Effect of the 35 nm and 70 nm Size Exclusion Chromatography (SEC) Column and Plasma Storage Time on Separated Extracellular Vesicles.

The technical difficulty of separating extracellular vesicles (EVs) from plasma proteins in human blood presents a significant hurdle in EV research, particularly during nano ultra-high-performance liquid chromatography-tandem mass spectrometric (UHPLC-MS/MS) analysis, where detecting "vesicular" proteins among abundant plasma proteins is challenging. Standardisation is a pressing issue in EV research, prompting collaborative global efforts to address it. While the MISEV guidelines offer valuable recommendations, unanswered questions remain, particularly regarding sample storage. We compared size exclusion chromatography (SEC) columns with pore sizes of 35 nm and 70 nm to identify fractions with minimal contaminating proteins and the highest concentration of small EVs (sEVs). Following column selection, we explored potential differences in the quality and quantity of sEVs isolated from platelet-free plasma (PFP) after long-term storage at -80 °C (>2.5 years) compared to freshly drawn blood. Our methodologically rigorous study indicates that prolonged storage, under correct storage and processing conditions, does not compromise sEV quality. Both columns effectively isolated vesicles, with the 70 nm column exhibiting a higher abundance of "vesicular" proteins. We propose a relatively rapid and moderately efficient protocol for obtaining a comparatively pure sEV fraction from plasma, facilitating sEV processing in clinical trials.

Vitrification preservation of good-quality blastocysts for more than 5 years reduces implantation and live birth rates.

STUDY QUESTION: Does vitrification cryopreservation of embryos for more than 5 years affect the pregnancy outcomes after frozen embryo transfer (FET)? SUMMARY ANSWER: Vitrification cryopreservation of good-quality blastocysts for more than 5 years is associated with a decrease in the implantation rate (IR) and live birth rate (LBR). WHAT IS KNOWN ALREADY: Previous studies have predominantly focused on embryos cryopreserved for relatively short durations (less than 5 years), yet the impact of extended cryopreservation duration on pregnancy outcomes remains a controversial issue. There is a relative scarcity of data regarding the efficacy and safety of storing embryos for 5 years or longer. STUDY DESIGN, SIZE, DURATION: This retrospective study involved 36 665 eligible vitrified-thawed embryo transfer cycles from 1 January 2016 to 31 December 2022, at a single fertility center in China. PARTICIPANTS/MATERIALS, SETTING, METHODS: Patients were divided into three groups according to embryo storage time: Group 1 consisted of 31 565 cycles, with storage time of 0-2 years; Group 2 consisted of 4458 cycles, with a storage time of 2-5 years; and Group 3 included 642 cycles, with storage time exceeding 5 years. The main outcome measures were IR and LBR. Secondary outcome variables included rates of biochemical pregnancy, multiple pregnancy, ectopic pregnancy, and miscarriage, as well as neonatal outcomes. Reproductive outcomes were analyzed as binary variables. Multivariate logistic regression analysis was used to explore the effect of preservation time on pregnancy outcomes after correcting for confounding factors. In addition, we also assessed neonatal outcomes, such as large for gestational age (LGA) and small for gestational age (SGA). MAIN RESULTS AND THE ROLE OF CHANCE: IRs in the three groups (0-2, 2-5, and >5 years) were 37.37%, 39.03%, and 35.78%, respectively (P = 0.017), and LBRs in the three groups were 37.29%, 39.09%, and 34.91%, respectively (P = 0.028). After adjustment for potential confounding factors, compared with the 0-2 years storage group, prolonged embryo vitrification preservation time (2-5 years or >5 years) did not affect secondary outcomes such as rates of biochemical pregnancy, multiple pregnancy, ectopic pregnancy, and miscarriage (P > 0.05). But cryopreservation of embryos for more than 5 years reduced the IR (adjusted odds ratio (aOR) 0.82, 95% CI 0.69-0.97, P = 0.020) and LBR (aOR 0.76, 95% CI 0.64-0.91, P = 0.002). Multivariate stratified analysis also showed that prolonging the cryopreservation time of blastocysts (>5 years) reduced the IR (aOR 0.78, 95% CI 0.62-0.98, P = 0.033) and LBR (aOR 0.68, 95% CI 0.53-0.87, P = 0.002). However, no effect on cleavage embryos was observed (P > 0.05). We further conducted stratified analyses based on the number and quality of frozen blastocysts transferred, and the results showed that the FET results after transfers of good-quality blastocysts in the >5 years storage group were negatively affected. However, the storage time of non-good-quality blastocysts was not significantly associated with pregnancy outcomes. Regarding the neonatal outcomes (of singletons), embryo vitrification preservation time had no effect on preterm birth rates, fetal birth weight, or neonatal sex ratios. However, as the storage time increased, rates of SGA (5.60%, 4.10%, and 1.18%) decreased, while rates of LGA (5.22%, 6.75%, and 9.47%) increased (P < 0.05). After adjusting for confounding factors, the increase in LGA and the decrease in SGA were significantly correlated with the duration of storage time. LIMITATIONS, REASONS FOR CAUTION: This was a retrospective study using data from a single fertility center, even though the data had been adjusted, our findings still need to be validated in further studies. WIDER IMPLICATIONS OF THE FINDINGS: With the full implementation of the two-child policy in China, there may be more patients whose embryos have been frozen for a longer time in the future. Patients should be aware that the IR and LBR of blastocysts are negatively affected when the cryopreservation time is longer than 5 years. Couples may therefore consider shortening the time until FET treatment. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by the National Nature Science Foundation of China (No. 82101672), Science and Technology Projects in Guangzhou (No. 2024A03J0180), General Guidance Program for Western Medicine of Guangzhou Municipal Health Commission (No. 20231A011096), and the Medical Key Discipline of Guangzhou (2021-2023). None of the authors have any conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Short communication: Storage time and temperature affect plasma osmolality values in field-collected blood samples.

As climate change alters the hydric regime of many habitats, understanding the hydric physiology of animals becomes increasingly important. Plasma osmolality is a popular metric to assess an organism's hydration, but samples often need to be stored before being analyzed, under varying conditions and for different lengths of time. Previous studies on plasma storage conditions, and how they impact sample integrity, are minimal and have focused more on clinical applications than field studies. We studied the stability of osmolality values from wild rattlesnake plasma samples stored in commonly used plastic snap-cap tubes under different time (0, 2, 3, 7, 29 days) and temperature (refrigerated at 2 °C and frozen at -18 °C) treatments. We hypothesized that frozen samples would remain more stable (e.g., retain osmolality values more similar to baseline values) than refrigerated samples because freezing the plasma would reduce evaporation. We found that osmolality of samples increased over time at both temperatures, becoming significantly higher than baseline after 7 days. Contrary to our prediction, osmolality increased more in frozen samples than in refrigerated samples. We discuss possible reasons for our results, along with their implications. To obtain the most accurate plasma osmolality values, we recommend refrigerating plasma samples for as short a time as possible, 3 days or fewer, before analyzing them on an osmometer.

Impact of prolonged storage time on homograft ultrastructures: an attempt to find optimal guidelines for homograft processing.

According to guidelines, total ischemic time for homografts at processing must be kept short to avoid degeneration. Many homografts are discarded due to practical inability to finish all steps from procurement to cryopreservation within the time limit. Although, several studies have shown that homografts with prolonged ischemic time show adequate quality and performance. Twenty aortic and 12 pulmonary homografts were collected and biopsies were retrieved at preparation (day 0) and after 1, 2, 3, 4, 7, 14, 21, 28, and 60 days in antibiotic decontamination at 4 °C. Biopsies were prepared for light microscopy (LM) and transmission electron microscopy (TEM). Assessment generated scores for cells, elastin, and collagen. Relative differences between times were compared with Wilcoxon signed rank test. Bonferroni corrected p value of 0.0056 was considered significant. LM could only reveal decrease in cell count at 60 days in aortic homografts, no other differences was detected. TEM showed affected cell appearance in day 3 and day 4 and beyond for aortic and pulmonary homografts respectively. Elastin appearance was affected at day 60 for aortic and day 21 for pulmonary homografts. Collagen appearance was affected at day 28 for aortic homografts, with no significant differences in pulmonary homografts. Cell degeneration starts early after homograft procurement, but elastic and collagen fibers are more resistant to degeneration. Overall structure integrity as seen in LM was not affected at all, while TEM could reveal small degeneration signs in individual elastic fibers and collagen bundles at 21 and 28 days respectively.

Effect of postharvest storage time on quality characteristics of explosion puffing dried whole shiitake mushroom (Lentinula edodes) crisps.

BACKGROUND: Non-fried shiitake mushroom (Lentinula edodes) crisps fabricated by explosion puffing drying (EPD) are receiving worldwide attention because of their crispness, convenience, nutrition and health functions. The quality of mushroom crisps varies with storage time of fresh L. edodes. Therefore, the effect of postharvest storage time (ranging from 0 to 14 days) of fresh L. edodes on quality characteristics of EPD- processed mushroom crisps was evaluated. RESULTS: The weight loss and total color difference of fresh L. edodes were increased to 2.95% and 24.66, but moisture content, firmness and lightness were reduced by 6.14%, 40.70% and 43.57%, respectively, after 14 days storage. The puffing degree of mushroom crisps was initially increased to its highest value (55.95%) on the 4th day storage and thereafter decreased. The highest rehydration ratio (2.36) and crispness (63.67), and lowest hardness (102.95 N) of mushroom crisps were fabricated with L. edodes on the 4th day of storage. Free water was predominant in fresh L. edodes, which was decreased for fresh L. edodes, whereas it increased initially to the maximum value and decreased thereafter for osmotic dehydrated and heat pump pre-dried L. edodes with increasing storage time. Principal component analysis and hierarchical cluster analysis confirmed that fresh L. edodes stored at different times had a remarkable effect on quality characteristics of mushroom crisps. CONCLUSION: Fresh L. edodes stored at 4 ± 1 °C for 4 days is recommended for fabrication of mushroom crisps with superior quality. This study provides a theoretical basis for selection of a suitable storage time for fresh L. edodes before EPD of crisps. © 2023 Society of Chemical Industry.

Long-term storage of vitrified oocytes does not affect pregnancy and live birth rates: analysis of 5362 oocyte donation cycles.

RESEARCH QUESTION: Does long-term storage of vitrified oocytes affect laboratory and reproductive outcomes after intracytoplasmic sperm injection? DESIGN: Retrospective cohort study including 41,783 vitrified-warmed oocytes from 5362 oocyte donation cycles between 2013 and 2021. Five categories of storage time were established to analyse its effect on clinical and reproductive outcomes (≤1 year [reference group], 1-2 years, 2-3 years, 3-4 years and >4 years). RESULTS: The mean number of warmed oocytes was 8.0 ± 2.5 oocytes. Oocyte storage time ranged from 3 days to 8.2 years (mean: 0.7 ± 0.9). Mean oocyte survival (90.2% ± 14.7% overall) did not significantly decrease with longer storage time after adjusting for confounders (88.9% for time >4 years, P = 0.963). A linear regression model did not show a significant effect of oocyte storage time on fertilization rate (about 70% in all time categories) (P > 0.05). Reproductive outcomes after the first embryo transfer were statistically comparable across storage times (P > 0.05 for all categories). Longer term oocyte storage (>4 years) did not affect the chances of clinical pregnancy (OR 0.700, 95% CI 0.423 to 1.158, P = 0.2214) or live birth (OR 0.716, 95% CI 0.425 to 1.208, P = 0.2670). CONCLUSIONS: Oocyte survival, fertilization rate, pregnancy and live birth rates are not affected by the time spent by vitrified oocytes in vapour-phase nitrogen tanks.

Surface-enhanced Raman scattering integrating with machine learning for green tea storage time identification.

The rapid and accurate identification of complex samples still remains a great challenge, especially for those with similar compositions. In this work, we report an integration strategy consisting of surface-enhanced Raman scattering (SERS) and machine learning to discriminate complex and similar analytes, in this case green tea products with different storage times. Surface-functionalized Ag nanoparticles (NPs) were used as a SERS substrate to reveal the changes in the sensory components of green tea with variable storage time. Principal components analysis (PCA)-based support vector machine (SVM) classification was used to extract the key spectral features and identify green tea with different storage times. The results showed that such an integration strategy achieved high predictive accuracy on time tag discrimination for green tea. The multiclass SVM classifier successfully recognized green tea with different storage times at a prediction accuracy of 95.9%, sensitivity of 96.6%, and specificity of 98.8%. Therefore, this work illustrates that the SERS-based PCA-SVM platform might be a facile and reliable tool for the identification of complex matrices with subtle differentiations.

Effects of time and temperature of storage on chemical and nutritional characteristics of raw milk for Provolone Valpadana PDO cheesemaking: a multivariate approach.

We evaluated the possibility of increasing the storage temperature of raw milk for Provolone Valpadana cheesemaking, to identify the most suitable conditions of time and temperature for a pre-maturation process. We used Principal Component Analysis (PCA) to analyze the overall effects of different storage conditions on chemical, nutritional and technological characteristics of the raw milk. Four different thermal storage cycles, two at fixed temperature/time (6 and 12°C for 60 h) and two with two-phase thermal cycle (10 and 12°C for 15 h, followed by refrigeration at 4°C for 45 h) were studied. Although a moderate heterogeneity among raw milks from the 11 producers of Provolone Valpadana cheese was observed, PCA revealed the critical aspects of the extreme storage conditions (60 h of refrigeration). Some samples resulted in anomalous behaviors, probably related to unexpected fermentation phenomena occurring with increasing storage temperature. The acidification and the increase in the contents of lactic acid, soluble calcium, and degree of retinol isomerization observed in the anomalous samples can compromise the technological functionality of milk. Conversely, the storage with a two-phase thermal cycle did not lead to variations in any measured characteristic, suggesting that mild refrigeration conditions (10 or 12°C for 15 h followed by 4°C for 45 h) could be a good compromise in favoring milk pre-maturation without altering its quality characteristics.

Influence of Pre-Analytic Conditions on Quantity of Lymphocytes.

Lymphocytes are key players in the pathogenesis of multiple sclerosis and a distinct target of several immunomodulatory treatment strategies. In this study, we aim to evaluate the effect of various pre-analytic conditions on immune cell counts to conclude the relevance for clinical implications. Twenty healthy donors were assessed for the effects of distinct storage temperatures and times after blood draws, different durations of tourniquet application, body positions and varying aspiration forces during blood draws. Immune cell frequencies were analyzed using multicolor flowcytometry. While storage for 24 h at 37 °C after blood draws was associated with significantly lower cell counts, different durations of tourniquet application, body positions and varying aspirations speeds did not have significant impacts on the immune cell counts. Our data suggest that immune cell counts are differently affected by pre-analytic conditions being more sensitive to storage temperature. Pre-analytic conditions should be carefully considered when interpreting the laboratory values of immune cell subpopulations.

The Conditions Matter: The Toxicity of Titanium Trisulfide Nanoribbons to Bacteria E. coli Changes Dramatically Depending on the Chemical Environment and the Storage Time.

In this work, we present an analysis of the antibacterial activity of TiS3 nanostructures in water and 0.9% NaCl solution suspensions. TiS3 nanoribbons 1-10 µm long, 100-300 nm wide, and less than 100 nm thick were produced by the direct reaction of pure titanium powder with elemental sulphur in a quartz tube sealed under vacuum. For the toxicity test of a bioluminescent strain of E. coli we used concentrations from 1 to 0.0001 g L-1 and also studied fresh suspensions and suspensions left for 24 h. The strongest toxic effect was observed in freshly prepared water solutions where the luminescence of bacteria decreased by more than 75%. When saline solution was substituted for water or when the solutions were stored for 24 h it resulted in a considerable decrease in the TiS3 antibacterial effect. The toxicity of TiS3 in water exceeded the toxicity of the reference TiO2 nanoparticles, though when saline solution was used instead of water the opposite results were observed. In addition, we did not find a relationship between the antibacterial activity of water suspensions of nanoribbons and the stability of their colloidal systems, which indicates an insignificant contribution to the toxicity of aggregation processes. In 0.9% NaCl solution suspensions, toxicity increased in proportion to the increase in the zeta potential. We suppose that the noted specificity of toxicity is associated with the emission of hydrogen sulphide molecules from the surface of nanoribbons, which, depending on the concentration, can either decrease or increase oxidative stress, which is considered the key mechanism of nanomaterial cytotoxicity. However, the exact underlying mechanisms need further investigation. Thus, we have shown an important role of the dispersion medium and the period of storage in the antibacterial activity of TiS3 nanoribbons. Our results could be used in nanotoxicological studies of other two-dimensional nanomaterials, and for the development of novel antibacterial substances and other biomedical applications of this two-dimensional material.

Differentiation of Goat Meat Freshness Using Gas Chromatography with Ion Mobility Spectrometry.

To investigate the flavor changes in goat meat upon storage, the volatile components observed in goat meat after different storage periods were determined using gas chromatography-ion mobility spectrometry (GC-IMS). A total of 38 volatile organic compounds (VOCs) were determined from the goat meat samples, including alcohols, ketones, aldehydes, esters, hydrocarbons, ethers, and amine compounds. 1-Hexanol, 3-Hydroxy-2-butanone, and Ethyl Acetate were the main volatile substances in fresh goat meat, and they rapidly decreased with increasing storage time and can be used as biomarkers for identifying fresh meat. When combined with the contents of total volatile basic-nitrogen (TVB-N) and the total numbers of bacterial colonies observed in physical and chemical experiments, the characteristic volatile components of fresh, sub-fresh, and spoiled meat were determined by principal component analysis (PCA). This method will help with the detection of fraudulent production dates in goat meat sales.

Effect of Gaseous Ozone and Storage Time on Polyphenolic Profile and Sugar Content in Fruits of Selected Vaccinium corymbosum L. Genotypes.

One of the best sources of antioxidant and health-promoting bioactive substances is the fruit of V. corymbosum. A potent oxidizing agent, ozone (O3), can effectively eliminate bacteria. The application of ozone gas to V. corymbosum fruit during storage had a favorable impact on the fruit's phenolic component and sugar content in the current investigation. After 7 days of storage, phenolic content in all highbush blueberry cultivars and clones tested increased on average by 28.60%, including anthocyanins by 34%. After 14 days of storage, an average increase of 16.50% in phenolic compounds was observed, including a 20.53% increase in anthocyanins. Among all the tested varieties, clone BOR-21 treated with a dose of 0.01 mL·L-1 ozone for 30 min after 14 days had the highest TPC-143.73 mg·100 g-1 f.w. The sugar content of berries treated with a dose of 0.01 mL·L-1 ozone for 30 min, on day 7 and day 14 of storage increased by 9.2% and 6.3%, respectively. On day 7, the highest amount of total sugar (22.74 g·100 g-1) was observed in Duke cultivar after being exposed to 0.01 mL·L-1 ozone for 15 min. The ozonation treatments enhanced the fruit's saturation with nutrients, which raises the fruit's value as food.

Impacts of polyphenols on laying hens' productivity and egg quality: A review.

There has been a rapid increase in the world's output of main poultry products (meat and eggs). This reflects customer desire for these high-quality and safe products and the comparatively low price. Recently, natural feed additives, plants and products have been increasingly popular in the poultry and livestock industries to maintain and improve their health and production. Polyphenols are a type of micronutrient that is plentiful in our diet. They are phytochemicals that have health benefits, notably cardiovascular, cognitive function, antioxidant, anti-mutagenic, anti-inflammatory, antistress, anti-tumour, anti-pathogen, detoxification, growth-promoting and immunomodulating activities. On the other hand, excessive polyphenol levels have an unclear and sometimes negative impact on gastrointestinal tract health, nutrient digestion, digestive enzyme activity, vitamin, mineral absorption, laying hens performance and egg quality. As a result, this review illuminated polyphenols' various sources, classifications, biological activities, potential usage restrictions and effects on poultry, layer productivity and egg external and internal quality.

I3 - /I- Redox Reaction-mediated Organic Zinc-Air Batteries with Accelerated Kinetics and Long Shelf Lives.

The storage time of Zn-air batteries (ZABs) for practical implementation have been neglected long-lastingly. ZABs based on organic solvents promise long shelf lives but suffer from sluggish kinetics. Here, we report a longly storable ZAB with accelerated kinetics mediated by I3 - /I- redox. In the charge process, the electrooxidation of Zn5 (OH)8 Cl2 ⋅H2 O is accelerated by I3 - chemical oxidation. In the discharge process, I- adsorbed on the electrocatalyst changes the energy level of oxygen reduction reaction (ORR). Benefitting from these advantages, the prepared ZAB shows remarkably improved round-trip efficiency (56.03 % vs. 30.97 % without the mediator), and long-term cycling time (>2600 h) in ambient air without replacing any components or applying any protective treatment to Zn anode and electrocatalyst. After resting for 30 days without any protection, it can still directly discharge continuously for 32.5 h and charge/discharge very stably for 2200 h (440 cycles), which is evidently superior to aqueous ZABs (only 0/0.25 h, and 50/25 h (10/5 cycles) by mild/alkaline electrolyte replenishment). This study provides a strategy to solve both storage and sluggish kinetics issues that have been plaguing ZABs for centuries, opening up a new avenue to the industrial application of ZABs.

Physicochemical characteristics and shelf-life stability of soya bean oil-based shortening.

The use of animal fats as raw material for shortening production has been avoided because of low supply, and religious restrictions of certain beliefs. The use of hydrogenated vegetable oils is also avoided because that may induce cardiovascular diseases. Palm oils and soya bean oil are theoretically potentials to be used as raw materials for shortening manufacturing due to their triacylglycerols composition and these oils can be easily modified to achieve desirable plasticity. In this study, shortening was produced by formulating a blend of palm stearin and soya bean oil in varying proportions. Physicochemical properties, product stability, and sensory acceptability of the processed shortening were determined. Stability tests of the processed shortening were determined for 6 months at two months intervals. The acidity, peroxide value, and free fatty acid values were increased with storage time and storage temperature. The physicochemical properties of the processed shortening samples were within the requirements of the food domain. The samples stored at 37 °C exhibited the highest acid, peroxide, and free fatty acid values throughout storage time. In conclusion, shortening produced from 60% palm stearin (S60) and stored at room temperature has shown a good physicochemical characteristic and is well accepted for different sensory attributes.

The effect of different staining solutions on the color stability of temporary crown materials.

Background: The esthetic expectations of patients are increasing by the day. That is why it is important to minimize the color changes in the oral cavity in both the temporary and permanent restorations. Aim: This study was carried out to compare the time-dependent color changes of polished and unpolished temporary crown materials prepared by different methods in various solutions. Materials and Methods: Half of the two different temporary restoration materials prepared with a diameter of 10 mm and a thickness of 2 mm were polished, and half were not polished. The ΔE* values of the samples kept in various solutions were recorded. Data were statistically evaluated by using variance analysis (ANOVA) and a Tukey HSD multiple comparison test. Results: It was determined that the material type, the solution, the interaction between the material types and the surface treatment, and the interaction between the surface treatment and the solution were statistically significant for color change (p < 0.001). Conclusion: The most significant color change in the inter-material evaluation was observed in chemically polymerized polymethyl methacrylate. In the evaluation between beverages, the highest color change was found in sugared coffee, and the lesser color change was observed in polished samples.

Differential expression of exosomal microRNAs in fresh and senescent apheresis platelet concentrates.

Patients have a high risk of suffering adverse reactions after receiving platelet products stored for 5 days. Bioactive exosomes in platelet products can be accumulated during storage, which is associated with adverse reactions. MicroRNAs are one of the critical cargoes in exosomes, which participate in cell differentiation, metabolism, and immunomodulation. This study intends to elucidate and analyze the differential expression of exosomal microRNAs in apheresis platelet concentrates during storage and predict the potential functions of target genes. Apheresis platelet concentrates were used to isolate exosomes by ultracentrifugation. Exosomes were phenotyped by western blot, transmission electron microscopy, and nano flow cytometry. The differential expression of the exosomal microRNAs was obtained by a microarray test using four bags of apheresis platelets stored for 5 days compared with 1 day. The differentially expressed microRNAs between the two time points were identified, and their target genes were analyzed by miRWalk and miRDB. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to predict the target genes' functions. Fifteen bags of apheresis platelet concentrates stored for 1 day and 5 days were used to verify the microarray results by quantitative reverse transcription-polymerase chain reactions (qRT-PCR). There were 134 microRNAs in total expressed differently in the two groups (day 1 and day 5), with 57 microRNAs up-regulated and 77 down-regulated (|fold change| > 2.0 and P < .05). Thirteen up-regulated microRNAs (hsa-miR-22-3p, hsa-miR-223-3p, hsa-miR-21-5p, hsa-miR-23a-3p, hsa-miR-320b, hsa-let-7a-5p, hsa-miR-25-3p, hsa-miR-126-3p, hsa-miR-320c, hsa-miR-342-3p, hsa-miR-320d, hsa-miR-328-3p, and hsa-miR-320e) detected in all samples were selected to validate the results. The qRT-PCR results showed that five (hsa-miR-22-3p, hsa-miR-223-3p, hsa-miR-21-5p, hsa-miR-23a-3p, and hsa-miR-320b) of them were increased more than 10-fold (P < .001); four (hsa-let-7a-5p, hsa-miR-25-3p, hsa-miR-126-3p, hsa-miR-320c) more than five-fold (P < .001); two (hsa-miR-342-3p and hsa-miR-320d) more than two-fold (P < .05); and two (hsa-miR-328-3p and hsa-miR-320e) more than two-fold (P > .05). Specifically, hsa-miR-22-3p increased 14.6-fold; hsa-miR-223-3p increased 13.0-fold; and hsa-miR-21-5p increased 12.0-fold. Based on bioinformatics functional analysis, target genes of top nine microRNAs (hsa-miR-22-3p, hsa-miR-223-3p, hsa-miR-21-5p, hsa-miR-23a-3p, hsa-miR-320b, hsa-let-7a-5p, hsa-miR-25-3p, hsa-miR-126-3p, and hsa-miR-320c) were annotated with positive regulation of cell proliferation and nervous system development, and mainly enriched in regulating pluripotency of stem cells signaling pathway, prolactin signaling pathway, and FoxO signaling pathway, etc. The prolactin, FoxO, ErbB, and TNF signaling pathway were relevant to immunomodulation. In particular, hsa-miR-22-3p expression was the most different during storage, with a fold change of 14.6, which might be a key mediator.

What is the context? Platelet transfusion is a widely used clinical treatment, but it is not totally safe. Side effects may happen to patients who receive the "older" platelet products. Exosomes in platelet products can be transfused to patients while receiving blood. Exosomes are accumulated in platelet products during storage. MicroRNAs are one of the important cargoes in exosomes, which can be delivered to the target cells, thus affecting their functions.This study aims to investigate and analyze the differential expression profiling of exosomal microRNAs in apheresis platelet concentrates during storage, and predict the potential function of the target genes. We found out the top nine differentially expressed microRNAs got involved in positive regulation of cell proliferation and nervous system development, and mainly enriched in regulating pluripotency of stem cells signaling pathway, prolactin signaling pathway, and FoxO signaling pathway, etc.What's new? Our study is the first one to test the exosomal microRNAs in the apheresis platelet concentrates. The apheresis platelet concentrates were stored for 1 day and 5 days. Compared the two time points, we obtained the differential expression profiling of exosomal microRNAs. Based on bioinformatics analyses and qRT-PCR results, we provided nine up-regulated microRNAs which might be critical mediators to communicate with target cells after transfusing.What's the impact? This study expands our knowledge of exosomal microRNA expression profiling from apheresis platelet concentrates along different storage periods. This might be relevant to immunomodulation in post transfusion situations.

Long-term porosity and retreatability of oval-shaped canals obturated using two different methods with a novel tricalcium silicate sealer.

OBJECTIVE: To evaluate the percentage volume of voids and gaps in oval-shaped canals obturated using two different methods with a tricalcium silicate-based sealer after short- or long-term storage. The long-term effect of storage on the efficiency of removing filling material was also investigated. MATERIALS AND METHODS: Forty premolar teeth with oval-shaped canals were instrumented to Reciproc R25 and obturated using single cone obturation (SCO) or warm vertical compaction (WVC) techniques with gutta-percha and HiFlow sealer. The specimens were stored at 100% humidity and 37°C for 2 weeks or 6 months and scanned using micro-computed tomography. Initial retreatment was performed up to a Reciproc R40, and the operating time was recorded. The residual material in the canal received a supplementary procedure using XP-endo Finisher R (XPFR) files. After each retreatment procedure, the specimens were rescanned. RESULTS: The percentage volume of voids and gaps in the SCO group was higher than that of the WVC group at both 2 weeks and 6 months (P < 0.05). The percentage volume of the filling material removed after initial retreatment and XPFR cleaning was significantly higher in the 6-month group than in the 2-week groups (P < 0.05). The proportion of the residual material decreased significantly when XPFR files were used, compared to the initial retreatment group (P < 0.05) in both storage times. CONCLUSION: The efficiency of retreatment in the oval-shaped canal was closely related to the storage time rather than the filling technique using a tricalcium silicate sealer. The XPFR instrument proved effective in the removal of the remaining materials from the oval-shaped canal. CLINICAL RELEVANCE: Obturation of the oval-shaped canal with TSBS using the SCO technique in the coronal area needs to be optimized. The retreatment was less efficacious in freshly filled canals than aged filled canals.

Freshness Identification of Oysters Based on Colorimetric Sensor Array Combined with Image Processing and Visible Near-Infrared Spectroscopy.

Volatile organic compounds (VOCs) could be used as an indicator of the freshness of oysters. However, traditional characterization methods for VOCs have some disadvantages, such as having a high instrument cost, cumbersome pretreatment, and being time consuming. In this work, a fast and non-destructive method based on colorimetric sensor array (CSA) and visible near-infrared spectroscopy (VNIRS) was established to identify the freshness of oysters. Firstly, four color-sensitive dyes, which were sensitive to VOCs of oysters, were selected, and they were printed on a silica gel plate to obtain a CSA. Secondly, a charge coupled device (CCD) camera was used to obtain the "before" and "after" image of CSA. Thirdly, VNIS system obtained the reflected spectrum data of the CSA, which can not only obtain the color change information before and after the reaction of the CSA with the VOCs of oysters, but also reflect the changes in the internal structure of color-sensitive materials after the reaction of oysters' VOCs. The pattern recognition results of VNIS data showed that the fresh oysters and stale oysters could be separated directly from the principal component analysis (PCA) score plot, and linear discriminant analysis (LDA) model based on variables selection methods could obtain a good performance for the freshness detection of oysters, and the recognition rate of the calibration set was 100%, while the recognition rate of the prediction set was 97.22%. The result demonstrated that the CSA, combined with VNIRS, showed great potential for VOCS measurement, and this research result provided a fast and nondestructive identification method for the freshness identification of oysters.

A unified operating procedure is crucial to evaluate sludge dewaterability, taking the setup of refrigerated storage time as an example.

Although extensive efforts have been carried out to study sludge dewatering mechanism, the lack of universal operating procedures makes it never be satisfactorily explained. This study evaluated the impact of a unified operating procedure on waste activated sludge (WAS) dewaterability by taking the setup of refrigerated storage time as an example. It was found that storage time played an important role in determining WAS dewaterability and sampled WAS should be refrigerated within 2 days. The results showed that after 2-d storage, sludge filterability was deteriorated significantly while the extent of dewatering efficiency had little change. Meanwhile, increasing storage time greatly increased the release of extracellular polymeric substances (EPS) and heavy metals, decreased sludge viscosity and weakened its network strength, but had little impact on the floc size and zeta potential of the sludge samples. It can hardly reveal the mechanism of storage time on sludge dewaterability due to the non-uniformity of operating procedures in literatures, which is normally ignored. This study emphasizes a unified operating procedure is crucial to evaluate WAS dewaterability. Therefore, more efforts shall be focused on establishing the uniform operating procedure while advancing applied research in the field of sludge dewatering.