Freezing Conditions Research Articles

Synergistic mechanism of steam blanching and freezing conditions on the texture of frozen yellow peaches based on macroscopic and microscopic properties.

Freezing and blanching are essential processing steps in the production of frozen yellow peaches, inevitably leading to texture softening of the fruit. In this study, the synergistic mechanism of stem blanching, freezing conditions (-20°C, -40°C, -80°C, and liquid nitrogen [-173°C]), and sample sizes (cubes, slices, and half peaches) on macroscopic properties of texture, cellular structure, and ice crystal size distribution of frozen yellow peaches were measured. Blanching enhanced the heat and mass transfer rates in the subsequent freezing process. For nonblanched samples, cell membrane integrity was lost at any freezing rate, causing a significant reduction in textural quality. Slow freezing further exacerbated the texture softening, while the ultra-rapid freezing caused structural rupture. For blanched samples, the half peaches softened the most. The water holding capacity and fracture stress were not significantly affected by changes in freezing rate, although the ice crystal size distribution was more susceptible to the freezing rate. Peach cubes that had undergone blanching and rapid freezing (-80°C) experienced 4% less drip loss than nonblanched samples. However, blanching softened yellow peaches more than any freezing conditions. The implementation of uniform and shorter duration blanching, along with rapid freezing, has been proven to be more effective in preserving the texture of frozen yellow peaches. Optimization of the blanching process may be more important than increasing the freezing rate to improve the textural quality of frozen yellow peaches.

Comparative Efficacy of the Dimethyl Sulfoxide, Glycerol and Methanol on the Post-Thaw Cell Viability of HeLa Cells

Cryoprotectants are used to protect cells during freezing. The concentration, type, and freeze-thaw conditions of these substances vary depending on the type of cell to be used. It is very important to determine the appropriate cryopreservation method for the particular cell. This study aims to provide insights into the optimal cryopreservation method for HeLa cells by comparing the performance of different cryoprotectants and evaluating their effectiveness under various freezing and storage conditions. Cell suspensions were frozen with a freezing media composed of cryoprotectant + fetal bovine serum + medium at a ratio of 5:10:85 (v:v:v) and stored under the following conditions: 3 months (-20 °C), 1 month (-80 °C), and 6 months (-80 °C). Cell viability and recovery rates were analyzed immediately post-thaw and after 48 h using the trypan blue dye exclusion assay. In 3 months (-20 °C), viability and recovery rates were higher in the methanol group. Glycerol showed better performance in 1 month (-80 °C). DMSO was the most efficient in 6 months (-80 °C). Methanol failed at -80 °C storage temperature. This study demonstrates the effect of these cryoprotectants in HeLa cells on cell viability and cell recovery rates immediately after thawing and after 48 hours of cultivation.

Microplate-Based Cryopreservation of Adherent-Cultured Human Cell Lines Using Amino Acids and Proteins.

With the progression of regenerative medicine and cell therapy, the importance of cryopreservation techniques for cultured cells continues to rise. Traditional cryoprotectants, such as dimethyl sulfoxide and glycerol, are effective in cryopreserving suspended cells, but they do not demonstrate sufficient efficacy for two-dimensional (2D)-cultured cells. In the past decade, small molecules and polymers have been studied as cryoprotectants. Some L-amino acids have been reported to be natural and biocompatible cryoprotectants. However, the cryoprotective effects of D-amino acids have not been investigated for such organized cells. In the present study, the cryoprotective effects of D- and L-amino acids and previously reported cryoprotectants were assessed using HepG2 cells cultured on a microplate without suspending the cells. d-Proline had the highest cryoprotective effect on 2D-cultured cells. The composition of the cell-freezing solution and freezing conditions were then optimized. The d-proline-containing cell-freezing solution also effectively worked for other cell lines. To minimize the amount of animal-derived components, fetal bovine serum in the cell freezing solution was substituted with bovine serum albumin and StemFit (a commercial supplement for stem cell induction). Further investigations on the mechanism of cryopreservation suggested that d-proline protected enzymes essential for cell survival from freeze-induced damage. In conclusion, an effective and xeno-free cell-freezing solution was produced using d-proline combined with dimethyl sulfoxide and StemFit for 2D-cultured cells.

Construction of a soybean protein isolate/polysaccharide-based whole muscle meat analog: Physical properties and freeze-thawing stability study

A growing interest has arisen in recreating real meat by mimicking its texture characteristics and muscle fiber structure. Our previous work successfully created meat analog fiber based on soybean protein isolate (SPI) and sodium alginate (SA) with the wet-spinning method. In this work, we analyzed the microstructure, texture profile, and water retainability of the assembled plant-based whole muscle meat analog (PMA) made of SPI/SA-based meat analog fiber and systematically studied the effect of different combinations and contents of transglutaminase (TG), salt, and soybean oil on the rheological behavior of the formulated adhesive. The estimated optimal condition that has the most similar texture characteristic with real chicken breast meat is: for every 1:1 mass ratio of simulated plant meat fibers to the adhesive, add 0.1 % TG enzyme addition in the adhesive and 100 mM NaCl addition. The physical behavior of PMA during cryopreservation was investigated through freeze-thaw cycles and freezing times. The addition of a small amount of oil and salt can efficiently prevent the PMA through freezing conditions which is comparable with the addition of D-Trehalose (TD). Overall, this study not only created a plant-based whole muscle meat analog product that is similar in texture to real chicken breast meat but also provided a new direction for constructing fiber-rich structure protein-based muscle meat analogs and their further commercialization.

Impact of cryopreservation on elastomuscular artery mechanics

Low temperatures slow or halt undesired biological and chemical processes, protecting cells, tissues, and organs during storage. Cryopreservation techniques, including controlled media exchange and regulated freezing conditions, aim to mitigate the physical consequences of freezing. Dimethyl sulfoxide (DMSO), for example, is a penetrating cryoprotecting agent (CPA) that minimizes ice crystal growth by replacing intracellular water, while polyvinyl alcohol (PVA) is a nonpenetrating CPA that prevents recrystallization during thawing. Since proteins and ground substance dominate the passive properties of soft biological tissues, we studied how different freezing rates, storage temperatures, storage durations, and the presence of cryoprotecting agents (5% [v/v] DMSO + 1 mg/mL PVA) impact the histomechanical properties of the internal thoracic artery (ITA), a clinically relevant blood vessel with both elastic and muscular characteristics. Remarkably, biaxial mechanical analyses failed to reveal significant differences among the ten groups tested, suggesting that mechanical properties are virtually independent of the cryopreservation technique. Scanning electron microscopy revealed minor CPA-independent delamination in rapidly frozen samples, while cryoprotected ITAs had better post-thaw viability than their unprotected counterparts using methyl thiazole-tetrazolium (MTT) metabolic assays, especially when frozen at a controlled rate. These results can be used to inform ongoing and future studies in vascular engineering, physiology, and mechanics.

Mouse Model of Chronic Social Stress-Induced Excessive Pavlovian Aversion Learning-Memory.

Increased experience of aversive stimuli/events is a psychological-neurobiological state of major importance in psychiatry. It occurs commonly in generalized anxiety disorder, post-traumatic stress disorder, and major depression. A sustained period of exposure to threat (chronic stressor) is a common risk factor, and a major symptom is generalized excessive perception of, and reactivity to, aversive stimuli. In rodents, Pavlovian aversion learning and memory (PAL, PAM), quantified in terms of the conditioned defensive behavior freezing, is an extensively studied behavioral paradigm, and well understood in terms of underlying neural circuitry. In mice, chronic social stress (CSS) is a 15-day resident-intruder paradigm in which C57BL/6 adult males are exposed continuously and distally to dominant-aggressive CD-1 male mice (sustained threat) interspersed with a brief daily period of proximal attack (acute threat). To ensure that physical wounding is minimized, proximal attacks are limited to 30 to 60 s/day and lower incisor teeth of CD-1 mice are blunted. Control (comparison) mice are maintained in littermate pairs. The CSS and CD-1 mice are maintained in distal contact during subsequent behavioral testing. For PAL, CSS and control (CON) mice are placed in a conditioning chamber (context) and exposed to a tone [conditioned stimulus (CS)] and mild, brief foot shock [unconditioned stimulus (US)]. For PAM, mice are placed in the same context and presented with CS repetitions. The CSS mice acquire (learn) and express (memory) a higher level of freezing than CON mice, indicating that CSS leads to generalized hypersensitivity to aversion, i.e., chronic social aversion leads to increased aversion salience of foot shock. Distinctive features of the model include the following: high reproducibility; rare, mild wounding only; male specificity; absence of "susceptible" vs "resilient" subgroups; behavioral effects dependent on continued presence of CD-1 mice; and preclinical validation of novel compounds for normalizing aversion hypersensitivity with accurate feedforward prediction of efficacy in human patients. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Chronic social stress (CSS) Basic Protocol 2: Pavlovian aversion learning and memory (PALM).

Assessment of the long-term stability of pesticide residues under freezing conditions in brown rice and soybean certified reference materials.

The objective of this study was to assess the long-term stability of pesticide residues in brown rice and soybean. The long-term stability of pesticide residues in brown rice and soybean was assessed for 5415 days (over 14 years) and 1801 days (about 5 years), respectively. The samples-certified reference materials (CRMs) 7504-a (brown rice) and 7509-a (soybean) -were prepared by freeze-pulverization. Two target pesticides (etofenprox and fenitrothion) were selected for brown rice and four (chlorpyrifos, diazinon, fenitrothion, and permethrin) for soybean. Our analytical results for long-term stability based on highly reliable isotope dilution mass spectrometry were in the range of expanded uncertainty (k=2) for the certified values of each CRM. The concentration showed a decreasing trend in none of the target pesticides when the samples were stored at temperatures between -20 °C and -30 °C, which indicated that the target pesticides were stable for the tested long terms.

Experimental evidence, global patterns of treeline position and climate provide no substance for a lignin limitation hypothesis of tree growth

While the position of alpine and arctic treelines can be predicted by climatic data, the underlying biological mechanisms are still unclear. In a recent paper in this journal (Körner C, Lenz A, Hoch G (2023) Chronic in situ tissue cooling does not reduce lignification at the Swiss treeline but enhances the risk of 'blue' frost rings. Alpine Botany https://doi.org/10.1007/s00035-023-00293-6) we presented results of an in situ stem-cooling experiment at a Swiss treeline site. The experiment provided answers to two entirely different questions, related to xylogenesis at treeline: (a) the absence of chronic effects of low temperature on lignification, and (b) a high time resolution insight into the rare occurrence of damages in young, still undifferentiated, and thus, non-lignified cells at the occasion of an exceptional early season frost event. In the last issue of Alpine Botany (August 7, 2023), our data had been re-interpreted by (Büntgen, Alpine Botany, 2023) by confusing (b) with (a). Cell death before secondary wall formation interrupts all metabolism, and thus, cannot exert a specific limitation of lignification. For the xylem to lignify, it requires a secondary cell wall in the first place. A frost damage in young tracheid cells is unsuitable for a dendrological treeline hypothesis based on a low-temperature threshold for lignification. Generally, the global pattern of treeline position is not associated with local freezing conditions.

Herbage accumulation, nutritive value, and fatty acid profile of bermudagrass cultivars grown in different latitudes

AbstractThe geographical distribution of bermudagrass cultivars is limited by freezing events (<0°C), commonly associated with higher latitudes in the northern hemisphere. The lipid composition in the plant cell membranes is critical to promote persistence under freezing conditions. This study evaluated herbage accumulation (HA), nutritive value, and lipid biosynthesis of different bermudagrass cultivars in Ona, FL (27°26' N 82°55' W) and Marianna, FL (30o52' N 85o11' W), in 2 years. Treatments were five bermudagrass cultivars (Coastal, Tifton 44, Tifton 85, Jiggs, and Mislevy) distributed in a randomized complete block design with four replicates. HA was greater in Marianna than in Ona for all cultivars (p < 0.05). Mislevy and Jiggs had greater HA in Marianna than the others (p < 0.05), while all cultivars were similar in Ona (p > 0.05). There was no difference between locations or among the cultivars for crude protein and neutral detergent fibre, but bermudagrass cultivars had greater in vitro digestibility of organic matter (IVDOM) in Marianna than in Ona (p < 0.05). The concentration of unsaturated fatty acids increased in the northern latitude (Marianna; p < 0.05). Coastal had greater unsaturated fatty acid concentration than other cultivars (p < 0.05). Geographical location affects HA, IVDOM, and unsaturated fatty acid concentration of bermudagrass cultivars. The unsaturated fatty acid concentration plays a key role in acclimating bermudagrass to different thermal environments. Coastal showed great potential to tolerate chilling stress in Florida. Further studies may be developed to identify promising molecular markers for cold tolerance in bermudagrass.

Fear extinction is impaired in aged rats.

Normal aging is accompanied by broad loss of cognitive function in humans and rodents, including declines in cognitive flexibility. In extinction, a conditional stimulus (CS) that was previously paired with a footshock is presented alone. This procedure reliably reduces conditional freezing behavior in young adult rats. Here, we aimed to investigate how normal aging affects extinction learning. Using young (3months) and aged (20months) male and female Long Evans rats, we compared extinction (using 20 CS-alone presentations) to a no extinction control (equal exposure to the conditioning chamber without CS presentations) following delay fear conditioning. We found that young animals in the extinction group showed a decrease in freezing following extinction; aged animals did not. We next examined changes in neural activity using expression of the immediate early gene zif268. In young animals, extinction corresponded with decreased expression of zif268 in the basolateral amygdala and anterior retrosplenial cortex; this was not observed in aged animals. Further, aged animals showed increased zif268 expression in each region examined, suggesting that dysfunction in neural activity precedes cognitive deficits. These results demonstrate that aging impacts both extinction learning and neural activity.

Freezing stress response of wild and cultivated chickpeas.

Chickpea is an economically and nutritionally important grain legume globally, however, cold stress has adverse effects on its growth. In cold countries, like Canada where the growing season is short, having cold stress-tolerant varieties is crucial. Crop wild relatives of chickpea, especially Cicer reticulatum, can survive in suboptimal environments and are an important resource for crop improvement. In this study, we explored the performance of eleven C. reticulatum wild accessions and two chickpea cultivars, CDC Leader and CDC Consul, together with a cold sensitive check ILC533 under freezing stress. Freezing tolerance was scored based on a 1-9 scale. The wild relatives, particularly Kesen_075 and CudiA_152, had higher frost tolerance compared to the cultivars, which all died after frost treatment. We completed transcriptome analysis via mRNA sequencing to assess changes in gene expression in response to freezing stress and identified 6,184 differentially expressed genes (DEGs) in CDC Consul, and 7,842 DEGs in Kesen_075. GO (gene ontology) analysis of the DEGs revealed that those related to stress responses, endogenous and external stimuli responses, secondary metabolite processes, and photosynthesis were significantly over-represented in CDC Consul, while genes related to endogenous stimulus responses and photosynthesis were significantly over-represented in Kesen_075. These results are consistent with Kesen_075 being more tolerant to freezing stress than CDC Consul. Moreover, our data revealed that the expression of CBF pathway-related genes was impacted during freezing conditions in Kesen_075, and expression of these genes is believed to alleviate the damage caused by freezing stress. We identified genomic regions associated with tolerance to freezing stress in an F2 population derived from a cross between CDC Consul and Kesen_075 using QTL-seq analysis. Eight QTLs (P<0.05) on chromosomes Ca3, Ca4, Ca6, Ca7, Ca8, and two QTLs (P<0.01) on chromosomes Ca4 and Ca8, were associated with tolerance to freezing stress. Interestingly, 58 DEGs co-located within these QTLs. To our knowledge, this is the first study to explore the transcriptome and QTLs associated with freezing tolerance in wild relatives of chickpea under controlled conditions. Altogether, these findings provide comprehensive information that aids in understanding the molecular mechanism of chickpea adaptation to freezing stress and further provides functional candidate genes that can assist in breeding of freezing-stress tolerant varieties.

Advanced testing and characterization of low-temperature cracking in bitumen and mastic

Low-temperature cracking is one of the most common failures in asphalt pavements, especially in cold regions. Accordingly, considerable amount of research has been performed in order to understand the low-temperature cracking mechanisms and to propose test methods for characterizing and determining cracking performance of bitumen and asphalt mixtures under freezing conditions. The existing test methods, however, require expensive equipment and skilled technicians; they are thus not well suited for routine tests. As a contribution to mitigate this situation, this study intends to investigate experimentally and characterize numerically the low-temperature cracking behavior of bitumen and mastic materials using a refined thermal cracking test method. The proposed method, the annular restrained cold temperature induced cracking (ARCTIC) test, allows to determine the low-temperature cracking properties of the mastic and bitumen with a relatively simple setup. In this paper, finite element (FE) modeling is used for evaluating the effect of test parameters on the temperature, stress and strain gradients induced in the specimen during the test. The ARCTIC test is employed to measure cracking temperatures of two bitumen and two mastic materials. The measurements repeatability is examined and the effect of bitumen type on the thermal cracking potential of bitumen and mastic is evaluated. FE modeling is employed to examine the effect of thermomechanical parameters on thermal cracking performance of the materials and to back-calculate fracture stress and strain from measurements. The results highlight the potential of the proposed test and analysis method for evaluation of low-temperature cracking in bitumen and asphalt mastic.

Cryogenic vs. Mechanical freezing impact on the quality of the sea bass (Dicentrarchus labrax) during long-term storage

In this study, we compared the long-term impact of cryogenic and mechanical freezing on sea bass quality. Mechanical freezing (MF) and cryogenic freezing (CF) were conducted at -40 °C and -80 °C, respectively, with air velocities of 2 m s-1 and 0.5 m s-1. Frozen samples were stored at -20 °C for 60 days. We assessed quality attributes, including exudate loss, texture, TBA value, color, enthalpy, microstructure, and protein denaturation, over this period. Exudate loss increased significantly during storage, with no significant difference between freezing methods. The cutting force, L*, b*, chroma (C*), and total color difference (ΔE) were mainly influenced by storage time rather than freezing methods, as p values were >0.05. MF samples were notably less green (p < 0.05) than CF samples, as indicated by higher a* value. Lipid oxidation occurred post-freezing and during storage, with the TBA value at the 0th month being 11 % lower for cryogenic freezing compared to mechanical freezing, reducing to just 2 % after the 2nd month. CF exhibited slightly higher enthalpy than MF, although not significant. Furthermore, the deconvolution data suggested that protein denaturation was impacted not only by freezing storage but also by the freezing treatment and freezing methods to some extent. Microstructure analysis showed that CF produced smaller ice crystals and maintained tissue integrity (inter-fibre distance) better than MF (p value = 0.000), a benefit that persisted during storage (p value = 0.009). Overall, fish quality attributes were more affected by the freezing treatment and storage conditions than by the freezing methods.

Top-down control of flight by a non-canonical cortico-amygdala pathway.

Survival requires the selection of appropriate behaviour in response to threats, and dysregulated defensive reactions are associated with psychiatric illnesses such as post-traumatic stress and panic disorder1. Threat-induced behaviours, including freezing and flight, are controlled by neuronal circuits in the central amygdala (CeA)2; however, the source of neuronal excitation of the CeA that contributes to high-intensity defensive responses is unknown. Here we used a combination of neuroanatomical mapping, in vivo calcium imaging, functional manipulations and electrophysiology to characterize a previously unknown projection from the dorsal peduncular (DP) prefrontal cortex to the CeA. DP-to-CeA neurons are glutamatergic and specifically target the medial CeA, the main amygdalar output nucleus mediating conditioned responses to threat. Using a behavioural paradigm that elicits both conditioned freezing and flight, we found that CeA-projecting DP neurons are activated by high-intensity threats in a context-dependent manner. Functional manipulations revealed that the DP-to-CeA pathway is necessary and sufficient for both avoidance behaviour and flight. Furthermore, we found that DP neurons synapse onto neurons within the medial CeA that project to midbrain flight centres. These results elucidate a non-canonical top-down pathway regulating defensive responses.

Evaluation of the Tensile and Puncture Properties of Geotextiles Influenced by Soil Moisture under Freezing Conditions.

Freezing conditions under different humidity will influence the mechanical properties of geotextiles, leading to the gradual fracture of geotextiles. It brings hidden danger to the whole isolation, reinforcement and protection of rock and soil. It is particularly important to study the tensile and puncture properties of geotextiles considering low temperature and moisture content. In this paper, a series of tensile and puncture tests of geotextiles are performed under different low temperatures (0, -3, -6, -9, and -12 °C) and at different moisture content levels (0, 5, 10, 30, 50, and 80%). From the microscopic perspective, the failure mechanism considering the low temperature and moisture content was explained comprehensively. Experimental results indicate that with a decrease in freezing temperature, the tensile strength of geotextiles increases as a parabolic function while the elongation at failure decreases as an exponential function. Additionally, the puncture strength of geotextiles presented a parabolic increase with the decreasing temperature. Under the freezing temperature environment, the higher moisture content of geotextiles can generate a higher puncture strength increment. This research contributes to a more comprehensive understanding of the tensile and puncture properties of geotextile materials considering low temperature and moisture content. It can provide important guidance for the design of slopes, the reinforcement of earthen dams, and roadbed reinforcement with geotextiles in cold regions.

Wave-absorbing performance of alumina thin-walled hollow particles under freezing condition

The reliability of the absorbing layer is crucial for realizing protective engineering’s protection function. However, the typical wave-absorbing material, sand, is unable to fulfill its intended wave-absorbing function in areas with seasonally frozen soil. This is because the internal pores of the material become filled with ice and the particles freeze. To address this issue, alumina thin-walled hollow particles were chosen as a new wave-absorbing material. These particles can introduce the gas phase into the absorbing layer which is essential for attenuating the stress waves and its wave-absorbing capacity under freezing conditions was investigated by the split Hopkinson bar (SHPB) test. According to the test data, the alumina thin-walled hollow particles are less dense than sand and have a lower wave impedance, allowing them to reflect more incident energy. Moreover, these particles have a better capacity for dissipating the absorbed energy, as compared to sand. Under freezing circumstances, the average transmittance coefficient of alumina thin-walled hollow particles is only 21.95% to 49.30% of ordinary sand. Additionally, the particle size positively correlates with the capacity for wave-absorption. The capacity of alumina thin-walled hollow particles to shatter and release the gas phase under impact stress significantly increases the compressibility of the absorbing layer under freezing conditions, which accounts for their enhanced wave-absorbing effectiveness. The stress-strain curve specifically manifests as a smoother curve and a longer stage of plastic energy dissipation. Other than that, the dynamic deformation modulus of the material and peak stress is lower, while the peak strain is larger. The findings of this study provide a low-cost, high-reliability solution to the problem of frost damage in the absorbing layer in regions with seasonal freezing.

Comprehensive ethological analysis of fear expression in rats using DeepLabCut and SimBA machine learning model.

Defensive responses to threat-associated cues are commonly evaluated using conditioned freezing or suppression of operant responding. However, rats display a broad range of behaviors and shift their defensive behaviors based on immediacy of threats and context. This study aimed to systematically quantify the defensive behaviors that are triggered in response to threat-associated cues and assess whether they can accurately be identified using DeepLabCut in conjunction with SimBA. We evaluated behavioral responses to fear using the auditory fear conditioning paradigm. Observable behaviors triggered by threat-associated cues were manually scored using Ethovision XT. Subsequently, we investigated the effects of diazepam (0, 0.3, or 1 mg/kg), administered intraperitoneally before fear memory testing, to assess its anxiolytic impact on these behaviors. We then developed a DeepLabCut + SimBA workflow for ethological analysis employing a series of machine learning models. The accuracy of behavior classifications generated by this pipeline was evaluated by comparing its output scores to the manually annotated scores. Our findings show that, besides conditioned suppression and freezing, rats exhibit heightened risk assessment behaviors, including sniffing, rearing, free-air whisking, and head scanning. We observed that diazepam dose-dependently mitigates these risk-assessment behaviors in both sexes, suggesting a good predictive validity of our readouts. With adequate amount of training data (approximately > 30,000 frames containing such behavior), DeepLabCut + SimBA workflow yields high accuracy with a reasonable transferability to classify well-represented behaviors in a different experimental condition. We also found that maintaining the same condition between training and evaluation data sets is recommended while developing DeepLabCut + SimBA workflow to achieve the highest accuracy. Our findings suggest that an ethological analysis can be used to assess fear learning. With the application of DeepLabCut and SimBA, this approach provides an alternative method to decode ongoing defensive behaviors in both male and female rats for further investigation of fear-related neurobiological underpinnings.

Physicochemical studies and electrophoretic profile of freeze-dried quail eggs

The aim of the research is a comparative physicochemical and electrophoretic analysis of lyophilized egg white (albumen), egg yolk and whole egg (mélange) from quail eggs, obtained by processing at three different freezing temperatures before lyophilization. The conditions of freezing and the parameters of lyophilization, in which products with the best physicochemical properties are obtained, have been established. The protein content varies from 87.76% (egg white) to 31.09% (yolk), and the amount of lipids is 0.21% and 52.94%, respectively. All egg powders have low residual moisture – from 1.92% to 3.49%. The content of essential amino acids is significantly higher in the lyophilized egg white - 44.07g/100g than in the lyophilized yolk - 16.29g/100g. The results of the electrophoretic analysis (SDS–PAGE) showed a reliable match in the protein profile of the raw and freeze-dried quail eggs. Therefore, the proteins in the egg white and yolk remain unchanged after freeze-drying. This technology is a suitable approach for the utilization of unrealized quail eggs while preserving the composition, nutritional value and beneficial biological qualities. In addition to the functional advantages, freeze-dried egg powders are convenient to transport and have a significantly extended shelf life.

Evaluation of Physicochemical Changes in Hard-Boiled Eggs Stored at Different Temperatures.

Eggs that have been hard-boiled are frequently used as ready-to-eat food. Refrigerated and frozen storage of hard-boiled eggs causes issues, such as customer rejection owing to textural changes. The objective of this research is to ascertain how storage temperature affects hard-boiled eggs' alteration in texture over time. Medium-sized brown shell eggs were acquired from a local market, boiled at 100°C for 15 min, and then stored at room temperature (25°C), refrigeration (4°C), and freezing (-18°C) conditions for 0, 12, 24, and 48 h. Fourier transform infrared spectroscopy (FTIR), texture profile, visual observation using a gemological microscope, free amino acid content, and color were measured. Freezing had a substantial impact on the eggs' hardness, gumminess, chewiness, and cohesiveness (p<0.05). The FTIR spectrums confirmed the textural changes in bonds of amide A (3,271 cm-1), amide I (1,626.2 cm-1), amide II (1,539.0 cm-1), C=O stretch of COO- (1,397 cm-1), asymmetric PO2- stretch (1,240 cm-1). Microscopic images confirmed structural changes in eggs stored at -18°C. The free amino acid content was lower in fresh and frozen eggs than in the rest (p<0.05). However, there was no discernible variation in the egg white's color when eggs were kept at 4°C (p>0.05). Salmonella spp. was found exclusively in eggs kept at room temperature. In conclusion, hard-boiled eggs did not exhibit structural or chemical changes when stored at 4°C for up to 48 h compared to freezing and room temperature conditions.

Characterization of Soybean Events with Enhanced Expression of the Microtubule-Associated Protein 65-1 (MAP65-1).

Microtubule-associated protein 65-1 (MAP65-1) protein plays an essential role in plant cellular dynamics through impacting stabilization of the cytoskeleton by serving as a crosslinker of microtubules. The role of MAP65-1 in plants has been associated with phenotypic outcomes in response to various environmental stresses. The Arabidopsis MAP65-1 (AtMAP65-1) is a known virulence target of plant bacterial pathogens and is thus a component of plant immunity. Soybean events were generated that carry transgenic alleles for both AtMAP65-1 and GmMAP65-1, the soybean AtMAP65-1 homolog, under control of cauliflower mosaic virus 35S promoter. Both AtMAP65-1 and GmMAP65-1 transgenic soybeans are more resistant to challenges by the soybean bacterial pathogen Pseudomonas syringae pv. glycinea and the oomycete pathogen Phytophthora sojae, but not the soybean cyst nematode, Heterodera glycines. Soybean plants expressing AtMAP65-1 and GmMAP65-1 also display a tolerance to the herbicide oryzalin, which has a mode of action to destabilize microtubules. In addition, GmMAP65-1-expressing soybean plants show reduced cytosol ion leakage under freezing conditions, hinting that ectopic expression of GmMAP65-1 may enhance cold tolerance in soybean. Taken together, overexpression of AtMAP65-1 and GmMAP65-1 confers tolerance of soybean plants to various biotic and abiotic stresses. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.