Storage Of Biological Samples Research Articles

Achieving temperature stability for storage of biological samples in an autodefrost freezer.

To assess the temperature stability of an autodefrost freezer commonly used in veterinary practices, whether the use of a Styrofoam cooler within the freezer provides temperature stability, and the ease of use of a remote monitoring system for the notification of temperature elevations. None. Temperature in the freezer and 2 Styrofoam coolers were assessed with remote monitoring thermometers every 15 minutes. Temperature values were monitored from October 11 to December 18, 2023 (for a 68-day period). Data analysis focused on temperatures for the freezer exceeding 0 °C and the elevations in temperatures within the coolers relative to the freezer. The freezer had an increase in temperature approximately every 16 hours. Over 68 days, the freezer had a temperature greater than 0 °C 27 times, representing 26 separate elevations. The Styrofoam coolers within the freezer never registered a temperature higher than -5 °C. Elevations in temperature within the freezer were larger in magnitude than temperature elevations within the coolers, which showed smaller-magnitude changes in temperature. The temperature stability provided by the Styrofoam cooler would avoid potential freeze-thaw cycles of any stored biological samples. Additionally, the remote temperature monitoring system is easy to install and monitor, providing peace of mind to practice management.

Analysis of Informed Consent Forms Submitted to Institutional Ethics Committee of a Medical Institute in Southern India: A Cross-sectional Observational Study

Introduction: Informed consent is an essential component in research involving human participants. However, the informed consent obtained may be incomplete and not fulfill the essential criteria of Informed Consent Forms (ICFs). Although the guidelines for developing ICFs have been clearly spelled out by various research bodies, these guidelines are not followed completely. Aim: To analyse the ICFs submitted to the Institutional Ethics Committee (IEC) of a Medical Institute in Southern India. Materials and Methods: The present study was a crosssectional observational study analysing ICFs submitted to the IEC of McGann District Teaching Hospital, Shimoga Institute of Medical Sciences (SIMS), Shivamogga, Karnataka, India, for the period 2014 to January 2023. All research projects containing ICFs during the study period were included in the study. Of the research projects submitted, only 70 had ICFs, and these were subjected to analysis as per Indian Council of Medical Research (ICMR) guidelines criteria. The criteria for ICFs were: statement of research, purpose/methods of the study, duration/frequency of the study, benefits to participants/community, foreseeable risks, discomfort/inconvenience, confidentiality, payment/ reimbursement for participation. In addition to these, ICFs were also analysed for additional elements as per ICMR criteria for tissue and blood samples. The results were then subjected to descriptive statistical analysis and presented as mean and percentages. Results: Many of the required essential elements were present in nearly 50% of ICFs submitted to the IEC, which include information on the basic purpose/methods of the study 70 (100%), identity of the principal investigator/research team 57 (81.42%), freedom to participate/withdraw from the study 55 (78.57%), confidentiality of records 54 (77.14%), and foreseeable risks, discomfort, and inconvenience to participants 35 (50%). Other essential elements like benefits were present to participants/community 28 (40%), payment/reimbursement for participation 28 (40%), duration and frequency 12 (17.14%), statement of research 9 (12.85%), treatment/compensation for injury 4 (5.71%). Regarding additional elements of ICFs for biological samples, ICFs adhered to the ICMR requirements except none of the submitted forms had any information on the period of storage of biological samples. Conclusion: The ICF is an essential requirement for conducting research. Ensuring adherence of ICF to guidelines is important from a research perspective. The present study concludes that the majority of the essential elements were present in ICFs with a few exceptions like study as research and information on the storage of biological samples, which was nil.

Biobank as a legal category

The development of medical technologies increases the need for legal regulation of relations on the formation of biobanks. The comparative legal method and the formal legal method were used. Currently, the term biobank is used to denote both the totality of objects of rights and to denote the entity carrying out the storage. The conclusion is made about the need for additional regulation of the concept of biobank. To protect the rights and legitimate interests of the patient, it is necessary to take into account his consent or lack of consent both for the removal of cells and other biological materials, and for their storage, transportation, cryopreservation. The conditions of consent may provide for the destruction of biological material upon the occurrence of certain conditions, for example, in the event of the death of a patient. Consent to the processing of personal data should be obtained separately. For more complete regulation, it is necessary to determine the features of the legal personality of organizations engaged in professional activities for the storage of biological samples and related data. It should be recognized that it is expedient to preserve the rules on licensing the activities of such organizations.

Industrial Biotechnology Conservation Processes: Similarities with Natural Long-Term Preservation of Biological Organisms

Cryopreservation and lyophilization processes are widely used for conservation purposes in the pharmaceutical, biotechnological, and food industries or in medical transplantation. Such processes deal with extremely low temperatures (e.g., -196 °C) and multiple physical states of water, a universal and essential molecule for many biological lifeforms. This study firstly considers the controlled laboratory/industrial artificial conditions used to favor specific water phase transitions during cellular material cryopreservation and lyophilization under the Swiss progenitor cell transplantation program. Both biotechnological tools are successfully used for the long-term storage of biological samples and products, with reversible quasi-arrest of metabolic activities (e.g., cryogenic storage in liquid nitrogen). Secondly, similarities are outlined between such artificial localized environment modifications and some natural ecological niches known to favor metabolic rate modifications (e.g., cryptobiosis) in biological organisms. Specifically, examples of survival to extreme physical parameters by small multi-cellular animals (e.g., tardigrades) are discussed, opening further considerations about the possibility to reversibly slow or temporarily arrest the metabolic activity rates of defined complex organisms in controlled conditions. Key examples of biological organism adaptation capabilities to extreme environmental parameters finally enabled a discussion about the emergence of early primordial biological lifeforms, from natural biotechnology and evolutionary points of view. Overall, the provided examples/similarities confirm the interest in further transposing natural processes and phenomena to controlled laboratory settings with the ultimate goal of gaining better control and modulation capacities over the metabolic activities of complex biological organisms.

National Blood Foundation 2021 Research and Development summit: Discovery, innovation, and challenges in advancing blood and biotherapies.

National Blood Foundation 2021 Research and Development summit: Discovery, innovation, and challenges in advancing blood and biotherapies.

Training in biobanking in the context of modern medical education

The development of biomedical research based on predictive, preventive and personalized medicine has served as a challenge to the formation and rapid development of a novel interdisciplinary scientific area — biobanking, the main goal of which is the long-term proper storage of biological samples and related data for use in scientific and clinical research. Qualified personnel, along with the creation and development of biobank infrastructure, provide high-quality results required for biomedicine. Education and training must reflect the changing scope of knowledge and adapt to biobanking challenges. In this regard, there is an increasing need to develop and implement educational programs for staff working in and managing biobanks, as well as researchers, doctors and students who are not familiar with this area, but plan research using bioresources. The aim of this review is to present an analysis of the main biobanking directions and training programs in the world and in Russia in order to assess the existing problems and needs.

How Can Biobanks Help You in Your Research Projects?

How Can Biobanks Help You in Your Research Projects?

Robust and Long-Term Cellular Protein and Enzymatic Activity Preservation in Biomineralized Mammalian Cells.

Preservation of evolved biological structure and function in robust engineering materials is of interest for storage of biological samples before diagnosis and development of vaccines, sensors, and enzymatic reactors and has the potential to avoid cryopreservation and its associated cold-chain issues. Here, we demonstrate that "freezing cells in amorphous silica" is a powerful technique for long-term preservation of whole mammalian cell proteomic structure and function at room temperature. Biomimetic silicification employs the crowded protein microenvironment of mammalian cells as a catalytic framework to proximally transform monomeric silicic acid into silicates forming a nanoscopic silica shell over all biomolecular interfaces. Silicification followed by dehydration preserves and passivates proteomic information within a nanoscale thin silica coating that exhibits size selective permeability (<3.6 nm), preventing protein leaching and protease degradation of cellular contents, while providing access of small molecular constituents for cellular enzymatic reaction. Exposure of dehydrated silicified cells to mild etchant or prolonged hydrolysis removes the silica, completely rerevealing biomolecular components and restoring their accessibility and functionality.

Creation of a collection of different biological sample types from elderly patients to study the relationship of clinical, systemic, tissue and cellular biomarkers of accumulation of senescent cells during aging

With aging, tissue homeostasis and their effective recovery after damage is violated. It has been shown that this may be due to the excessive accumulation of senescent (SC) cells in various tissues, which leads to the activation of chronic sterile inflammation, tissue dysfunction and, as a result, to the development of age-related diseases. To assess the contribution of SC cells to human body aging and pathogenesis of such diseases, relevant biomarkers are studied. For successful translation into clinical practice of approaches aimed at regulating the SC cell content in various tissues, it is necessary to study the relationship between the established clinical biomarkers of aging and age-related diseases, systemic aging parameters, and SC biomarkers at the tissue and cellular levels.Aim. To develop and describe action algorithms for creating a biobank of samples obtained from patients aged &gt;65 years in order to study biomarkers of SC cell accumulation.Material and methods. To collect samples, an interaction system was built between several research, clinical and infrastructure departments of a multidisciplinary medical center. At the stage of preanalytical training, regulatory legal acts were developed, including informed consent for patients, as well as protocols for each stage of the study.Results. A roadmap was formed with action algorithms for all participants in the study, as well as with a convenient and accessible system of annotations and storage of biological samples. To date, the collection includes biological samples of 7 different types (peripheral blood serum, formalin-fixed tissue samples and formalin fixed paraffin embedded tissue specimens, samples of different cells isolated from peripheral blood, skin and adipose tissue, samples of deoxyribonucleic and ribonucleic acids, cell secretome conditioned media) obtained from 82 patients. We accumulated relevant anamnestic, clinical and laboratory data, as well as the results of experimental studies to assess the SC cell biomarkers. Using the collection, the relationship between clinical, tissue and cellular biomarkers of SC cell accumulation was studied.Conclusion. The creation of a collection of biological samples at the molecular, cellular, tissue and organism levels from one patient provides great opportunities for research in the field of personalized medicine and the study of age-related disease pathogenesis.

Guidelines for collection of biological samples for giant otters (Pteronura brasiliensis Gmelin, 1788) and Neotropical otters (Lontra longicaudis Olfers, 1818)

To optimize the health evaluation of Giant and Neotropical otters in the wild and under human care and to guarantee the quality of the information collected and maximize efforts in research projects, Brazilian veterinarians developed two protocols: 1) Protocol for collection, processing and storage of biological samples from live Giant and Neotropical otters, 2) Protocol for collection of biological material during necropsy.

Assessing consent for and response to health survey components in an era of falling response rates: National Health and Nutrition Examination Survey, 2011-2018.

Response rates for national population-based surveys have declined, including the National Health and Nutrition Examination Survey (NHANES). Declining response to the initial NHANES interview may impact consent and participation in downstream survey components such as record linkage, physical exams, storage of biological samples and phlebotomy. Interview response rates dropped from 68% in 2011-2012 to 53% in 2017-2018 for adults age 18 and older. Response was higher for children (1-17 years) but with a similar downward trend (2011-2012, 81%; 2017-2018, 65%). Despite declining interview response rates, changes in consent and response rates for downstream components over time have been mixed. Among those interviewed, the examination response rate was over 93%, consent for record linkage was over 90%, and consent for storage of specimens for future research was over 99%. The availability of a blood sample for storage ranged between 60%-65% for children and 78%-85% for adults.

Preparation of High-Temperature Sample Grids for Cryo-EM.

The sample grids for cryo-electron microscopy (cryo-EM) experiments are usually prepared at a temperature optimal for the storage of biological samples, mostly at 4 °C and occasionally at room temperature. Recently, we discovered that the protein structure solved at low temperature may not be functionally relevant, particularly for proteins from thermophilic archaea. A procedure was developed to prepare protein samples at higher temperatures (up to 70 °C) for cryo-EM analysis. We showed that the structures from samples prepared at higher temperatures are functionally relevant and temperature dependent. Here we describe a detailed protocol for preparing sample grids at high temperature, using 55 °C as an example. The experiment made use of a vitrification apparatus modified using an additional centrifuge tube, and samples were incubated at 55 °C. The detailed procedures were fine-tuned to minimize vapor condensation and obtain a thin layer of ice on the grid. Examples of successful and unsuccessful experiments are provided.

Assessment of the viability of cultures of lactic acid microorganisms during their freezing and low-temperature storage

Relevance. A significant increase in microbiome-associated diseases, closely related to violations of the bacterial diversity and functions of the normal intestinal microbiota, dictates the need to develop and implement measures for the long-term preservation of individual representatives of the normal microbiota in order to create new strategies for modifying the composition of microbiomes.Methods. The influence of the technology of deep freezing and storage of intestinal isolates of lactic acid bacteria of 2 taxonomic groups isolated from poultry in the conditions of the Low-temperature automated storage of biological samples of the Departmental Collection of useful microorganisms for Agricultural purposes of the Russian Agricultural Academy (VKSM) on the MRS culture medium using 10 and 20% glycerin or 10 and 20% sucrose as cryopreservants was studied. The suspensions of the isolates were frozen at - 150 °C for 18 hours and then placed in an automated cryopreservation at -80 °C. Control of samples for safetyResults. The technology of cryofreezing of lactic acid bacteria on MRS-broth using 10 and 20% glycerin or 10 and 20% sucrose as cryopreservants allows preserving the viability, physiological and biochemical properties of intestinal isolates of lactic acid bacteria when stored for 18 months. All the protective media used (MRS-broth with glycerin 10 and 20%, MRS -broth with sucrose 10 and 20%) showed comparable results in the preservation of viability and acid-forming activity of Lactobacillus fermentum-2, Pediococcus pentosaceus 6p-3, Pediococcus pentosaceus 28p-1 isolates. Then the storage of Pediococcus pentosaceus isolate (28p-1) in a given parameter on a protective medium with 10 and 20% sucrose led to a decrease in the activity of acid formation.

Evaluation of different skin regions derived from a postmortem jaguar, Panthera onca (Linnaeus, 1758), after vitrification for development of cryobanks from captive animals.

Biological resource banks represent valuable tools for the conservation of species vulnerable to extinction, such as the jaguar. Cryobanks of skins have the potential to safeguard rare genotypes, allowing the potential exploitation of biological samples in animal multiplication technologies and the study of genetic variability. Determination of the most suitable skin regions for tissue conservation can help increase the efficiency of cryobanks and the storage of biological samples. To this end, we evaluated the effects of vitrification of skin tissues from the ear, caudal, and femoral regions of a post-mortem jaguar belonging to a zoo in Brazil. Non-vitrified and vitrified samples were evaluated and compared using quantitative methods, focusing on skin thickness, cell quantification, number of perinuclear halos, collagen and elastic density, and proliferative activity. No differences were observed in skin thickness, number of perinuclear halos, elastic density, and proliferative activity between non-vitrified and vitrified tissues in skin from any region. However, vitrified tissues derived from femoral skin showed a reduction in the number of fibroblasts, epidermal cells and collagen density compared to non-vitrified tissues. In summary, the ear and caudal regions provided the best conservation of somatic tissues derived from jaguars, and skin samples from these regions are therefore the most suitable for the formation of cryobanks.

Design of a parallel robot with additively manufactured flexure hinges for a cryogenic work environment

Automation is ubiquitous in today’s industrial landscape and is finding its way into more and more highly specialised applications - also in the field of cryopreservation. The extreme work conditions in cryobanks place exceptionally high demands on the mechanical and electronic components used. The preservation and storage of biological samples take place at temperatures between -130 °C and -196 °C using liquid nitrogen as a cooling medium. The bearings and joints used in industrial parallel kinematic robots (for example, ball bearings or Cardan joints) jam at these ambient parameters and are unsuitable for an application within a cryobank. We, therefore, develop methods and technologies to enable fully automated handling of biological samples under cryogenic working conditions. The basis for this is a parallel kinematic robot structure that allows the drives to be placed outside the cold environment. In contrast, the rest of the robot structure can be actuated in a cryogenic container.In this context, the passive joints for this parallel robot are designed as additively manufactured monolithic flexure hinges. This paper presents the design, simulation, and construction of the parallel robot and focuses on the flexure hinges fabricated using the selective laser melting process (SLM). We describe the design of the flexure hinges, their intended use in the robot, and the experimental setup used for their validation. We also compare the operating parameters recorded in experiments (such as bending angle, bending moment) with the data obtained in finite element method simulations (FEM). In addition, we describe the geometric constraints and deviations of the manufactured joints due to the manufacturing process.

Influence of exogenous and endogenous factors on the quality of the preanalytical stage of laboratory tests (review of literature).

A literature review in the article presents an analysis of the influence of endogenous and exogenous factors on quality of preanalytical phase of laboratory testing. The review shows significance of external and internal factors influencing blood samples at preanalytical phase of laboratory testing. Among the exogenous factors considered: phlebotomy, test tubes for samples, transportation and storage. A number of factors exist at this phase that significantly affect test results. We examined these aspects of phlebotomy process: staff training, disinfectant contamination, needle diameter, needle material contamination. The review considers possible contamination with tube components and the importance of choosing the right anticoagulants and excipients. Transportation and storage of biological samples can be a source of errors at the preanalytical phase of laboratory testing. We analyzed the problem of determining the stability of analytes during storage and aspects of transportation samples by modern means. Among the endogenous factors considered: hemolysis, lipemia, icterricity, cell metabolism.. Hemolysis is one of the most frequent consequences of errors at the preanalytical phase. We analyzed importance of choosing a method for identifying hemolized tubes and the heterogeneity of bias results on different analytical systems. The review shows contribution of various classes of lipoproteins to turbidity of sample, possible preanalytical errors and impact on analytical tests. We examined possible effects of high bilirubin concentrations on analyte measurements. In the review, we also examined metabolism of some cells and its effect on samples.

An alternative method for determining the body condition index of the free-living South American coati

Assessing and monitoring the welfare of free-living mammals is not a usual process due to the logistical complications associated with their capture and sedation, collection and storage of biological samples and their release. In this context, non-invasive methods for monitoring wildlife constitute a good alternative approach for in situ conservation. Body condition index, as a measurement of health status, has been used in free-living mammals; its low value may be associated with negative effects on reproduction and survival. The present study aimed to generate an alternative and reliable non-invasive method and then determine the body condition index, based on previously-collected biometric measurements, without the need to capture and immobilise the animals. A total of 178 free-living Nasua nasua Linnaeus, 1766 were trapped, weighed and measured. Statistical methods were used, based on Boosted Regression Trees (BRT) using body mass, biometric measurements (body length, height and chest girth) and gender as explanatory variables. To assess the agreement between the real Body Condition Indices (BCIs) and the predicted values of BCIs, we explored the correlation between each model using the Bland-Altman method. This method showed a strong agreement between the predictive BRT models proposed (standardised residuals from a linear regression between body length and chest girth) and standardised residuals (linear regression between body mass and body length). The results obtained herein showed that BRT modelling, based on biometrical features, is an alternative way to verify the body conditions of coatis without the need to capture and immobilise the animals.

ISO 20387 biobanking standard. Analysis of requirements and experience of implementation.

Currently one of the most important problems facing biobanking specialists is the standardization of biobanks operation. Close attention is paid to this issue by international biobanking organizations, such as ISBER and BBMRI-ERIC, which develop regulatory documentation in this area. The article provides examples of standardization tools - implementation of the ISO 9001 quality standard and ISBER Best Practices. General information about the development, scope, and structure of the ISO 20387 standard is provided. The standard does not provide ready-made solutions and does not contain specific requirements for storage temperature or biosamples processing in biobanks, allowing each biobank to adapt its own management system to existing conditions and needs. The standard contains requirements for both the organization of the biobanking and the supporting processes - personnel competence; requirements for biological safety; infrastructure management, including equipment used by the biobank, environmental parameters that affect the storage of biomaterial. The standard contains requirements for the quality management system of biobank, as a necessary element of the organization of any biorepository. At the initiative of the Russian National Association of biobanks and biobanking specialists (NASBIO), development of the Russian standard GOST R ISO 20387 «Biotechnology. Collection and storage of biological samples in biobanks. General requirements» is included in the plan of the National Standardization Program for 2020 by order of Rosstandart No. 2612 of 11/01/2019. Implementing quality standards is a long and painstaking process that requires the involvement of all employees and certain resources. However, the effectiveness of strict compliance exceeds the cost of developing, implementing and maintaining management systems, as it significantly increases the confidence of researchers in the work of biobanks, guarantees high quality of biospecimens and associated data, and creates opportunities for cooperation, both at the national and international level, based on the application of common quality standards in the work.

Enrolment of children in clinical research: Understanding Ghanaian caregivers’ perspectives on consent/assent procedures, and their attitudes towards storage of biological samples for future use

Child assent is recommended in addition to parental consent when enrolling children in clinical research; however, appreciation and relevance ascribed to these concepts vary in different contexts, and information on attitudes towards storage of biological samples for future research is limited, especially in developing countries. We assessed caregivers’ understanding and appreciation of consent and assent procedures, and their attitudes towards use of stored blood samples for future research prior to enrolling a child in clinical research. A total of 17 in-depth interviews were conducted with primary caregivers of children (fathers [n = 3], mothers [n = 12], and grandmothers [n = 2]) at enrolment or on the immediate follow-up date. All caregivers recalled significant amount information from the study information sheet and were able to appropriately link such information to the consent process. While all participants confirmed information received prior to blood sampling as adequate, a few noted that the purpose was not sufficiently well communicated. Caregivers felt children were cognitively vulnerable, and prone to decisions that were not necessarily in their best interest. Nearly all caregivers felt it was their right and responsibility to overrule objections from their ward’s regarding enrolment into specific study or receipt of a therapeutic procedure. There were no objections or concerns regarding use of stored biological samples for future research purposes. There is thus, a need to improve understanding of caregivers on the information provided during the informed consent process. Context-specific studies on the age of assent in specific populations are needed.

Guidance to post-mortem collection and storage of biological specimens for the diagnosis of Covid-19 infection

The following document is based on the current knowledge of SARS-CoV-2 and the related disease (COVID-19). The Italian Institute of Health (ISS) is monitoring the virus spread throughout the country through daily reports sent by individual regions and regional reference laboratories. The diagnosis of infection is based on the use of standardized and validated molecular tests for the search for viral RNA in different sets of biological samples, from patients suspected of having contracted the infection. This document provides a specific guidance for the collection and storage of biological samples from deceased persons. It also provides recommendations on safety practices to be adopted during both collection and handling of specimens, and during autopsy procedures. With regard autopsy, it is recommended to follow the procedures for the execution of diagnostic findings in patients died with SARS-CoV-2 infection issued by ISS Working Group on Causes of Death from COVID-19, published on March 27, 2020.