Common Storage Conditions Research Articles

Particle formation in response to different protein formulations and containers: Insights from machine learning analysis of particle images

Subvisible particle count is a biotherapeutics stability indicator widely used by pharmaceutical industries. A variety of stresses that biotherapeutics are exposed to during development can impact particle morphology. By classifying particle morphological differences, stresses that have been applied to monoclonal antibodies (mAbs) can be identified. This study aims to evaluate common biotherapeutic drug storage and shipment conditions that are known to impact protein aggregation. Two different studies were conducted to capture particle images using micro-flow imaging and to classify particles using a convolutional neural network. The first study evaluated particles produced in response to agitation, heat, and freeze-thaw stresses in one mAb formulated in five different formulations. The second study evaluated particles from two common drug containers, a high-density polyethylene bottle and a glass vial, in six mAbs exposed solely to agitation stress. An extension of this study was also conducted to evaluate the impact of sequential stress exposure compared to exposure to one stress alone, on particle morphology. Overall, the convolutional neural network was able to classify particles belonging to a particular formulation or container. These studies indicate that storage and shipping stresses can impact particle morphology according to formulation composition and mAb.

Short- and long-term stability of synthetic cathinones and dihydro-metabolites in human urine samples

PurposeSynthetic cathinones constitute the second largest group of new psychoactive substances, which are often used for recreational purposes and reported in toxicological analysis. Various factors may influence the stability of synthetic cathinones between sampling and analysis, and therefore, stability studies are required to determine the best storage conditions as well as extend the period of detection.MethodsThis study involved sixteen synthetic cathinones and ten dihydro-metabolites spiked in human urine to evaluate the stability under common storage conditions to imitate real forensic toxicology samples. The samples were stored at either room temperature (22–23 °C) for up to 3 days, refrigerated (4 °C) for up to 14 days or frozen (–40 °C) for up to 12 months, and analyzed in triplicate using a validated liquid chromatography–tandem mass spectrometry method.ResultsAnalytes’ concentrations decreased over time, although slower when stored frozen. All analytes remained stable (> 80%) for 1 month when stored frozen before losses in content were more apparent for some compounds, depending on their chemical structure. Under all storage conditions, the highest instability was observed for analytes containing halogens (i.e., chlorine or fluorine). Thus, halogenated analytes were further investigated by using liquid chromatography coupled to quadruple time-of-flight mass spectrometry to attempt identifying degradation products.ConclusionsIrrespective of parent analytes, dihydro-metabolites had improved stability at each tested temperature, which highlights their importance as appropriate urine biomarkers when retesting is required after a long period of storage.

Human Health Risk Assessment of Phthalate Esters and Antimony Levels in Beverages Packaged in Polyethylene Terephthalate under Different Storage Conditions

The main objective of this study was to examine how common storage conditions affect the migration of phthalate esters (PAEs) and antimony (Sb) from polyethylene terephthalate (PET) bottles into bottled drinks. The drinks were collected from local supermarkets in Turkey, including different types such as bottled drinking water (DW), mineral water (MW), cola (C), orange flavored carbonated drink (OFCD), fermented turnip drink (FTD), and lemonade (L). Additionally, a deterministic model was utilized to evaluate the potential risk to human health associated with this migration process. The performance and optimization of the LC-MS/MS and ICP-MS instruments for the analysis of PAEs and Sb were also evaluated. The limits of detection (LODs) for PAEs and Sb ranged from 0.004 to 1.88ng/mL, while the limits of quantification (LOQs) ranged from 0.013 to 6.27ng/mL. The analysis results revealed the presence of PAEs at varying concentrations. Di-isodecyl phthalate (DIDP) and di-isononyl phthalate (DINP) exhibited the highest level of migration to drink samples. The type of beverage also affected the migration of PAEs and Sb. The highest migration levels were detected in MWs. Storage temperature (4 °C, 25 °C, and 40 °C) and storage duration (365 days) had an impact on the release of PAEs and Sb into beverages from PET bottles. The Hazard Quotients (HQs) calculated for PAEs and Sb were significantly lower than 1 in all scenarios, indicating that no adverse health effects are anticipated due to exposure to PAEs and Sb through beverage consumption.

The forgotten variable? Does the euthanasia method and sample storage condition influence an organisms transcriptome – a gene expression analysis on multiple tissues in pigs

BackgroundTranscriptomic studies often require collection of fresh tissues post euthanasia. The chosen euthanasia method might have the potential to induce variations in gene expressions that are unlinked with the experimental design. The present study compared the suitability of ‘nitrogen gas in foam’ (ANOXIA) in comparison to a non-barbiturate anaesthetic, T-61® (T61), for euthanizing piglets used in transcriptome research. Further, the effect of common tissue storage conditions, RNAlater™ (RL) and snap freezing in liquid nitrogen (LN2), on gene expression profiles were also analysed.ResultsOn comparison of the 3’mRNA-Seq data generated from pituitary, hypothalamus, liver and lung tissues, no significant differential expression in the protein coding genes were detected between the euthanasia methods. This implies that the nitrogen anoxia method could be a suitable alternative for euthanasia of piglets used in transcriptomic research. However, small nuclear RNAs (snRNAs) that constitute the eukaryotic spliceosomal machinery were found to be significantly higher (log2fold change ≥ 2.0, and adjusted p value ≤ 0.1) in pituitary samples collected using ANOXIA. Non-protein coding genes like snRNAs that play an important role in pre-mRNA splicing can subsequently modify gene expression. Storage in RL was found to be superior in preserving RNA compared to LN2 storage, as evidenced by the significantly higher RIN values in representative samples. However, storage in RL as opposed to LN2, also influenced differential gene expression in multiple tissues, perhaps as a result of its inability to inhibit biological activity during storage. Hence such external sources of variations should be carefully considered before arriving at research conclusions.ConclusionsSource of biological variations like euthanasia method and storage condition can confound research findings. Even if we are unable to prevent the effect of these external factors, it will be useful to identify the impact of these variables on the parameter under observation and thereby prevent misinterpretation of our results.

Survival of Listeria monocytogenes and Salmonella in citrus storage waxes or on lemons held under common commercial storage conditions

To prolong cold storage, diluted storage waxes are applied to washed lemons after harvest and before packing, without drying steps, to reduce premature rotting and water loss. The survival of Listeria monocytogenes and Salmonella in undiluted and diluted storage waxes (S1–S4), and on lemon surfaces under common commercial storage were investigated. Populations of L. monocytogenes declined more slowly than Salmonella in undiluted storage waxes over 24 h of storage at 4 or 22 °C. L. monocytogenes (inoculated at ∼6 log CFU/mL) was detected by enrichment in undiluted waxes S2, S3, and S4 after 75–135 days at 4 °C but not after 30, 10, or 105 days, respectively at 22 °C. L. monocytogenes survived better in diluted than in undiluted storage waxes at 22 °C. Populations of L. monocytogenes (∼6 log CFU/lemon) declined by 0.64–1.62 log on lemon surfaces right after waxing. Populations of L. monocytogenes decreased to <1.30 log CFU/lemon after 28 days (1:9 S1) or 75 days (other treatments) at 12 °C and ≥93% RH. Except for 1:9 S1, L. monocytogenes was detected by enrichment in all lemon samples over 87 days of storage. Packinghouses should consider the survival of L. monocytogenes and Salmonella in citrus storage waxes in their food safety programs.

Pre-analytical and analytical variability of reticulocyte counts in dogs.

Automated fluorescence-based haematology analysers are now available for reticulocyte enumeration in veterinary medicine, but manual counting is still largely used. This study aimed to evaluate potential sources of analytical and pre-analytical errors when performing automatedand manual counts. Automated and two-operator double-blind manual reticulocyte counts were performed on 15 blood samples. The intra-assay variation of the automated and manual counts and the interoperator variationin the manual counts were then calculated. In addition, the effects of storage were evaluated using samples refrigerated at 4°C or stored at room temperature for 2, 4, 12, 24, 48 or 72 hours after sampling. Intra-assay coefficients of variation were lower for automated counts than for manual counts. Comparison between automated and mean total manual reticulocyte count showed no significant differences. In both refrigerated samples and those stored at room temperature, an increase in reticulocyte count was recorded only after 72 hours. Staining artefacts occurred only in one stored sample counted manually. The presence of cytoplasmic particles other than RNA can cause misinterpretation of cells, leading to an erroneous reticulocyte count. The use of an automated analyser is preferable for reticulocyte enumeration in dogs. Common storage conditions seem to minimally affect reticulocyte evaluation; however, it is recommended to perform the analysis as soon as possible after sampling.

Effect of surface functionalization on the moisture stability and sorption properties of porous boron nitride

Porous boron nitride (BN) is a promising adsorbent owing to its high surface area and porosity, as well as thermal and oxidative stability. It has been explored in the past decade for applications in gas and liquid separations, such as CO2 capture and water cleaning. However, the material has displayed hydrolytic instability. Owing to the presence of moisture in most industrial settings, whether it is for storage or cyclic adsorption processes, ensuring the moisture stability of an adsorbent is crucial. While this topic has been researched for other adsorbents such as zeolites and metal organic frameworks (MOFs), little is known on controlling the hydrolytic stability of porous BN. In this study, we propose a method to enhance porous BN's hydrolytic stability via surface functionalization using a fluoroalkylsilane. We explored two different routes of functionalization: (i) functionalization of porous BN powder followed by pelletization (route 1) and (ii) coating of porous BN pellets with fluoroalkylsilane (route 2). Spectroscopic, analytical and imaging techniques confirmed the functionalization process qualitatively and quantitatively. We subjected the functionalized samples to moisture exposure at 54% RH (similar to common storage conditions) and 92% RH (similar to flue gas stream conditions with high moisture content), and characterized them to probe their resistance to moisture. We also investigated their equilibrium and kinetic sorption properties in the context of CO2/N2 separation. Both routes produced materials with enhanced moisture stability. However, we noted differences between both functionalization routes. Route 2 produced a sample with a higher grafting yield and hydrophobic nature, and therefore better resistance to moisture exposure than route 1. From a sorption point of view, despite reduced porosity, the functionalized samples maintain reasonable CO2 uptakes. The functionalization led to changes in the textural features of the samples, which caused differences in the mass transfer. This work shows that functionalization could be used to protect porous BN upon moisture exposure.

New protocol for kinetic assay seeding ability recovery "KASAR" from formalin-fixed paraffin-embedded tissues.

The real-time quaking-induced conversion (RT-QuIC) alpha-synuclein (aSyn) protein kinetic seeding assay has been very useful for detecting pathological aggregates in various synucleinopathies including Parkinson's disease (PD). This biomarker assay relies on fresh frozen tissue to effectively seed and amplify aSyn aggregating protein. With vast repositories of formalin-fixed paraffin-embedded (FFPE) tissues, it is paramount to harness the power of kinetic assays to unlock the diagnostic potential of archived FFPE biospecimens. However, the major challenge posed by significantly reduced amplification of formalin-fixed tissues in the assay suggests that formalin fixation deterred monomer interaction with the sample seed and depressed subsequent protein aggregation. To overcome this challenge, we developed a kinetic assay seeding ability recovery (KASAR) protocol to maintain the integrity of the tissue and seeding protein. For this, we implemented a series of heating steps with the brain tissue suspended in a buffer composed of 500mM tris-HCl (pH 7.5) and 0.02% SDS after the standard deparaffinization of the tissue sections. Initially, samples from seven human brain samples, including four samples from patients diagnosed with dementia with Lewy bodies (DLB) and three samples from healthy controls without DLB, were compared to fresh frozen samples under three different, but clinically common sample storage conditions: formalin-fixed, FFPE, and FFPE slices cut 5µm thick. The KASAR protocol was able to recover seeding activity for all positive samples in all storage conditions. Next, 28 FFPE samples from the submandibular gland (SMG) of patients diagnosed with PD, incidental Lewy body disease (ILBD), or healthy controls were tested with 93% of results replicating when blinded. With samples of only a few milligrams, this protocol recovered the same quality of seeding in formalin-fixed tissue as fresh frozen tissue. Moving forward, protein aggregate kinetic assays, in conjunction with the KASAR protocol, can be used to understand and diagnose neurodegenerative diseases more comprehensively. Overall, our KASAR protocol unlocks and restores the seeding ability of formalin-fixed paraffin-embedded tissues for the amplification of biomarker protein aggregates in kinetic assays.

Shelf-life of uranium oxide microparticle reference materials and possible implications for the identification of optimal storage conditions

During their inspections of nuclear facilities, inspectors of the International Atomic Energy Agency (IAEA) collect environmental swipe samples containing uranium dust particles. As reference materials, well-characterized uranium microparticles are required that are suitable for quality control, analytical refinement and method development tasks. At Forschungszentrum Jülich (FZJ), uranium oxide reference microparticles are produced using an aerosol-based process. A core requirement for distribution of a potential reference material is to guarantee a practical shelf-life. Previous studies of structure and shape of uranium oxide microparticles demonstrated possible alteration leading to the formation of uranium hydroxides such as schoepite. A systematic shelf-life study exploring storage under different environmental conditions was launched in late 2021. Uranium microparticles were stored in three environments simulating potential long-term storage conditions, whereas a fourth experiment used an unrealistically harsh environment to assess the effects of accelerated alteration. At the time of writing, only the uranium microparticles intentionally stored under these extreme conditions have shown significant signs of alteration. This implies that synthetic uranium oxide microparticles under common storage conditions have minimal shelf-lives of at least 1 year, and likely much longer.Graphical abstract

Establishing preanalytical stability of vitamin A and vitamin E

Vitamin A and Vitamin E, a group of lipid-soluble vitamins, can be degraded by photolysis and photooxidation after exposure to light. As an essential area of the preanalytical stage, inappropriate storage conditions of patient samples could lead to inaccurate results. In this study, we evaluated three of the most common preanalytical storage conditions (RT, +4°C, and −20 °C) in the workflow in the clinical laboratory setting using both clear and amber Eppendorf tubes. Analyte stability in each storage condition was tested with five patient pools. After 24 hrs at RT, a significant decrease in vitamin A and E was observed for samples stored in clear Eppendorf tubes (19.2 % and 17.9 %, for vitamin A and E, respectively), compared with samples stored in amber Eppendorf tubes (9.1 % and 7.3 %), respectively. After the specimen was stored at 4 °C for 7 days, vitamin A concentrations decreased in clear tubes by 14.7 % and in amber tubes by 12.7 %. Similarly, vitamin E concentrations decreased in both clear and amber tubes by 11.2 % and 13.4 %, respectively. Both vitamin A and vitamin E demonstrated acceptable stability in clear and amber Eppendorf tubes at −20 °C for up to 22 days. It is preferable to use amber tubes to protect vitamin A and E from light during short term storage. Storage of vitamin A and E specimens beyond 7 days should be at −20 °C.

Evaluation of Storage Quality of Hardy Kiwifruit (Actinidia arguta): Effect of 1-MCP and Maturity Stage

Hardy kiwifruit fits into consumer expectations, especially in terms of taste and nutritional value. The main reason for the loss of quality is that the fruit softens too quickly, but storage at low temperatures (0–1 °C) is not effective enough in maintaining high fruit quality. Two cultivars were evaluated for storage quality, i.e., ‘Geneva’ and ‘Ananasnaya’. Minikiwi were harvested at two stages of maturity. The fruit were treated with 1-methylcyclopropene at a concentration of 0.65 µL/L and then stored under common cold storage, ULO, and CA conditions. Fruit evaluation was carried out for 12 weeks, and firmness, soluble solids content, and titratable acidity were evaluated. Post-harvest treatments, as well as the conditions under which the fruit is stored, determine its quality after storage. The application of 1-MCP decisively inhibits the softening of mini kiwi even under normal atmospheric conditions.

Bionomics of Callosobruchus analis (F.) in ten common food legumes

Range of edible legumes suffer huge post-harvest deterioration due to infestation by Callosobruchus spp. (Coleoptera: Chrysomelidae). Among the bruchid species, despite wider distribution in India and its destructive potential, Callosobruchus analis (F.) has been largely overlooked. Hence, we report comparative bionomics of C. analis, influence of seed traits and seed losses in ten commonly cultivated food legumes namely, mungbean, urdbean, chickpea-desi, chickpea-kabuli, cowpea, field pea, pigeonpea, rajmash, moth bean and lentil. Oviposition behaviour, developmental rate of insect and seed losses varied significantly under the influence of seed physico-chemical traits under free-choice and no-choice conditions. Cowpea, pigeonpea and rajmash harboured higher number of eggs when choice for legume was offered, however, none of the tested legume was completely avoided for egg laying. The survivability was found negligible on rajmash and field pea. Developmental time was 1.3–1.5 times longer on urdbean (45.97 days) in comparison to other legumes, indicating the role of antibiosis factors. More progenies were produced on pigeonpea, cowpea, moth bean, chickpea-desi and mungbean leading to high seed losses. Seed properties namely, seed surface area, hardness, seed coat thickness, 100-seed weight, seed moisture and phenol contents significantly influenced oviposition behaviour and growth of the insects. Further studies are needed to elucidate exact mode of action of seed traits and their interaction on the insect bionomics. The legumes those suffered considerable seed losses by C. analis warrant adequate preventative measures to avoid cross-infestation under common storage conditions and reduce post-harvest deterioration.

A Shelf Stable Fmoc Hydrazine Resin for the Synthesis of Peptide Hydrazides.

C-terminal hydrazides are an important class of synthetic peptides with an ever expanding scope of applications, but their widespread application for chemical protein synthesis has been hampered due to the lack of stable resin linkers for synthesis of longer and more challenging peptide hydrazide fragments. We present a practical method for the regeneration, loading, and storage of trityl-chloride resins for the production of hydrazide containing peptides, leveraging 9-fluorenylmethyl carbazate. We show that these resins are extremely stable under several common resin storage conditions. The application of these resins to solid phase peptide synthesis (SPPS) is demonstrated through the synthesis of the 40-mer GLP-1R agonist peptide "P5". These studies support the broad utility of Fmoc-NHNH-Trt resins for SPPS of C-terminal hydrazide peptides.

Survival kinetics, membrane integrity and metabolic activity of Salmonella enterica in conventionally and osmotically dehydrated coconut flakes

Many outbreaks involving Salmonella enterica in dehydrated coconut have been reported. Little is known about the survival of S. enterica in dehydrated coconut flakes at common retail or domestic storage conditions. This study evaluated the behavior of a S. enterica cocktail (S. Enteritidis PT4, S. Typhimurium PT4, S. Bredeney, S. Muenster and S. Agona) in conventionally and osmotically dehydrated coconut flakes under four storage regimes: 25 °C for 120 days, 25 °C for 30 days then 7 °C for 90 days, 7 °C for 30 days then 25 °C for 90 days, and 7 °C for 120 days. S. enterica membrane integrity (using with propidium iodide and bis-1,3-dibutylbarbutyric acid) and metabolic activity (using 5-cyano-2,3-ditolyl tetrazolium chloride) were assessed by flow cytometry analysis after dehydration and storage at 7 °C or 25 °C for 120 days. Lower S. enterica inactivation rates (kmax 0.02 to 0.04 1/days) were observed in conventionally dehydrated coconut flakes compared to osmotically dehydrated coconut flakes (kmax 0.16 to 0.20 1/days). Changes in storage temperature did not affect the behavior of S. enterica in conventionally or osmotically dehydrated coconut flakes. Results show that S. enterica inactivation in conventionally dehydrated coconut flakes could be described by log-linear with tail models. S. enterica inactivation in osmotically dehydrated coconut flakes could be described by log-linear with shoulder and tail models. Subpopulations of S. enterica cells with damaged membranes and without metabolic activity were larger in conventionally (32.1% and 90.9%, respectively) than osmotically dehydrated coconut (18.5% and 82.2%, respectively) flakes at the beginning of the storage. Subpopulations of S. enterica cells with damaged membrane decreased by 9.4–14.4%, while cells with membrane potential and intact membrane increased by 23.7 and 24.2% in conventionally dehydrated coconut flakes after 120 days of storage at 7 °C or 25 °C, respectively. Subpopulations of S. enterica with damaged membranes did not change significantly in osmotically dehydrated coconut flakes. Our findings suggest that S. enterica populations decline during storage occurs due in part to membrane integrity losses. These data can contribute to the development of risk management strategies for S. enterica in dehydrated coconut flakes.

A New In Vitro Blood Hyperviscosity Model.

We developed a model of blood hyperviscosity avoiding extreme impact on the blood. The model shows reproducibility in rat blood under common storage conditions (4±1°C; stabilization with citrate-phosphate-glucose additive solution). Storage of rat blood under these condition leads to impairment of its rheological properties, which manifested in an increase in blood viscosity in a wide range of shear rates (3-300 sec-1). An increase in blood viscosity appeared the first day of storage and reached a maximum on the third day. During further 11-day storage, the blood viscosity did not change significantly. A hybrid macromolecular compound O-(3-(3,5-di-tert-butyl-4-hydroxyphenyl)propanoyl)-(1→6)-α-D-glucan improved the hemorheological properties during storage. The most pronounced effect was observed on the third day of storage and manifested in a decrease in blood viscosity in the range of shear rates of 50-300 sec-1. Thus, storage of rat blood with citrate-phosphate-glucose additive solution for 3 days at 4±1°C reproduces the phenomenon of blood hyperviscosity; this model can be used to screen agents with hemorheological activity.

(IMCS First Place Best Poster Award) Encapsulation through Photoinitiated Chemical Vapor Phase Deposition (piCVD) for Obtaining Antifouling and Stabilized Biosensing Interface

Biorecognition elements based biosensing devices have been applied in a broad range of clinical applications and are considered of great potential for being transformed into on-site portable techniques due to their high selectivity and sensitivity. Yet, given the success of immobilization methodologies adapted for various sensing platforms, the drive for enhanced surface microenvironments including constant reaction conditions and suppression of nonspecific binding remains. Considering the complexity of the real sensing scenario with various biofoulants (proteins, cells, polysaccharides, etc.), the fast analytes build-up at the interface would cause sensing accuracy to be either false positive or false negative in a short time. While the concern may be alleviated regarding disposable usage, the fragile biocomponent may degrade even on common storage conditions since the temperature, humidity, as well as air quality, keep varying.Great emphasis has been put on encapsulation techniques to create surface barriers that may simultaneously produce a stable inner environment at the proper humidity and temperature zone and less complicated outer surroundings arising from fouling. However, to compensate for deactivation induced by surface pretreatments and organic solvent used in many widely adopted antifouling techniques, such as spin-coating, layer-by-layer assembly (LbL), surface grafting, etc., high biocomponent loadings are always required. Besides, without precise control over obtaining thin and uniform encapsulation layers, such techniques usually fail to ensure quick mass transfer and accurate data acquisition. Chemical vapor deposition (CVD), on the other hand, is a solvent-free process in which vapor-phase precursors chemically react on a substrate surface to generate uniform films with varying thickness. Previous studies have demonstrated that specific protein-resistant hydrogels can be synthesized via photoinitiated chemical vapor deposition (piCVD) on optical sensors without sacrificing sensitivity and accuracy. However, antifouling layers deposited directly on immobilized biomolecules and used for screening more complicated analytes, such as lysate, have not been investigated.Herein, antifouling and stabilized interfaces capable of sensitively assaying cell lysate were constructed by depositing ultrathin encapsulation layers on silicon wafers with immobilized nanobodies via piCVD. Since an antifouling mechanism can be either interpreted as solvation barrier induced by hydrophilic groups or presented as easy foulants removal given by the low surface energy nature of hydrophobic materials, poly(hydroxyethyl acrylate) (pHEA) and poly(1H,1H,2H,2H-perfluorooctyl acrylate) (pTFOA) are selected and studied as the representatives of the two groups due to their biocompatibility and multiple synthetic routes. Interestingly, despite significant fouling reduction and stabilized local environments under harsh conditions were achieved by both materials, the compatibility of pTFOA with piCVD was much better than pHEA, which was observed as being more sensitive to the environmental humidity and thus exhibited poor repeatability. Consequently, the piCVD was successfully applied to form an encapsulation layer on “sensitive” substrates to reject unwanted binding and provide constant storage microenvironments, indicating great promise for enhanced biosensing interface fabrication

MAGEA4 Coated Extracellular Vesicles Are Stable and Can Be Assembled In Vitro.

Extracellular vesicles (EVs) are valued candidates for the development of new tools for medical applications. Vesicles carrying melanoma-associated antigen A (MAGEA) proteins, a subfamily of cancer-testis antigens, are particularly promising tools in the fight against cancer. Here, we have studied the biophysical and chemical properties of MAGEA4-EVs and show that they are stable under common storage conditions such as keeping at +4 °C and −80 °C for at least 3 weeks after purification. The MAGEA4-EVs can be freeze-thawed two times without losing MAGEA4 in detectable quantities. The attachment of MAGEA4 to the surface of EVs cannot be disrupted by high salt concentrations or chelators, but the vesicles are sensitive to high pH. The MAGEA4 protein can bind to the surface of EVs in vitro, using robust passive incubation. In addition, EVs can be loaded with recombinant proteins fused to the MAGEA4 open reading frame within the cells and also in vitro. The high stability of MAGEA4-EVs ensures their potential for the development of EV-based anti-cancer applications.

Effect of Storage on the Nutritional Quality of Queen Garnet Plum.

Due to high perishability, plums are harvested at an early stage of maturity to extend postharvest storage life. Storage time and temperature can significantly affect the phytochemical and sugar composition of plums, altering their palatability and nutritional quality. In this study, variations in physiochemical properties (total soluble solids (TSS), titratable acidity (TA), color (chroma and hue angle)), phytochemical composition (total phenolic content (TPC), total anthocyanin content (TAC), and carotenoids), and sugars in three different tissues of the Queen Garnet plum (QGP) during storage at two common domestic storage temperatures (4 and 23 °C) were evaluated. There was an increase (p > 0.05) in TSS and a reduction (p < 0.05) in TA of the outer flesh at 23 °C. Chroma values of all the tissues reduced (p < 0.05) at 23 °C. At 4 °C, chroma values fluctuated between storage days. The TAC of the peel was the highest (p < 0.05) among the different tissues and continued to increase up to 10 days of storage at 23 °C (3-fold increase). At 4 °C, the highest (p < 0.05) TAC (peel) was observed after 14 days of storage (1.2-fold increase). TPC showed similar results. The highest (p < 0.05) TPC was recorded in the peel after 10 days of storage at 23 °C (2.3-fold increase) and after 14 days of storage at 4 °C (1.3-fold increase), respectively. Total carotenoids in the flesh samples at both storage temperatures were reduced (p < 0.05) after 14 days. Total sugars also decreased during storage. The results of the present study clearly showed that common domestic storage conditions can improve the nutritional quality of plums by increasing the content of bioactive anthocyanins and other phenolic compounds. However, the increase in phytochemicals needs to be counterbalanced with the decrease in total sugars and TA potentially affecting the sensory attributes of the plums.

Formation and Stabilization of W1/O/W2 Emulsions with Gelled Lipid Phases.

Water-in-oil-in-water (W1/O/W2) emulsions are emulsion-based systems where the dispersed phase is an emulsion itself, offering great potential for the encapsulation of hydrophilic bioactive compounds. However, their formation and stabilization is still a challenge mainly due to water migration, which could be reduced by lipid phase gelation. This study aimed to assess the impact of lipid phase state being liquid or gelled using glyceryl stearate (GS) at 1% (w/w) as well as the hydrophilic emulsifier (T80: Tween 80 or lecithin) and the oil type (MCT:medium chain triglyceride or corn oil (CO) as long chain triglyceride) on the formation and stabilization of chlorophyllin W1/O/W2 emulsions. Their colloidal stability against temperature and light exposure conditions was evaluated. Gelling both lipid phases (MCT and CO) rendered smaller W1 droplets during the first emulsification step, followed by formation of W1/O/W2 emulsions with smaller W1/O droplet size and more stable against clarification. The stability of W1/O/W2 emulsions was sensitive to a temperature increase, which might be related to the lower gelling degree of the lipid phase at higher temperatures. This study provides valuable insight for the formation and stabilization of W1/O/W2 emulsions with gelled lipid phases as delivery systems of hydrophilic bioactive compounds under common food storage conditions.

Study on Post Harvest Losses in Potato in Different Storage Conditions

The study aimed to conduct a comparative study of three most common potato storage conditions (in-house store, in-basket store and cold store) and to find the most suitable storage condition for farmers. The experimental work involved post-harvest loss assessment and study of pattern of respiratory and reducing sugar level for sixty days storage period. Total loss was lowest (4.38%) for cold stored potatoes and highest (13.04%) for in-house stored potatoes. Reducing sugar accumulation was least (0.65%) and remained almost constant throughout study period for in-house stored potatoes. Reducing sugar accumulation gradually increased and was maximum (1.04%) for cold stored potatoes. Respiratory rate was least in cold stored potatoes which gradually decreased and reached 3.17mg CO2/kg/hr at the end of sixty days storage. Respiratory rate was maximum for in-basket stored potatoes which reached up to 6.55 mg CO2/kg/hr at the end of storage. Storage loss and respiratory rate are minimum for cold stored potatoes but showed high sugar accumulation. In-house storage do not suffer from excessive sugar accumulation but storage loss is maximum of all. In case of in-basket storage, besides being cheap and feasible, there is no problem of sugar accumulation and storage losses was also significantly lower than in-house storage.