Biopreservation and Biobanking最新文献

Aim: Ethylene glycol (EG) has been employed as a cryoprotectant for many years in mammalian semen cryopreservation but not assessed for birds except for its recently illustrated beneficial effects on commercial chicken lines. The Indian red jungle fowl is facing trouble in its native range due to human encroachment. Therefore, the present study was designed to elucidate the cryoprotective effect of different EG concentrations (5%, 10%, 15%, and 20%) on frozen Indian red jungle fowl semen. Materials and Methods: Semen was collected from 20 cocks, and qualifying ejaculates (>70% motility) were pooled and diluted (15) with red fowl extender. EG was added to the four samples and 20% glycerol in control at 4°C. Samples were equilibrated and cryopreserved in LN₂. Semen quality and biochemical activity were assessed at various stages of cryopreservation. Results: Sperm motility, viability, plasma membrane and acrosomal integrity, chromatin integrity, and mitochondrial activity were recorded highest (p < 0.05) with 20% EG at the post-equilibration and post-thaw stages. Lipid peroxidation was recorded lowest (p < 0.05) with 20% EG compared with other concentrations and control at the post-equilibration and post-thaw stages. Conclusions: It is concluded that 20% EG exhibits cryoprotective properties in terms of regulating morphological and biochemical traits of frozen Indian red jungle fowl sperm.

Biobanking is crucial for advancing medical research and personalized medicine, offering high-quality biospecimens for studies on biomarkers, drug development, and diagnostics. Despite its global potential, challenges such as fragmented governance and varying standards hinder biorepository collaboration, particularly in South Africa (SA). A unified national biobank network could enhance research and healthcare by improving biospecimen access, ethical governance, and collaboration. Global biobank networks offer models for standardization, data sharing, and international cooperation. SA can benefit from these models by creating a centralized biobank platform, promoting capacity building, and fostering regional and global partnerships. To address the challenges SA faces regarding biobanking, the Medical Biobanks Cluster established a network named Medical Biorepositories of SA (MBirSA), which seeks to build a cohesive network of medical biorepositories in SA. Through this network, it plans to foster an inclusive culture of biospecimen and data protocol harmonization, while encouraging adherence to legal, ethical, and quality best practices and standards. The network aims to bring stakeholders together, increasing visibility and transparency, and encouraging sector-wide collaboration. MBirSA also aims to offer training to build capacity in global best practices, aid in the development of dependable biorepository infrastructure, and promote research partnerships to enhance healthcare advancements.

Introduction: Rare disease research in South Africa (SA) faces significant challenges, including limited prioritization and awareness, which hinder advancements in patient care and scientific discovery. This article explores the recruitment strategies employed by the Centre for Human Metabolomics (CHM) Biobank, a nonhospital-based academic rare disease biobank, to address these challenges. Methods: We explain the consent process and documents as well as the three recruitment models employed, namely (1) Recruitment via referring clinician, (2) implementation of monthly diagnostic follow-up sessions, and (3) recruitment of patients for specific projects through clinic-based recruitment drives. Discussion: We discuss the benefits as well as the challenges of each model. Challenges included clinician and patient time constraints, distrust from current consent practices, and limited public awareness. We elaborate on future strategies to address these gaps such as simplifying consent, expanding recruitment sites, collaborating with clinical, academic and public institutions, and raising public awareness of the role of the CHM Biobank. Conclusions: From the models employed over the past 5 years, it is evident that recruitment is most effective when patients perceive a direct benefit, such as involvement in active projects. These strategies outlined in the discussion are crucial for ensuring the CHM Biobank's sustainability, diversity, and its impact on scientific research and patient outcomes in SA.

Cryopreservation of boar semen is essential for maintaining genetic diversity and improving reproductive efficiency. However, optimizing semen extenders and packaging methods is crucial to enhancing sperm quality and cryotolerance. Equex-STM is commonly used as a surfactant to enhance cryoprotective properties by stabilizing sperm membranes during freezing and thawing. A total of 24 ejaculates (n = 24), six from each of four Hampshire crossbred boars, aged between 18 and 24 months were used in the present study. Semen was collected twice weekly by gloved hand technique. The sperm-rich fraction showing more than 70% initial motility was considered for further processing and freezing. The fresh semen was diluted in Beltsville thawing solution (BTS) at a 1:1 ratio and further processed with the addition of Fraction I and Fraction II extenders supplemented with and without 1.5% Equex-STM paste {Control (C)-no Equex; Treatment (T-1.5% Equex-STM supplementation}. The extended semen was packaged in 0.25 and 0.5 mL straws to check the effect of straw size (T₁-0.25 mL straw and T₂-0.50 mL straw). Straws were cryopreserved in liquid nitrogen (LN₂) for post-thaw sperm quality evaluation. Results showed that the sperm motility, viability, plasma membrane integrity (PMI), DNA integrity, and mitochondrial membrane potential (MMP) were significantly (p < 0.001) higher in the T group in comparison with control (C). Sperm motility, PMI, and DNA integrity were highly significant (p < 0.001) and sperm viability and MMP were significant (p < 0.05) in group T₂ in comparison with T₁.

Background: Biological material and health information from patients are valuable for medical research. Under a "broad" consent model, hospital patients in Norway can consent to their biological material and health information being stored in research biobanks and used for "specific, broadly defined research purposes" within a specified medical research area but not for medical research in general. Patients are asked to provide new consent each time researchers wish to use their material in a different medical research area. This study investigated patient representatives' views on having a general consent for medical research without limitation to specific research purposes. We also investigated preferences for the storage of biological samples, the process of consent collection, and factors motivating or hindering consent. Method: An online, anonymous survey was shared with patient representatives from patient advisory councils at hospitals in Norway, who answered the survey on behalf of patients. A total of 157 representatives completed the survey (response rate of 41%). Results: A majority (66.2%) supported general consent for medical research and the use of surplus material for medical research in general (63.7%) without limitation to specific research purposes. A minority (35%) supported the use of surplus material without being informed. Sixty-five percent agreed that biological samples could be stored with no time limitation. Over half (56%) preferred to ask patients to consent prior to a hospital visit, and the majority (70.7%) supported the possibility of choosing between digital or paper consent. Factors motivating consent included the desire to contribute to medical research (89.8%) and faith in scientific progress (24.2%). Main hindrances included the fear that health information may be used for other purposes than research (49%), uncertainty regarding research uses (43.9%), and lack of information (31.8%). Conclusion: A move toward a general consent for medical research may better comply with patients' wishes and maximize research potential.

Background: Biobanking of prostate cancer tissue is crucial for advancing biomarker-guided precision medicine. However, there is no standardized optimal protocol for biobanking prostatectomy specimens. This study aims to compare the representativeness and sustainability of two biobanking protocols-"Punch" and "Slice"-currently used in Norway. Methods: Fresh frozen tissue from 40 radical prostatectomy specimens was biobanked using both the Punch and Slice protocols. Following macroscopic evaluation, a 2 mm thick transverse slice of the prostate (Slice protocol) was collected and stored in an ultra-freezer for future drill biopsy subsampling, guided by histopathological assessment of adjacent formalin-fixed, paraffin-embedded tissue sections. After the slice was collected, five cylindrical tissue samples were punched from the cut surfaces (Punch protocol). Statistical analyses were conducted to compare the sampling precision and time consumption of both protocols. Results: Cancerous tissue was successfully sampled in 87.5% of cases using the Punch protocol and 75% of cases using the Slice protocol. Both methods yielded comparable results in terms of the number of cancerous cores and the ability to sample tissue representing the highest Gleason grade. The mean biobanking time of tissue slices was 4.9 minutes compared to 15.1 minutes for the ready-to-use tissue punches. Both methods have previously been shown to provide high-quality RNA extracts. Conclusion: Both biobanking protocols are effective for sampling prostate cancer tissue, with no significant difference in precision or quality. The choice between protocols should consider factors such as resource availability, tissue quantity, and specific research needs. The Punch protocol is less resource-intensive overall, while the Slice protocol collects vastly more tissue, has a shorter period of ischemia, and provides detailed mapping of biobanked components, allowing for further subsampling at multiple time points.

The endoplasmic reticulum (ER) is the organelle responsible for protein folding in the cell. The damage that may occur during the freezing process of the sperm can exceed the protein loading capacity in the ER. Antioxidants, such as coenzyme Q₁₀ (CoQ₁₀), are added to freezing media to protect sperm cells. In this study, the aim was to investigate the expression levels of ER stress-related genes (protein kinase-like ER kinase [PERK], activating transcription factor 4 [ATF4], CCAAT-enhancer-binding-protein homologous protein [CHOP], and nuclear factor erythroid 2-related factor 2 [NRF2]) and quality parameters (viability, motility, acrosome status, and plasma membrane integrity) of mice sperm after freezing with an extender containing CoQ₁₀. Male BALB/c mouse spermatozoa were cryopreserved using a combination of 18% raffinose + 3% skimmed milk and 50 µM CoQ₁₀. The combination of 18% raffinose + 3% skimmed milk without CoQ₁₀ was used as the control group. The results showed that post-thaw sperm motility, viability, plasma membrane integrity, and intact acrosome rates were significantly higher in the CoQ₁₀-supplemented group compared with the control (untreated) group (p < 0.05). The expression of ER stress-related genes was then analyzed to investigate whether CoQ₁₀ attenuates ER stress in frozen-thawed sperm. The results significantly revealed that the addition of 50 µM CoQ₁₀ to the extender increased PERK, ATF4, and CHOP mRNA levels compared with the control group (p < 0.001). Next, NRF2 gene expression was analyzed to investigate whether CoQ₁₀ affects the antioxidant mechanism of post-thaw sperm. It was revealed that the expression of the NRF2 gene significantly increased in the CoQ₁₀ group compared with the control group (p < 0.001). Collectively, these results suggest that the freeze-thaw process induces ER stress in mouse sperm, and the supplementation of CoQ₁₀ to the cryoprotectant agent reduces ER stress-related genes, activates the gene related to the antioxidant defense system, and improves post-thaw sperm quality parameters.

The COVID-19 pandemic, spanning from early 2020 to late 2022, posed unprecedented challenges for global public health. However, it also spurred innovative approaches to pandemic management, notably the development of pathogen detection in wastewater. It was successfully demonstrated that wastewater analysis can not only reflect ongoing COVID-19 infections but also serve as an early indicator of disease prevalence within communities. Recognizing the value of longitudinal analyses of various pathogens, we identified the need for wastewater biobanking. This practice allows for the retrospective analysis of samples, offering critical public health insights at the population level. Moreover, the potential to transport and store biobanked samples at ambient temperature or in a dry state could greatly enhance the utility of this technology, especially in resource-limited settings such as low- and middle-income countries. This article also addresses the ethical considerations and public health implications of wastewater-based epidemiology. While this approach holds significant potential beyond pathogen detection, it is essential to evaluate the benefits and potential risks carefully.

Objectives: Biobanks are crucial for clinical biomarker research, but pre-analytical processing can impact sample suitability. Insufficient reporting of pre-analytical methods can lead to invalid conclusions. Methods: This study evaluated the pre-analytical reporting quality in 294 clinical biomarker studies using biobank plasma and serum samples published from 2018 to 2023, based on the Sample PREanalytical Code and the Biospecimen Reporting for Improved Study Quality guidelines. Results: The analysis revealed frequent incomplete reporting. Key underreported elements included fasting time (reported in 31% of articles), freeze-thaw cycles (22.8%), internal transport (8.5%), and centrifugation settings (20%-35%). Demographic data (96.9%), storage temperatures (81%), and blood tube additives (82.7%) were more consistently reported. No significant correlation was found between reporting quality and journal impact factor. Conclusions: Our findings highlight the need for standardized pre-analytical reporting to improve research replicability and biomarker study quality.

Introduction: Long-term storage of bee semen by freezing is a critical process for both the preservation of the genetic material and the sustainability of beekeeping activities. It has been observed that low-density lipoproteins (LDLs) increase sperm quality after freezing and thawing. Although studies have been conducted on the use of LDL for this purpose in different animal species, no research has been conducted on honeybee semen to date. Objectives: This study aimed to evaluate the potential effects of using LDL instead of egg yolk (EY) on sperm quality and fertilization rate by examining the effects of different LDL ratios (2.5%, 5%, 10%, 25%) on bee semen. Methods: Sperm collection was conducted using a Schley-type device, resulting in six distinct groups, including both no-supplemented and experimental groups. In the first experiment, sperm collected from 36 drones were diluted with varying LDL concentrations before being frozen and thawed; motility, membrane integrity, viability, and longevity were measured. In the second experiment, a total of 56 virgin sister queens, 8 from each group, were inseminated. Results: In the group containing 25% LDL, a significant increase was observed in the motility, membrane integrity, and viability rates of frozen-thawed honeybee sperm. In the group containing 25% EY, there was a clear decrease in these parameters; moreover, the lifespan of the sperm was significantly reduced. In the groups, the highest value in terms of fertility was observed in the 25% LDL group, and the lowest value was determined in the 25% EY-added group. Conclusion: The findings demonstrated that the addition of 25% LDL significantly enhanced both sperm quality and fertility rate in honeybees.