In vitro Hemostatic Functions of Cold-Stored Platelets.

Background: Transfusion of platelets is a life-saving medical strategy used worldwide to treat patients with thrombocytopenia as well as platelet function disorders. Summary: Until the end of 1960s, platelets were stored in the cold because of their superior hemostatic functionality. Cold storage of platelets was then abandoned due to better posttransfusion recovery and survival of room temperature (RT)-stored platelets, demonstrated by radioactive labeling studies. Based on these findings, RT became the standard condition to store platelets for clinical applications. Evidence shows that RT storage increases the risk of septic transfusion reactions associated with bacterial contamination. Therefore, the storage time is currently limited to 4-7 days, according to the national guidelines, causing a constant challenge to cover the clinical request. Despite the enormous efforts made to optimize storage conditions of platelets, the quality and efficacy of platelets still decrease during the short storage time at RT. In this context, during the last years, cold storage has seen a renaissance due to the better hemostatic functionality, reduced risk of bacterial contamination, and potentially longer storage time. Key Messages: In this review, we will focus on the impact of cold storage on the in vitro platelet functions as promising alternative storage temperature for future medical applications.

Donor Lung Preservation at 10°C: Clinical and Logistical Impact.

INTRODUCTION: Cold static donor lung preservation at 10°C appears to be a promising method to safely extend the cold ischemic time (CIT) and improve lung transplant (LTx) logistics. METHODS: LTx from November 2021 to February 2023 were included in this single institution, prospective, non-randomized study comparing prolonged preservation at 10°C versus standard preservation on ice. The inclusion criteria for 10°C preservation were suitable grafts for LTx without any donor retrieval concerns. PRIMARY ENDPOINT: primary graft dysfunction (PGD) grade-3 at 72-h. Secondary endpoints: clinical outcomes, cytokine profile and logistical impact. RESULTS: Thirty-three out of fifty-seven cases were preserved at 10°C. Donor and recipient characteristics were similar across the groups. Total preservation times (h:min) were longer (p<0.001) in the 10°C group [1st lung: median 12:09 (IQR 9:23-13:29); 2nd: 14:24 (12:00-16:20)] vs. standard group [1st lung: median 5:47 (IQR 5:18-6:40); 2nd: 7:15 (6:33-7:40)]. PGD grade-3 at 72-h was 9.4% in 10°C group vs. 12.5% in standard group (p=0.440). Length of mechanical ventilation (MV), ICU and hospital stays were similar in both groups. Thirty and ninety-day mortality rates were 0% in 10°C group (vs. 4.2% in standard group). IL-8 concentration was significantly higher 6-h post-LTx in the standard group (p=0.025) and IL-10 concentration was increased 72-h post-LTx in the 10°C group (p=0.045). CONCLUSIONS: Preservation at 10°C may represent a safe and feasible strategy to intentionally prolong the CIT. In our center, extending the CIT at 10°C may allow for semi-elective LTx and improve logistics with similar outcomes compared to the current standard preservation on ice.

Preparation and Performance Study of n-Undecane Phase Change Cold Storage Material.

With the fast development of the cold chain transportation industry, the traditional refrigeration method results in significant energy consumption. To address the national call for energy saving and emission reduction, the search for a new type of energy storage material has already become a future development trend. According to the national standard GB/T28577 for the classification and basic requirements of cold chain logistics, the temperature in frozen logistics is typically below -18 °C. In this study, n-undecane with a phase change temperature of -26 °C is chosen as the core material of microcapsules. Poly(methyl methacrylate) is applied as the shell material, with n-undecane microcapsules being prepared through suspension polymerization for phase change cold storage materials (MEPCM). Using characterization techniques including SEM, DSC, FTIR, and laser particle size analysis, the effects of three types of emulsifiers (SMA, Tween-80, Tween-80/span-80 (70/30)), SMA emulsifier dosage, core-shell ratio, and emulsification rate on the thermal performance and micro-surface morphology of n-undecane/PMMA microcapsules were studied. The results indicate that when comparing SMA, Tween-80, and Tween-80/span-80 (70/30) as emulsifiers, the dodecane/PMMA microcapsules prepared with SMA emulsifier exhibit superior thermal performance and micro-surface morphology, possessing a complete core-shell structure. The optimal microstructure and the highest enthalpy of phase change, measuring 120.3 kJ/kg, are achieved when SMA is used as the emulsifier with a quantity of 7%, a core-to-wall ratio of 2.5:1, and an emulsification speed of 2000 rpm. After 200 hot and cold cycles, the enthalpy of phase change decreased by only 18.6 kJ/kg, indicating the MEPCM thermal performance and cycle life. In addition, these optimized microcapsules exhibit favorable microstructure, uniform particle size, and efficient energy storage, making them an excellent choice for the refrigeration and freezing sectors.

Review for special issue: Corneal lamellar surgery: Present outcomes and future perspectives.

Since the establishment of the first eye bank in the 1940s, their role has evolved to face new challenges. With the recent development of lamellar keratoplasties, eye banks play an even bigger role in the selection and preparation of donor tissues. The increasing number of keratoplasty techniques and the high demand for "ready-to-use" tissues are challenging eye banks to improve and develop new preparation techniques. Besides necessary examinations, new approaches of tissue analysis in eye banks allow a better/optimized selection of corneal tissues. These new challenges in tissue preservation, preparation, and selection are propelling eye banks into a new era of modern eye banking.

Warmer autumns and winters could reduce honey bee overwintering survival with potential risks for pollination services.

Honey bees and other pollinators are critical for food production and nutritional security but face multiple survival challenges. The effect of climate change on honey bee colony losses is only recently being explored. While correlations between higher winter temperatures and greater colony losses have been noted, the impacts of warmer autumn and winter temperatures on colony population dynamics and age structure as an underlying cause of reduced colony survival have not been examined. Focusing on the Pacific Northwest US, our objectives were to (a) quantify the effect of warmer autumns and winters on honey bee foraging activity, the age structure of the overwintering cluster, and spring colony losses, and (b) evaluate indoor cold storage as a management strategy to mitigate the negative impacts of climate change. We perform simulations using the VARROAPOP population dynamics model driven by future climate projections to address these objectives. Results indicate that expanding geographic areas will have warmer autumns and winters extending honey bee flight times. Our simulations support the hypothesis that late-season flight alters the overwintering colony age structure, skews the population towards older bees, and leads to greater risks of colony failure in the spring. Management intervention by moving colonies to cold storage facilities for overwintering has the potential to reduce honey bee colony losses. However, critical gaps remain in how to optimize winter management strategies to improve the survival of overwintering colonies in different locations and conditions. It is imperative that we bridge the gaps to sustain honey bees and the beekeeping industry and ensure food and nutritional security.

Changes in the bacterial communities of Harmonia axyridis (Coleoptera: Coccinellidae) in response to long-term cold storage and progressive loss of egg viability in cold-stored beetles.

Bacteria have a profound influence on life history and reproduction of numerous insects, while the associations between hosts and bacteria are substantially influenced by environmental pressures. Cold storage is crucial for extending the shelf life of insects used as tools for biological control, but mostly causes detrimental effects. In this study, we observed a great decrease in egg hatch rate of cold-stored Harmonia axyridis during the later oviposition periods. Furthermore, most eggs produced by their F1 offspring exhibited complete loss of hatchability. We hypothesized that long-term exposure to cold may greatly alter the bacterial community within the reproductive tracts of H. axyridis, which may be an important factor contributing to the loss of egg viability. Through sequencing of the 16S rRNA gene, we discovered considerable changes in the bacterial structure within the reproductive tracts of female cold-stored beetles (LCS_F) compared to non-stored beetles (Control_F), with a notable increase in unclassified_f_Enterobacteriaceae in LCS_F. Furthermore, in accordance with the change of egg hatchability, we observed a slight variation in the microbial community of eggs produced by cold-stored beetles in early (Egg_E) and later (Egg_L) oviposition periods as well as in eggs produced by their F1 offspring (Egg_F1). Functional predictions of the microbial communities revealed a significant decrease in the relative abundance of substance dependence pathway in LCS_F. Moreover, this pathway exhibited relatively lower abundance levels in both Egg_L and Egg_F1 compared to Egg_E. These findings validate that long-term cold storage can greatly modify the bacterial composition within H. axyridis, thereby expanding our understanding of the intricate bacteria-insect host interactions.

Ultraviolet irradiation as alternative non-thermal cold pasteurization to improve quality and microbiological parameters of mango juice during cold storage.

The objectives of this research were to study the effect of UV irradiation on quality characteristics of mango juice during cold storage. Mango juice exposed to UV radiation was also used to determine zero-order and first-order kinetic models of microbial (total plate count, yeast and mold count, and Escherichia coli) reduction. According to the microbiological results, UV light at 120 J/cm2 caused a 5.19 log reduction. It was found that microbial inactivation of all tested microorganisms followed first-order kinetic model. The treatments did not differ significantly in terms of the quality metrics. L*, b*, pH, total soluble solid, total phenolic compound, total flavonoid content, and antioxidant activity as measured by the DPPH and FRAP assay all tended to decline during storage at 4 °C, whereas a*, ∆E, titratable acidity, total plate count, yeast and mold count, as well as the total plate count, had an increasing trend. During storage at 4 °C, UV irradiation increased the shelf life of mango juice by about 14 days compared to the control sample. In conclusion, this study demonstrated the potential of UV treatment as an alternative to thermal pasteurization for preserving mango juice quality and safety while also prolonging shelf life.

High-concentration bovine serum albumin enhances fertilization ability of cold-stored rat sperm.

Cold transport of the cauda epididymides is a useful technique for shipping laboratory rat sperm. Cold transport of rat sperm avoids potential risks of microbiological infection, animal escape or death, and animal welfare issues. Previously, we reported that a cold-storage solution containing dimethyl sulfoxide and quercetin maintained the fertility of cold-stored rat sperm. However, cold-stored rat sperm exhibited a decreased fertilization rate after 24-h storage. To recover the fertility of cold-stored sperm, we focused on the effects of bovine serum albumin (BSA), a cholesterol acceptor that induces sperm capacitation. We sought to determine the optimal concentration of BSA in fertilization medium based on the fertility of cold-stored rat sperm. High concentrations of BSA (40 mg/ml) enhanced the fertilization rate of cold-stored rat sperm and maintained sperm fertility for 144 h. Embryos derived from cold-stored and BSA-treated sperm normally developed into pups after embryo transfer. In summary, high BSA concentrations enhanced the fertility of cold-stored rat sperm and prolonged the storage period to 144 h, thereby expanding the transportable region for genetically engineered rats.

The Predictive Value of Graft Viability and Bioenergetics Testing Towards the Outcome in Liver Transplantation.

Donor organ biomarkers with sufficient predictive value in liver transplantation (LT) are lacking. We herein evaluate liver viability and mitochondrial bioenergetics for their predictive capacity towards the outcome in LT. We enrolled 43 consecutive patients undergoing LT. Liver biopsy samples taken upon arrival after static cold storage were assessed by histology, real-time confocal imaging analysis (RTCA), and high-resolution respirometry (HRR) for mitochondrial respiration of tissue homogenates. Early allograft dysfunction (EAD) served as primary endpoint. HRR data were analysed with a focus on the efficacy of ATP production or P-L control efficiency, calculated as 1-L/P from the capacity of oxidative phosphorylation P and non-phosphorylating respiration L. Twenty-two recipients experienced EAD. Pre-transplant histology was not predictive of EAD. The mean RTCA score was significantly lower in the EAD cohort (-0.75 ± 2.27) compared to the IF cohort (0.70 ± 2.08; p = 0.01), indicating decreased cell viability. P-L control efficiency was predictive of EAD (0.76 ± 0.06 in IF vs. 0.70 ± 0.08 in EAD-livers; p = 0.02) and correlated with the RTCA score. Both RTCA and P-L control efficiency in biopsy samples taken during cold storage have predictive capacity towards the outcome in LT. Therefore, RTCA and HRR should be considered for risk stratification, viability assessment, and bioenergetic testing in liver transplantation.

Thyroid hormone protects human lung epithelial cells from cold preservation and warm reperfusion-induced injury.

BACKGROUND: Cellular stress associated with static-cold storage (SCS) and warm reperfusion of donor lungs can contribute to ischemia-reperfusion (IR) injury during transplantation. Adding cytoprotective agents to the preservation solution may be conducive to reducing graft deterioration and improving post-transplant outcomes. METHODS: SCS and warm reperfusion were simulated in human lung epithelial cells (BEAS-2B) by exposing cells to low potassium dextran glucose solution at 4 °C for different periods and then switching back to serum-containing culture medium at 37 °C. Transcriptomic analysis was used to explore potential cytoprotective agents. Based on its results, cell viability, caspase activity, cell morphology, mitochondrial function, and inflammatory gene expression were examined under simulated IR conditions with or without thyroid hormones (THs). RESULTS: After 18 h SCS followed by 2 h warm reperfusion, genes related to inflammation and cell death were upregulated, and genes related to protein synthesis and metabolism were downregulated in BEAS-2B cells, which closely mirrored gene profiles found in thyroid glands of mice with congenital hypothyroidism. The addition of THs (T3 or T4) to the preservation solution increases cell viability, inhibits activation of caspase 3, 8 and 9, preserves cell morphology, enhances mitochondrial membrane potential, reduces mitochondrial superoxide production, and suppresses inflammatory gene expression. CONCLUSION: Adding THs to lung preservation solutions may protect lung cells during SCS by promoting mitochondrial function, reducing apoptosis, and inhibiting pro-inflammatory pathways. Further in vivo testing is warranted to determine the potential clinical application of adding THs as therapeutics in lung preservation solutions.

Impact of controlled hypothermic preservation on outcomes following heart transplantation.

BACKGROUND: Severe primary graft dysfunction (PGD) is a major cause of early mortality after heart transplant, but the impact of donor organ preservation conditions on severity of PGD and survival has not been well characterized. METHODS: Data from US adult heart-transplant recipients in the Global Utilization and Registry Database for Improved Heart Preservation-Heart Registry (NCT04141605) were analyzed to quantify PGD severity, mortality, and associated risk factors. The independent contributions of organ preservation method (traditional ice storage vs controlled hypothermic preservation) and ischemic time were analyzed using propensity matching and logistic regression. RESULTS: Among 1,061 US adult heart transplants performed between October 2015 and December 2022, controlled hypothermic preservation was associated with a significant reduction in the incidence of severe PGD compared to ice (6.6% [37/559] vs 10.4% [47/452], p = 0.039). Following propensity matching, severe PGD was reduced by 50% (6.0% [17/281] vs 12.1% [34/281], respectively; p = 0.018). The Kaplan-Meier terminal probability of 1-year mortality was 4.2% for recipients without PGD, 7.2% for mild or moderate PGD, and 32.1%, for severe PGD (p < 0.001). The probability of severe PGD increased for both cohorts with longer ischemic time, but donor hearts stored on ice were more likely to develop severe PGD at all ischemic times compared to controlled hypothermic preservation. CONCLUSIONS: Severe PGD is the deadliest complication of heart transplantation and is associated with a 7.8-fold increase in probability of 1-year mortality. Controlled hypothermic preservation significantly attenuates the risk of severe PGD and is a simple yet highly effective tool for mitigating post-transplant morbidity.

The Impact of Rootstock on "Big Top" Nectarine Postharvest Concerning Chilling Injury, Biochemical and Molecular Parameters.

Peaches and nectarines have a short shelf life even when harvested at appropriate physiological maturity. Market life is increased by storage at low temperatures. However, chilling injury symptoms can appear, causing physiological disorders and limiting shipping potential. The rootstock effect on the post-harvest quality has hardly been explored. Thus, the principal aim of this work was to study the influence of seven different Prunus rootstocks on the "Big Top" nectarine cv, considering harvest and post-harvest quality parameters and their correlation with chilling injury disorders. Basic fruit quality traits, individual sugars and organic acids analyzed by HPLC and other biochemical compounds such as relative antioxidant capacity, total phenolics content, flavonoids, anthocyanins, vitamin C and related enzyme activities (PAL, POD, PPO) were considered. In addition, correlations with possible candidate genes for chilling injury (CI) tolerance were searched by qPCR. Although a low susceptibility to CI symptoms has been found in "Big Top", rootstocks "PADAC 9902-01", "PADAC 99-05" and "ReplantPAC" exhibited lower CI symptoms. A statistically significant influence of the evaluated rootstocks was found concerning the parameters of this study. Phenols and anthocyanins seem to be important parameters to be considered in the prevention of chilling injury disorders. Moreover, PAL1, PPO4, PG2 and LDOX genes relative expressions were positively associated with chilling injury susceptibility. This study opens new perspectives for understanding peach fruit adaptation and response to cold storage temperatures during the post-harvest period.

Validity of cold storage and heat treatment on the deactivation of Vibrio parahaemolyticus isolated from fish meat markets.

Vibrio parahaemolyticus is a zoonotic disease transmitted to humans when handling or consuming improperly cooked fish meat. This study aimed to evaluate the effect of thermal treatment on V. parahaemolyticus isolates. Different heat treatment methods are used to determine the best methods for controlling V. parahaemolyticus, isolated from fish meat, which include microwave, low-temperature long-time, and high-temperature short-time methods. The V. parahaemolyticus isolates significantly declined in bacteria count when they were kept at 4°C, and 25°C for a long time, and the V. parahaemolyticus isolates significantly declined in bacteria count manner when they were kept at -20°C for a long time. The high temperature and long-time exposure at 75°C/25 minutes by moist heat, 87°C/5 minutes by dry heat, and 70°C/20 minutes by frying heat were enough to kill V. parahaemolyticus isolates. This work can be useful to decrease the hazards of infections related to V. parahaemolyticus and reduce the causes of fishborne pathogens.

Impact of the Temperature Reconditioning of Cold-Stored Potatoes on the Color of Potato Chips and French Fries.

The effect of temperature reconditioning on cold-stored potato tubers was investigated for three popularly consumed potato varieties (Shangi, Unica, and Dutch robjin) grown in Kenya. The potatoes were stored at 4 °C for 30 days, followed by removal and storage at 22 ± 3 °C for 9 days during which changes in sugar concentration were evaluated every two days. In parallel, potato chips and French fries were processed, and their colors were determined. The results showed that sugar content decreased significantly with increasing reconditioning time. The relative decrease in fructose content was the highest (p < 0.05) in Dutch robjin (57.49%), followed by Shangi (49.22%) and Unica (38.18%). Glucose content decreased by 54.1% in Dutch robjin, 49.5% in Shangi, and 50.8% in Unica. The lightness (L*) of French fries and chips increased significantly (p < 0.05) with reconditioning time while the redness (a*) values decreased significantly (p < 0.05) across all varieties. The correlation between lightness and the total reducing sugar content of the potatoes was r < -0.93, indicating a strong negative correlation for both products. The coefficient of determination showed that the glucose content of the tubers accounted for 80.5-97.6% of the lightness of French fries and 88.4-94.2% for potato chips. The critical glucose content range for acceptable products in French fries and chips based on the color (L* and a*) values was 12-22 mg/100g and 8-14 mg/100g, respectively, for the varieties in this study.

The Potential of 3D Printing in Thermal Insulating Composite Materials-Experimental Determination of the Impact of the Geometry on Thermal Resistance.

This paper focuses on the analysis of the thermal properties of prototype insulation structures produced using SLS and SLA additive technologies. There is a noticeable lack of analysis in the scientific literature regarding the geometry of 3D-printed structures in terms of their thermal properties. The aim of this paper was to analyze printed samples of prototype thermal insulation composite structures and their potential for use in building applications. The research material consisted of closed and open cell foams of varying structural complexity. Increasing the complexity of the composite core structure resulted in a statistically significant decrease in the value of the thermal conductivity coefficient λ and the heat transfer coefficient U, and an increase in the thermal resistance Rc. The experimental results showed that the geometric structure of the air voids in the material is a key factor in regulating heat transfer. The control of porosity in materials produced by additive technology can be an effective tool for designing structures with high insulation efficiency. The best performance of the prototype materials produced by the SLS method was a three-layer cellular composite with a gyroid core structure. It was also shown that the four-layer gyroid structure panels with an outer layer of metallized polyethylene film produced using 3D SLA printing had the best thermal insulation. As a result, the analysis confirmed the possibility of producing energy-efficient insulation materials using 3D printing. These materials can be used successfully in construction and other industries. Further research will significantly improve the quality, accuracy, and speed of printing insulation materials, reduce the negative impact on the natural environment, and develop intelligent adaptive solutions.

Normal Proteasome Function Is Needed to Prevent Kidney Graft Injury during Cold Storage Followed by Transplantation.

Kidney transplantation is the preferred treatment for end-stage kidney disease (ESKD). However, there is a shortage of transplantable kidneys, and donor organs can be damaged by necessary cold storage (CS). Although CS improves the viability of kidneys from deceased donors, prolonged CS negatively affects transplantation outcomes. Previously, we reported that renal proteasome function decreased after rat kidneys underwent CS followed by transplantation (CS + Tx). Here, we investigated the mechanism underlying proteasome dysfunction and the role of the proteasome in kidney graft outcome using a rat model of CS + Tx. We found that the key proteasome subunits ß5, α3, and Rpt6 are modified, and proteasome assembly is impaired. Specifically, we detected the modification and aggregation of Rpt6 after CS + Tx, and Rpt6 modification was reversed when renal extracts were treated with protein phosphatases. CS + Tx kidneys also displayed increased levels of nitrotyrosine, an indicator of peroxynitrite (a reactive oxygen species, ROS), compared to sham. Because the Rpt6 subunit appeared to aggregate, we investigated the effect of CS + Tx-mediated ROS (peroxynitrite) generation on renal proteasome assembly and function. We treated NRK cells with exogenous peroxynitrite and evaluated PAC1 (proteasome assembly chaperone), Rpt6, and ß5. Peroxynitrite induced a dose-dependent decrease in PAC1 and ß5, but Rpt6 was not affected (protein level or modification). Finally, serum creatinine increased when we inhibited the proteasome in transplanted donor rat kidneys (without CS), recapitulating the effects of CS + Tx. These findings underscore the effects of CS + Tx on renal proteasome subunit dysregulation and also highlight the significance of proteasome activity in maintaining graft function following CS + Tx.

Evaluating the Effects of Kidney Preservation at 10 °C with Hemopure and Sodium Thiosulfate in a Rat Model of Syngeneic Orthotopic Kidney Transplantation.

Kidney transplantation is preferred for end-stage renal disease. The current gold standard for kidney preservation is static cold storage (SCS) at 4 °C. However, SCS contributes to renal graft damage through ischemia-reperfusion injury (IRI). We previously reported renal graft protection after SCS with a hydrogen sulfide donor, sodium thiosulfate (STS), at 4 °C. Therefore, this study aims to investigate whether SCS at 10 °C with STS and Hemopure (blood substitute), will provide similar protection. Using in vitro model of IRI, we subjected rat renal proximal tubular epithelial cells to hypoxia-reoxygenation for 24 h at 10 °C with or without STS and measured cell viability. In vivo, we preserved 36 donor kidneys of Lewis rats for 24 h in a preservation solution at 10 °C supplemented with STS, Hemopure, or both followed by transplantation. Tissue damage and recipient graft function parameters, including serum creatinine, blood urea nitrogen, urine osmolality, and glomerular filtration rate (GFR), were evaluated. STS-treated proximal tubular epithelial cells exhibited enhanced viability at 10 °C compared with untreated control cells (p < 0.05). Also, STS and Hemopure improved renal graft function compared with control grafts (p < 0.05) in the early time period after the transplant, but long-term function did not reach significance. Overall, renal graft preservation at 10 °C with STS and Hemopure supplementation has the potential to enhance graft function and reduce kidney damage, suggesting a novel approach to reducing IRI and post-transplant complications.

Refrigerated storage and cryopreservation of hormonally induced sperm in the threatened frog, Litoria aurea.

As sperm cryopreservation and other assisted reproductive technologies (ARTs) advance in common amphibian species, focus on applying non-lethal sperm collection methods to the conservation and genetic management of threatened species is imperative. The goal of this study was to examine the application of logistically practical ART protocols in a threatened frog (Litoria aurea). First, we tested the efficacy of various concentrations of human chorionic gonadotropin (hCG) (20, 40 IU/g bodyweight) and Gonadotropin releasing hormone antagonist (0.25 µg/g and 0.5 µg/g body weight GnRH-a) on the induction of spermatozoa. Using the samples obtained from the previous trials, we tested the effect of cold storage and cryopreservation protocols on long-term refrigerated storage and post-thaw sperm recovery. Our major findings include: (1) high quality sperm were induced with 20 and 40 IU/g bodyweight of (hCG); (2) proportions of live, motile sperm post-thaw, were recovered at higher levels than previously reported for L. aurea (>50%) when preserved with 15% v/v DMSO and 1% w/v sucrose; and (3) spermic urine stored at 5 °C retained motility for up to 14 days. Our findings demonstrate that the protocols developed in this study allowed for successful induction and recovery of high-quality spermatozoa from a threatened Australian anuran, L. aurea, providing a prime example of how ARTs can contribute to the conservation of rare and threatened species.

Variations of the fungal microbiome in Corydalis Rhizoma with different collection areas, processing methods, and storage conditions.

Corydalis Rhizoma (CR, Yanhusuo in Chinese) has been widely used as an analgesic in herbal medicine and functional food. Cases of fungal and mycotoxin contamination in CR have been reported. In this study, the composition and diversity of fungal microbiome in CR samples from four herbal markets and two processing methods were investigated by DNA metabarcoding. Variations of the fungal microbiome in CR during cold and conventional storage were monitored. Results showed that Aspergillus was the dominant genus and saprotroph was the dominant trophic mode. Six potential toxigenic fungi, namely, Aspergillus fumigatus, Aspergillus ostianus, Aspergillus terreus, Penicillium citrinum, Penicillium oxalicum, and Trichothecium roseum, were detected. Differences in fungal composition and diversity among various groups based on collection areas and processing methods were also observed. Moreover, the relative abundance of dominant genera in CR samples stored at different temperatures was significantly different and changed with storage time. This study is the first to reveal the influence of collection areas, processing methods, and storage conditions on the fungal microbiome in CR, which was expected to provide a basis for control strategies of fungal contamination in the industrial chain of CR.

3-O-Methyl-D-Glucose Blunts Cold Ischemia Damage in Kidney via Inhibiting Ferroptosis.

BACKGROUND: The glucose derivative 3-O-methyl-D-glucose (OMG) is used as a cryoprotectant in freezing cells. However, its protective role and the related mechanism in static cold storage (CS) of organs are unknown. The present study aimed to investigate the effect of OMG on cod ischemia damage in cold preservation of donor kidney. METHODS: Pretreatment of OMG on kidney was performed in an isolated renal cold storage model in rats. LDH activity in renal efflux was used to evaluate the cellular damage. Indicators including iron levels, mitochondrial damage, MDA level, and cellular apoptosis were measured. Kidney quality was assessed via a kidney transplantation (KTx) model in rats. The grafted animals were followed up for 7 days. Ischemia reperfusion (I/R) injury and inflammatory response were assessed by biochemical and histological analyses. RESULTS: OMG pretreatment alleviated prolonged CS-induced renal damage as evidenced by reduced LDH activities and tubular apoptosis. Kidney with pCS has significantly increased iron, MDA, and TUNEL+ cells, implying the increased ferroptosis, which has been partly inhibited by OMG. OMG pretreatment has improved the renal function (p <0.05) and prolonged the 7-day survival of the grafting recipients after KTx, as compared to the control group. OMG has significantly decreased inflammation and tubular damage after KTx, as evidenced by CD3-positive cells and TUNEL-positive cells. CONCLUSION: Our study demonstrated that OMG protected kidney against the prolonged cold ischemia-caused injuries through inhibiting ferroptosis. Our results suggested that OMG might have potential clinical application in cold preservation of donor kidney.