The widespread use of multipotent mesenchymal stromal cell-derived secretome (MSC-sec) requires optimal preservation methods. Lyophilization offers benefits like concentrating the secretome, reducing the storage volume, and making storage conditions more flexible. This study evaluated the influence of storage duration and temperature on lyophilized MSC-sec. The conditioned medium from Wharton's jelly MSCs was stored at - 80 °C or lyophilized with or without trehalose. Lyophilized formulations were kept at - 80 °C, - 20 °C, 4 °C, or room temperature (RT) for 3 and 30 months. After storage and reconstitution, the levels of growth factors and cytokines were assessed using multiplex assay. The storage of lyophilized MSC-sec at - 80 °C ensured biomolecule preservation for 3 and 30 months. Following 3 month storage at 4 °C and RT, a notable decrease occurred in BDNF, bNGF, and sVCAM-1 levels. Prolonged 30 month storage at the same temperatures significantly reduced BDNF, bNGF, VEGF-A, IL-6, and sVCAM-1, while storage at - 20 °C decreased BDNF, bNGF, and VEGF- A levels. Trehalose supplementation of MSC-sec improved the outcome during storage at 4 °C and RT. Proper storage conditions were crucial for the preservation of lyophilized MSC-sec composition. Short-term storage at various temperatures maintained over 60% of the studied growth factors and cytokines; long-term preservation was only adequate at -80 °C.
- MeSH
- Cytokines metabolism MeSH
- Cryopreservation methods MeSH
- Culture Media, Conditioned chemistry MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Freeze Drying * MeSH
- Mesenchymal Stem Cells * metabolism cytology MeSH
- Secretome metabolism MeSH
- Temperature MeSH
- Trehalose metabolism pharmacology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
In this research, influence of storage conditions on properties of oxidized cellulose was studied with respect to its haemostatic function. The aim was to examine changes of the properties of oxidized cellulose stored properly and that stored at laboratory conditions for 2 years. We studied surface morphology and chemical composition, as well as absorption of the simulated body fluid, behaviour in aqueous environment via potentiometric measurement of pH, and antimicrobial activity in vitro on the S. epidermidis bacteria. It was found out that the material properties of oxidized cellulose did not deteriorate. Higher absorption of simulated body fluid, lower pH in water and simulated body fluid represented positive changes with respect to the haemostatic function. Due to the acidic nature of the mate-rial, degraded oxidized cellulose preserved its antibacterial properties.
- MeSH
- Anti-Bacterial Agents analysis MeSH
- Cellulose, Oxidized * analysis MeSH
- Microscopy, Electron methods MeSH
- Photoelectron Spectroscopy methods MeSH
- Hemostatics analysis MeSH
- Quality Control MeSH
- Drug Storage MeSH
- Environmental Exposure prevention & control MeSH
- Publication type
- Clinical Study MeSH
- Research Support, Non-U.S. Gov't MeSH
- MeSH
- Diagnosis, Differential MeSH
- Hemochromatosis diagnosis genetics MeSH
- Humans MeSH
- Adolescent MeSH
- Mutation MeSH
- Pathological Conditions, Signs and Symptoms MeSH
- Fatigue etiology MeSH
- Iron metabolism MeSH
- Check Tag
- Humans MeSH
- Adolescent MeSH
- Female MeSH
- Publication type
- Case Reports MeSH
BACKGROUND: Human milk harbors diverse bacterial communities that contribute to infant health. Although pumping and storing milk is a common practice, the viable bacterial composition of pumped milk and the impact of storage practice on these bacteria remains under-explored. This metagenomic observational study aimed to characterize viable bacterial communities in freshly pumped human milk and its changes under different storage conditions. METHODS: In 2023, twelve lactating mothers from the CELSPAC: TNG cohort (Czech Republic) provided freshly pumped milk samples. These samples were stored under various conditions (refrigeration for 24 h, 48 h, or freezing for six weeks) and treated with propidium monoazide (PMA) to selectively identify viable cells. The DNA extracted from individual samples was subsequently analyzed using 16S rRNA amplicon sequencing on the Illumina platform. RESULTS: The genera Streptococcus, Staphylococcus, Diaphorobacter, Cutibacterium, and Corynebacterium were the most common viable bacteria in fresh human milk. The median sequencing depth and Shannon index of fresh human milk samples treated with PMA (+ PMA) were significantly lower than in untreated (-PMA) samples (p < 0.05 for all), which was true also for each time point. Also, significant changes in these parameters were observed between fresh human milk samples and their paired frozen samples (p < 0.05), while no differences were found between fresh human milk samples and those refrigerated for up to 48 h (p > 0.05). Of specific genera, only + PMA frozen human milk samples showed a significant decrease in the central log-ratio transformed relative abundances of the genera Diaphorobacter and Cutibacterium (p < 0.05) in comparison to + PMA fresh human milk samples. CONCLUSIONS: The study demonstrated that the bacterial profiles significantly differed between human milk samples treated with PMA, which represent only viable bacteria, and those untreated. While storage at 4 °C for up to 48 h did not significantly alter the overall diversity and composition of viable bacteria in human milk, freezing notably affected both the viability and relative abundances of some bacterial genera.
- MeSH
- Azides MeSH
- Bacteria * isolation & purification genetics classification MeSH
- Refrigeration MeSH
- Adult MeSH
- Humans MeSH
- Milk, Human * microbiology MeSH
- Microbiota * MeSH
- Propidium analogs & derivatives MeSH
- RNA, Ribosomal, 16S MeSH
- Food Storage * methods MeSH
- Freezing MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Observational Study MeSH
The aim of this study was to compare hygiene status of wild boar meat (shoulder and leg) stored up to 21days at 0°C, 7°C or 15°C. The microbial counts increased gradually in the expected sequence of increasing storage temperatures, with TVC at the end of storage ranging from approx. 2logCFU/g (0°C) to 5logCFU/g (15°C). The lactic acid bacteria and psychrotrophic microflora didn't exceed 2logCFU/g and 2.5logCFU/g, respectively. Whereas odor of the meat stored at 0°C and 7°C was still acceptable at the end of storage, the odor of the meat stored at 15°C was barely acceptable after only 7d of storage and also the content of ammonia was significantly higher. Game meat obtained from animals hunted in the correct way and stored at low temperatures had good microbiological and hygiene status which could be maintained for more than 15days of storage.
- MeSH
- Ammonia analysis MeSH
- Color MeSH
- Red Meat analysis MeSH
- Taste * MeSH
- Hydrogen-Ion Concentration MeSH
- Food Contamination MeSH
- Lactobacillaceae growth & development isolation & purification MeSH
- Humans MeSH
- Microbial Viability MeSH
- Cold Temperature MeSH
- Odorants analysis MeSH
- Food Microbiology MeSH
- Food Storage * MeSH
- Sus scrofa MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Makana Local Municipality is located in the Eastern Cape Province of South Africa. The area is water-scarce and has been undergoing aridification in recent years, ie, there has been a 7-year long drought. At the same time, there has been a breakdown in provision of municipal services, such as drinking water, to the population since at least 2008. Mi-crobial water quality has been a result of this, and monitoring has been a challenge. Given the drought and the prob-lems with drinking water delivery, it was necessary to conduct this study to investigate the microbial quality of alterna-tive water resources that the Makana population can use during the municipal water outage. The microbial water quality of alternative sources of drinking water in the Makana Local Municipality was examined using the H2S test kit and enumeration of the fecal coliforms. Storage of the collected water was examined for potential factors influencing the microbial water quality of the alternative sources of drinking water. The costs of the water provision from the most suitable alternative sources of potable water were calculated. There was a general correlation between the H2S test kit results and the fecal coliform concentrations, with the latter values ranging from <0 to 23 ± 7 colony-forming units/100 mL. The bottled water from two retail outlets was provided the best alternative source of potable/drinking water, which is microbially safe, for the Makana population. If the consumption of the drinking water from an alternative source takes place within 24 hours of collection, then the Fairview spring could also be used as a source of drinking water for Maka-na residents. The total cost per 1 L of drinking water from alternative sources was estimated to be from 1.51 to 5.81 ZAR. Therefore, the maximum cost of daily provision of drinking water from alternative sources would account for a maximum of 0.88 percent of the monthly household expenditure in the middle-to-high-income household. However, the daily costs of such provision of drinking water would account for a maximum of 8.11 percent of the monthly household expenditure in the low-income household. Provision of the drinking water from the alternative sources would have a minor impact of the monthly ex-penditure in the middle-to-high-income households. However, it is likely that the low-income households would not be able to sustain their water supply from alternative sources for longer than 24 hours, during a municipal outage in the drinking water supply.
- Publication type
- Journal Article MeSH
The purpose of this study was to improve the survival of Bifidobacterium animalis subsp. lactis 10140 during freeze-drying process by microencapsulation, using a special pediatric prebiotics mixture (galactooligosaccharides and fructooligosaccharides). Probiotic microorganisms were encapsulated with a coat combination of prebiotics-calcium-alginate prior to freeze-drying. Both encapsulated and free cells were then freeze-dried in their optimized combinations of skim milk and prebiotics. Response surface methodology (RSM) was used to produce a coating combination as well as drying medium with the highest cell viability during freeze-drying. The optimum encapsulation composition was found to be 2.1 % Na-alginate, 2.9 % prebiotic, and 21.7 % glycerol. Maximum survival predicted by the model was 81.2 %. No significant (p > 0.05) difference between the predicted and experimental values verified the adequacy of final reduced models. The protection ability of encapsulation was then examined over 120 days of storage at 4 and 25 °C and exposure to a sequential model of infantile GIT conditions including both gastric conditions (pH 3.0 and 4.0, 90 min, 37 °C) and intestinal conditions (pH 7.5, 5 h, 37 °C). Significantly improved cell viability showed that microencapsulation of B. lactis 10140 with the prebiotics was successful in producing a stable symbiotic powdery nutraceutical.
- MeSH
- Bifidobacterium physiology radiation effects MeSH
- Time Factors MeSH
- Gastrointestinal Tract microbiology MeSH
- Infant MeSH
- Humans MeSH
- Freeze Drying * MeSH
- Microbial Viability radiation effects MeSH
- Infant Formula MeSH
- Colony Count, Microbial MeSH
- Drug Compounding MeSH
- Probiotics radiation effects MeSH
- Drug Storage methods MeSH
- Temperature MeSH
- Check Tag
- Infant MeSH
- Humans MeSH
- Publication type
- Journal Article MeSH
The rate of beer aging is affected by storage conditions including largely time and temperature. Although bottled beer is commonly stored for up to 1 year, sensorial damage of it is quite frequent. Therefore, a method for retrospective determination of temperature of stored beer was developed. The method is based on the determination of selected carbonyl compounds called as "aging indicators", which are formed during beer aging. The aging indicators were determined using GC-MS after precolumn derivatization with O-(2,3,4,5,6-pentaflourobenzyl)hydroxylamine hydrochloride, and their profile was correlated with the development of old flavor evolving under defined conditions (temperature, time) using both a mathematical and statistical apparatus. Three approaches, including calculation from regression graph, multiple linear regression, and neural networks, were employed. The ultimate uncertainty of the method ranged from 3.0 to 11.0 °C depending on the approach used. Furthermore, the assay was extended to include prediction of beer tendency to sensory aging from freshly bottled beer.
- MeSH
- Time Factors MeSH
- Taste * MeSH
- Flavoring Agents analysis MeSH
- Humans MeSH
- Beer analysis MeSH
- Gas Chromatography-Mass Spectrometry methods MeSH
- Food Storage MeSH
- Temperature MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
The crucial requirement of molecular genetic methods is high-quality input material. The key question is "how to preserve DNA during long-term storage." Biobanks are recommended to aliquot isolated DNA into provided volumes. The aim of this study was to analyse the effect of repeated freezing and thawing on the genomic DNA integrity, quality and concentration. The aliquoted DNA isolated from blood cells using the automatic MagNA system and manual salting out method underwent freeze/thaw cycles at different storage conditions (-20 °C, -80 °C and liquid nitrogen). The average initial concentrations were 270.6 ng/μl (salting out method) and 125.0 ng/μl (MagNA). All concentration deviations relative to the concentration after the first freeze/ thaw cycle were less than 5 % for -20 °C and -80 °C cycling with both isolation methods. The average percentage differences of liquid nitrogen samples were higher, and the MagNA isolation method showed significant differences. There were no significant changes in the DNA purity or quality. The repeating freeze/ thaw up to 100 cycles (through -20 °C and -80 °C, respectively) did not significantly influence the integrity, concentration, or purity of genomic DNA, suggesting that storage of samples in high-volume pools without multiple aliquoting is possible. Storage in a freezer seems to be the most suitable way of long-term DNA preservation, because liquid nitrogen storage leads to formation of DNA clumps.
- MeSH
- DNA * MeSH
- Genomics * MeSH
- Freezing MeSH
- Publication type
- Journal Article MeSH
