and quality degradation during freeze-thaw cycles.

The storage and processing of Litopenaeus vannamei are often challenged by the freeze-thaw (F-T) cycle phenomenon. This study delved into the influence of pretreatment with l-arginine (Arg) and l-lysine (Lys) on the myofibrillar proteins oxidation and quality of shrimp subjected to F-T cycles. Arg and Lys pretreatment notably improved water-holding capacity (WHC), textural integrity as well as the myofibrillar structure of the shrimps. A lesser reduction in the amounts of immobile and bound water was found in the amino acid-treated groups, and the oxidation of lipids and proteins were both decelerated. Molecular simulation results indicated that Arg and Lys could form hydrogen and salt-bridge bonds with myosin, enhancing the stability of Litopenaeus vannamei. The study concludes that Arg and Lys are effective in alleviating the adverse effects of F-T cycles on the quality of Litopenaeus vannamei, and provides a new solution for the quality maintenance during storage and processing.

Advances in Highly Ductile Concrete Research.

In recent years, high-ductility concrete (HDC) has gradually become popular in the construction industry because of its excellent ductility and crack resistance. Concrete itself is a kind of building material with poor tensile properties, and it is necessary to add a large number of steel bars to improve its tensile properties, which increases the construction cost of buildings. However, most of the research studies on high-ductility concrete are scattered. In this paper, the basic mechanical properties of high-ductility concrete and the effects of dry and wet cycles, freeze-thaw cycles, and salt erosion on the durability of high-ductility concrete are obtained by comprehensive analysis. The results show that the tensile properties of HDC can be significantly improved by adding appropriate fiber. When the volume fraction of steel fiber is 2.0%, the splitting tensile strength of concrete is increased by 98.3%. The crack width threshold of concrete chloride erosion is 55-80 µm, and when the crack width threshold is exceeded, the diffusion of CL-1 will be accelerated, and the HDC can control the crack within the threshold, thereby improving the durability of the concrete. Finally, the current research status of high-ductility concrete is analyzed, and the future development of high-ductility concrete is proposed.

Durability Performance of CGF Stone Waste Road Base Materials under Dry-Wet and Freeze-Thaw Cycles.

The disposal of stone waste derived from the stone industry is a worldwide problem. The shortage of landfills, as well as transport costs and environmental pollution, pose a crucial problem. Additionally, as a substitute for cement that has high carbon emissions, energy consumption, and pollution, the disposal of stone wastes by utilizing solid waste-based binders as road base materials can achieve the goal of "waste for waste". However, the mechanical properties and deterioration mechanism of solid waste-based binder solidified stone waste as a road base material under complex environments remains incompletely understood. This paper reveals the durability performance of CGF all-solid waste binder (consisting of calcium carbide residue, ground granulated blast furnace slag, and fly ash) solidified stone waste through the macro and micro properties under dry-wet and freeze-thaw cycling conditions. The results showed that the dry-wet and freeze-thaw cycles have similar patterns of impacts on the CGF and cement stone waste road base materials, i.e., the stress-strain curves and damage forms were similar in exhibiting the strain-softening type, and the unconfined compressive strengths all decreased with the number of cycles and then tended to stabilize. However, the influence of dry-wet and freeze-thaw cycles on the deterioration degree was significantly different; CGF showed excellent resistance to dry-wet cycles, whereas cement was superior in freeze-thaw resistance. The deterioration grade of CGF and cement ranged from 36.15 to 47.72% and 39.38 to 47.64%, respectively, after 12 dry-wet cycles, whereas it ranged from 57.91 to 64.48% and 36.61 to 40.00% after 12 freeze-thaw cycles, respectively. The combined use of MIP and SEM confirmed that the deterioration was due to the increase in the porosity and cracks induced by dry-wet and freeze-thaw cycles, which in turn enhanced the deterioration phenomenon. This can be ascribed to the fact that small pores occupy the largest proportion and contribute to the deterioration process, and the deterioration caused by dry-wet cycles is associated with the formation of large pores through the connection of small pores, while the freeze-thaw damage is due to the increase in medium pores that are more susceptible to water intrusion. The findings provide theoretical instruction and technical support for utilizing solid waste-based binders for solidified stone waste in road base engineering.

Screening and characterization of an antifreeze peptide from sea cucumber intestinal protein hydrolysates.

Protein deterioration caused by ice crystals is an important factors affecting the frozen storage of fish. In this study, antifreeze peptides extracted from hydrolysates of sea cucumber intestinal protein with inhibition of protein denaturation were screened and characterized. The peptide Leu-Pro-Glu-Phe-Thr-Glu-Glu-Glu-Lys (LPEFTEEEK), derived from neutral protease hydrolysates of sea cucumber intestinal protein, was investigated for its potential to enhance the quality of salmon fillets during three freeze-thaw cycles. The results showed that the application of LPEFTEEEK effectively maintained the texture of fish fillets, as well as the oxidative and conformation stability of myofibrillar protein during the freezing process. Additionally, molecular dynamics simulations verified that LPEFTEEEK could bind to ice crystals and inhibit their recrystallization, thus preventing organisms from being damaged by freezing. This suggests that LPEFTEEEK holds significant promise as a novel cryoprotective agent for marine-derived antifreeze peptides.

Mechanistic study on the effect of hydroxypropyl corn starch, guar gum and compound phosphates on the freeze-thaw quality of quick-frozen kuey teow.

Kuey teow is one of the delicacies of Guangdong, China and is a gluten-free noodle dish made from rice. It has a short storage period and extending the shelf life by quick freezing induces quality deterioration due to temperature fluctuations. To improve its freeze-thaw frozen storage quality, this paper examined the effects of hydroxypropyl corn starch (HCS), guar gum (GG), and compound phosphates (CP) on the quality of quick-frozen kuey teow during freeze-thaw cycles. The mechanism was investigated by identifying changes in the moisture status, aging degree of the starch, and textural and cooking characteristics. The results showed that all three additions improved the toughness, chewiness and steaming characteristics of the kuey teow, with CP significantly enhancing chewiness. XRD and FTIR results revealed that GG more significantly inhibited the decrease of starch crystallinity, while HCS inhibited starch aging. GG, HCS and CP all improved the hydration characteristics and water holding capacity of rice starch. GG enhances the ability of starch to bind more tightly with water, resulting in a more uniform water distribution and a more continuous and tight structure of the kuey teow. This study will provide a theoretical basis for compounding and optimizing the quick-freezing of kuey teow.

mRNA Fragmentation Pattern Detected by SHAPE.

The success of messenger RNA (mRNA) vaccines in controlling COVID-19 has warranted further developments in new technology. Currently, their quality control process largely relies on low-resolution electrophoresis for detecting chain breaks. Here, we present an approach using multi-primer reverse transcription sequencing (MPRT-seq) to identify degradation fragments in mRNA products. Using this in-house-made mRNA containing two antigens and untranslated regions (UTRs), we analyzed the mRNA completeness and degradation pattern at a nucleotide resolution. We then analyzed the sensitive base sequence and its correlation with the secondary structure. Our MPRT-seq mapping shows that certain sequences on the 5' of bulge-stem-loop structures can result in preferential chain breaks. Our results agree with commonly used capillary electrophoresis (CE) integrity analysis but at a much higher resolution, and can improve mRNA stability by providing information to remove sensitive structures or sequences in the mRNA sequence design.

Low-carbon treatment and remediation of oil sludge in mid-to-high latitude regions: A coupled approach of freeze-thaw and supercritical CO2 extraction.

The oil sludge produced while extracting large oil and gas fields in the middle and high latitude regions has caused serious pollution to the surrounding soil. The key to solving this problem in the future is to unify the remediation of soil and the treatment of oil sludge. This study uses supercritical carbon dioxide(scCO2) technology to construct a low-carbon method, providing a new approach to achieve this goal. The study determines the optimal extraction conditions for black calcareous soil with 15% oil content to be 55 °C, 25 MPa, and 90 min through single factor and response surface experiments. Experiments on the scCO2 extraction coupled with freeze-thaw cycles show that oil sludge with a water content of 10% can improve the extraction efficiency of scCO2 by about 2.69% after less than five freeze-thaw cycles. The study also compares the extraction efficiency of the four soils, with a difference of 6.03% observed under the same conditions. Additionally, we analyze the impact of the extraction process on changes in the properties of the oil and soil in the oil sludge. Comprehensive tests, including scanning electron microscope (SEM), nutrient detection, X-ray powder diffractometer (XRD), fourier transform infrared spectroscopy (FTIR), and Gas Chromatography (GC), have been conducted. Results show that standalone scCO2 extraction can remove up to 98.2% of petroleum hydrocarbons from the oil sludge, while simultaneously causing small changes to the soil microstructure and the crystal structure of the oil sludge. Furthermore, this process does not lead to a significant depletion of key nutrients or the generation of new pollutants.

Colloid mobilization and transport in response to freeze-thaw cycles: Insights into the heavy metal(loid)s migration at a smelting site.

Smelting sites often exhibit significant heavy metal(loid)s (HMs) contamination in the soil and groundwater, which are inevitably subjected to environmental disturbances. However, there is limited information available regarding the migration behaviors of HMs in a disturbed scenario. Thus, this work explored the migration of HMs-bearing colloids in response to freeze-thaw treatments by laboratory simulation and pore-scale study. Ultrafiltration results of soil effluents revealed that 61.5 %, 47.6 %, 68.0 %, and 59.2 % of Zn, Cd, Pb, and As were present in colloidal phase, and co-transported during treatments. Nanoparticle tracking analysis (NTA) further confirmed that freeze-thaw cycles were conducive to the generation of colloidal particles and showed the heteroagglomeration among different particles. Pore-network model (PNM) was used to quantify the soil macropore characteristics (macropore diameter, macropore number, coordination number, and Euler value) after treatments. It is evident that freeze-thaw cycles induced the formation of larger macropores while simultaneously enhancing macropore connectivity, thereby establishing an optimal pathway for colloid migration. These findings underscored the importance of environmental disturbances as a trigger for the release and migration of HMs in the smelting site, offering valuable insights for controlling HMs pollution. ENVIRONMENTAL IMPLICATION: The contaminated site has been subjected to prolonged environmental disturbances, causing the exacerbation of pollutants leaching and frequent occurrences of unstable pollution situations. This work explored the migration of HMs-bearing colloids in response to freeze-thaw treatments by laboratory simulation and pore-scale study. The distinct effects of freeze-thaw treatment on colloidal particle number concentration and macropore characteristics may explain the generation and migration of colloid-associated HMs driven by environmental disturbances. This work revealed the underlying mechanisms driving the redistribution of HMs under freeze-thaw cycles, offering valuable insights for risk assessment of soil and groundwater associated with HMs migration.

Quality deterioration of mashed potatoes during the freeze-thaw cycle: From the perspective of moisture and microstructure.

This study aimed to simulate cold chain sales temperatures to predict the effects of temperature fluctuations on the physicochemical properties, moisture distribution, microstructure, and flavor of mashed potatoes. The results showed a decline in the hardness and chewability of mashed potatoes alongside the migration of water from bound water states to weakly bound states under freeze-thaw cycles (FTC) conditions. Microstructural analysis indicated that the adhesive forces between proteins and starch granules were weakened, and the structure of mashed potatoes particles was destroyed following FTC. The oxidation and degradation of fat induced by FTC increased the content of key compounds such as octanal and nonanal, thereby contributing to an overall deterioration in the flavor of mashed potatoes. This study elucidates the effects of FTC on water migration, microstructure, and flavor characteristics of mashed potatoes, thereby providing a theoretical foundation for improving the quality of prefabricated frozen mashed potatoes dishes.

Analysis of Freeze-Thaw Damage of Cement Mortars Doped with Polyethylene Glycol-Based Form Stable Phase Change Materials.

The development of construction materials with the integration of phase change materials (PCMs) has been a topic of wide interest in the scientific community, especially in recent years, due to its positive impact on temperature regulation inside buildings. However, little is known about the behavior of materials doped with PCMs when exposed to accidental or severe environments. Currently, a large area of the planet experiences seasonal freeze-thaw effects, which impact the durability and performance of construction materials. Accordingly, the main objective of this study was to evaluate the damage caused by cyclic freeze-thaw actions on the behavior of a cement mortar, including a PEG-based form-stable PCM. An experimental methodology was developed based on the physical and mechanical characterization of mortars under normal operating conditions and after being subjected to freeze-thaw cycles. The results indicated that, under normal exposure conditions, the incorporation of aggregate functionalized with PCM led to a decrease in the mortar's water absorption capacity, compressive strength, and adhesion. However, its applicability has not been compromised. Exposure to freeze-thaw cycles caused a loss of mass in the specimens and a decrease in the compressive strength and adhesion capability of the mortar.

Macro-micro correlation analysis on the loess from Ili River Valley subjected to freeze-thaw cycles.

The Ili River Valley in Xinjiang, China, is a typical seasonal frozen area where loess landslide disasters have become increasingly common during the freeze-thaw periods in recent years. This study analyzed the macroscopic mechanical strength and microstructure changes of the Ili loess under different freeze-thaw cycles (FTCs) through the post-freeze-thaw triaxial compression test on the unsaturated soil in laboratory. Apart from the scanning electron microscopy (SEM), and the nuclear magnetic resonance (NMR), the macro-micro correlation analysis and the cluster-principal component analysis were applied for the theoretical discussion. The results indicated that the cohesive force of the loess exhibits an initial decreases, followed by the increases, and eventually keep stable after various FTCs, while the internal friction angle showed the opposite developing trend before the final constant. Similar to the strong correlation between the cohesive force and the particle abundance, the internal friction angle is also closely related to the abundance and orientation fractal dimension of the loess particles. However, the principal component analysis results showed that cohesive force strongly correlates with the average maximum pore size and the pore size fractal dimension, for which the internal friction angle most strongly affected by the average maximum particle size. The possible reason is that the extracted principal components represent a class of microscopic parameters with the same or similar change trend, although there may be a certain offset between them. The mechanical deterioration of loess is attributed to the repeated frost heaving force and the migration potential caused by FTCs. The alterations of the microstructure accelerated the deterioration of the macroscopic mechanical properties of the loess, which further widens the understanding of the mechanism behind the deterioration of loess mechanical strength in the Ili River Valley under FTCs, and contributes to the prevention and management of the local landslide disasters.

Dynamic responses of soil microbial communities to seasonal freeze-thaw cycles in a temperate agroecosystem.

Soil freeze-thaw cycles (FTCs) are common in temperate agricultural ecosystems during the non-growing season and are progressively influenced by climate change. The impact of these cycles on soil microbial communities, crucial for ecosystem functioning, varies under different agricultural management practices. Here, we investigated the dynamic changes in soil microbial communities in a Mollisol during seasonal FTCs and examined the effects of stover mulching and nitrogen fertilization. We revealed distinct responses between bacterial and fungal communities. The dominant bacterial phyla reacted differently to FTCs: for example, Proteobacteria responded opportunistically, Actinobacteria, Acidobacteria, Choroflexi and Gemmatimonadetes responded sensitively, and Saccharibacteria exhibited a tolerance response. In contrast, the fungal community composition remained relatively stable during FTCs, except for a decline in Glomeromycota. Certain bacterial OTUs acted as sensitive indicators of FTCs, forming keystone modules in the network that are closely linked to soil carbon, nitrogen content and potential functions. Additionally, neither stover mulching nor nitrogen fertilization significantly influenced microbial richness, diversity and potential functions. However, over time, more indicator species specific to these agricultural practices began to emerge within the networks and gradually occupied the central positions. Furthermore, our findings suggest that farming practices, by introducing keystone microbes and changing interspecies interactions (even without changing microbial richness and diversity), can enhance microbial community stability against FTC disturbances. Specifically, higher nitrogen input with stover removal promotes fungal stability during soil freezing, while lower nitrogen levels increase bacterial stability during soil thawing. Considering the fungal tolerance to FTCs, we recommend reducing nitrogen input for manipulating bacterial interactions, thereby enhancing overall microbial resilience to seasonal FTCs. In summary, our research reveals that microbial responses to seasonal FTCs are reshaped through land management to support ecosystem functions under environmental stress amid climate change.

Damage evolution characteristics of siliceous slate with varying initial water content during freeze-thaw cycles.

Comprehensive studies on the freeze-thaw (F-T) damage mechanism in siliceous slates are lacking. In this study, we investigated the evolutionary characteristics of F-T damage in siliceous slates. To this end, scanning electron microscopy, X-ray diffraction, X-ray fluorescence, and uniaxial compression tests were used to analyze the microstructure, phase composition, porosity, and macroscopic mechanical parameters of siliceous slate with varying initial water content during F-T cycles. The results revealed several insights. (1) The microstructure of siliceous slate undergoes significant change with respect to increasing water content and number of F-T cycles. The rock surface changed from smooth to rough, and the arrangement of the mineral particles changed from tight to loose. (2) More than 80 % of the contents of siliceous slate comprise oxygen, aluminum, silicon, potassium, and iron. In particular, siliceous slate comprises muscovite, quartz, clinochlore, and kaolinite. Both the clinochlore and kaolinite are unstable clay minerals. As clay minerals exhibit strong water absorption and expansion characteristics, kaolinite undergoes strong hydration reactions. Compared to rock samples without F-T cycles in the dry state, the clay mineral content of siliceous slate decreased by nearly 50 %, from 28.8 % to 15.5 %, after 30 F-T cycles in the saturated state. (3) The mechanical parameters of siliceous slates with varying water content decreased exponentially with the number of F-T cycles, while their porosity exhibited a positive correlation with the number of F-T cycles. The degree of deterioration in both increased with increasing water content. Both the number of F-T cycles and the initial water content were observed to wield a significant effect on the deterioration of siliceous slates. (4) The evolution curve of F-T load damage in siliceous slate exhibited characteristics of transitioning from gentle to concave and then to a convex stage of growth. Our results are expected to provide theoretical guidance for the evaluation and prevention of F-T disasters in cold regions.

The influence of various sample storage conditions and sample bacterial contamination on concentrations of routine biochemical parameters.

Background: The pre-analytical (PA) phase is the most vulnerable phase of the laboratory testing procedure, with critical procedures-collection, handling, sample transport, and time and temperature of sample storage. This study aimed to examine the stability of basic biochemical parameters depending on the samples' storage conditions and the number of freeze-thaw cycles (FTCs). In parallel, the presence of sample bacterial contamination during routine laboratory work was examined. Methods: Two plasma pools (ethylenediaminetetraacetic acid (EDTA), and sodium-fluoride/potassium oxalate plasma (NaF)) were stored at +4 ˚C/-20 ˚C. Total chole - sterol (TC), glucose, triglycerides (TG), urea, and albumin concentrations were measured using BioSystems reagents (cholesterol oxidase/peroxidase, glucose oxidase/per - oxidase, glycerol phosphate oxidase/peroxidase, urease/ salicylate, and bromcresol green method, respectively) on Ilab 300+. Sample bacterial contamination was determined by 16S rRNA sequence analysis. The expe - riment encompassed a 5 day-period: Day 1-fresh sample, Day 2-1st FTC, Day 3-2nd FTC, Day 4-3rd FTC, Day 5-4th FTC. The appearance of bacteria in two consecutive samples was the experiment's endpoint.

Characterization of gelatin-oxidized riclin cryogels and their applications as reusable ice cubes in shrimp preservation.

Traditional ice is usually employed to preserve food freshness and extend shelf life. However, ice cannot bear repeated freeze - thaw cycles during the transportation and retailing process, resulting in microbial cross-contamination and spoilage of foods. Herein, succinoglycan riclin was oxidated (RO) and crosslinked with gelatin (Ge), the Ge-RO cryogels were prepared via Schiff base reaction and three freeze - thaw cycles. The Ge-RO cryogels showed improved storage modulus (G') and thermal stability compared with pure gelatin hydrogel. The polymer framework of Ge-RO gels exhibited stable properties against ice crystals destructions during nine freeze - thaw treatments. During the storage and repeated freeze - thaw treatments of shrimps, Ge-RO cryogels exhibited a remarkable preservation effect on shrimps, and their freshness was evaluated using an electronic nose technique equipped with ten sensors. The results demonstrated that the shrimp muscle preserved in ice generated off-odors and resulted in high sensor responses. The sensor responses were reduced sharply of shrimps preserved in cryogels. Moreover, 1H NMR-based metabolomics analysis revealed that shrimps in Ge-RO cryogels group reversed the metabolic perturbations compared with the traditional ice group, the metabolic pathways were related to energy metabolism, nucleotide metabolism, and amino acid metabolism, which provide new clues to the freshness of shrimps. Furthermore, RO exhibited superior antimicrobial activity against E. coli and S. aureus microorganisms. Thus, the crosslinked cryogels are potentially applicable to food preservation, offering sustainable and reusable solutions against traditional ice.

Experimental Study of Recycled Concrete under Freeze-Thaw Conditions.

Recycled concrete is a new and environmentally friendly material for future construction. When applying recycled concrete to cold and severe regions, it is necessary to consider the freeze-thaw resistance of recycled concrete. Using an indoor freeze-thaw cycle test system, the rapid freezing method was used to conduct rapid freeze-thaw tests on recycled concrete specimens under set freeze-thaw cycle conditions. Relevant parameters (such as compressive strength, quality loss rate, rebound value) were tested on recycled concrete specimens that completed the set number of freeze-thaw cycles. The influence of various factors (freeze-thaw cycle number, replacement rate of recycled aggregates) on the compressive strength, quality loss rate, and rebound value of recycled concrete was analyzed. The results indicate that with the increase of freeze-thaw cycles and recycled aggregate content, the workability, rebound value, and compressive strength of the specimens decrease, and the quality loss rate increases. The changes in workability of concrete are sharp, with a slump difference of 21 mm between concrete with 100% recycled coarse aggregate and concrete using natural coarse aggregate. The rebound value of the specimen shows a decreasing trend overall, but the rebound value varies for different measuring points on the same specimen. The attenuation of compressive strength is significant. When fully using recycled aggregates to prepare concrete, the compressive strength after 30 freeze-thaw cycles decreases by 49.42% compared to the compressive strength after 0 freeze-thaw cycles. The overall quality loss rate shows a decreasing trend, but the quality loss is not severe.

A green extraction method for agar with improved thermal stability and water holding capacity.

The conventional agar extraction method has drawbacks such as high energy consumption, low yield, poor quality, and possible residual harmful factors, which greatly limit its application in high-end fields such as biomedicine and high-end materials. This work explored a new freezing-thawing-high-temperature coupling technique for agar extraction. It increased the yield and the strength of agar by 10.6 % and 13.7 %, respectively, as compared to direct high-temperature extraction of agar (HA). The greater molecular weight and lower sulfate content of agar obtained from freeze-thaw cycles combined with high temperature extraction (FA) may be attributed to the desulfurization effect caused by freeze-thaw cycles and the preservation of the molecular chain structure. The reduction in sulfate content decreases the steric hindrance resistance of the polysaccharide chains, enhances their interactions, and promotes the regularity and density of the agar structure, while also improving its water retention and thermal stability. In conclusion, this research can offer a theoretical basis and guidance for the eco-friendly extraction of agar with improved agar characteristics and expended its applications.

Effects of storage conditions on the stability of qPCR reagents: implications for environmental DNA detection.

OBJECTIVE: Environmental DNA (eDNA) detection is a transformative tool for ecological surveys which in many cases offers greater accuracy and cost-effectiveness for tracking low-density, cryptic species compared to conventional methods. For the use of targeted quantitative PCR (qPCR)-based eDNA detection, protocols typically require freshly prepared reagents for each sample, necessitating systematic evaluation of reagent stability within the functional context of eDNA standard curve preparation and environmental sample evaluation. Herein, we assessed the effects of long-term storage and freeze-thaw cycles on qPCR reagents for eDNA analysis across six assays. RESULTS: Results demonstrate qPCR plates (containing pre-made PCR mix, primer-probe, and DNA template) remain stable at 4 °C for three days before thermocycling without fidelity loss irrespective of qPCR assay used. Primer-probe mixes remain stable for five months of - 20 °C storage with monthly freeze-thaw cycles also irrespective of qPCR assay used. Synthetic DNA stocks maintain consistency in standard curves and sensitivity for three months under the same conditions. These findings enhance our comprehension of qPCR reagent stability, facilitating streamlined eDNA workflows by minimizing repetitive reagent preparations.

PON1 enzyme activity assays for serum and heparinized plasma in horses and stability evaluation of the enzyme activity over different freeze-thaw cycles and mimic transportation.

Consistent information and standardization procedures regarding the time of storage for frozen samples and the effects of storage time on enzyme activity are still missing in the literature. Thus, we evaluated the effects of different storage temperatures (-20 °C and - 80 °C), three repetitive freeze/thaw cycles, and 24-h mimic transportation on the activities of PON1 (paraoxonase and arylesterase), enzymes involved in the protection and detoxification processes of reactive molecules. PON1 enzymes' activity was validated on serum and heparinized plasma in horses. The results revealed that conditions and time of storage of blood samples for PON1 analyses altered the activities of both enzymes in both sample types, evidencing that these conditions can lead to protein degradation or general alteration. Specifically, paraoxonase and arylesterase activities significantly decreased among storage temperatures, with major effects detected at -20 °C. The repeated freeze/thaw cycles at -20 °C and 24-h mimic transport conditions also generated an expected degradation of the arylesterase in both serum and heparinized plasma while freeze/thaw cycles at -80 °C caused an increase of both arylesterase and paraoxonase activities on both sample types. In general, similar enzyme responses were detected between serum and heparinized plasma.

Insight into the role of lipids in odor changes of frozen grass carp (Ctenopharyngodon idella) based on lipidomics and GC-MS analysis: Impact of freeze-thaw cycles and heat treatment.

The role of lipids in changes of volatile organic compounds (VOCs) in grass carp during 1 month of frozen storage with different freeze-thaw cycles and subsequent heat treatment was investigated. Sixty VOCs were identified in all groups by SPME-GC-MS. Odor contents fluctuated along with the freeze-thaw cycles and heat treatment, and the highest odor content was observed in frozen sample without freeze-thaw cycles. Freeze-thaw and heat treatment significantly promoted the lipid oxidation and hydrolysis for all the groups(p<0.05). Lipid metabolites were analyzed using non-targeted lipidomics and could be well distinguished among different freeze-thaw groups and heat-treatment groups. A total of 10 key differential lipid molecules were annotated, involving 4 metabolic pathways related to lipid degradation and odor formation. Spearman correlation analysis showed that these key differential lipids were significantly related to the formation of key VOCs (p<0.05).