Preservation Period Research Articles

The Effect Of Fresh Human Amniotic Membrane Storage Time On Growth Factor Levels (EGF, TGF-B, bFGF) in Freeze-Dried Human Amniotic Membranes

Background: Each preservation method can cause different levels of damage to tissue components and can affect the regenerative performance of the amniotic membrane. The concentrations of most of the cytokines were found to decrease during the preservation period. There are still no studies regarding the effect of fresh human amnion storage time on growth factor levels in freeze dried human amnion products. Method: This study used an experimental post test group design. Result: There was no significant difference in EGF levels of freeze-dried human amniotic membrane products resulting from 3 types of fresh human amniotic membrane storage (1 week storage with 8.49 ± 5.16 pg/mL, 3 months with 4.55 ± 1.98 pg/mL, and 9 months with levels of 4.04 ± 2.12 pg/mL). There was no significant difference in bFGF values between freeze-dried human amniotic membrane products resulting from 3 types of storage duration of fresh human amniotic membrane (1 week storage with levels of 97.86 ± 113.98 pg/mL, 3 months with levels of 109.69 ± 113.39 pg/mL, and 9 months with levels of 117.53 ± 73.46 pg/mL). There was a significant difference in the levels of TGF-b, namely in the product from storage for 1 week (97.93 ± 18.27 pg/mL) and 3 months (55.94 ± 28.67 pg/mL). Whereas TGF-b levels in freeze dried human amniotic membrane from the results of fresh human amniotic membrane storage at 1 week and 9 months, and 3 months and 9 months were found to have no significant difference. Conclusion: There was no significant difference in EGF and bFGF levels in this study. There was a significant difference in the levels of TGF-b freeze-dried human amniotic membrane produced from fresh human amniotic membrane with higher growth factor levels at 1 week of storage compared to 3 months.

СОСТОЯНИЕ ЖЕНСКОЙ ГЕНЕРАТИВНОЙ СФЕРЫ СОСНЫ ОБЫКНОВЕННОЙ В ДЕЙСТВУЮЩЕМ ОЧАГЕ КОРНЕВОЙ ГУБКИ (HETEROBASIDION ANNOSUM (FR.) BREF.)

Evaluation of the manifestation of Scots pine trees reaction to the weakening effect of Heterobasidion annosum (Fr.) Bref. is relevant from the perspective of the potential opportunities for further natural regeneration of the forest. The main reason for the decrease in the biological stability of the pine plantation was the development of the root sponge focus. Over the three-year period 2017-2019 the weighted average tree health category in the sample area increased from 2.21 to 2.52. The morphometric parameters of cones turned out to be more stable in the female generative sphere. Greater variability was noted for full-grain seed yield and the number of sterile apex scales. Relatively high preservation of seed ovules was found in the first and second vegetative periods of development within the boundaries of average values. For trees of different status categories during the whole period of observations it was 70.60-81.77 and 79.07-88.35% respectively, which indicated sufficient quality of pollination and marked compatibility of male and female genotypes against the background of reduced viability of mother trees. Embryo survival in the experiment reached 63.44-81.48%. The sensitivity of the generative sphere of Scots pine to the stress caused by the development of the disease has been proved. A significant decrease in the average length and diameter of mature cones and a relative increase in female reproduction rates were confirmed with the weakening and dying out of pine trees infected with root sponge. The possibility of identifying separate periods of preservation of ovules during gametophytic and embryonic cycles and the annual regularity of seed production allow us to recommend Scots pine for monitoring the state of the female generative sphere and assess the potential of the species to survive the action of aggressive stressors, such as Heterobasidion annosum (Fr.) Bref.

Postharvest treatments to optimize long-term storage of cut peony flowers

Cut peonies have limited time during which they can be harvested and commercialized. Interest in extending this period through long-term storage, both in dry and wet conditions, to obtain more profits is constantly growing. Sixty cut peony stems (cv. Alertie) were pulse-treated 24 h after harvest with 10 μM thidiazuron, 500 μL L-1 1-methylcyclopropene, 10 mM glycerol, and a combination of them (All) in a refrigerated cell at 4 °C, where the flowers were stored with wet preservation for 15 and 30 days. After these periods, flowers were maintained at 20 °C, and their quality was assessed through in vivo and destructive analyses at 0, 4, 12, and 14 days. Thirty days of storage strongly inhibited flower opening, in contrast to the shorter preservation period. After 15 days of storage and during preservation at room temperature, flowers treated with glycerol showed less water loss and senescence symptoms. Nitrate and phenol concentrations increased in all treatments after storage. In conclusion, wet storage at 4 °C for 15 days was more suitable for the cultivar tested in the present study and it also allowed the identification of treatments that involved the use of glycerol as promising for long-term preservation of peonies and their vase-life.

Preparation of a novel glucose oxidase-N-succinyl chitosan nanospheres and its antifungal mechanism of action against Colletotrichum gloeosporioides

In this work, a new glucose oxidase-N-succinyl chitosan (GOD-NSCS) nanospheres was prepared through the immobilization of glucose oxidase (GOD) on N-succinyl chitosan (NSCS) nanospheres. Compared to the free GOD, GOD-NSCS nanospheres demonstrated the excellent anti-Colletotrichum gloeosporioides activity with the EC50 values of 211.2 and 10.7 μg/mL against mycelial growth and spores germination. The computational biology analysis demonstrated that the substrate presented the similar binding free energy with GOD-NSCS nanospheres (−27.64 kcal/mol) compared with the free GOD (−24.04 kcal/mol), indicating that GOD-NSCS nanospheres had the same oxidation efficiency and produced more H2O2. Moreover, the enzyme activity stability of GOD-NSCS nanospheres could be prolonged to 10 d. The cell membrane was destructed by the treatment of H2O2 produced by GOD, leading to the cell death. In vivo test, GOD-NSCS nanospheres treatment significantly prolonged the preservation period of mangoes 2-fold. Collectively, these results suggested that GOD-NSCS nanospheres suppresses anthracnose in postharvest mangoes by inhibiting the growth of C. gloeosporioides and might become a potential natural preservative for fruits and vegetables.

Divya Iratchakar’s Pillai Thamizh

Just as the Pallavar period is said to be the period of devotional literature and the medieval Chola period is said to be the period of preservation, so the period of Nayak who ruled Tamil Nadu from 1350 AD can be said to be the period of minor literature. Pallu, Ula, Thootu, Kalambagam, Pillaitamil etc. are minor literary works that are written to entertain the rich people and kings. There are 96 types of minor literary work called Prabandhams. In the book Sathuragarathi written by Veeramamunivar, mentions 96 types of Prabandhams. Pillai Thamizh ia also known as Pilliakavi, Pillaipaatu, and Seithamil. Some of these Pillai Thamizh books have been composed with changes in the structural and cultural elements. Thus, the article examines the overall changes that took place in Divya Iratchakar’s Pillai Thamizh.

Evaluation of 10°C as the optimal storage temperature for aspiration-injured donor lungs in a large animal transplant model.

Our recent work has challenged 4°C as an optimal lung preservation temperature by showing storage at 10°C to allow for the extension of preservation periods. Despite these findings, the impact of 10°C storage has not been evaluated in the setting of injured donor lungs. Aspiration injury was created through bronchoscopic delivery of gastric juice (pH: 1.8). Injured donor lungs (n=5/group) were then procured and blindly randomized to storage at 4°C (on ice) or at 10°C (in a thermoelectric cooler) for 12 hours. A third group included immediate transplantation. A left lung transplant was performed thereafter followed by 4 hours of graft evaluation. After transplantation, lungs stored at 10°C showed significantly better oxygenation when compared to 4°C group (343 ± 43 mm Hg vs 128 ± 76 mm Hg, p=0.03). Active metabolism occurred during the 12 hours storage period at 10°C, producing cytoprotective metabolites within the graft. When compared to lungs undergoing immediate transplant, lungs preserved at 10°C tended to have lower peak airway pressures (p=0.15) and higher dynamic lung compliances (p=0.09). Circulating cell-free mitochondrial DNA within the recipient plasma was significantly lower for lungs stored at 10°C in comparison to those underwent immediate transplant (p=0.048), alongside a tendency of lower levels of tissue apoptotic cell death (p=0.075). We demonstrate 10°C as a potentially superior storage temperature for injured donor lungs in a pig model when compared to the current clinical standard (4°C) and immediate transplantation. Continuing protective metabolism at 10°C for donor lungs may result in better transplant outcomes.

Effects of different preservation methods of human iliac veins.

With the progress of vascular anastomosis technology, the radical resection surgery of cancer combining with vascular resection and reconstruction has been focused by surgeon. As a natural substitute material for blood vessel, vascular allografts have good vascular compliance and histocompatibility. Generally, the donated veins could not be used immediately, and need to be well preserved. So, it is greatly significant to do research in the preservation effects of different preservation methods on veins. In this study, the effects of different preservative methods of human iliac veins were compared and analyzed in terms of cell viability, vascular wall structure and tension resistance. The donated human iliac veins were randomly divided into three groups: Cold Storage Group (4 °C) (CSG), Frozen Storage Group (-186 °C) (FSG)and Fresh Control Group (FCG). Six detection time-points of preservation for 1, 3, 5, 7, 14, 28days were set respectively. There are ten samples in each group and each time-point separately. Survival and apoptosis of vascular cell were evaluated by MTT assay and Tunel fluorescence staining. Tensile test was used to evaluate mechanical properties of vessels. The changes of vascular endothelial cells, smooth muscle cells, collagen fibers and elastic fibers were evaluated by HE staining, Masson staining and EVG staining. Furthermore, the changes of organelles were observed by transmission electron microscope. With the extension of preservation period, the vascular cell viability and tension resistance of two groups decreased, and the apoptotic cells increased gradually. The apoptosis index of CSG was higher than FSG at each time point (P < 0.05). In terms of cell viability, CSG was higher within 3days (P < 0.05), both groups were same between 3 and 14days, and then CSG lower than FSG after 14days (P < 0.05). In terms of tension resistance, CSG was stronger than FSG (P < 0.05) in first 7days, both groups were same in 2nd week, and then CSG was weaker in 4th week (P < 0.05). In terms of vascular wall structure, in CSG, vascular endothelial cells were damaged and shed, smooth muscle cells were edema after 14days, but the cell membrane and intercellular connection were still intact. In 4th week, endothelial cells were completely damaged and shed, the boundary of smooth muscle cell membrane was unclear, intercellular connection was damaged. Moreover, organelles were destroyed and disappeared, perinuclear condensation of chromatin was observed, and some cells had incomplete nuclear membrane or nuclear fragmentation; However, there were no obvious changes in the FSG within 28days. Finally, local exfoliation and destruction of endothelial cells and edema-like changes of organelles were observed; the collagen fibers and elastic fibers of blood vessels in the two groups had no obvious damage and change within 28days. For excised human iliac vein, cold and frozen storage can effectively preserve the cell viability, wall structure and tension resistance of blood vessels. With the extension of preservation time, the related performance of vessels declined in varying degrees. Within first week, the effect of cold storage is better than frozen storage, but frozen storage is significantly better than cold storage after 2weeks.

Prolonged Controlled Oxygenated Rewarming Improves Immediate Tubular Function and Energetic Recovery of Porcine Kidneys During Normothermic Machine Perfusion.

Normothermic machine perfusion (NMP) is typically performed after a period of hypothermic preservation, which exposes the kidney to an abrupt increase in temperature and intravascular pressure. The resultant rewarming injury could be alleviated by gradual rewarming using controlled oxygenated rewarming (COR). This study aimed to establish which rewarming rate during COR results in the best protective effect on renal rewarming injury during subsequent NMP. Twenty-eight viable porcine kidneys (n = 7/group) were obtained from a slaughterhouse. After these kidneys had sustained 30 min of warm ischemia and 24 h of oxygenated HMP, they were either rewarmed abruptly from 4-8 °C to 37 °C by directly initiating NMP or gradually throughout 30, 60, or 120 min of COR (rate of increase in kidney temperature of 4.46%/min, 2.20%/min, or 1.10%/min) before NMP. Kidneys that were rewarmed during the course of 120 min (COR-120) had significantly lower fractional excretion of sodium and glucose at the start of NMP compared with rewarming durations of 30 min (COR-30) and 60 min (COR-60). Although COR-120 kidneys showed superior immediate tubular function at the start of normothermic perfusion, this difference disappeared during NMP. Furthermore, energetic recovery was significantly improved in COR-30 and COR-120 kidneys compared with abruptly rewarmed and COR-60 kidneys. This study suggests that a rewarming rate of 1.10%/min during COR-120 could result in superior immediate tubular function and energetic recovery during NMP. Therefore, it may provide the best protective effect against rewarming injury.

THE ROLE AND IDENTITY OF LATVIAN EXILE ART IN THE UNITED STATES OF AMERICA: EXAMPLE OF THE STUDENT FRATERNITY “ DZINTARZEME ” OF THE ART ACADEMY OF LATVIA

The year 1944/45 is inscribed on the pages of Latvia’s history as the time of the second occupation by the USSR, as a result hundreds of thousands of Latvian citizens left Latvia as refugees. Since the 1945 the development of contemporary Latvian art was tragically divided between the occupied native land and the free world. The period in exile from 1945 to 1952 can be called the “restoration period”, or the period of preservation of Latvian national values and art, when the continuation of the form of artistic expression during the period of Latvia’s independence took place in refugee camps in Germany. The issue of national art became more problematic after 1952, when the exile lasted and Latvian artists were scattered on several continents of the world, the question of the fate of the expression of Latvian national art became more topical. In November 1958, with an art exhibition in New York (USA), the Art Academy of Latvia student fraternity “Dzintarzeme” (“Amberland”), which was banned in July 1940 by the USSR, was renewed. There is reason to assume that “Dzintarzeme” can be called as one of the most purposeful associations of Latvian artists in exile in the efforts to preserve and popularize Latvian national art in the USA until 1973. The main purpose of this research is through the example of student fraternity “Dzintarzeme” to clarifiy the concept – what is Latvian national art in exile, what we can define as Latvianness in Latvian exile art, and also look at art development and its role in the society of exile Latvians in the United States of America where the most active community of “Dzintarzeme” was located.

Bioactive metabolite profile and antioxidant properties of brown juice, a processed alfalfa (Medicago sativa) by-product

Recently, leaf protein concentrate (LPC) has gained increased attention in response to the constantly growing protein demand. Green biorefineries can become more economical by valorizing their by-products and reducing environmental risks. The current study describes the variations in the antioxidant capacity and phytochemical composition of a liquid by-product (referred to as brown juice (BJ)) obtained during the extraction of leaf protein concentrate (LPC) from the fresh biomass of alfalfa (Medicago sativa L.). Four varieties of alfalfa were investigated during three harvest times, i.e., August 2017 (first harvest), September 2017 (second harvest), and June 2018 (third harvest). Also, the fresh BJ was lacto-fermented to extend its preservation period but also modifying its composition. The results of different general phytochemical composition analyses and antioxidant assays revealed similar tendencies across different alfalfa varieties and harvest times. Most of the phytochemicals in the BJ identified by HPLC-MS/MS can be classified as flavonoids/flavonoid derivatives, e.g., apigenin, naringenin, luteolin, formononetin. Substantially, the lacto-fermentation process induced a switch into aglycones, e.g., apigenin content increased by an order of magnitude, while apigenin-7-O-glucuronide content was halved after lacto-fermentation. Additionally, several B vitamins were detected, including B2, B3, and B7. These results could provide a basis for various ways of industrial valorization but need to be strengthened by data generated from large-scale production.

Insights on the potential of natural deep eutectic solvents (NADES) to fine-tune durian seed gum for use as edible food coating

Approximately 75,000 tonnes of durian seeds are disposed to the environment every year and the amount will continue to increase due to the escalating demand for durians. The long-term accumulation of the wastes will lead to destruction of the ecological environment, and the disposal requires considerable effort. In fact, valuable products including durian seed gum and flour could be processed from the discarded durian seeds and hence, they should not be regarded as wastes. At the same time, the value of food wasted every year exceeds 1 trillion USD, of which 20% is caused by storage or shelf-life issues. After the outbreak of COVID-19, consumers tend to reduce the number of trips required to get fresh foods from the markets. Eventually, storage of larger amount of foods at extended period of time is crucial. Thus, safe and effective food preservation materials extracted from these biomasses could potentially help in reducing food wastes. Natural deep eutectic solvent (NADES) is a green solvent composed of primary metabolites (PRIM). When combined with gum into the form of eutectogel, the characteristics are tunable indicating that the properties of natural durian seed gum can be further improved for specific application. This paper discusses the potential of eutectogels formulated using NADES and durian seed gum for use as food coating . Since the coated gel is edible, potentially low in cost, and strongly sustainable, the food preservation period can be lengthened at higher commercial value. • Eutectogel can be formulated using durian seed gum and natural deep eutectic solvents. • Eutectogel produced by deep eutectic solvents is tunable based on specific requirements. • Eutectogel is highly promising for use as edible gum-based food coatings. • Computational tools play an important role in the formulation of NADES and eutectogel.

Hydrocolloid edible coatings extend shelf life, reduce postharvest decay, and maintain keeping quality of mango fruits (Mangifera indica L.) under ambient storage.

Mango fruit exhibit high postharvest losses due to physiological, biochemical, and pathological deterioration during storage. Edible coatings such as hydrocolloids (HC) bear promising potential for fruit quality preservation at ambient storage due to its triple action (physiological, biochemical and pathological) on fruit and thus widely researched in recent years. This study demonstrates the influence of health and eco-safe hydrocolloid edible coatings such as "Carboxymethyl cellulose" (CMC) (1%), "Guar gum" (1.5%), "Gum Arabica" (10%), and "Xanthan gum" (0.3%) as dip treatment to enhance the postharvest quality and storage life of mangoes at ambient storage (25 ± 4°C and 65 ± 5% RH). "Xanthan gum" (0.3%) treatment exhibited the highest efficacy in reducing the decay loss by more than threefold and physiological loss by twofold over control fruit. It lowered the physiological and fruit softening enzyme activities (PG, PME, and LOX), while maintaining the biochemicals. Moreover, it maintained both internal as well as external (consumer preference) quality of fruit and extended 6 days shelf life on the physiological loss standard basis (≤10%) than that of the control. The results recommend the application of "Xanthan gum" (0.3%) as an efficacious ecological, sustainable, and health-friendly surface edible coating for quality preservation and storage period extension of mango fruit under ambient storage. PRACTICAL APPLICATIONS: The selected hydrocolloid edible coatings dip treatment showed promising potential in controlling the physiological, biochemical, and pathological deterioration of mango fruit stored under ambient condition. The selected treatments extended the shelf life without diminishing fruit quality. However, among the attempted HC treatments, the "Xanthan gum" (0.3%) (XG) coating displayed the excellent results. It added the storage life of mango fruit by 6 days over the control. XG treated fruit displayed the excellent results in terms of storage period extension, quality retention, consumer preference, and control over the fruit decay and softening enzymes activities. Postharvest preservation of mango fruit using HC is nonchemical, cost-effective approach which is GRAS (generally recognized as safe), health, and eco safe.

A simple culture technique of Rhodobacter azotoformans EBN-7 for public use: application to NH4+-N removal in shrimp aquaculture water

Photosynthetic bacteria (PSB) attract considerable interest as useful microorganisms; nevertheless, a generalized culture technique has not been previously reported owing to difficulty in their cultivation. Therefore, a simple culture technique suitable for public use was investigated. Among the PSB tested, the strain Rhodobacter azotoformans EBN-7 was the most suitable for scaleup production because it showed the highest specific growth rate (0.20 h−1) on basal medium. In scale-up cultivation (500 L), R. azotoformans EBN-7 showed 4.50 × 1010 colony-forming units mL−1 (number of viable cells), dry cell weight of 26.8 g/L, and a specific growth rate of 0.15 h−1. Cultivation using this final culture broth (as seed culture) in a 15 L simple reactor was successful, with maintenance of cell activity evident. For use as seed culture, the maximum allowable preservation period of R. azotoformans EBN-7 at 4°C was 3 months. When R. azotoformans EBN-7 cultivated in a simple technique was applied to shrimp aquaculture water, NH4+-N was reduced from 0.61 mg/L to 0.24 mg/L (by 60.7%) in 4 days in comparison with the control. Thus, this simple culture technique using R. azotoformans EBN-7 has the potential for a good removal efficiency of NH4+-N, making seed culture easier and suitable for public use.

Dynamic and Static Switching in ITO/SnOx/ITO and Its Synaptic Application.

The attempts to devise networks that resemble human minds are steadily progressing through the development and diversification of neural networks (NN), such as artificial NN (ANN), convolution NN (CNN), and recurrent NN (RNN). Meanwhile, memory devices applied on the networks are also being studied together, and RRAM is the one of the most promising candidates. The fabricated ITO/SnOX/TaN device showed two forms of current–voltage (I-V) curves, classified as dynamic and static. It was triggered from the forming process, and the difference between the two curves resulted from the data retention measured at room temperature for 103 s. The dynamic curve shows a time-dependent change in the data, and the cause of the data preservation period was considered through X-ray photoelectron spectroscopy (XPS) and linear fitting in conduction mechanisms. To confirm whether the memory performance of the device may be implemented on the synapse, the change in the plasticity was confirmed using a rectangular-shaped pulse. Paired-pulse facilitation (PPF) was implemented, and the change from short-term potentiation (STP) to long-term potentiation (LTP) was achieved.

P12.06: Effect of Supplementing Taurine to the Developed Lowtemperature Islet Preservation Solution

Introduction: Pancreatic islet transplantation has recently emerged as one of the most promising therapeutic approaches for improving glycemic control in type 1 diabetes patients. However, one of the problems with islet transplantation is that it is impossible to culture the isolated islet for extended periods to while recipients are selected, tested and prepared for surgery. To address this problem, we developed the islet preservation solution, and to improve the function of the preservation solution, we added taurine and conducted a preservation experiment. Taurine is antioxidant activity and it is not a classical free radical scavenger. Therefore, its mechanism remains unclear but it controls osmotic pressure. Our hypothesis is that taurine supplementation can improve islet recovery and enhance islet function after low temperature preservation. Methods: CMRL 1066 medium (Corning) was used as a control group, and a comparison group was prepared some compounds OPTI(Optipharm presevation) solution (with taurine) and OPTI-T solution (without taurine). To confirm the preservation effect of these solutions, we isolated islet from adult Yucatan pig’s pancreas using standard enzymatic digestion (Nordmark) followed by Ficoll purification, these islets were preserved in a humidified tissue culture incubator with preservation solutions and preserved in 4℃ for up to 4 weeks. We compared islet survival in groups of islets. To islet viability and islet functionality, we utilized Glucose Stimulated Insulin Secretion (GSIS) for measuring insulin release, calculated stimulation index (SI) and AO/PI viability staining for during the preservation period. Results: Highest islet recovery was preserved in the group of pig islets cultured with Taurine as compared to the group of islets hypothermically preserved in standard tissue culture media over the 4 week preservation period (p<0.001 using ANOVA). The porcine islet viability/cell number ratio in the OPTI solution maintained a high survival rate for up to 2 weeks (94%). In contrast, porcine islet in OPTI-T solution and CMRL 1066 supplemented media significantly reduced viability after 1 weeks. OPTI solution (with taurine) was significantly higher than OPTI-T solution (without taurine) and CMRL 1066 during 4 weeks after preservation. Glucose stimulation insulin secretion test was performed for functional analysis. As a result, a level of porcine insulin was higher during 2 weeks in the OPTI solution than OPTI-T solution and CMRL 1066. But after 3 weeks preservation, insulin level in high glucose, significantly decreased. Therefore, the viability and functionality of islets were improved by adding taurine to the developed preservation solution. Conclusion: Taurine supplementation has been shown to highest islet recovery, viability and insulin secretion level for 2 weeks. Based on this data, we have performed 2 weeks cold storage of isolated islets for current clinical islet transplantation. Ministry of trade industry and energy, KEIT 20011276, NTIS 1415172664.

Normothermic liver machine perfusion as a dynamic platform for regenerative purposes: What does the future have in store for us?

Liver transplantation has become an immense success; nevertheless, far more recipients are registered on waiting lists than there are available donor livers for transplantation. High-risk, extended criteria donor livers are increasingly used to reduce the discrepancy between organ demand and supply. Especially for high-risk livers, dynamic preservation using machine perfusion can decrease post-transplantation complications and may increase donor liver utilisation by improving graft quality and enabling viability testing before transplantation. To further increase the availability of donor livers suitable for transplantation, new strategies are required that make it possible to use organs that are initially too damaged to be transplanted. With the current progress in experimental liver transplantation research, (long-term) normothermic machine perfusion may be used in the future as a dynamic platform for regenerative medicine approaches, enabling repair and regeneration of injured donor livers. Currently explored therapeutics such as defatting cocktails, RNA interference, senolytics, and stem cell therapy may assist in the repair and/or regeneration of injured livers before transplantation. This review will provide a forecast of the future utility of normothermic machine perfusion in decreasing the imbalance between donor liver demand and supply by enabling the repair and regeneration of damaged donor livers.

Temperature effects on the disappearance and reappearance of corneal-endothelium primary cilia.

To elucidate the specific functions of the primary cilia in corneal endothelial cells (CECs) by investigating the histological changes of corneal endothelium exposed at low temperature. Experimental study. This study involved corneas freshly obtained from Japanese white rabbits preserved in Optisol™-GS (Bausch & Lomb) corneal storage medium at 4°C for 0, 1, and 7days. Corneas preserved for 7days were also incubated at 37°C in culture media for an additional 2days. A rabbit CEC line was also preserved in Optisol™-GS at 4°C for 0 and 1day. The corneal endothelium specimens and CECs were then assessed by immunostaining and scanning electron-microscopy (SEM). Immediately post isolation, the CECs of the specimens showed positive immunostaining for primary cilia (i.e., approximately 20%) via anti-acetylated alpha Tubulin antibody and SEM observation. Primary cilia were found to have attenuated/disappeared on the corneal endothelium specimens preserved for 1 or 7days at 4°C. After an additional 2-day incubation at 37°C, primary cilia reappeared on the corneal endothelium specimens (approximately 20%). The disappearance of cilia during the preservation period was also observed in the immortalized CECs. The findings in this study using rabbit corneas indicate that the primary cilia of corneal endothelium preserved at low temperature disappeared, then reappeared after returning to body temperature, suggesting that temperature has a direct effect on the primary cilia of corneal endothelium.

Extending the utility of anterior corneal buttons through refrigeration and glycerol cryopreservation: utility rate and outcome analysis

Background/aimsTo evaluate the utility rate, indication, outcome, and cost of refrigeration and glycerol cryopreservation for storing anterior corneal buttons during endothelial keratoplasty for subsequent use in tectonic lamellar patch grafting.MethodAnterior...

Influence of different types of fermentation-produced chymosin on quality of soft cheeses

The disadvantage of soft cheeses is their short shelf life. In soft cheeses with a high moisture content, proteolysis occurs at a high rate, as a result of which the cheeses quickly overripe. The main proteolytic agent in soft cheeses is the milk-clotting enzyme (MCE). Increasing the shelf life of cheeses can be achieved by using MCE types having low proteolytic activity (PA). We have studied the effect of MCE based on different types of fermentation-produced chymosin: Chy-max® Extra (bovine chymosin), Chy-max® M (camel chymosin), Chy-max® Supreme (“modified” chymosin) on the dynamics of proteolysis in soft cheeses and related changes in the structure of cheeses during their storage. All 3 types of studied MCEs have different levels of nonspecific PA. The higher the level of nonspecific PA of the used MCE, the higher the rate of the proteolysis process in the resulting cheeses. Increasing the dose of MCE also increases the rate of proteolysis in cheeses. To increase the shelf life of soft cheeses, which depends on the period of preservation of a dense consistency, it is promising to use MCE with low PA based on camel chymosin (Chy-max® M) and “modified” chymosin (Chy-max® Supreme).

Antifungal activity and molecular mechanisms of mulberrin derivatives against Colletotrichum gloeosporioides for mango storage

In this work, by using high throughput virtual screening and bioactivity assays, this work revealed that three natural compounds, mulberrin (Mul) exhibiting the highest anti-CYP51 activity, isoxanthohumol and (s)-isopsoralen markedly inhibited 14α-demethylase (a pivotal biosynthetic enzyme involved in the biosynthesis of ergosterol) in Colletotrichum gloeosporioides. Results of computational biology analysis demonstrated that, among the three inhibitors bound to the catalytic pocket of CYP51, Mul showed a closer distance with heme in CYP51 and a stronger binding free energy with CYP51. In vitro tests, Mul demonstrated excellent anti-Colletotrichum gloeosporioides activity by inhibiting CYP51 activity. Notably, Mul treatment decreased the bioactivity of CYP51, thereby increasing cell membrane permeability and cell death. Moreover, Mul treatment significantly prolonged the preservation period of fruits. These results suggest that Mul suppresses anthracnose in postharvest mango by inhibiting the growth of Colletotrichum gloeosporioides and can be used as a potential natural preserving agent.