Biopreservation and Biobanking最新文献

This study aimed to evaluate whether the addition of vitamins E and C as two conventional antioxidants improves the cryotolerance of preantral follicles enclosed in ovine ovarian tissue slices. For this purpose, ovarian slices were obtained from abattoired juvenile lambs and randomly distributed to the following groups: fresh, toxicity, vitrified (control), and three treatment groups in two experiments. Vitamin E, vitamin C, or vitamin E + C was added to the vitrification media alone in the first experiment and added to all vitrification, warming, and culture media in the second experiment. Finally, the treated tissues were cultured in vitro for 12 hours. The histological analysis showed that single or combined use of vitamins E and C increases intact preantral follicles in comparison to the control in two experiments (p < 0.05), and simultaneous use of vitamins E and C had a synergistic effect on increasing the percentage of normal preantral follicles in experiment 2 (p < 0.05). Due to the better results in Experiment 2, stromal cell density, antioxidant activity, and molecular evaluation were followed only in this experiment. The vitamin E + C group had higher stromal cell density compared with control group (p < 0.05). Vitamin E strengthened antioxidant capacity compared with the control and vitamin C groups (p < 0.05). This effect was exacerbated when used in combination with vitamin C (p < 0.05). The expression of all evaluated genes (BMP4, BMP15, GDF9, and KITLG) was significantly increased in ovarian tissue treated with vitamin E + C compared with the control group (p < 0.05). This increase was also observed in BMP4, GDF9, and KITLG genes compared with the vitamin C group (p < 0.05). In conclusion, this study revealed the positive effects of vitamins E and C on preantral follicle viability and to some extent a synergistic action of vitamin C on the protective effects of vitamin E against preantral follicle degeneration and increasing antioxidant capacity and development of preantral follicles after ovine ovarian tissue vitrification.

Objectives: To evaluate the population health returns from investment in the Victorian Cancer Biobank (VCB), a research consortium including five hospital-integrated sample repositories located in Melbourne, Australia. Methods: This economic evaluation assigned monetary values to the health gains attributable to VCB-supported research. These were then compared with the total investment in VCB infrastructure since inception (2006-2022) to determine the return on investment (ROI). A time lag of 40 years was incorporated, recognizing the delay from investment to impact in scientific research. Health gains were therefore measured for the years 2046-2066, with a 3% discount rate applied. Health gains were measured in terms of disability-adjusted life years (DALYs) attributable to VCB-associated research, with monetary cost assigned via the standardized value of a statistical life year (AU$227,000). The age-standardized DALY rate attributable to cancer was modeled for two standpoints (1) extrapolating the current decreasing trajectory and (2) assuming nil future improvement from current rates, with 33% of the difference attributed to scientific innovation. The proportion of the aggregate health gain attributable to VCB-supported research was estimated from the number of VCB-credited scientific publications as a proportion of total oncology publications over the same period. Results: The AU$32,628,016 of public funding invested in VCB activities over the years 2006-2022 is projected to generate AU$84,561,373 in total (discounted) savings. ROI was AU$1.59 for each AU$1 invested. Conclusions: The VCB offers a strong ROI in terms of impacts on health, justifying the expenditure of public funds and supporting the use of biobanks to advance scientific research.

Importance of Study: Semen cryopreservation results in sperm damage due to lipid peroxidation or oxidative stress, leading to a decrease in conception rate. The sperm damage during cryopreservation can be minimized with the use of suitable antioxidant supplements in semen diluent. Some herbs have potent antioxidant potential and can be used in semen diluent to protect the spermatozoa. Objective: Hence, the investigation was planned to evaluate the effect of Asparagus racemosus (A. racemosus) aqueous extract on buck semen quality during cryopreservation. Methodology: In the current study, semen was collected from eight Sirohi bucks, and from each buck, 8 ejaculates were collected. Good-quality semen samples were pooled during each collection. Pooled semen samples were then divided into four equal parts and diluted in TRIS buffer containing different concentrations of A. racemosus aqueous extract (different groups, i.e., G I -5 mg, G II -2.5 mg, G III -1.25 mg, and G IV -0 mg of A. racemosus aqueous extract in 1 mL TRIS buffer). All the diluted semen samples were kept at equilibration temperature (5°C) for 2 hours and then cryopreserved by the manual method. Semen samples were evaluated for various sperm characteristics and antioxidant status before and after cryopreservation. Results: Asparagus racemosus aqueous extract showed significant (p < 0.05) enhancement of sperm viability, sperm motility, acrosomal integrity, and plasma membrane integrity, whereas it reduced sperm abnormality. Furthermore, in the experimental groups, the antioxidant gene expression was found to be increased compared to that of the treatment group. G III (p < 0.05) showed significantly better results in terms of sperm viability, sperm motility, acrosomal integrity, and plasma membrane integrity. Conclusion: Asparagus racemosus aqueous extract has the antioxidant potential to protect buck spermatozoa during semen cryopreservation.

Biobanks are important resources for improving public health and individual care. Some legal frameworks can be more or less conducive to advancing the potential benefits of biobanks. The purpose of this article is to assess biobanking legislation and practices in Spain to determine how well they fare in such a regard. We focus here on some of the primary ethical values that ground relevant legislation and that we believe are consistent with promoting biobanking benefits: the value of scientific research; efficient use of scarce resources; and respect for the dignity of donors. We argue that although Spanish regulations advance these values in important ways, they also have provisions that undermine them and thus risk limiting the potential benefits of biobanks. We offer some suggestions for improvement.

Objectives: The aim of this study was to determine the cause of elevated serum potassium levels when blood collection tubes containing separating gel are stored under refrigeration. Methods: Fifty-seven hospitalized patients and 11 healthy volunteers were recruited. Venous blood samples were obtained using Insepac II, Neotube, and Venoject^® II, without anticoagulant. After centrifugation under different processing conditions, the capped tubes were stored at 4°C without aliquoting, and serum potassium levels were measured for up to 14 days. Correlation between the increase in potassium levels and blood cell counts was assessed. Furthermore, serum was replaced with a saline solution and potassium levels were determined after refrigeration. Results: Refrigerated samples stored in Insepac II tubes had significantly higher serum potassium levels on day 14 than on the day of blood collection. The increase in serum potassium levels was positively correlated with the number of red blood cells, but not white blood cells and platelets in venous blood. Furthermore, potassium levels were elevated when serum was replaced with a saline solution. Using Venoject II, which has a larger tube diameter and thicker separating gel than those of Insepac II and Neotube, did not increase serum potassium levels after storage. Increase in the serum potassium level was markedly suppressed by centrifugation at 2330 g for 15 minutes relative to other processing conditions. Conclusions: Potassium levels increase when serum is refrigerated in collection tubes containing separating gel. This can be attributed to contamination of the serum layer by blood cell components beyond the separating gel.

In this study, the effects of ferulic acid (0.1, 1, ve 10 mM), tryptophan (5, 25, ve 50 mM), and L-glutamine (10, 50, ve 100 mM) at different doses added to 18% raffinose + 3% skimmed milk powder sperm extender on the freezing of mouse spermatozoa in liquid nitrogen were investigated. The combination of 18% raffinose + 3% skimmed milk powder without additives was used as the control group. Frozen spermatozoa were thawed in a 37°C water bath for 30 seconds. After freeze-thawing, motility, dead spermatozoa ratio, plasma membrane integrity, abnormal acrosome ratio, motility endurance (for 4 hours), and cell apoptosis tests were performed in Human Tubal Fluid (HTF). Compared with the control group after freezing and thawing, the highest motility and plasma membrane integrity were obtained in the 10 mM L-glutamine group with 56.6% ± 2.11% and 77.8% ± 0.87%, respectively (p < 0.05). In addition, when compared to the control group, the lowest rate of dead spermatozoa and abnormal acrosome was found in the 10 mM L-glutamine group as 26.0% ± 1.46% and 6.3% ± 1.09%, respectively (p < 0.05). The highest motility values for spermatozoa endurance were determined in the 10 and 50 mM L-glutamine groups up to the 4th hour compared to the control group (p < 0.05). In the evaluation of apoptosis in semen samples, there was no significant difference between the control, 0.1 mM ferulic acid, and 10 mM L-glutamine groups (p > 0.05). As a result, it was determined that the addition of 10 mM L-glutamine to the spermatozoa extender increased the motility, viable spermatozoa, functional membrane integrity, intact acrosome ratios, or motility endurance after freeze-thawing and could be used successfully in the freezing extender of mouse spermatozoa.

Development of novel biomarkers for diagnosis of disease and assessment of treatment efficacy utilizes a wide range of biospecimens for discovery research. The fitness of biospecimens for the purpose of biomarker development depends on the clinical characteristics of the donor and on a number of critical and potentially uncontrolled pre-analytical variables. Pre-analytical factors influence the reliability of the biomarkers to be analyzed and can seriously impact analytic outcomes. Sample quality stratification assays and tools can be utilized by biorepositories to minimize bias resulting from samples' inconsistent quality. In this study, we evaluated the quality of biobanked specimens by comparing analytical outcomes at 1, 5, and 10 years after collection. Our results demonstrate that currently available assays and tools can be used by biobank laboratories to support objective biospecimen qualification. We have established a workflow to monitor the quality of different types of biospecimens and, in this study, present the results of a qualification exercise applied to fluid samples and their derivatives in the context of urological diseases.

This study examined the influence of heat exposure on DNA samples during polymerase chain reaction (PCR) detection. In this study, λDNA samples, as model DNA, were exposed to 105°C for 3-90 minutes or to 105°C-115°C for 15 minutes by autoclaving. The exposed samples were subjected to real-time PCR using nine primer sets with amplicon sizes of 45-504 bp. Regarding DNA samples exposed to 105°C by autoclaving, the data showed negative correlations between the logarithm of λDNA concentration (log λDNA) calculated using real-time PCR and exposure duration and a good relationship between the slope of the regression line and amplicon size. Regarding λDNA samples exposed to heat for 15 minutes, the data showed negative correlations between the log λDNA and exposure temperature and a good relationship between the slope of the regression line and amplicon size. These results showed that the equations used in this study could predict the degree of degradation in λDNA samples by autoclaving, and the PCR detection levels of the DNA at each amplicon size.