Effect Of Prolonged Storage Research Articles

Assessing the impact of long-term storage on the quality and integrity of biological specimens in a reproductive biobank.

Biobanks hold a pivotal role in facilitating translational and clinical research endeavors. However, the effects of prolonged storage on frozen blood samples analytes are not well defined yet. The aim of this study was to investigate the long-term stability of the quality of DNA, RNA, and endocrine markers within blood samples amassed from the biobank over the past 11 years. The results show that the overall quality and integrity of DNA remained not significantly influenced. However, RNA integrity and purity displayed substantial deterioration as storage duration increased, to ensure high-quality RNA for downstream analyses, advised to prioritize using blood samples stored within 3 years. Furthermore, the study examined the influence of storage time on endocrine markers. Through repeated measures ANOVA and linear regression analyses, it was evident that storage duration significantly influenced the levels of endocrine markers. This insight aids researchers in selecting appropriate markers for their investigations and augments the precision and dependability of results when dealing with long-term stored samples.

Germination and viability of wild sunflower species seeds stored at room temperature and low humidity for 38 years

The effects of prolonged storage on the germination and viability of seeds of sunflower crop wild relatives stored under less-than-optimal conditions in working genebanks is not known. Seeds of two wild annual species, Helianthus annuus (common wild sunflower) and annual H. petiolaris (prairie sunflower), were stored at room temperature (20 to 22°C) and low humidity of ∼ 22% in the laboratory for 38 years. Germination of stored annual sunflower seeds was 5.5%, while that of prairie sunflower seeds was 0.75%, compared with the germination of freshly harvested seeds of 34.7 and 18.5%, respectively. Tetrazolium, a vital stain, indicated that the seeds were alive. The viability equations were used to predict final viability of the stored seeds. Using the three tests of viability (germination with and without gibberellic acid, and tetrazolium staining), all appeared to underestimate the actual condition of the stored seeds of annual sunflower, but provided better predictions for prairie sunflower. This study indicated that wild sunflower seeds can be stored at less than ideal conditions in working genebanks where viability decreases over time reducing germination, but gibberellic acid treatment can sustainably increase germination to produce a number of plants to maintain the genetic integrity of the original stored seeds often used in long-term prebreeding programmes.

Effect of prolonged storage on quality characteristics of recovered plasma: Is there an expiration date?

Although there are guidelines on industrial manufacture of plasma-derived medicinal products, there are no clear recommendations about plasma intended for fractionation, as there is no expiry time and the effect of prolonged storage on the activity of coagulation factors is unknown. A total of 237 units of plasma stored at -30°C in the National Blood Transfusion Centre for 1year (62 units), 5 years (75 units), and 10 years (100 units) were studied. The effect of storage time was investigated by determining the activity of clotting factors FII, FV, FVII, FVIII, FIX, FX, FXI, FXII, FXIII using coagulometric methods and antithrombin III and fibrinogen with chromogenic assays, using System BCSR > XP (Siemens Healthcare diagnostics Marburg, Germany). Albumin was measured by Medilyzer (BX, Medicon). ABO blood group was recorded and correlated with the levels of FVIII. Comparison of values between one and five, 1 and 10 and 5 and 10 years of storage was performed via the SAS for Windows 9.4 software platform (SAS Institute Inc., NC, U.S.A.). Albumin, AT III, fibrinogen, FIX, FXI, FXII, and FXIII remain rather stable even after 10 years of storage. Levels of FII, FV, FVII, FVIII, and FX decreased after 5 years of storage. Our study is in agreement with all the previous studies and concludes that there is a putative usability of recovered plasma and some of its coagulation factors after many years of storage at the recommended temperature.

Formation and characterisation of high-internal-phase emulsions stabilised by high-pressure homogenised quinoa protein isolate

High pressure homogenised (HPH) quinoa protein isolates (QPI) suspensions (1, 3, and 5 wt%) were used to prepare high-internal-phase emulsions (HIPEs) (ɸ=0.8). The viscoelastic behaviours of HIPEs were characterised by oscillatory rheology, while the oil droplet size distribution and microstructural characteristics were examined by static light scattering and confocal laser scanning microscopy (CLSM). At all QPI concentrations, HIPEs behaved as viscoelastic gels due to network formation as a result of interactions between jammed oil droplets and protein aggregates. As the QPI concentration increased, the oil droplet size decreased from ~80 to~40 µm and the elastic modulus G´(1 Hz) increased from ~100 to~580 Pa. Effects of prolonged storage (30 days) and heat treatment (80 °C, 30 min) on the rheological behaviour and microstructures of HIPEs were also investigated. All emulsions showed excellent stabilities against phase separation, oil coalescence, and creaming during storage or under heat treatment. Prolonged storage induced a substantial increase in G´, while no significant change in oil droplet size was observed. CLSM revealed that the interfacial layer became thicker and large protein aggregates were formed during storage, which led to a more dense and compact packing of droplets and greater gel strength. Heat treatment induced a 3–5 folds enhancement in the gel strength due to protein denaturation and proteins and oil droplets aggregation. These findings provide useful information for the design and preparation of HIPEs stabilised by quinoa proteins.

P–194 Impact of cryopreservation duration on pregnancy outcomes of vitrified-warmed blastocysts transfer using an open-device system

Abstract Study question Is there a negative effect of long-term cryopreservation upon pregnancy outcomes after transfer of vitrified-warmed blastocysts stored in an open-device system? Summary answer Prolonged cryopreservation of vitrified blastocysts up to 24 months increased the incidences of clinical pregnancy, ongoing pregnancy, and live birth, while decreased early miscarriage rate. What is known already Vitrification is adopted as the dominant approach for cryopreservation of human oocytes and embryos. However, little is known about the potential effect of prolonged storage after vitrification on the genomic integrity and metabolism of embryos. Several studies have sought to decipher the effect of cryopreservation duration on IVF pregnancy outcomes, but few were confined to vitrification and the results were inconsistent. Study design, size, duration This retrospective study included 6722 patients undergoing their first vitrified-warmed blastocyst transfer (VBT) cycles from January 2015 to June 2019 in a single fertility center in South China. The study was approved by the hospital’s Ethics Committee. Participants/materials, setting, methods A total of 6722 eligible patients were divided into five groups according to the storage duration after vitrification: Group I: 0–3 months; Group II: 3–6 months; Group III: 6–12 months; Group IV: 12–24 months; Group V: 24–36 months. The IVF pregnancy outcomes were compared between groups. Multivariate logistic regression was conducted to evaluate the independent effect of storage duration on pregnancy outcomes. Main results and the role of chance The odds of clinical pregnancy outcomes were similar from Group 1 to 4. However, the chance of clinical pregnancy (Group 1 as reference; Group 2: adjusted odds ratio (aOR)= 1.04, 95% CI 0.93–1.17; Group 3: aOR = 1.02, 95% CI 0.84–1.25; Group 4: aOR = 0.93, 95% CI 0.66–1.31; Group 5: aOR = 0.54, 95% CI 0.38–0.76) , ongoing pregnancy (Group 2: aOR=0.99, 95% CI 0.89–1.11; Group 3: aOR = 0.94, 95% CI 0.77–1.14; Group 4: aOR = 0.87, 95% CI 0.62–1.22; Group 5: aOR = 0.41, 95% CI 0.29–0.60), and live birth rate (Group 2: aOR=1.00, 95% CI 0.89–1.12; Group 3: aOR = 0.98, 95% CI 0.81–1.19; Group 4: aOR = 0.91, 95% CI 0.65–1.27; Group 5: aOR = 0.46, 95% CI 0.32–0.66) significantly decreased, while the early miscarriage rate (Group 2: aOR=1.11, 95% CI 0.92–1.35; Group 3: aOR =1.25, 95% CI 0.92–1.70; Group 4: aOR = 1.33, 95% CI 0.77–2.31; Group 5: aOR = 2.42, 95% CI 1.36–4.31) significantly increased as the storage duration increased up to 24–36 months. Limitations, reasons for caution The primary limitation of this study was its retrospective nature. Besides, as all these data come from a single IVF treatment center, the results should be confirmed by a larger multicenter study. Wider implications of the findings: Our study provides more evidence about the negative impact of long-term storage of vitrified embryos on the clinical outcome. Clinicians should adapt FET strategies based on the embryo storage duration. Trial registration number Not applicable

A physicochemical assessment of the thermal stability of dextrin\u2013colistin conjugates

Attachment of polysaccharide carriers is increasingly being used to achieve precision delivery and improved effectiveness of protein and peptide drugs. Although it is clear that their clinical effectiveness relies on the purity and integrity of the conjugate in storage, as well as following administration, instability of polysaccharide-based conjugates can reduce the protective efficacy of the polymer, which may adversely affect the bioactive’s potency. As a model, these studies used dextrin–colistin conjugates, with varying degrees of polymer modification (1, 2.5 and 7.5 mol% succinoylation) to assess the effect of storage temperature (− 20, 4, 21 and 37 °C) and duration (up to 12 months) on saccharide and colistin release and antimicrobial activity. Estimation of the proportion of saccharide release (by comparison of area under the curve from size exclusion chromatograms) was more pronounced at higher temperatures (up to 3 and 35% at − 20 °C and 37 °C, respectively after 12 months), however, repeated freeze–thaw did not produce any measurable release of saccharides, while addition of amylase (20, 100, 500 IU/L) caused rapid release of saccharides (> 70% total within 24 h). At all temperatures, conjugates containing the lowest degree of succinoylation released the highest proportion of free colistin, which increased with storage temperature, however no trend in saccharide release was observed. Despite the clear physical effects of prolonged storage, antimicrobial activity of all samples was only altered after storage at 37 °C for 12 months (> threefold decreased activity). These results demonstrate significant release of saccharides from dextrin–colistin conjugates during prolonged storage in buffered solution, especially at elevated temperature, which, in most cases, did not affect antimicrobial activity. These findings provide vital information about the structure–activity relationship of dextrin–colistin conjugates, prior to full-scale commercial development, which can subsequently be applied to other polysaccharide-protein and -peptide conjugates.

Effect of prolonged storage at 2°C-6°C for 120 h on the coagulation factors of thawed cryoprecipitate: Can we extend its shelf life post thaw beyond 4 h?

BACKGROUND:Cryoprecipitate helps in replenishing important coagulation factors like fibrinogen, Factor VIII and von Willebrand factor without running the risk of volume overload. It is very useful in the treatment of trauma patients with active bleeding and works best when administered early. Extending the shelf life of thawed cryoprecipitate beyond 4 hours enables us to manage inventory better, reduces the burden of demand vs supply as well as minimizes wastage. It can also help in logistically supporting the transfusion services in making cryoprecipitate readily available in mass casualty scenarios (war, natural calamity) in remote locations by reducing the time required for thawing cryoprecipitate and the need for costly storage equipment. AIM: The aim of this study was to compare the levels of Factor VIII, Fibrinogen and von Willebrand factor on thawed cryoprecipitate after prolonged storage for 5 days at a temperature of 2-6°C.METHODOLOGY:The above mentioned coagulation factors were analyzed in cryoprecipitate at the time of product thaw and again after 120 hours of 2 to 6°C storage using fully automated coagulation analyser (STA Compact Max). All parameters were expressed as Mean ± Standard deviation and were analyzed using paired t-test with level of significance, P < 0.05.RESULTS:There was a significant decrease in the level of Factor VIII, whereas the levels of fibrinogen and von Willebrand Factor remained stable during the storage period. All the cryoprecipitate units retained factor activities above therapeutic range even after 5 days of storage at 2-6°C.CONCLUSION:Although the levels of clotting factors are reduced during storage, they are still maintained above the therapeutic range. In scenarios where maintaining frozen cryoprecipitate inventory is a logistical challenge and emergency massive demands of cryoprecipitate are foreseen, the use of pre-thawed cryoprecipitate can be considered as a viable option.

Dehiscence and prolonged storage of 'Kerman' Pistachios: Effects on morphometry and nutraceutical value.

'Kerman' pistachios (KP; Pistacia vera L.) are an important crop for several countries but their commercial value is diminished by their shell dehiscence status and prolonged storage in popular marketplaces. The aim was to evaluate the independent/synergistic effect of prolonged storage (1-4year) and dehiscence status (split/unsplit) on KP's morphometry and chemical composition. Whole nut's and kernel's length, width, thickness, surface area, and volume were more affected by dehiscence (split > unsplit; p ≤ 0.01) than storage time; Kernel's mass, macronutrient composition and tocopherols (T)/tocotrienols (T3) were not much affected by dehiscence but time-trend correlations were observed with macronutrient composition (split/unsplit; ρ = - 0.57-0.42) and T + T3 (unsplit; ρ = 0.81). Specific/total fatty acids were affected by a complex dehiscence × storage time interaction, and they linearly correlated with certain morphometric characteristics (r ≥ 0.6). Shell dehiscence status more than prolonged storage substantially modifies KP's quality.

Effect of Storage Temperature and Time on Stability of Liver Enzymes in Blood Serum.

It is increasingly common to collect and store specimens for future unspecified research. However, the effects of prolonged storage on the stability and quality of analytes in serum have not been well investigated. We aimed to determine whether the stability of liver enzymes extracted from frozen bio-samples stored at the baseline is affected by storage conditions. A total of four liver enzymes in the sera of 400 patients were examined following storage. After deter-mining the baseline measurements, the serum of each patient was aliquoted and stored at -70°C for three and six months, as well as one, two, and five years after collecting the original sample. The percent change from baseline measurements was calculated both statistically and clinically. Linear models were also used to correct the results of the samples based on the time they were frozen. In almost all samples, liver enzymes were detectable until two years after the baseline, while in a signifi-cant proportion of samples, enzymes were not ultimately detectable five years after the baseline. Linear regression analysis on log-transformed levels of enzymes shows that the performance is acceptable until one year after the baseline. The performance of the prediction model declines substantially two and five years after the baseline, except for GGT. Long-term storage of serum samples significantly decreases the concentration of the liver enzymes from the baseline, except for GGT. It is not recommended to store samples for more than two years, as liver en-zymes are not detectable afterwards.

Influence of post-condensation on the crystallization kinetics of PA12: From virgin to reused powder

The effect of prolonged storage at elevated temperatures on the isothermal crystallization kinetics of Polyamide 12 (PA12), the most used polymer in selective laser sintering (SLS), has been studied by means of Flash DSC experiments. The experimental results are described and modelled with a full multi phase crystallization model which allowed us to estimate the growth and nucleation kinetics for every PA12 crystal phase. Virgin PA12 powder has been annealed at different temperatures for different times to quantify the molecular weight evolution due to post condensation. To check the effect of thermal treatments on the final viscosity, rheology measurements have been carried out. We show that thermal annealing results in a significant increase in molecular weight and viscosity which lead to a reduction of the crystallization rate in a limited temperature range. At very low temperature the crystallization kinetics, expressed in terms of peak-time of crystallization, appears to be independent on the annealing treatment while in the temperature range between 100∘C and 150∘C a significant rate reduction is observed. We demonstrate that the crystallization kinetics in this temperature range is governed by the crystal growth process, i.e. the increase in molecular weight due to post-condensation causes a reduction of the growth rate of the main crystalline phases of PA12, namely α' and γ-phase. Similar to the low temperature crystallization, at temperature higher than 150∘C the annealing does not noticeably affect the growth rates and the crystallization kinetics of virgin and annealed PA12 overlap. These results can provide a better insight of the effect of reusing PA12 powder in the SLS process.

Effects of prolonged storage of blood in EDTA on blood cells morphology in peripheral blood smears

Effects of prolonged storage of blood in EDTA on blood cells morphology in peripheral blood smears

The effect of prolonged storage and disinfection on the dimensional stability of 5 vinyl polyether silicone impression materials.

PURPOSEVinyl polyether silicone (VPES) has a different composition from other elastomeric impression materials as it combines vinyl polysiloxane (VPS) and polyether (PE). Therefore, it is important to study its properties and behavior under different test conditions. This study investigated the dimensional stability of 5 VPES consistencies when stored for up to 2 weeks, with and without using a standard disinfection procedure.MATERIALS AND METHODS40 discs of each VPES consistency (total 200) were made using a stainless steel die and ring as described by ANSI /ADA specification No. 19. 20 discs of each material were immersed in a 2.5% buffered glutaraldehyde solution for 30 minutes. Dimensional stability measurements were calculated immediately after fabrication and repeated on the same discs after 7 and 14 days of storage. The data was analyzed using two-way ANOVA with a significance level set at α = 0.05.RESULTSThe discs mean contraction was below 0.5% at all test times ranging from 0.200 ± 0.014 to 0.325 ± 0.007. Repeated measures ANOVA showed a statistically significant difference after 2-week storage between the disinfected and non-disinfected groups (P < .001). Although there was no statistically significant difference between the materials at the time of fabrication, the contraction of the materials increased with storage for 1 and 2 weeks.CONCLUSIONThe dimensional changes of VPES impression discs after disinfection and prolonged storage complied with ANSI/ADA standard. The tested VPES impression materials were dimensionally stable for clinical use after disinfection for 30 minutes in glutaraldehyde and storage for up to 2 weeks.

Effect of prolonged-storage on tensile bond strength of adhesive systems

Effect of prolonged-storage on tensile bond strength of adhesive systems

Time Dependent Assessment of Morphological Changes: Leukodepleted Packed Red Blood Cells Stored in SAGM.

Usually packed red blood cells (pRBCs) require specific conditions in storage procedures to ensure the maximum shelf life of up to 42 days in 2–6°C. However, molecular and biochemical consequences can affect the stored blood cells; these changes are collectively labeled as storage lesions. In this study, the effect of prolonged storage was assessed through investigating morphological changes and evaluating oxidative stress. Samples from leukodepleted pRBC in SAGM stored at 4°C for 42 days were withdrawn aseptically on day 0, day 14, day 28, and day 42. Morphological changes were observed using scanning electron microscopy and correlated with osmotic fragility and hematocrit. Oxidative injury was studied through assessing MDA level as a marker for lipid peroxidation. Osmotic fragility test showed that extended storage time caused increase in the osmotic fragility. The hematocrit increased by 6.6% from day 0 to day 42. The last 2 weeks show alteration in the morphology with the appearance of echinocytes and spherocytes. Storage lesions and morphological alterations appeared to affect RBCs during the storage period. Further studies should be performed to develop strategies that will aid in the improvement of stored pRBC quality and efficacy.

Effect of prolonged storage in controlled atmospheres on the conservation of the onion (Allium cepa L)quality

The onion is a low perishability product, but in long-term storage, postharvest losses occur due to some phytopathological and sprouting factors. The aim of this study was to evaluate the effect of storage in controlled atmospheres (CA) on onion cultivar “Sierra Blanca” for seven months. The onions were subjected to three treatments: 1 % O2 + 1 % CO2 (T1), 3 % O2 + 5 % CO2 (T2), both at 2.5 °C and 60-75 % relative humidity (RH), and in environmental atmosphere (control treatment, CT) at 2.5 °C without controlled RH. Changes in enzymatic pyruvic acid (EPA) content, total soluble solids (TSS), dry matter (DM), firmness and sprouting during storage were evaluated. The results showed effect (p≤0.05) of treatment and storage time on all parameters. Changes in preserved bulbs in CA were minimal compared to those in the CT. The EPA was reduced on T1 onions while in T2 and CT onions it was increased at the end of storage. The TSS and firmness decreased in the three treatments, DM remained constant in T1 and T2 onions and in CT onions decreased in the sixth month by 19 %. T1 and T2 onions showed no sprouting, while CT onions, presented internal sprouts in 20 % of sampled bulbs, reaching 100 % from the third month till the end of storage. In conclusion, storage in CA is the best way to preserve the quality of onion “Sierra Blanca” for seven months, which is evidenced by the positive changes in quality.

Quality assurance in the pre-analytical phase of human urine samples by 1H NMR spectroscopy

Metabolomic approaches investigate changes in metabolite profiles, which may reflect changes in metabolic pathways and provide information correlated with a specific biological process or pathophysiology. High-resolution 1H NMR spectroscopy is used to identify metabolites in biofluids and tissue samples qualitatively and quantitatively. This pre-analytical study evaluated the effects of storage time and temperature on 1H NMR spectra from human urine in two settings. Firstly, to evaluate short time effects probably due to acute delay in sample handling and secondly, the effect of prolonged storage up to one month to find markers of sample miss-handling. A number of statistical procedures were used to assess the differences between samples stored under different conditions, including Projection to Latent Structure Discriminant Analysis (PLS-DA), non-parametric testing as well as mixed effect linear regression analysis. The results indicate that human urine samples can be stored at 10 °C for 24 h or at −80 °C for 1 month, as no relevant changes in 1H NMR fingerprints were observed during these time periods and temperature conditions. However, some metabolites most likely of microbial origin showed alterations during prolonged storage but without facilitating classification. In conclusion, the presented protocol for urine sample handling and semi-automatic metabolite quantification is suitable for large-scale epidemiological studies.

The effect of prolonged storage on the virulence of isolates of Bacillus anthracis obtained from environmental and animal sources in the Kars Region of Turkey.

The stability of the plasmid-mediated virulence factors of Bacillus anthracis, a tripartite toxin located on pXO1 and an antiphagocytic capsule encoded by genes located on pXO2, following long-term storage was investigated. A collection of 159 isolates of B. anthracis were collected from the Kars region of Turkey between 2000 and 2013 and stored at -20°C in Brucella broth supplemented with 20% glycerine. A total of 142 isolates were recovered of which one failed to express a capsule upon primary culture. A further 35 isolates yielded a mixture of mucoid and non-mucoid colonies; the majority of which had lost the pXO2 plasmid as determined by PCR analysis. Results would suggest that pXO2 is more unstable than pXO1 and that this instability increases with the length of storage. It is possible that the pXO2-deficient isolates of B. anthracis described here could be developed into a vaccine to treat at risk animals in the Kars region as many animal vaccines are based upon pXO2 deficiency.

Towards microfluidic-based depletion of stiff and fragile human red cells that accumulate during blood storage.

In this study, the effects of prolonged storage on several biophysical properties of red blood cells (RBCs) were investigated. Single cell deformability was used as an important criterion in determining subgroups of RBCs evolved during storage lesion. A deformability-based microfluidic cell sorting technology was applied, which demonstrates the ability to enrich and separate the less deformable subpopulations of stored blood. These less deformable RBC subpopulations were then associated with other important markers such as osmotic fragility indicating cell integrity as well as microparticle content. This work demonstrates a systematic methodology to both monitor and improve banked blood quality, thereby reducing risks related to blood transfusion.

Effect of modified pre-milking sanitizing practices on raw milk quality obtained from the dairy farmers of Tawau, East Malaysia

The aim of the study was to investigate the raw milk hygiene and quality among the small holder dairy farmers in Tawau area, Sabah. A total of 216 samples were collected from the respective dairy farmers and milk collecting centre located at Mile 15, Tawau. Preliminary results indicated that the quality of the raw milks obtained at farm level were inferior with high bacteria load (> than 107 CFU/mL). The total coliform (2.9–3.8 CFU/mL) and Staphylococcus count (2.3–3.6 CFU/mL) were relatively high in certain samples. However, none of the food borne pathogens was found. Trace back study revealed that the causes of contamination were attributed by poor hygienic handling among the dairy farmers. A significant reduction in bacteria load was observed if the raw milk was chilled immediately at farm. The implementation of modified pre-milking sanitizing practices improved the microbiological quality of the raw milks obtained from respective dairy farms. Future studies will be emphasized on effect of prolonged storage on microbiological quality of raw milk that undergo two days collection system.

Clinical effects of transfusing older red cell concentrates: an updated overview

The literature on possible effects of prolonged storage of red cells is growing fast. Recent studies still come up with conflicting results. This review describes some of the underlying mechanisms which may explain the opposing results from the different studies.