Search Results (29)

The supplementation of freezing medium with crocin results in an amelioration of post-thawing sperm quality, as determined by motility and viability. This study aimed to examine the molecular mechanisms underlying the ameliorative effect of crocin. Bovine spermatozoa were cryopreserved in a freezing medium supplemented with 0, 0.5, or 1 mM of crocin. Sperm lysates were evaluated for their redox status and the expression of proteins implicated in the heat stress response (HSR) and apoptosis. Crocin protected spermatozoa from the accumulation of superoxide anion and ameliorated their post-thawing antioxidant capacity in terms of ROS scavenging activity and glutathione content. Moreover, crocin decreased the levels of inducible nitric oxide synthase (iNOS), while it increased superoxide dimsutase-1 (SOD-1) levels. These effects were associated with an inhibition of apoptosis, as evidenced by a decreased Bax/Bcl-2 protein ratio and decreased levels of caspase-cleaved substrates. Finally, crocin affected the heat shock response of spermatozoa, since it upregulated the levels of heat shock proteins (Hsp) 60, 70, and 90. In conclusion, the addition of crocin to the freezing medium ensured controlled amounts of ROS, enhanced the antioxidant capacity of spermatozoa, and upregulated the anti-apoptotic proteins and Hsps, thus contributing to the maintenance of cellular homeostasis. Full article

The budding yeast Saccharomyces cerevisiae is a widely utilized model system with myriad applications in terms of industrial, biotechnology, and synthetic biology purposes. One such application is the biosynthesis of commercially and medically important bioactive compounds and their precursors, which oftentimes require culturing conditions at low temperatures to optimize production yield rather than cellular fitness. To lend insight into genetic modifications that may assist this goal, this work focuses on a systematic analysis of the genes that result in an increase in survival following freezing. At present, these genes have been identified in a wide variety of S. cerevisiae wild-type backgrounds—that vary significantly in their properties and behaviors—and in the conditions that led to the annotation of the freeze–thaw survival phenotype. In this work, we report a complete characterization of the thermal tolerance and viability for the freeze–thaw gene family following a standardized protocol within a unified genetic background, the extensively used BY4741 laboratory strain. Our results reveal that five out of these six genes are linked to increased viability in response to both freeze–thaw stress as well as enhanced survival during a heat shock stressor. Follow-up analysis characterized the local spatial effects that gene modification at each locus causes when utilizing the common kanamycin resistance cassette (KanMX6) for the creation of mutant strains and engineering purposes. Full article

Low temperatures can significantly affect the growth of ornamental plants, emphasizing the importance of improving their cold tolerance. However, comparative studies on the photosynthetic responses of sun and shade plants to low temperatures remain limited. In this study, gas exchange, chlorophyll fluorescence in Photosystem II (PSII) and Photosystem I (PSI), the antioxidant system, the osmoregulator substance, and lipid peroxidation were investigated in the shade plant Helleborus viridis (Hv) and the sun plant Lupinus polyphyllus (Lp) during cold acclimation (CA) and the freezing–thaw recovery (FTR). The CA treatment significantly declined the net photosynthetic rate (Pn) and the maximum photochemical efficiency of PSII (Fv/Fm) in Hv and Lp, indicating the photoinhibition occurred in both species. However, Hv exhibited a much better photosynthetic stability to maintain Pn, Fv/Fm, and carboxylation efficiency (CE) than Lp during CA, suggesting that Hv had a greater photosynthetic resilience compared to Lp. Furthermore, Hv preferred to maintain Pn, Fv/Fm, the actual photosynthetic efficiency of PSII (Y(II)), and the actual photosynthetic efficiency of PSI (Y(I)) to consistently provide the necessary energy for the carbon assimilation process, while Lp tended to divert and dissipate excess energy by thermal dissipation and cyclic electron flow during CA. Moreover, there were higher soluble sugar contents in Hv in comparison to Lp. These traits allowed Hv to recover photosynthetic efficiency and maintain cellular integrity better than Lp after the freezing stress. In conclusion, CA significantly reduced the photosynthetic capacity and led to the divergent photosynthetic strategies of both species, which finally resulted in a different freezing tolerance after the freezing–thaw recovery. These findings provide insights into the divergent photoprotective strategies of sun and shade plants in response to cold temperatures. Full article

In railway engineering research, there is a notable gap as existing studies often focus separately on train-induced vibrations or freeze–thaw cycle impacts on subgrades, lacking a comprehensive analysis of their combined effects on subgrade dynamic responses. This study developed a three-dimensional finite-element model of a double-track ballastless track railway subgrade. The model considers various conditions, including train speeds of 180 km/h, 200 km/h, and 220 km/h, and soil temperatures of 5 °C, −5 °C, and −15 °C, with typical subgrade materials. The results show that under train load, the maximum vertical displacement of the subgrade decreases as train speed increases. Conversely, the maximum vertical stress and acceleration are directly proportional to the train speed. When the train speed rises from 180 km/h to 220 km/h, the maximum vertical stress of the subgrade increases by 1.1% and 3.1%, respectively. As the soil temperature drops from 5 °C to −15 °C, the maximum vertical displacement of the subgrade decreases. The displacement reduces with increasing distance from the train load. At a specific point A, the maximum vertical stress increases by 2.02% and 1.43%, respectively. Additionally, the deformation of the railway subgrade is directly proportional to the temperature difference. These findings are valuable for understanding subgrade behavior and guiding railway construction in freeze–thaw-affected areas. Full article

The excavation of the rock mass at the tunnel entrance in regions characterized by high altitudes and elevated stress levels results in the direct exposure of the surrounding rock to atmospheric conditions. This surrounding rock is subjected to the compounded effects of excavation-induced unloading damage and freeze–thaw erosion, which contribute to the degradation of its mechanical properties. Such deterioration has a negative impact on production and construction operations. Following tunnel excavation, the lateral stress exerted by the surrounding rock at the tunnel face is reduced, leading to a predominance of uniaxial compressive stress. As a result, the failure mode and mechanical behavior of the rock exhibit characteristics similar to those observed in uniaxial loading tests conducted in controlled laboratory environments. This study conducts laboratory-based uniaxial loading and unloading tests, as well as freeze–thaw tests, to examine the strength, deformation characteristics, and fracture attributes of unloading sandstone subjected to freeze–thaw erosion. A damage deterioration model for unloading sandstone under uniaxial conditions is developed, and the patterns of damage response are further analyzed through the identification of compaction points and the definition of damage response points. The results indicate that (1) as the degree of freeze–thaw erosion increases, the failure threshold of the sandstone significantly decreases, with the residual rock fragments on the fracture surface transitioning from hard and sharp to soft and sandy; (2) freeze–thaw erosion has a pronounced negative impact on the cohesion of the sandstone, while the reduction in the internal friction angle is relatively moderate; and (3) the strain induced by damage following three, six, and nine freeze–thaw cycles exhibits a gradual decline and appears to reach a state of stabilization when compared to conditions without freeze–thaw exposure. Investigating the mechanical properties and deterioration mechanisms of the rock in this specific context is crucial for establishing a theoretical foundation to assess the stability of the tunnel’s surrounding rock and determine the necessary support measures. Full article

Protein-based hydrogels with stretchability and conductivity have potential applications in wearable electronic devices. However, the development of protein-based biocomposite hydrogels is still limited. In this work, we used natural ferritin to develop a PVA/ferritin biocomposite hydrogel by a repetitive freeze–thaw method. In this biocomposite hydrogel, ferritin, as a nano spring, forms a hydrogen bond with the PVA networks, which reduces the crystallinity of PVA and significantly improves the stretchability of the hydrogel. The fracture strain of the PVA/ferritin hydrogel is 203%, and the fracture stress is 112.2 kPa. The fracture toughness of the PVA/ferritin hydrogel is significantly enhanced to 147.03 kJ/m³, more than 3 times that of the PVA hydrogel (39.17 kJ/m³). In addition, the free residues and iron ions of ferritin endow the biocomposite hydrogel with enhanced ionic conductivity (0.15 S/m). The strain sensor constructed from this hydrogel shows good sensitivity (gauge factor = 1.7 at 150% strain), accurate real-time resistance response, and good long cyclic working stability when used for joint motion monitoring. The results indicate that a PVA/ferritin biocomposite hydrogel prepared by a facile method has enhanced stretchability and conductivity for flexible strain sensors. This work develops a new method for the preparation of protein-based hydrogels for wearable electronic devices. Full article

Freeze tolerance is a remarkable adaptive trait exhibited by wood frogs (Rana sylvatica) during their hibernation period. To show the epigenetic mechanisms that contribute to kidney protection during freezing stress, this present study provides the first investigation of the role and dynamics of histone arginine methylation and the expression of protein arginine methyltransferases (PRMTs) in a freeze-tolerant vertebrate. Kidney samples from three groups were assessed: (a) control frogs acclimated at 5 °C, (b) 24 h frozen frogs, and (c) 8 h thawed frogs. Our findings revealed significant downregulation of PRMT1, PRMT3, and PRMT5 in kidneys from frozen wood frogs compared to the control group. This downregulation indicates a potential role for PRMT enzymes in the regulation of arginine methylation under freezing stress. In addition, we observed distinct changes in histone marks. H3R17me2a showed significant upregulation after 24 h of freezing, potentially indicating its involvement in the activation of genes related to freezing survival. By contrast, H3R26me2a was downregulated after both 24 h freezing and 8 h thawing, whereas H3R8me2a showed sustained levels after freezing but was downregulated after thawing. These findings highlight the dynamic nature of histone arginine methylation and PRMT expression in wood frog kidneys during freezing–thawing. Our results indicate that epigenetic modifications play a crucial role in shaping the adaptive responses of wood frog kidneys to freezing stress and contribute new information on the underlying biochemical modifications that support vertebrate freeze tolerance. Full article

Large volumes of waste tires are generated due to the rapid growth of the transportation industry. An effective method of recycling waste tires is needed. Using rubber from tires to improve problematic soils has become a research topic. In this paper, the dynamic response of rubber fiber-reinforced expansive soil under freeze–thaw cycles is investigated. Dynamic triaxial tests were carried out on rubber fiber-reinforced expansive soil subjected to freeze–thaw cycles. The results showed that with the increase in the number of freeze–thaw cycles, the dynamic stress amplitude and dynamic elastic modulus of rubber fiber-reinforced expansive soils first decrease and then increase, and the damping ratio first increases and then decreases, all of which reach the turning point at the 6th freeze–thaw cycle. The dynamic stress amplitude and dynamic elastic modulus decreased by 59.4% and 52.2%, respectively, while the damping ratio increased by 99.8% at the 6th freeze–thaw cycle. The linear visco-elastic model was employed to describe the hysteretic curve of rubber fiber-reinforced expansive soil. The elastic modulus of the linear elastic element and the viscosity coefficient of the linear viscous element first decrease and then increase with the increase in the number of freeze–thaw cycles; all reach the minimum value at the 6th freeze–thaw cycle. The dynamic stress–dynamic strain curve calculation method is established based on the hyperbolic model and linear visco-elastic model, and the verification shows that the effect is better. The research findings provide guidance for the improvement of expansive soil in seasonally frozen regions. Full article

The deformation and damage to seasonal permafrost roadbeds, as seasons shift, stems from the intricate interplay of temperature, moisture, and stress fields. Fundamentally, the frost heave and thaw-induced settlement of soil represent a multi-physics coupling phenomenon, where various physical processes interact and influence each other. In this investigation, a comprehensive co-coupling numerical simulation of both the temperature and moisture fields was successfully executed, utilizing the secondary development module within the finite element software, COMSOL Multiphysics 6.0. This simulation inverted the classical freezing–thawing experiment involving a soil column under constant temperature conditions, yielding simulation results that were in excellent agreement with the experimental outcomes, with an error of no more than 10%. Accordingly, the temperature, ice content, and liquid water content distributions within the seasonal permafrost region were derived. These parameters were then incorporated into the stress field analysis to explore the intricate coupling between the moisture and temperature fields with the displacement field. Subsequently, the frost heave and thaw settlement deformations of the roadbed were calculated, accounting for seasonal variations, thereby gaining insights into their dynamic behavior. The research results show that during the process of freezing and thawing, water migrates from the frozen zone towards the unfrozen zone, with the maximum migration amount reaching 20% of the water content, culminating in its accumulation at the interface separating the two. Following multiple freeze–thaw cycles, this study reveals that the maximum extent of freezing within the roadbed reaches 2.5 m, while the road shoulder experiences a maximum freezing depth of 2 m. A continuous trend of heightened frost heave and thaw settlement deformation of the roadbed is observed in response to temperature fluctuations, leading to the uneven deformation of the road surface. Specifically, the maximum frost heave measured was 51 mm, while the maximum thaw settlement amounted to 13 mm. Full article

Epigenetic regulation, notably histone post-translational modification (PTM), has emerged as a major transcriptional control of gene expression during cellular stress adaptation. In the present study, we use an acid extraction method to isolate total histone protein and investigate dynamic changes in 23 well-characterized histone methylations/acetylations in the brains of wood frogs subject to 24-h freezing and subsequent 8-h thawed recovery conditions. Our results identify four histone PTMs (H2BK5ac, H3K14ac, H3K4me3, H3K9me2) and three histone proteins (H1.0, H2B, H4) that were significantly (p < 0.05) responsive to freeze-thaw in freeze-tolerant R. sylvatica brains. Two other permissive modifications (H3R8me2a, H3K9ac) also trended downwards following freezing stress. Together, these data are strongly supportive of the proposed global transcriptional states of hypometabolic freeze tolerance and rebounded thawed recovery. Our findings shed light on the intricate interplay between epigenetic regulation, gene transcription and energy metabolism in wood frogs’ adaptive response to freezing stress. Full article

Ultralow-temperature fluids (such as liquid nitrogen, liquid CO₂) are novel waterless fracturing technologies designed for dry, water-sensitive reservoirs. Due to their ultralow temperatures, high compression ratios, strong frost heaving forces, and low viscosities, they offer a solution for enhancing the fracturing and permeability of low-permeability reservoirs. In this study, we focus on the combined effects of high-pressure fluid rock breaking, low-temperature freeze-thaw fracturing, and liquid-gas phase transformation expansion on coal-rock in low-permeability reservoirs during liquid nitrogen fracturing (LNF). We systematically analyze the factors that limit the LNF effectiveness, and we discuss the pore fracture process induced by low-temperature fracturing in coal-rock and its impact on the permeability. Based on this analysis, we propose a model and flow for fracturing low-permeability reservoirs with low-temperature fluids. The analysis suggests that the Leidenfrost effect and phase change after ultralow-temperature fluids enter the coal support the theoretical feasibility of high-pressure fluid rock breaking. The thermal impact and temperature exchange rate between the fluid and coal determine the temperature difference gradient, which directly affects the mismatch deformation and fracture development scale of different coal-rock structures. The low-temperature phase change coupling fracturing of ultralow-temperature fluids is the key to the formation of reservoir fracture networks. The coal-rock components, natural fissures, temperature difference gradients, and number of cycles are the key factors in low-temperature fracturing. In contrast to those in conventional hydraulic fracturing, the propagation and interaction of fractures under low-temperature conditions involve multifield coupling and synergistic temperature, fluid flow, fracture development, and stress distribution processes. The key factors determining the feasibility of the large-scale application of ultralow-temperature fluid fracturing in the future are the reconstruction of fracture networks and the enhancement of the permeability response in low-permeability reservoirs. Based on these considerations, we propose a model and process for LNF in low-permeability reservoirs. The research findings presented herein provide theoretical insights and practical guidance for understanding waterless fracturing mechanisms in deep reservoirs. Full article

The results of artificial insemination (AI) are adversely affected by changes in sperm motility and function throughout the cryopreservation procedure. The proteome alterations of frozen–thawed spermatozoa with various levels of freezability in dairy goats, however, remain largely unknown. To discover differentially expressed proteins (DEPs) and their roles in dairy goat sperm with high or low freezability (HF or LF), we conducted 4D-DIA quantitative proteomics analysis, the results of which are presented in this work. Additionally, we explored the underlying processes that may lead to the variations in sperm freezing resistance. A total of 263 DEPs (Fold Change > 2.0, p-value < 0.05) were identified between the HF group and LF group in frozen–thawed dairy goat spermatozoa. In our Gene Ontology (GO) enrichment analysis, the DEPs were mostly associated with the regulation of biological processes, metabolic processes, and responses to stress and cellular component biogenesis. Our Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis also revealed that the DEPs were predominantly engaged in oxidative phosphorylation, N-Glycan biosythesis, and cysteine and methionien metabolism. A protein–protein interaction (PPI) network analysis revealed 14 potential proteins (NUDFB8, SDHC, PDIA4, HSPB1, etc.) that might influence the freezability of dairy goat sperm. These findings shed light on the processes underlying alterations in the proteome and sperm freezability, aiding further research on sperm cryopreservation. Full article

Although the theoretical pavement structure design method (TPSDM) is widely used for designing asphalt pavements in Japan, it still exhibits certain limitations, such as not considering the variation in moduli of the base and subgrade layers due to water contents, freeze–thaw action, and stress states. This study aims to enhance Japanese TPSDM’s accuracy by considering variations in the resilient modulus of environmental impacts, pavement materials, pavement structure, and traffic load actions to accurately calculate the mechanical responses and predict pavement fatigue life. Firstly, the study develops a 3D Thermo-Hydro-Mechanical (THM) model using the finite element method (FEM) to investigate temperature and moisture distributions of the pavement with time. Then, based on the numerical results of the moisture, temperature, and stress state obtained from the THM analysis, the constant resilient modulus of the base and subgrade layers in the Japanese TPSDM is replaced with a resilient modulus that considers the stress state and the combined effects of water content fluctuations and freeze–thaw action. Finally, the fatigue life of the pavement is calculated based on the obtained mechanical response in THM analysis. The reliability and validity of the proposed fatigue life prediction method are well verified by comparing the calculated with the actual pavement fatigue life. Results indicate that the modifications improve the Japanese TPSDM by considering the environmental impacts, traffic load actions, pavement materials, and pavement structure, thereby improving the accuracy of predicting the fatigue life of asphalt pavements, particularly in cold regions. Full article

The anti-Müllerian hormone (AMH) is a glycoprotein that plays an important role in prenatal sex differentiation. It is used as a biomarker in polycystic ovary syndrome (PCOS) diagnostics, as well as for estimating an individual’s ovarian reserve and the ovarian response to hormonal stimulation during in vitro fertilization (IVF). The aim of this study was to test the stability of AMH during various preanalytical conditions that are in accordance with the ISBER (International Society for Biological and Environmental Repositories) protocol. Plasma and serum samples were taken from each of the 26 participants. The samples were then processed according to the ISBER protocol. AMH levels were measured in all the samples simultaneously using the chemiluminescent kit ACCESS AMH in a UniCel^® DxI 800 Immunoassay System (Beckman Coulter, Brea, CA, USA). The study proved that AMH retains a relatively high degree of stability during repeated freezing and thawing in serum. AMH was shown to be less stable in plasma samples. Room temperature proved to be the least suitable condition for the storage of samples before performing the biomarker analysis. During the testing of storage stability at 5–7 °C, the values decreased over time for all the plasma samples but remained stable in the serum samples. We proved that AMH is highly stable under various stress conditions. The anti-Müllerian hormone retained the greatest stability in the serum samples. Full article

Advancements in detection instruments have enabled the real-time acquisition of water information during plant growth; however, the real-time monitoring of freeze–thaw information during plant overwintering remains a challenge. Based on the relationship between the change in the water–ice ratio and branch impedance during freezing, a miniature noninvasive branch volume ice content (BVIC) sensor was developed for monitoring real-time changes in volumetric ice content and the ice freeze-thaw rate of woody plant branches during the overwintering period. The results of the performance analysis of the impedance measurement circuit show that the circuit has a lateral sensitivity range, measurement range, resolution, measurement accuracy, and power consumption of 0–35 mm, 0–100%, 0.05%, ±1.76%, and 0.25 W, respectively. The dynamic response time was 0.296 s. The maximum allowable error by the output voltage fluctuation, owing to the ambient temperature and humidity, was only ±0.635%, which meets the actual use requirements. The calibration curve fit coefficients were >0.98, indicating a significant correlation. The ice content of plant branches under cold stress was measured for indoor and field environments, and the sensors could effectively monitor changes in the branch ice content in plants exposed to cold stress. Additionally, they can differentiate between plants with different cold resistances, indicating the reliability of the BVIC sensor. Full article