PNIPAAm-based temperature responsive ionic conductive hydrogels for flexible strain and temperature sensing.

Conductive hydrogels have received much attention in the field of flexible wearable sensors due to their outstanding flexibility, conductivity, sensitivity and excellent compatibility. However, most conductive hydrogels mainly focus on strain sensors to detect human motion and lack other features such as temperature response. Herein, we prepared a strain and temperature dual responsive ionic conductive hydrogel (PPPNV) with an interpenetrating network structure by introducing a covalent crosslinked network of N-isopropylacrylamide (NIPAAm) and 1-vinyl-3-butylimidazolium bromide (VBIMBr) into the skeleton of the hydrogel composed of polyvinylalcohol (PVA) and polyvinylpyrrolidone (PVP). The PPPNV hydrogel exhibited excellent anti-freezing properties (-37.34 °C) and water retention with high stretchability (∼930 %) and excellent adhesion. As a wearable strain sensor, the PPPNV hydrogel has good responsiveness and stability to a wide range of deformations and exhibits high strain sensitivity (GF=2.6) as well as fast response time. It can detect large and subtle body movements with good signal stability. As wearable temperature sensors, PPPNV hydrogels can detect human physiological signals and respond to temperature changes, and the volumetric phase transition temperature (VPTT) can be easily controlled by adjusting the molar ratio of NIPAAm to VBIMBr. In addition, a bilayer temperature-sensitive hydrogel was prepared with the temperature responsive hydrogel by two-step synthesis, which shows great promising applications in temperature actuators.

An adhesive, stretchable, and freeze-resistant conductive hydrogel strain sensor for handwriting recognition and depth motion monitoring.

Wearable electronics based on conductive hydrogels (CHs) offer remarkable flexibility, conductivity, and versatility. However, the flexibility, adhesiveness, and conductivity of traditional CHs deteriorate when they freeze, thereby limiting their utility in challenging environments. In this work, we introduce a PHEA-NaSS/G hydrogel that can be conveniently fabricated into a freeze-resistant conductive hydrogel by weakening the hydrogen bonds between water molecules. This is achieved through the synergistic interaction between the charged polar end group (-SO3-) and the glycerol-water binary solvent system. The conductive hydrogel is simultaneously endowed with tunable mechanical properties and conductive pathways by the modulation caused by varying material compositions. Due to the uniform interconnectivity of the network structure resulting from strong intermolecular interactions and the enhancement effect of charged polar end-groups, the resulting hydrogel exhibits 174 kPa tensile strength, 2105 % tensile strain, and excellent sensing ability (GF = 2.86, response time: 121 ms), and the sensor is well suited for repeatable and stable monitoring of human motion. Additionally, using the Full Convolutional Network (FCN) algorithm, the sensor can be used to recognize English letter handwriting with an accuracy of 96.4 %. This hydrogel strain sensor provides a simple method for creating multi-functional electronic devices, with significant potential in the fields of multifunctional electronics such as soft robotics, health monitoring, and human-computer interaction.

Self-assembled high-entropy Prussian blue analogue nanosheets enabling efficient sodium storage.

High entropy material (HEM) has emerged as an appealing material platform for various applications, and specifically, the electrochemical performances of HEM could be further improved through self-assembled structure design. However, it remains a big challenge to construct such high-entropy self-assemblies primarily due to the compositional complexity. Herein, we propose a bottom-up directional freezing route to self-assemble high-entropy hydrosols into porous nanosheets. Taking Prussian blue analogue (PBA) as an example, the simultaneous coordination-substitution reactions yield stable high-entropy PBA hydrosols. During subsequent directional freezing process, the anisotropic growth of ice crystals could guide the two-dimensional confined assembly of colloidal nanoparticles, resulting in high-entropy PBA nanosheets (HE-PBA NSs). Thanks to the high-entropy and self-assembled structure design, the HE-PBA NSs manifests markedly enhanced sodium storage kinetics and performances in comparison with medium/low entropy nanosheets and high entropy nanoparticles.

Biopsy vitrification: New tool for endometrial tissue cryopreservation for research applications.

Patient-derived endometrial biopsies serve as a crucial source for molecular studies, highlighting the necessity for tissue cryopreservation methods that preserve cell viability and tissue morphology with minimal to no impact. The passive slow freezing (PSF) protocol has demonstrated efficacy for cryopreserving endometrial biopsies, allowing for the subsequent isolation of viable epithelial and stromal cells. Vitrification (VT) enables the avoidance of ice crystal formation and could therefore potentially prevent mechanical injury to tissues. In this study, PSF and VT techniques were applied to endometrial biopsies, and the effects of cryopreservation on tissue samples were evaluated using traditional histology. In addition, transmission electron microscopy (TEM), gene expression profiling analyses, the viability of endometrial cells, and the ability to form epithelial organoids were compared between PSF and VT endometrial biopsies in a subset of samples. The histology and TEM studies demonstrated relatively mild cellular and sub-cellular damage in both cryopreservation protocols which did not affect tissue functionality and the formation of the organoids. Additionally, the cryopreservation methodology did not affect the gene expression profile of the 68 endometrial-receptivity associated genes studied. In conclusion, our findings indicate that although current cryopreservation methodologies need further improvements, they still allow us to achieve acceptable cell viability and functionality, showing promising potential for facilitating the utilization of cryopreserved endometrial tissue samples for research purposes.

Integrative indexes reveal the tolerance of winter wheat to different overwinter freezing injury.

Winter wheat (Triticum aestivum L.) is a crucial crop that guarantees food supply in the North China Plain (NCP). As the frequency of extreme cold events increases, it is necessary to explore the freezing resistance of different wheat varieties in order to clarify planting boundaries and help with risk assessment. In this study, 2-year controlled experiments were conducted to explore the effect of freezing temperatures (T air) and freezing durations on three winterness types. A set of indexes were used to characterize the subfreezing stress on wheat tiller, leaf, and final yield. Logistical regressions were used to quantify the temperature threshold for 10%, 30%, and 50% of freezing injury. The results showed that the lower temperature threshold of tiller (LT) varied from -9.6 to -15.9°C, -10.7 to -19.1°C and -11.4 to -21.2°C for LT10, LT30, and LT50, respectively. The difference between LT and yield loss (YL) indexes reduced with decreased winterness types and was -0.1 to 3.4°C, -0.7 to 2.1°C, and 0.3 to 0.9°C higher compared with YL thresholds for winterness, semi-winterness, and weak-winterness types, respectively. The average minimum soil temperature was 7.5, 4.8, and 4.2°C higher than T air for 1-, 2-, and 3-day treatment, respectively. Soil effective negative accumulated temperature hours (TSEh) ranged from 6.9 to 12.0, 48.4 to 6.9, and 84.7 to 106.9°C·h for 10%, 30%, and 50% tiller mortality, respectively. Freezing treatment with T air < -12, -9, and -8°C obviously decreased leaf Fv/Fm for the three varieties and Fv/Fm declined obviously after 5 days of recovery under field conditions. Our results provided multiple indexes for quantifying subfreezing damage in practical wheat production and could shed light on future risk assessment.

Comprehensive evaluation of freezing tolerance in prickly ash and its correlation with ecological and geographical origin factors.

Low temperatures are a key factor affecting the growth, development, and geographical distribution of prickly ash. This study investigated the impact of ecological and geographical factors on the freezing tolerance of prickly ash germplasm. Thirty-seven germplasm samples from 18 different origins were collected, and their freezing tolerance was comprehensively evaluated. The correlation between freezing tolerance and the ecological and geographical factors of their origins was also analyzed. Significant differences in freezing tolerance were observed among germplasm from different origins. The semi-lethal temperature of the germplasm ranged from - 12.37 to 1.08 °C. As temperatures decreased, the relative conductivity (REC) and catalase (CAT) activity of the germplasm gradually increased, while soluble sugar (SS), soluble protein (SP), free proline (Pro), and Peroxidase (POD) activities decreased and then increased. Superoxide dismutase (SOD) activity initially increased and then decreased. A comprehensive evaluation of freezing tolerance was conducted using a logistic equation, membership function, and cluster analysis. Germplasm from Tongchuan and Hancheng (Shaanxi Province, China), Asakura (Japan), and Yuncheng (Shanxi Province, China) exhibited the highest freezing tolerance, whereas those from Rongchang (Chongqing Municipality, China), Qujing (Yunnan Province, China), and Honghe (Yunnan Province, China) had the lowest. The correlation analysis revealed a significant positive correlation between freezing tolerance and latitude, and a significant negative correlation with the temperature of origin. Germplasm from higher latitudes showed higher SS content, SOD and CAT activities, stronger antioxidant enzyme activity, and better freezing tolerance compared to those from lower latitudes. REC was lower in germplasm originating from low-temperature areas than in those from high-temperature areas. Additionally, SP, Pro content, SOD, and POD activities were higher, indicating effective scavenging of active oxygen free radicals. No significant correlation was found between altitude and longitude of origin and freezing tolerance. However, at similar latitudes, prickly ash from higher altitudes displayed higher antioxidant enzyme activity and stronger freezing tolerance compared to those from lower altitudes. These findings provide a scientific basis for breeding prickly ash cultivars suited to different ecological regions.

A simple and practical approach to elective egg freezing to control costs and expand access to care.

Social elective egg freezing (EEF) is now widely used globally but in many countries is unaffordable to many women because of high costs and lacking insurance coverage. Efforts to reduce costs, therefore, are of importance. Surprisingly, a simple, well-defined and practical approach ensuring optimal outcomes for EEF has, however, so-far not been published. We, therefore, conducted a narrative review of the literature for relevant articles regarding the different steps of ovarian stimulation (OS) in the EEF process, in order to define such a standard protocol. This review revealed that in order to maximize oocyte yields with minimal number of OS cycles - while ensuring patient safety - a multiple-dose GnRH antagonist protocol with a daily gonadotropin dose of 300 IU appears best, unless patients demonstrate a polycystic ovarian phenotype, suggestive of likely high responses. The initial gonadotropin should be recFSH, while LH supplementation should be co-administered with the addition of GnRH antagonist. Final follicular maturation should be triggered by GnRH agonist trigger, with a dual trigger (1000-1500 IU hCG) considered for suboptimal responders to GnRH agonist trigger, optionally with Cabergoline to mitigate ovarian hyperstimulation syndrome (OHSS) in high responders.

Greenhouse gas emissions from Boreal Lakes: Highlighting the impact of salinity and freezing period on emission dynamics.

Lakes and ponds in boreal regions are considerable natural sources of greenhouse gases (GHGs), including carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O). Although the seasonal variability of GHG emissions from boreal lakes is crucial for improving global emission models, emissions during the freezing period have not received sufficient attention. Focusing on two representative boreal lakes in China-Ulansuhai and Daihai-this study investigated variations in GHG emissions during both the non-freezing and freezing periods. The concentrations of CO2 and CH4 in lake porewater during the non-freezing period were observed to be 50 to 74 times higher than those during the freezing period. In both lakes, CO2 and CH4 emissions predominantly occurred at the water-air interface, with N2O absorption. The Global Warming Potential (GWP) of GHGs in Ulansuhai was 234.35×104 kg/yr, with CO2, CH4, and N2O contributing 12.0 %, 87.4 %, and 0.6 %, respectively. In Daihai, the GWP was 40.47×103 kg/yr, with CO2 CH4, and N2O contributing 40.4 %, 24.5 %, and 35.1 %, respectively. Notably, the GHG 'storage' capacities of Ulansuhai and Daihai were 227.51 × 105 kg/yr and 9.23 × 102 kg/yr, respectively. In both lakes, dissolved organic carbon and total nitrogen in the porewater exhibited a negative relationship with GHG concentrations. Compared to lake Ulansuhai, salinity exhibited a stronger correlation with GHGs in lake Daihai, which has high salinity. Our research reveals that the freezing period and the salinity (in high salinity lakes) have distinct impacts on GHG emissions in boreal lakes. The findings are crucial for understanding the contributions of boreal lakes to GHG emissions and their potential impact on climate change, and provide vital information for developing conservation and management strategies regarding these ecosystems.

Effects of astaxanthin supplementation during vitrification and liquid nitrogen vapor freezing on motility, morphology, survival, reactive oxygen species (ROS), and DNA fragmentation of post-cryopreserved human sperm.

OBJECTIVE: To investigate the effect of astaxanthin supplementation in cryopreservation media on post-thawed sperm motility, viability, morphology, reactive oxygen species (ROS), and DNA fragmentation in two cryopreservation techniques using vitrification and liquid nitrogen vapor freezing. METHODS: Thirty normozoospermic semen samples were used in the study. Post-prepared semen samples were divided into 1) non-cryopreserved control, 2) and 3) vitrified without (V) and with astaxanthin 0.5 µM (V+ATX), 4) and 5) frozen in liquid nitrogen vapor without (L) and with astaxanthin 0.5 µM (L+ATX). RESULTS: Cryopreservation using vitrification and liquid nitrogen vapor freezing significantly decreased sperm motility and viability and increased ROS levels. However, no changes were seen in sperm morphology or DNA fragmentation. The addition of astaxanthin in cryopreservation media significantly increased post-thawed motility in both vitrification (77.6±8.9% vs. 69.0±9.5% in V+ATX and V) and vapor freezing (57.0±13.3% vs. 47.7±14.6% in L+ATX and L); it significantly increased sperm viability in vitrification (75.0±11.9% vs. 65.9±11.1% in V+ATX and V), and significantly decreased ROS level in both vitrification (4.7 (2.6-8.3) RLU/sec/106 vs. 10.6 (9.4-16.0) RLU/sec/106 in V+ATX and V) and vapor freezing (4.6 (3.3-10.5) RLU/sec/106 vs. 10.3 (7.9-18.6) RLU/ sec/106 in L+ATX and L). Astaxanthin supplementation in cryopreservation media did not affect sperm morphology or DNA fragmentation. CONCLUSIONS: Astaxanthin supplementation improved post-cryopreserved sperm motility, decreased ROS levels in both vitrification and liquid nitrogen vapor freezing and improved sperm viability only in the vitrification technique.

Distribution of mercury and methylmercury in ice-water-sediments in lakes during the freezing period under the influence of ice cover.

The presence of lake ice cover alters the subglacial water environment, thereby influencing the migration and transformation of mercury (Hg) and methylmercury (MeHg) within the ice-water-sediment media of lakes. This study investigated the occurrence characteristics of mercury and methylmercury in various environmental compartments within lakes located at high latitudes in cold regions during the freezing period. To this end, Wuliangsuhai Lake, the largest freshwater lake situated at 40°N in China, was selected as the study site. The contents of mercury and methylmercury in lake ice were determined for the first time. The percentage of methylmercury (MeHg%) and ice-water partition coefficient were analyzed. The pollution situation and health risk were evaluated by single factor pollution index. The results show that the ice body and water body of Wuliangsuhai are not polluted by mercury and methylmercury, but some sampling points in the sediment are slightly polluted. The mercury content in sediment is negatively correlated with the ice thickness, and the methylmercury content in water is positively correlated with the methylmercury content in sediment, but negatively correlated with the ice thickness. The migration ability of methylmercury in ice-water system is stronger than that of mercury. The MeHg% of water in ice period is higher than that in non-freezing period, which is different from other lakes without ice sheet. The results show that in the dynamic equilibrium of methylation and demethylation in the high-latitude lake water, the methylation is higher in the ice period than in the non-freezing period due to the influence of light intensity, while the mercury in the non-freezing period is more susceptible to the demethylation.

Knowledge, Attitudes, and Perceptions of Women of Reproductive Age Regarding Fertility and Elective Oocyte Cryopreservation: A Study from the Al-Qassim Region.

BACKGROUND: Oocyte cryopreservation is used for fertility preservation, medical reasons, and social reasons to overcome physiological decline in fertility with age. There was no research to find the knowledge, attitude, and perception of women regarding oocyte cryopreservation in our region. This study thus aimed to investigate the knowledge, attitudes, and perceptions of women of reproductive age in Al-Qassim, Saudi Arabia, regarding fertility and elective oocyte cryopreservation. METHODS: An Arabic-language online survey was conducted over six months targeting women aged 18-45 years in the Al-Qassim region. A total of 612 participants completed the questionnaire, which covered topics such as reproductive aspirations, knowledge of fertility, knowledge of elective oocyte cryopreservation, and concerns regarding fertility preservation. Data analysis was performed using IBM SPSS Statistics for Windows, Version 21.0 (Released 2012; IBM Corp., Armonk, New York, United States). RESULTS: The majority of participants (47.1%) were aged 18-25 years. Participants' perceptions towards fertility preservation and social egg freezing were mixed, indicating varying levels of awareness about fertility issues. A statistically significant relationship was found between education level and knowledge of oocyte cryopreservation (p-value=0.004). Women with higher education levels (bachelor's, master's, and PhD degrees) demonstrated significantly greater knowledge (p-value=0.004) about oocyte cryopreservation compared to those with lower education levels. The study highlights the influence of educational attainment on knowledge of oocyte cryopreservation. CONCLUSION: These findings align with broader trends in health education, suggesting that higher educational attainment correlates with better fertility health literacy. Enhancing fertility health literacy through educational programs may lead to more proactive health behaviours and better awareness about available assisted reproductive technologies.

Cryoprotective effect of wheat starch granular surface proteins on frozen HMW and LMW glutenins: Structure, property and functionality across length scales.

Although frozen dough technology has demonstrated significant benefits, the mechanisms underlying dough deterioration during freezing remain unclear. To overcome this obstacle, the effect of freezing-induced deteriorations of wheat starch granular surface proteins (SGSPs)-high/low molecular weight (HMW/LMW) glutenins complexes were analyzed from the molecular to macroscopic scales. After 7 cycles of freezing/thawing treatment, SGSPs-LMW complex showed a higher antifreeze stability than SGSPs-HMW complex. The freezable water content of SGSPs-HMW increased from 32 % to 39 %, indicating a marked migration and recrystallization of ice. In this situation, the interactions of SGSPs-HMW complex were affected and destabilized, leading to partially denatured and depolymerized molecular structures. Furthermore, the bulk protein aggregation network was also dissociated under the ice tearing and splitting, which irreversibly collapsed to small molecular protein particles. In comparison, the resistance of SGSPs-LMW complex on continued network disruption appear to be the key to maintain the quality of frozen dough.

Auditory stimuli suppress contextual fear responses in safety learning independent of a possible safety meaning.

Safety learning allows the identification of non-threatening situations, a learning process instrumental for survival and psychic health. In contrast to fear learning, in which a sensory cue (conditioned stimulus, CS) is temporally linked to a mildly aversive stimulus (US), safety learning is studied by presenting the CS and US in an explicitly unpaired fashion. This leads to conditioned inhibition of fear responses, in which sensory cues can acquire a safety meaning (CS-). In one variant of safety learning, an auditory CS- was shown to reduce contextual fear responses during recall, as measured by freezing of mice. Here, we performed control experiments to test whether auditory stimuli might interfere with freezing by mechanisms other than safety learning, a phenomenon also called external inhibition. Surprisingly, when auditory stimulation was omitted during training (US-only controls), such stimuli still significantly suppressed contextual freezing during recall, indistinguishable from the reduction of freezing after regular safety training. The degree of this external inhibition was positively correlated with the levels of contextual freezing preceding the auditory stimulation. Correspondingly, in fear learning protocols which employ a new context during recall and therefore induce lower contextual freezing, auditory stimuli did not induce significant external inhibition. These experiments show that in safety learning protocols that employ contextual freezing, the freezing reduction caused by auditory stimuli during recall is dominated by external inhibition, rather than by learned safety. Thus, in safety learning experiments extensive controls should be performed to rule out possible intrinsic effects of sensory cues on freezing behavior.

Effect of chronic alcohol exposure and single-prolonged stress on conditioned fear behavior.

The present study investigated the impact of chronic intermittent ethanol (CIE) exposure and single-prolonged stress (SPS) on the acquisition of fear memories in both male and female Wistar rats. Adult rats were first subjected to CIE by vapor inhalation followed by SPS. Following a subsequent 8-day incubation period, the rats underwent a Pavlovian fear conditioning procedure (tone-shock pairings) followed by cued-tone extinction training, and then testing of extinction recall memory and fear renewal memory. In control animals that had not been exposed to either CIE or SPS, female rats exhibited significantly lower levels of freezing compared to male rats during tone-shock pairings. This lower level of freezing in female rats during conditioning was associated with an increased speed of movement compared to males. Also compared to males, female rats exhibited lower levels of fear extinction, recall, and renewal. Exposure to CIE, SPS, or CIE+SPS had no effect on freezing during the cued-conditioning, extinction, or extinction recall phases of the testing procedure in either sex. In fear renewal, CIE exposure decreased freezing in male but not female rats, while SPS increased freezing in female but not male rats. CIE exposure significantly reduced freezing during the fear renewal phase. Taken together, these results provide further evidence that male and female rats adopt different avoidance strategies for threat responding. These results also revealed that prior exposure to CIE, SPS, or CIE+SPS had minimal effects on threat responding using conditioned freezing as an indicator of fear responsivity.

Boostering motor imagery processing to improve gait in patients with Parkinson disease and freezing of gait: A pilot study.

BACKGROUND: Given that patients with Parkinson's Disease (PD) and Freezing of Gait (FoG) may lack the cognitive resources necessary to activate the motor imagery (MI) process, investigating how to boost MI vividness and accuracy could be a valuable therapeutic strategy in MI Practice (MIP). OBJECTIVE: We aim to evaluate the priming effect of visual, or auditory, or attentional stimuli in enhancing MI ability by using quantitative data on gait and turning performance. METHODS: Nineteen PD participants with FoG underwent four one-week sessions of MIP, with pre and post clinical assessments. Each session included MI alone or one of three booster MI tasks (Attentional, Action observation, or Auditory) before imagining and executing walking straight and performing a 180° turn. Gait and turning performances were evaluated using six inertial sensors before and after each session. RESULTS: Our findings showed that both MI and boosted MI induced similar improvement in gait (speed and stride length) and 180° (step number and velocity) and 360° turning (velocity, angle) parameters compared to baseline. When differences among "booster" tasks were analyzed, results showed that Auditory and Attentional boosted MI were superior to MI alone in some gait and turning parameters. At the end of the 4 sessions, MI ability measured by means of Kinesthetic and Visual Imagery Questionnaire and Gait Imagery Questionnaire and FoG symptoms were also improved. CONCLUSION: Our preliminary results suggest that boosting MI is a feasible strategy for enhancing MI ability and addressing FoG symptoms. Auditory and Attentional conditions appear to enhance the priming effect of MI on gait and turning performance more effectively.

Weighing the risks and benefits of flowering early in the spring for the woody perennial Prunus pumila.

PREMISE: There are advantages to flowering early in the spring, including greater pollinator fidelity and longer fruit maturation time. But plant phenology has advanced in recent years, making many plants vulnerable to freezing damage from late frosts. METHODS: To determine the costs and benefits of flowering early in the growing season, we exposed Prunus pumila plants to two freezing treatments and a delayed flowering treatment in subsequent years. Data were collected on ovary swelling, fruit production, and pollinator visitation on hand- and open-pollinated plants in all treatments. We also measured tissue damage after freeze events. RESULTS: Our results suggest that flowering time and temperature affect reproductive success, with fewer fruits produced after hard freezes. The same was not true for light freezes, which had minimal impact on reproduction. Freezing damage to plants after a hard freeze did affect the number of dipteran pollinators but not the overall pollinator visitation rate. Despite the clear impact of freezing temperatures on plant reproduction, flowering early provided an advantage in that reproductive output decreased with delayed flowering. CONCLUSIONS: Our findings suggest that Prunus pumila will retain the ability to attract pollinators and produce viable seeds if exposed to false spring conditions that involve a light freeze, but hard freezes may reduce yield by an order of magnitude. Although the advantages to flowering early may outweigh the risk of freezing damage under current conditions, it is possible that flower viability may be constrained under continued climate warming.

Histological outcome evaluation of selected brain preparation protocols for white fiber dissection.

BACKGROUND: White fiber dissection is essential for studying brain connections.　However, preparation protocols have not been validated. METHODS: We microstructurally analyzed Klingler's brain preparation method and freezing process and assessed changes under two protocols:freeze-only and freeze-thaw.　The microstructure changes of these protocols were evaluated by measuring the ratio of the total gap area to the white matter area and determining the mean eccentricity value to assess the degree of anisotropy. RESULTS: Sixty hemispheres were allocated to ten different freezing protocols.　In the freeze-only protocols, the total gap area ratio was significantly higher compared to that of specimens fixed with only formaldehyde, particularly after continuous freezing for 3-4 weeks;however, the difference in eccentricity was not significant.　In the freeze-thaw protocols, both the area ratio and eccentricity were significantly higher compared to the freeze-only.　The optimum degree of fiber separation in the freeze-thaw protocols reached its peak with four cycles of 1-week freezing periods interrupted by six hours of thawing. CONCLUSION: The Klingler method assists in the separation of the white fibers through the gaps formed by ice crystals, but an appropriate degree of anisotropy is reached when the freezing protocol is interrupted by at least four thawing cycles.

Use of electrolyte leakage to assess floral damage after freezing.

Premise: With growing interest in the impact of false springs on plant reproduction, there is the need to develop reliable, high-throughput methods for assessing floral freezing damage. Here we present a method for use with floral tissue that will facilitate more comparative work on floral freezing tolerance in the future. Methods and Results: We examined the effectiveness of a modified electrolyte leakage protocol to assess floral freezing damage. By comparing data from temperature response curves to an estimate of visual tissue damage, we optimized the protocol for different floral types and improved the signal-to-noise ratio for floral data. Conclusions: Our modified protocol provides a quick and straightforward method for quantifying floral freezing damage that can be standardized across floral types. This method allows for cross-species comparisons and can be a powerful tool for studying broad patterns in floral freezing tolerance.

An Exceptionally Salt Tolerant Copoly(Maleimide Sulfobetaine) - Structural Requirements for Ultra-Salt Tolerance.

Zwitterionic polymers are an important class of polymers with far-ranging applications. In the widely studied poly(meth)acrylate and poly(meth) acrylamide-based zwitterions, properties can be tuned by changing the nature of substituents attached to ammonium ions. However, these changes influenced salt tolerance of zwitterionic polymers only to a limited extent. Upon adding salt these polymers expanded in solution initially. Further increase in salt concentration caused the polymer chains to shrink similar to the common water soluble, uncharged polymers thereby deteriorating the viscosity of aqueous solutions. In contrast to the conventional poly(meth)acrylate and poly(meth)acrylamide-based zwitterions, zwitterionic copolymaleimides showed substituent dependent salt-tolerant nature. In the absence of any substituent on the polymer backbone such as zwitterionic poly(ethylene-alt-maleimide) (ZI-PEMA) the viscosity of salt solutions increased both with the increasing salt concentration as well as the concentration of polymer. This is likely due to the continuous expansion of polymer coil in salt solutions with increasing salt concentration caused primarily by the rigidity of the polymer backbone. ZI-PEMA also enhanced the saturation limit of mono- and divalent salts like sodium chloride and hydrated calcium bromide in water. This property is useful for various applications like fish curing, for making high-density fluids, refrigeration, etc. across various industrial sectors.

Controlled Ice Nucleation With a Sand-PDMS Film Device Enhances Cryopreservation of Mouse Preantral Ovarian Follicles.

Ovarian follicle cryopreservation is a promising strategy for fertility preservation; however, cryopreservation protocols have room for improvement to maximize post-thaw follicle viability and quality. Current slow-freezing protocols use either manual ice-seeding in combination with expensive programmable-rate freezers or other clinically incompatible ice initiators to control the ice-seeding temperature in the extracellular solution, a critical parameter that impacts post-cryopreservation cell/tissue quality. Previously, sand has been shown to be an excellent, biocompatible ice initiator, and its use in cryopreservation of human induced pluripotent stem cells enables high cell viability and quality after cryopreservation. This study applies sand as an ice initiator to cryopreserve multicellular microtissue, preantral ovarian follicles, using a simple slow-freezing protocol in the mouse model. Ovarian follicles cryopreserved using the sand partially embedded in polydimethylsiloxane (PDMS) film to seed ice in the extracellular solution exhibit healthy morphology, high viability, and the ability to grow similarly to fresh follicles in culture post-thaw. This sand-based cryopreservation strategy can facilitate convenient ovarian follicle cryopreservation using simple equipment, and this study further demonstrates the translatability of this strategy to not only single cells but also multicellular tissues.