Long-term Storage Conditions Research Articles

Long-term storage of seeds of plant genetic resources in permafrost conditions

Collections of plant genetic resources are the guarantor of the preservation of both natural biological diversity and breeding achievements. Basically, genetic collections consist of seeds and, proper storage in optimal conditions is a necessity (it frees them from frequent reproduction associated with material and labor costs). It is necessary to search for optimal natural conditions for long-term storage of seeds, because in case of emergency situations of a technogenic nature, they will be the most vulnerable. A long-term experiment of long-term storage of agricultural plant seeds in permafrost conditions is being carried out. The aim of research is to find optimal and safe conditions for long-term storage of seeds and grain for both sowing and food purposes, and to study the conditions of natural storage facilities for creation of strategic reserves of seeds and food. In 2010, consignments of seeds of the main grain crops were put into storage (spring wheat, spring barley, winter rye and oats); in 2016, a seed collection of 26 varieties of 8 types of vegetable crops was laid. Meteorological observations were carried out, thermographs and hygrographs were used in the storage. The first sample extraction took place after 6 years of storage. Laboratory analysis of seed quality was carried out using the following standard methods: GOST 12036-85; GOST 12038-84; GOST 12041-82; GOST 12042-80; and GOST R 52325-2005. The results showed high viability of seeds of grain and vegetable crops. Seeds of all samples of grain crops met the requirements for reproductive seeds (germination over 87 %) according to GOST R 52325-2005. Seeds of 17 samples of vegetable crops retained germination at the first-class level in accordance with the requirements of GOST 12038-84. Long-term meteorological observations show a gradual warming of the Arctic climate due to relatively high summer temperatures. In general, the continentality of the climate is growing. The experiment continues, most of the above seed batches remain in storage, further expeditions with the extraction of samples are planned for 2025, 2035 and 2050. The obtained weather data indicate an increase in the continentality of the Arctic climate and its general warming. The data obtained on the viability of cereal grains and vegetable seeds during long-term storage show the effectiveness of using permafrost conditions for this purpose.

Changes in Plant Genetic Resources in the Southeast Region of Poland from the 1980s to 2023

This paper presents data from sixteen collecting missions conducted over three decades (2000–2023) in the Lubelskie Voivodeship, located in Southeastern Poland. During these missions, 756 samples of functional plants were collected, along with accompanying species such as weeds, crop wild relatives, ornamental plants, spice plants, and grafts of fruit trees. These materials were compared with those collected during six expeditions in two earlier decades (1980–1999), which resulted in the collection of 665 genotypes. The samples were primarily obtained from local farmers or gathered directly from fields. This study aims to summarise and compare the results of the collecting missions across decades. The analysis indicates that the Lubelskie region remains a place where old varieties of annual vegetable plants, rare weed species (on a national scale), crop wild relatives, and old varieties of fruit trees can still be found. However, old cereal varieties have nearly disappeared. The collected materials were characterised and stored as a base collection under long-term storage conditions at the Plant Breeding and Acclimatization Institute—National Research Institute. Vegetable plant materials were deposited in the active collection at the National Institute of Horticultural Research (INHORT). Fruit tree collections were also deposited at INHORT, as well as in the Botanical Garden of the Polish Academy of Sciences in Poland.

Different mesoporous carbon carriers for the improvement of solubility and physical stability of poorly soluble drugs.

Mesoporous carriers have gained significant attention for enhancing the solubility and bioavailability of Biopharmaceutics Classification System (BCS) Class II drugs. However, the contribution of mesoporous carriers with varying morphologies to the physical stability of these drugs is not well-defined. In this work, mesoporous carbon nanoparticles (MCN) and hollow carbon mesoporous nanoparticles (HMC) were prepared, while the weakly acidic Indomethacin (IMC) and alkaline Celecoxib (CXB) were incorporated into these carriers in the amorphous state by the solvent evaporation method. Notably, HMC demonstrated superior drug loading efficiencies (approximately 43 %) for both IMC and CXB owning to its hollow structure. The mesoporous drug loading systems significantly enhanced dissolution rates in comparison with both self-made amorphous drugs and raw drugs. Furthermore, under accelerated and long-term storage conditions, the mesoporous carriers effectively prevented drugs loaded from crystallization, maintaining constant dissolution profiles for over 12 months. Intriguingly, CXB exhibited a slower rate of crystallization after loading into the mesoporous carriers, likely due to the formation of hydrogen bonds between the carbonaceous carrier material and the amino groups of CXB. Compared with loaded drugs, the self-made amorphous drugs exhibited a crystallinity increase beyond 60 % within the initial month. Collectively, these findings highlighted the potential of mesoporous carbon carriers to elevate the dissolution behaviors of BCS Class II drugs while preserving the physical stability of the loaded amorphous drugs.

Evaluation of Short-Term Stability of Different Nitazenes Psychoactive Opioids in Dried Blood Spots by Liquid Chromatography-High-Resolution Mass Spectrometry.

Nitazenes represent a new synthetic opioids sub-class belonging to new psychoactive substances (NPSs). Their high pharmacological potency has led to numerous intoxications and fatalities, even at minimum doses. The aim of this study was to assess the stability of four nitazenes (etazene, flunitazene, isotonitazene and protonitazene) in dried blood spot (DBS) samples at different storage temperatures (room temperature and 4 °C) and determine the optimal storage conditions. Moreover, we developed and validated a new and fast liquid chromatography-high-resolution mass spectrometry method by the optimization of chromatographic conditions with the use of a different chromatographic column and mobile phases. Two concentrations, 1 and 5 ng/mL, were chosen based on the available data on nitazenes-related intoxications and their stability was evaluated at days 0 (control), 1, 7 and 30. The results showed that all analytes at 1 ng/mL were not detectable after 30 days at room temperature; a similar pattern was observed for 1 ng/mL etazene and isotonitazene samples when stored at 4 °C, whereas flunitazene and protonitazene decreased to a mean of 66% and 69% initial concentrations, respectively, at day 30. Differently, all analytes at 5 ng/mL were quantified above 44% and 41% initial concentrations at room temperature and 4 °C, respectively, showing a higher stability. The study of nitazenes stability in DBSs represents an important tool to determine the optimal sample storage conditions, such as temperature and time between sample collection and analysis. In contrast to another study, our study showed distinct stability behaviors for every investigated analyte, which also depended on the concentration. Therefore, it is difficult to define an optimal storage condition acceptable for all nitazenes. Room temperature proved to be the best medium- and long-term storage conditions for the highest concentrations, but the stability of low levels of flunitazene and protonitazene improved at 4 °C.

Development of Long-Term Stability of Enveloped rVSV Viral Vector Expressing SARS-CoV-2 Antigen Using a DOE-Guided Approach.

Liquid formulations have been successfully used in many viral vector vaccines including influenza (Flu), hepatitis B, polio (IPV), Ebola, and COVID-19 vaccines. The main advantage of liquid formulations over lyophilized formulations is that they are cost-effective, as well as easier to manufacture and distribute. However, studies have shown that the liquid formulations of enveloped viral vector vaccines are not stable over extended periods of time. In this study, we explored the development of the liquid formulations of an enveloped recombinant Vesicular Stomatitis Virus (VSV) expressing the SARS-CoV-2 spike glycoprotein. To do so, we used a design of experiments (DOE) method, which allowed us to assess the stability dynamics of the viral vector in an effective manner. An initial stability study showed that trehalose, gelatin, and histidine were effective at maintaining functional viral titers during freeze-thaw stress and at different temperatures (-20, 4, 20, and 37 °C). These preliminary data helped to identify critical factors for the subsequent implementation of the DOE method that incorporated a stress condition at 37 °C. We used the DOE results to identify the optimal liquid formulations under the selected accelerated stress conditions, which then guided the identification of long-term storage conditions for further evaluation. In the long-term stability study, we identified several liquid formulations made of sugar (sucrose, trehalose, and sorbitol), gelatin, and a histidine buffer that resulted in the improved stability of rVSV-SARS-CoV-2 at 4 °C for six months. This study highlights an effective approach for the development of liquid formulations for viral vector vaccines, contributing significantly to the existing knowledge on enveloped viral vector thermostability.

Long-Life Inoculant: Bradyrhizobium Stored in Biodegradable Beads for Four Years Shows Optimal Cell Vitality, Interacts with Peanut Roots, and Promotes Early Growth.

Currently, bacterial inoculant technology focuses on improving long-term storage conditions to ensure adequate rhizobia numbers and their effectiveness as plant growth promoters. This study aimed to investigate whether storage at 4 °C for four years of alginate beads immobilizing Bradyrhizobium sp. SEMIA6144 maintains bacterial vitality, efficacy in growth promotion, and ability to establish early interactions with Arachis hypogaea L. The recovery of viable SEMIA6144 cells decreased over time (10% at six months, 1% at one year, and 0.01% at four years), while cell vitality remained high at 94.1%, 90.2%, and 93.4%, respectively. The unsaturated/saturated fatty acid ratio declined during storage, reducing membrane fluidity and metabolic activity. Mobility and root adhesion of SEMIA6144 decreased after one and four years. However, growth promotion in peanuts inoculated with SEMIA6144 beads was observed through increased biomass, total chlorophyll, leaf number, leaf area, and decreased chlorophyll fluorescence compared to non-inoculated plants. Although nodulation was low in plants inoculated with four-year-old beads, leghemoglobin levels were maintained. These results demonstrate that Bradyrhizobium sp. SEMIA6144 can be stored for four years in alginate beads at 4 °C, maintaining its vitality and ability to establish a symbiosis that stimulates early peanut growth. Understanding these physiological changes could be valuable for the future improvement of long-lasting inoculants.

Unveiling the secrets of lotus seed longevity: insights into adaptive strategies for extended storage.

Seed longevity is crucial for long-term storage, but prolonged unfavorable conditions can lead to viability loss. This study integrated theoretical and experimental techniques to elucidate the inherent mechanisms underlying the unique ability of lotus seed capacity to maintain stable viability even after enduring years. Transcriptome analysis and microscopy revealed the sturdy structure of the lotus seed pericarp, which predominantly expressed cellulose synthase genes involved in cell wall biogenesis. The cotyledon serves as a nutrient source for seeds during long-term storage. Additionally, the inactivation of chlorophyll degradation pathways may allow for the retention of chlorophyll in the lotus seed plumule, potentially enhancing the environmental adaptability of lotus seedlings. While the reduced abundance of transcripts corresponding to heat shock protein genes could impact protein processing and consequently diminish the vitality of aging lotus seeds. Moreover, an expansion in the number of seed maturation and defense response genes was observed in the lotus genome compared to other 11 species, which might represent an adaptive strategy against long-term adverse storage conditions. Overall, these findings are crucial for understanding the mechanisms underlying lotus seed longevity and may inform future improvements in the extended storage periods of seed crops.

Short- and long-term stability of SARS-CoV-2 antibodies on dried blood spots under different storage conditions.

Dried blood spots (DBS) are broadly used for different serological analyses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody assessment. In order to biobank DBS samples, an understanding of the preservation of SARS-CoV-2 antibodies is needed. Therefore, we assessed the stability of SARS-CoV-2 antibodies on DBS during short- and long-term storage under different storage temperatures. Seven sample donors were enrolled, each donating twenty 6 mm DBS to assess anti-spike (S1) SARS-CoV-2 IgG antibodies (EUROIMMUN). Baseline samples were analyzed on the day of collection. The remainder of the samples was stored in grip seal bags kept in a cryobox at room temperature/4°C until 2 months after collection and at -20°C until 2 years after collection. Samples were analyzed at regular intervals within the total storage duration and after one and five freeze-thaw cycles. A pooled coefficient of variation was calculated for each storage temperature. We found that anti-S1 SARS-CoV-2 antibodies collected on DBS saver cards remain stable during short-term storage at RT, 4°C, and -20°C (at least to 2 months) and long-term storage at -20°C (at least 2 years). Moreover, up to five freeze-thaw cycles can occur without impacting the anti-S1 SARS-CoV-2 antibody level. The inter-assay coefficient of variation lies between 10 and 15%. As DBS can be preserved for both shorter periods of time at RT and longer periods of time at -20°C, they are a perfect application for studies that require sample shipment by mail, self-sampling studies, studies in limited resource settings and biobanking.IMPORTANCEDried blood spots (DBS) are currently widely used as a microsampling technique for different qualitative and quantitative serological assessments. Yet, there is a lack of long-term stability and storage condition studies. In our study, first, we assessed the stability of SARS-CoV-2 antibodies on DBS up to 2 years post-collection. We believe that our data are not only important for future COVID-19 research but also for studies on other infections/diseases using DBS-based serology.

Research on the Stability of Salt Cavern Hydrogen Storage and Natural Gas Storage under Long-Term Storage Conditions

The stability of salt cavern storage during prolonged operation is a crucial indicator of its safety. This study focuses on an operational underground gas storage facility, conducting comparative numerical simulations for the storage of natural gas and hydrogen. We investigated the evolution of stability for natural gas and hydrogen storage under long-term storage conditions. The main conclusions are as follows: (1) A new equation for stress equilibrium and constitutive relations are derived. (2) At the same storage pressure, the effective stress at the same position in the interlayer is greater for hydrogen storage compared to natural gas storage, signifying a higher level of danger. (3) At the same storage pressure, the displacement at the cavity top for hydrogen storage is greater than that for natural gas storage. The displacement difference between the two is greatest at 9 MPa, amounting to 0.026 m. (4) Due to hydrogen’s lower dynamic viscosity and higher permeability, the depth and extent of the plastic zones within the interlayers are greater compared to natural gas. When the storage pressure is 15 MPa, the depth of the plastic zone within the interlayer can be up to 2.1 m greater than when storing natural gas, occurring in the third interlayer from the top. These research findings may serve as a valuable reference for determining the operational parameters of on-site salt cavern hydrogen storage facilities.

Effect of storage conditions on host egg suitability and the reproductive fitness of Oobius agrili (Hymenoptera: Encyrtidae), an egg parasitoid of the invasive emerald ash borer (Agrilus planipennis) (Coleoptera: Buprestidae).

Oobius agrili Zhang and Huang (Hymenoptera: Encyrtidae) is an important egg parasitoid of the emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae). Methods for laboratory-rearing O. agrili have been developed but its mass-production depends on the continuous production and storage of freshly laid EAB eggs as well as diapaused parasitoid progeny (inside parasitized EAB eggs). The purpose of this study was to determine optimal environmental conditions for long-term storage of host eggs as well as diapaused parasitoid progeny. Fresh host eggs and diapaused parasitoid progeny were stored at two low storage temperatures (1.7 and 12.8 °C) and three levels of relative humidity (low ~31%, medium ~74%, and high ~99.9%) for various length of time (15-270 days) and then evaluated for host egg suitability and the reproductive fitness of stored parasitoid progeny. EAB eggs were stored for approximately 30 days without significant reduction of their viability and suitability to O. agrili parasitism at low storage temperatures under high and medium relative humidity. Neither storage temperature or humidity had any significant effects on adult parasitoid emergence for storage durations of up to 270 days. When storage durations were over 120 days, however, both adult parasitoid longevity and fecundity declined approximately 20-30% across all temperature and humidity treatments. Relevance of findings to mass-production and storage of O. agrili for biocontrol is discussed.

Storage behaviour and seed longevity of traditional African vegetables: a review

Effective management of seed collections in genebanks requires knowledge of seed storage behaviour and species-specific longevity under conventional air-dry storage. This knowledge is lacking for traditional African vegetable (TAV) crops compared to cereals and conventional vegetables. This review aimed to assess the published literature until December 2023. The findings will guide post-harvest seed processing (cleaning and drying), storage (containers, storage parameters), viability monitoring and regeneration intervals, and to identify further research gaps that will inform data needs from the genebanks conserving TAV seeds. Our findings show that, compared to cereals and legumes, there is limited information available on vegetable crops and more acutely on TAVs. While conventional vegetable seeds benefited from research on their storage behaviour and longevity, there was significant neglect of TAV crops, which represented only 17% of the studies reviewed. Overall, genebanks involved in TAV germplasm conservation should invest more time and resources to investigate seed physiology and generate data that will inform on the storage behaviour and longevity of these crops under medium and long-term storage conditions. Such data are important to establish crop-specific knowledge on optimal conditions for high post-harvest seed quality, and for genebanks to plan seed regeneration.

An Overview of Pre-Analytical Factors Impacting Metabolomics Analyses of Blood Samples.

Discrepant sample processing remains a significant challenge within blood metabolomics research, introducing non-biological variation into the measured metabolome and biasing downstream results. Inconsistency during the pre-analytical phase can influence experimental processes, producing metabolome measurements that are non-representative of in vivo composition. To minimize variation, there is a need to create and adhere to standardized pre-analytical protocols for blood samples intended for use in metabolomics analyses. This will allow for reliable and reproducible findings within blood metabolomics research. In this review article, we provide an overview of the existing literature pertaining to pre-analytical factors that influence blood metabolite measurements. Pre-analytical factors including blood tube selection, pre- and post-processing time and temperature conditions, centrifugation conditions, freeze-thaw cycles, and long-term storage conditions are specifically discussed, with recommendations provided for best practices at each stage.

Feasibility study of geothermal assisted energy storage using hydraulic fracturing

Similar to wind and solar energy, geothermal energy, as a renewable energy source that is widely distributed, abundant, green, and low-carbon, has substantial potential. In this study, we introduce and validate the feasibility of a novel approach that employs hydraulic fracturing techniques to store electrical power in hydraulic fractures, while simultaneously extracting geothermal energy from the strata. The primary advantage of this methodology is its dual functionality (i.e., storing energy while harvesting heat). This approach outperforms conventional energy storage methods in terms of efficiency, in which the total energy storage efficiency is far greater than 1. Our study analyzed the factors influencing energy and efficiency, as well as the variations in energy and efficiency under long-term energy storage conditions. This study also provides a theoretical basis for optimizing the design of hydraulic fracturing energy storage assisted by geothermal energy.

Phenotypic patterns of antimicrobial resistance in Campylobacter spp. in Ukraine

Campylobacter spp. are major foodborne zoonotic pathogens that have recently become more resistant to fluoroquinolones and macrolides, which are broad-spectrum antibiotics used in both medicine and veterinary practice. Campylobacter is a commensal of the intestines of mammals and birds, which facilitates the transfer of antimicrobial resistance (AMR) determinants from other bacteria through horizontal gene transfer. The aim of this study was to establish the prevalence and determine the AMR phenotypes of Campylobacter species isolated in Ukraine. Using the disk diffusion method (DDM), 33 isolates of Campylobacter spp. isolated from animals and poultry on farms between May and September 2023 were tested. Additionally, an analysis of the resistance of 293 Campylobacter spp. isolates obtained from children with acute intestinal infections from 2020 to 2023 was conducted. The level of resistance of clinical Campylobacter spp. to ciprofloxacin (CIP) was found to be 83.3%, to tetracycline (TE) 53.6%, to erythromycin (E) 11.6%, and the isolates from farms showed resistance to ciprofloxacin (CIP) at 72.7%, to tetracycline (TE) at 60.6%, and to erythromycin (E) at 18.2%. In 4.1% of clinical isolates, multidrug resistance (MDR) (CIP/TE/E) was detected, with the most common AMR combination being (CIP/TE), reaching 42.7%. Resistance to at least one antibiotic was found in 37.8%. The isolates from animals and poultry had a multiple antibiotic resistance rate of 12.1%, with 42.4% being resistant to at least one antibiotic. This study provides insight into the relevance and importance of Campylobacter spp. resistance in Ukraine. It expands the understanding of the issue, which requires more detailed study, including the molecular mechanisms of resistance and the identification of genetic determinants shaping the epidemiology of antimicrobial resistance in Ukraine. For this purpose, a collection of isolates has been created, and optimal long-term storage conditions have been selected, which will allow the study of Campylobacter spp. decades from now.

Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap).

Proteins undergo reversible S-acylation via a thioester linkage in vivo. S-palmitoylation, modification by C16:0 fatty acid, is a common S-acylation that mediates critical protein-membrane and protein-protein interactions. The most widely used S-acylation assays, including acyl-biotin exchange and acyl resin-assisted capture, utilize blocking of free Cys thiols, hydroxylamine-dependent cleavage of the thioester and subsequent labeling of nascent thiol. These assays generally require >500 μg of protein input material per sample and numerous reagent removal and washing steps, making them laborious and ill-suited for high throughput and low input applications. To overcome these limitations, we devised "Acyl-Trap", a suspension trap-based assay that utilizes a thiol-reactive quartz to enable buffer exchange and hydroxylamine-mediated S-acyl enrichment. We show that the method is compatible with protein-level detection of S-acylated proteins (e.g., H-Ras) as well as S-acyl site identification and quantification using "on trap" isobaric labeling and LC-MS/MS from as little as 20 μg of protein input. In mouse brain, Acyl-Trap identified 279 reported sites of S-acylation and 1298 previously unreported putative sites. Also described are conditions for long-term hydroxylamine storage, which streamline the assay. More generally, Acyl-Trap serves as a proof-of-concept for PTM-tailored suspension traps suitable for both traditional protein detection and chemoproteomic workflows.

Screening of NIAS World Rice Core Collection for Seeds with Long Longevity as Useful Potential Breeding Materials Focusing on the Stability of Embryonic RNAs.

Seed longevity is a crucial trait for the seed industry and genetic resource preservation. To develop excellent cultivars with extended seed lifespans, it is important to understand the mechanism of keeping seed germinability long term and to find useful genetic resources as prospective breeding materials. This study was conducted to identify the best cultivars with a high and stable seed longevity trait in the germplasm of rice (Oryza sativa L.) and to analyze the correlation between seed longevity and embryonic RNA integrity. Seeds from 69 cultivars of the world rice core collection selected by the NIAS in Japan were harvested in different years and subjected to long-term storage or controlled deterioration treatment (CDT). The long-term storage (4 °C, RH under 35%, 10 years) was performed on seeds harvested in 2010 and 2013. The seeds harvested in 2016 and 2019 were used for CDT (36 °C, RH of 80%, 40 days). Seed longevity and embryonic RNA integrity were estimated by a decrease in the germination percentage and RNA integrity number (RIN) after long-term storage or CDT. The RIN value was obtained by the electrophoresis of the total RNA extracted from the seed embryos. Seeds of "Vandaran (indica)", "Tupa 729 (japonica)", and "Badari Dhan (indica)" consistently showed higher seed longevity and embryonic RNA integrity both under long-term storage and CDT conditions regardless of the harvest year. A strong correlation (R2 = 0.93) was observed between the germination percentages and RIN values of the seeds after the long-term storage or CDT among nine cultivars selected based on differences in their seed longevity. The study findings revealed the relationship between rice seed longevity and embryo RNA stability and suggested potential breeding materials including both japonica and indica cultivars for improving rice seed longevity.

Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap).

Proteins undergo reversible S-acylation via a thioester linkage in vivo. S-palmitoylation, modification by C16:0 fatty acid, is a common S-acylation that mediates critical protein-membrane and protein-protein interactions. The most widely used S-acylation assays, including acyl-biotin exchange and acyl resin-assisted capture, utilize blocking of free Cys thiols, hydroxylamine-dependent cleavage of the thioester and subsequent labeling of nascent thiol. These assays generally require >500 micrograms of protein input material per sample and numerous reagent removal and washing steps, making them laborious and ill-suited for high throughput and low input applications. To overcome these limitations, we devised "Acyl-Trap", a suspension trap-based assay that utilizes a thiol-reactive quartz to enable buffer exchange and hydroxylamine-mediated S-acyl enrichment. We show that the method is compatible with protein-level detection of S-acylated proteins (e.g. H-Ras) as well as S-acyl site identification and quantification using "on trap" isobaric labeling and LC-MS/MS from as little as 20 micrograms of protein input. In mouse brain, Acyl-Trap identified 279 reported sites of S-acylation and 1298 previously unreported putative sites. Also described are conditions for long-term hydroxylamine storage, which streamlines the assay. More generally, Acyl-Trap serves as a proof-of-concept for PTM-tailored suspension traps suitable for both traditional protein detection and chemoproteomic workflows.

O/W nanoemulsions encapsulated octacosanol: Preparation, characterization and anti-fatigue activity

Octacosanol has various biological effects such as antioxidant, hypolipidemic and anti-fatigue. However, poor solubility has limited the application of octacosanol in food. The aim of this study was to prepare octacosanol nanoemulsions with better solubility, stability and safety and to investigate in vivo anti-fatigue effect. The food-grade formulation of the octacosanol nanoemulsions consisted of octacosanol, olive oil, Tween 80, glycerol and water with 0.1 %, 1.67 %, 23.75 %, 7.92 % and 66.65 % (w/w), respectively. The nanoemulsions had an average particle size of 12.26 ± 0.76 nm and polydispersity index of 0.164 ± 0.12, and showed good stability under different pH, cold, heat, ionic stress and long-term storage conditions. The results of animal experiments showed that the octacosanol nanoemulsions significantly prolonged the fatigue tolerance time, alleviated the fatigue-related biochemical indicators, and weakened the oxidative stress. Meanwhile, octacosanol nanoemulsions upregulated hepatic glycogen levels. Taken together, these findings suggested that octacosanol nanoemulsions have promising applications as anti-fatigue functional foods.

Effects of Delaying the Storage of 'Hass' Avocados under a Controlled Atmosphere on Skin Color, Bioactive Compounds and Antioxidant Capacity.

During ripening, 'Hass' avocado skin changes from green to purple/black. Low-temperature storage with a controlled atmosphere (CA) is the most widely used method for avocado storage; however, few studies have simulated this technology and considered the days of regular air (RA) storage prior to CA storage. Herein, the effect of delaying the storage of 'Hass' avocado (>30% dry matter) in a CA was examined. Long-term storage conditions (5 °C for 50 days) corresponded to (i) regular air storage (RA), (ii) CA (4 kPa O2 and 6 kPa CO2) and (iii) 10 days in RA + 40 days in a CA and (iv) 20 days in RA + 30 days in a CA. Evaluations were performed during storage and at the ready-to-eat (RTE) stage. Skin color remained unchanged during storage, but at the RTE stage, more color development was observed for fruits stored under CA conditions, as these fruits were purple/black (>50%). At the RTE stage, the anthocyanin content increased, and compared to fruit under RA, fruit under a CA contained a five-fold greater content. A 20-day delay between harvest and CA storage increased the fruit softening rate and skin color development after cold storage, reducing the effectiveness of CA as a postharvest technology for extending storage life.

The association of protein-bound methionine sulfoxide with proteomic basis for aging in beech seeds

BackgroundEuropean beech (Fagus sylvatica L.) trees produce seeds irregularly; therefore, it is necessary to store beech seeds for forestation. Despite the acquisition of desiccation tolerance during development, beech seeds are classified as intermediate because they lose viability during long-term storage faster than typical orthodox seeds. In this study, beech seeds stored for short (3 years) or long (20 years) periods under optimal conditions and displaying 92 and 30% germination capacity, respectively, were compared.ResultsAged seeds displayed increased membrane damage, manifested as electrolyte leakage and lipid peroxidation levels. Analyses have been based on embryonic axes, which contained higher levels of reactive oxygen species (ROS) and higher levels of protein-bound methionine sulfoxide (MetO) in aged seeds. Using label-free quantitative proteomics, 3,949 proteins were identified, of which 2,442 were reliably quantified pointing to 24 more abundant proteins and 35 less abundant proteins in beech seeds under long-term storage conditions. Functional analyses based on gene ontology annotations revealed that nucleic acid binding activity (molecular function), ribosome organization or biogenesis and transmembrane transport (cellular processes), translational proteins (protein class) and membranous anatomical entities (cellular compartment) were affected in aged seeds. To verify whether MetO, the oxidative posttranslational modification of proteins that can be reversed via the action of methionine sulfoxide reductase (Msr) enzymes, is involved in the aging of beech seeds, we identified and quantified 226 MetO-containing proteins, among which 9 and 19 exhibited significantly up- and downregulated MetO levels, respectively, in beech seeds under long-term storage conditions. Several Msr isoforms were identified and recognized as MsrA1-like, MsrA4, MsrB5 and MsrB5-like in beech seeds. Only MsrA1-like displayed decreased abundance in aged seeds.ConclusionsWe demonstrated that the loss of membrane integrity reflected in the elevated abundance of membrane proteins had a higher impact on seed aging progress than the MetO/Msr system. Proteome analyses enabled us to propose protein Sec61 and glyceraldehyde-3-phosphate dehydrogenase as potential longevity modulators in beech seeds.