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Ballen, Karen K., Juliet N. Barker, Susan K. Stewart, Michael F. Greene, and Thomas A. Lane. "Collection and Preservation of Cord Blood for Personal Use." Biology of Blood and Marrow Transplantation 14, no. 3 (March 2008): 356–63. http://dx.doi.org/10.1016/j.bbmt.2007.11.005.
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Walter, Dagmar, Jawad Abousoud, Tingsheng Drennon, Connor Kunihiro, Kelly Martin, Yina Li, Nandhini Raman, Veronica Rodriguez, and Jill Lynden Herschleb. "Blood Collection Workflow for Interrogation of Translational Samples with Single Cell Technologies." Blood 142, Supplement 1 (November 28, 2023): 7182. http://dx.doi.org/10.1182/blood-2023-190485.
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Blood is a valuable and easily accessible sample type for clinical researchers that provides a snapshot of the organ systems at the time of sample collection. Readouts obtained from blood help inform clinical treatments, investigate disease mechanisms, evaluate drug response, and monitor outcomes. However, the impact of existing blood collection workflows on data output from new and high resolution technologies, like single cell RNA sequencing (scRNA-seq) remains a challenge. Recent scRNA-seq studies have shown that changes in gene expression can occur in blood cells in as little as a few hours post collection1. While bulk measurements rely on sample fixation to preserve the sample during transport, this approach does not preserve intact cells and hence, is not compatible with single cell technologies. With the recent availability of fixation compatible scRNA-seq assays comes the need to preserve blood while retaining single cell information. In this study, we evaluated various PBMC isolation and blood preservation methods which preserve single cell information while easing sample logistical constraints. Our initial experiments compared various PBMC isolation methods. scRNA seq analysis of these samples demonstrated that all isolation methods captured single cell gene expression with some variability in cell types along with user-variability. Changes in cell type proportions were observed in as little as 4 hrs post blood draw, emphasizing the need for an effective blood preservation method to retain single cells during transport and blood cell isolation. Next, we compared different methods of blood preservation including cryopreservation and fixation. Healthy and diseased whole blood samples were collected, preserved, stored followed by isolation of blood samples at different time points. scRNA-seq analysis of the samples demonstrated that blood can be preserved, transported, and stored (weeks to months) before blood cell isolation. Blood cells isolated from these cells can then be stored long term for scRNA-seq analysis. This blood collection and preparation workflow also eases blood transportation logistical constraints, allowing for batch shipping of samples from distributed collection sites. Coupled with automatable cell isolation methods and multiplexing single cell sequencing solutions, these workflows can be adapted for large scale translational research studies. References: Massoni-Badosa, R., Iacono, G., Moutinho, C. et al. Sampling time-dependent artifacts in single-cell genomics studies. Genome Biol 21, 112 (2020). Keywords: single cell RNA-seq, blood preservation
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Wilt, Kristen, Charis Ober, and John Garcia. "Abstract 15 A State-Run Public Cord Blood Collection Program." Stem Cells Translational Medicine 12, Supplement_1 (September 1, 2023): S17. http://dx.doi.org/10.1093/stcltm/szad047.016.
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Abstract Introduction The Arizona Public Cord Blood Program was created in 2011 by the Arizona Biomedical Research Centre, a sub-section of the Arizona Department of Health Services, to advance the collection and increase the number of racially and ethnically diverse cord blood units available for transplantation, as well as promote awareness of the benefits of cord blood stem cells through our educational partner, Save the Cord Foundation (STCF). Objectives Create a sustainable program for women of all racial and ethnic backgrounds to have the opportunity to donate cord blood, with the primary goal of transplantation, and secondary goal of providing non-transplantable cord blood units for research. A second objective was to educate the residents of the state of Arizona about cord blood stem cells and the need for their preservation. Methods A portion of state lottery funds supports the program monetarily. Those funds are provided to four collection hospitals who employ "cord blood consenters" whose responsibility it is to consent patients, assist delivery providers with collections, and package and ship cord blood units to our partner cord blood bank at MD Anderson Cancer Center. There are also two clinical coordinators who educate and train hospital staff on quality collection practices, with special emphasis on the importance of high volume, sterile collections. STCF provides education across the state to expectant parents, healthcare providers, schools, and the public about the need for cord blood stem cell donation for transplant and research. Results Since 2011, the Arizona Public Cord Blood Program has banked hundreds of racially and ethnically diverse cord blood units with the National Marrow Donor Program, and has had 86 life-saving cord blood units matched with patients in need around the globe. This innovative program has not only expanded cord blood awareness and promoted the preservation of cord blood; it has resulted in the creation of an economic engine for the state of Arizona that is an attractant for STEM based businesses and careers. Discussion A decade later, the Arizona Public Cord Blood Program has proven to be a sustainable model for collecting and providing suitable cord blood units for transplant to diverse patient populations.
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Spratt, John R., Lars M. Mattison, Paul A. Iaizzo, and Gabriel Loor. "The ABCs of autologous blood collection for ex vivo organ preservation." Journal of Thoracic and Cardiovascular Surgery 155, no. 1 (January 2018): 433–35. http://dx.doi.org/10.1016/j.jtcvs.2017.08.036.
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Wilt, Kristen, Charis Ober, John Garcia, and Jennifer Botsford. "Abstract 33 A Diverse and Sustainable State-Run Public Cord Blood Program." Stem Cells Translational Medicine 11, Supplement_1 (September 1, 2022): S39. http://dx.doi.org/10.1093/stcltm/szac057.033.
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Abstract Introduction The Arizona Public Cord Blood Program was created in 2011 by the Arizona Biomedical Research Centre, a subsection of the Arizona Department of Health Services, to advance the collection and increase the number of racially and ethnically diverse cord blood units available for transplantation, as well as to promote awareness of the benefits of cord blood stem cells through our educational partner, Save the Cord Foundation (STCF). Objective The main objective of this study was to create a sustainable program for women of all racial and ethnic backgrounds to have the opportunity to donate cord blood, with the primary goal of transplantation and the secondary goal of providing non-transplantable cord blood units for research. A second objective was to educate the residents of the state of Arizona about cord blood stem cells and the need for their preservation. Methods A portion of state lottery funds support the program monetarily. Those funds are provided to four partner collection hospitals employing "cord blood consenters," whose responsibility it is to consent patients, assist delivery providers with collections, and package and ship cord blood units to our partner cord blood bank at MD Anderson Cancer Center. There are also two clinical coordinators who educate and train hospital staff on quality collection practices, with special emphasis on the importance of high-volume, sterile collections. STCF provides education across the state to expectant parents, health care providers, schools, and the public about the need for cord blood stem cell donation for transplant and research. Results Since 2011, the Arizona Public Cord Blood Program has banked several hundreds of racially and ethnically diverse cord blood units with the National Marrow Donor Program (Figure 1) and has had 80 life-saving cord blood units matched with patients in need around the globe. This innovative program has expanded cord blood awareness and promoted the preservation of cord blood, and it also has resulted in the creation of an economic engine for the state of Arizona that is an attractant for STEM-based businesses and careers. Discussion A decade later, the Arizona Public Cord Blood Program has proven to be a sustainable model for collecting and providing suitable cord blood units for transplant to diverse patient populations.
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Okada, K., and J. Yasuda. "Methods for Collection and Preservation of Cattle Blood for Metabolic Profile Test." Japanese Journal of Veterinary Clinics 24, no. 1 (2001): 13–18. http://dx.doi.org/10.4190/jjvc2001.24.13.
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Martin, Kelly, Connor Kunihiro, Jawad Abousoud, Tingsheng Drennon, Yina Li, Sean Marrache, Nandhini Raman, et al. "Abstract 995: Preservation of patient whole blood preserves cancer immune cell biology at single cell resolution." Cancer Research 84, no. 6_Supplement (March 22, 2024): 995. http://dx.doi.org/10.1158/1538-7445.am2024-995.
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Abstract Blood is an easily accessible clinical sample type that helps inform treatments, investigate disease mechanisms, and monitor outcomes. Logistical challenges in the transportation and processing of blood samples impact data output from single cell RNA sequencing (scRNA-seq). Recent scRNA-seq studies have shown that significant changes in gene expression and immune cell populations occur in as little as a few hours post collection. Thus, there is a need for rapid preservation of blood upon collection before biological changes can occur. Existing preservation approaches compromise the cell membrane, making samples incompatible with single cell transcriptomic analyses. Here, we describe a novel workflow for fixation and long-term storage of whole blood, and isolation of peripheral blood mononuclear cells (PBMCs) from the fixed blood using a readily available, low-cost, high-throughput procedure. We then profile the gene expression of these cells using the fixation-compatible Single Cell Gene Expression Flex workflow. Importantly, this novel blood collection and preparation workflow eases blood transportation logistical constraints, allowing distributed sample collection and batched shipping of blood samples for scRNA-seq and potentially enabling large-scale translational research studies. We compared whole blood fixed immediately after collection using our workflow with clinical blood samples stored at ambient temperature. This revealed that immune cell populations and biological pathways shift significantly as early as five hours following sample collection, if not immediately stabilized. Across a time course of ambient temperature storage of whole blood without immediate stabilization, we observe significant upregulation of stress-related genes (FOS, JUN, CIRBP) in samples using Single Cell Gene Expression Flex profiling. Additionally, expression of a proto-oncogene TNFAIP3 was upregulated in whole blood stored at 20°C compared to samples stored at 4°C for the same lengths of time, unless samples were stabilized using the novel workflow shown here. This novel whole blood fixation method, coupled with automatable cell isolation methods and Single Cell Gene Expression Flex-enabled multiplexed single cell sequencing, will usher in a new age of economical, large-scale translational research studies that result in valuable clinical insights. Citation Format: Kelly Martin, Connor Kunihiro, Jawad Abousoud, Tingsheng Drennon, Yina Li, Sean Marrache, Nandhini Raman, Veronica Rodriguez, Paul Lund, Jens Durruthy Durruthy, Sarah Taylor, Andrew Kohlway, Peter Smibert, Dagmar Walter, Jill Herschleb. Preservation of patient whole blood preserves cancer immune cell biology at single cell resolution [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 995.
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Contreras Perea, Juan Carlos, Arturo Galindo Fraga, Martha Asunción Huertas Jiménez, Aurora Muñoz Pedraza, and Juan Miguel Terán Soto. "Guía de práctica clínica para toma de muestra de gases en sangre en México." Latin american journal of clinical sciences and medical technology 4, no. 1 (July 21, 2022): 121–33. http://dx.doi.org/10.34141/ljcs6842624.
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In order to promote preanalytic blood gas sample quality and to standardize sample collection, transport, and preservation protocols, a multidisciplinary experts group developed Guía de práctica clínica para toma de muestra de gases en México (Clinical practice guidelines for blood gas sample collection in Mexico). It includes recommendations for request, identification and patient´s preparation, sampling indications, contraindications, simple type selection, anatomical site, use and selection of the optimal sampling materials, operating room sampling, complications, handling, transport, and simple rejection criteria.
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Song, Jiaojiao, and Junmei Zhou. "Effects of preservation duration at 4 °C on the quality of RNA in rabbit blood specimens." PeerJ 8 (April 10, 2020): e8940. http://dx.doi.org/10.7717/peerj.8940.
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A prolonged preservation duration of blood specimens at 4 °C may occur due to the distance from collection points to storage facilities in many biobanks, especially for multicenter studies. This could lead to RNA degradation, affecting downstream analyses. However, effects of preservation durations at 4 °C on RNA quality in blood specimens need to be studied. We collected rabbit blood using EDTA tubes and stored them at 4 °C for different preservation durations. Then, we examined the quality of RNA from whole blood and leukocytes isolated from rabbit blood. Our results show that the purity of whole blood RNA and leukocyte RNA does not indicate significant change after rabbit blood is stored at 4 °C for different preservation durations (from 1 h to 7 days). The integrity of leukocyte RNA indicates the same result as above, but the integrity of whole blood RNA is significantly decreased after rabbit blood is stored at 4 °C for over 3 days. Moreover, expression of SMAD7, MKI67, FOS, TGFβ1 and HIF1α of whole blood RNA and leukocyte RNA remains basically stable, but PCNA expression of whole blood RNA or leukocyte RNA is significantly decreased after rabbit blood is stored at 4 °C for over 24 h or 7 days. Therefore, these results suggest that high-quality RNA is obtained from the fresher blood specimens and if blood specimens are stored for over 3 days at 4 °C, the quality of leukocyte RNA is more stable and of better quality than that of whole blood RNA.
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Nurunabi, Abu Sadat Mohammad, Miliva Mozaffor, Mohammad Tipu Sultan, Md Mozaharul Islam, and Kaisar Haroon. "Utilization of Brain Tissue as A Viable Postmortem Toxicological Specimen: A Review on Collection and Preservationof Samplefor Toxicological Analysis and Its Advantage Over Other Specimens." Bangladesh Journal of Neurosurgery 11, no. 2 (September 7, 2022): 114–17. http://dx.doi.org/10.3329/bjns.v11i2.61455.
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Collection of proper autopsy specimen and preservation are essential stepsfor the toxicological analysis in Forensic Medicine. Faulty collection and preservation of the specimens/samples can greatly alter or negate forensic chemical or toxicologicalexamination. In forensic toxicology practicein Bangladesh, postmortem specimen that is subjected to toxicological examinations generally focusing on mainly blood and sometimes urine or other fuds from different body cavities. Analysis of blood from different anatomical sites and tissue samples and urine may assist in the interpretation of the postmortem results. However, in many postmortem cases, there is little or no blood for quantitative drug analysis, or there might be such traumatic injury which led to significant blood loss or there is possibility of contamination form contents of the ruptured stomach. Besides, analysis of urine reveals negative result, if death occurs closely the time of intoxication. Given the circumstances, brain tissue may be a valuable specimen in postmortem toxicological analysis. The position of the brain in the body secures a tremendous protection and isolation which can eliminates or at least attenuates many of the interpretive challenges with postmortem blood, urine or other fluid specimens.This review paper is an update on the standard methods of brain tissue specimen collection and preservationprocedures for toxicological analysis and its value as well as advantages over other specimens, which might be of possible interest for forensic professionals in the country. Bang. J Neurosurgery 2022; 11(2): 114-117
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Azad, Nilima Rubaba, Md Mottaleb Ali, Md Iqramul Haque, Khaled Mahmud Sujan, Kazi Rafiqul Islam, Mohamma Alam Miah, and Md Kamrul Islam. "Influence of preservation length of the sample on the performance of complete blood count (CBC) in rats." Asian Journal of Medical and Biological Research 6, no. 1 (April 8, 2020): 22–26. http://dx.doi.org/10.3329/ajmbr.v6i1.46475.
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The performance of hematological tests deteriorates with the increase in the length of sample preservation. Therefore it has been an issue to characterize the maximum permissible period spent between blood collection and measurement to have the acceptable test report. From this view point, a study was undertaken to know about the effect of preservation length on complete blood count (CBC) in rat of Long Evans strain. A total of 30 samples were collected from 10 apparently healthy rats aged between 45-48 days and the blood samples were kept in commercial test tubes treated with EDTA. The test tubes containing whole blood samples were divided into three different groups based on preservation length and were allowed to keep at 4ºC for three different lengths of time viz. 2 hours, 4 hours and 6 hours until analysis. The samples were then analyzed for their complete blood count (TEC, TLC, Hb, PCV, DLC, Absolute Leukocyte Count, Red Cell Indices, RDW-SD, RDWCV, Platelet, MPV, PCT and PDW) using Sysmex XT-1800i auto hematological analyzer. Result showed that no significant change in CBC with the variation in preservation length. Based on these findings, it can be concluded that blood samples can be preserved for as long as 6 hours to have the same report obtainable when the samples are preserved at 4ºC in refrigerated condition for 2 or 4 hours.
Asian J. Med. Biol. Res. March 2020, 6(1): 22-26
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Kluge, Jonathan A., Adrian B. Li, Brooke T. Kahn, Dominique S. Michaud, Fiorenzo G. Omenetto, and David L. Kaplan. "Silk-based blood stabilization for diagnostics." Proceedings of the National Academy of Sciences 113, no. 21 (May 9, 2016): 5892–97. http://dx.doi.org/10.1073/pnas.1602493113.
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Advanced personalized medical diagnostics depend on the availability of high-quality biological samples. These are typically biofluids, such as blood, saliva, or urine; and their collection and storage is critical to obtain reliable results. Without proper temperature regulation, protein biomarkers in particular can degrade rapidly in blood samples, an effect that ultimately compromises the quality and reliability of laboratory tests. Here, we present the use of silk fibroin as a solid matrix to encapsulate blood analytes, protecting them from thermally induced damage that could be encountered during nonrefrigerated transportation or freeze–thaw cycles. Blood samples are recovered by simple dissolution of the silk matrix in water. This process is demonstrated to be compatible with a number of immunoassays and provides enhanced sample preservation in comparison with traditional air-drying paper approaches. Additional processing can remediate interactions with conformational structures of the silk protein to further enhance blood stabilization and recovery. This approach can provide expanded utility for remote collection of blood and other biospecimens empowering new modalities of temperature-independent remote diagnostics.
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Kostaman, Tatan, and S. Sopiyana. "Development and Conservation of Gonadal Primordial Germ Cells for Preservation of Local Chicken in Indonesia." Indonesian Bulletin of Animal and Veterinary Sciences 26, no. 3 (February 6, 2017): 125. http://dx.doi.org/10.14334/wartazoa.v26i3.1394.
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One of the <em>ex situ</em> conservation techniques for poultry that recently developed was to collect primordial germ cell (PGC) or gonadal primordial germ cell (gPGC) that isolated from embryo development. Primordial germ cells (PGC) are embryonic cells that migrate to the gonads and form the precursors of gametes. The unique nature and accessibility of PGC during the early development provides an opportunity to manipulate the poultry germplasm, for example by forming germline chimeras. There are some stages that must be done through isolation and collection of PGC from its resources i.e. blastoderm, embryonic circulation blood and gonad. PGC collection originating from the gonads is one of existing PGC resources and technologies. gonadal PGC have advantages compared with other sources, namely (1) A large number of gonadal PGC can be taken from an embryo; and (2) A collection of gonadal PGC can be used in developing management systems of local avian germplasm conservation. This review is intended to describe the usefulness of isolation and collection technology of gonadal PGC as the local poultry germplasm conservation in Indonesia.
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Penjišević, Aleksandra, and Branislav Sančanin. "KEY FACTORS FOR SAFE AND RATIONAL SECURITY OF BLOOD AND BLOOD COMPONENTS." MEDIS – International Journal of Medical Sciences and Research 2, no. 3 (September 18, 2023): 19–24. http://dx.doi.org/10.35120/medisij020319p.
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The use of human blood for therapeutic purposes requires the continuous construction of safety and quality systems, in order to minimize potential risks during the process of collection, processing, circulation and final use. Safety, security, rationality, and reliability are important prerequisites for the continuous provision of blood products, regardless of their purpose. Health institutions for blood and blood products should have an organized quality system that will be compatible with strategic goals, while at the same time emphasizing the self-sufficiency of a particular community. In this context, it is necessary to establish a stable and applicable mechanism for issuing permits and accreditation by certified institutions, from this field, which would ensure their operation in accordance with current regulations. Modern blood transfusion practice must provide guarantees to voluntary blood donors regarding the security and confidentiality of all data related to the health of each individual. The key factors, on which safe and rational provision of blood and blood products are based, are visible through continuous activities associated with the promotion of voluntary and unpaid blood donation, thus contributing, in a specific way, to raising and maintaining high standards. Medical and other personnel involved in the process of collection, examination, processing, storage and circulation of human blood and blood components should be qualified and should undergo continuous education. The future development of transfusiology will be based on teamwork and interdisciplinary cooperation, as well as continuous monitoring and application of regulations from this field. The implementation of the Type and Screen method, i.e., blood group typing and antibody screening, should become a standard in the rationalization of the need for blood, which ensures a greater number of available blood units, increases self-sufficiency and directly contributes to the preservation of the health of donors and recipients of blood and blood components.
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Alrahma, Mohammed, Hanan Almulla, Hanan Almulla, Maryam Almuhairi, Naima Aljanahi, Ayesha Alsabhan, and Hussain Alghanim. "Performance of Different Cotton and Nylon Swabs on DNA Recovery and Storage." Arab Journal of Forensic Sciences and Forensic Medicine 5, no. 2 (September 13, 2023): 135–43. http://dx.doi.org/10.26735/yqmy9190.
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Touch DNA samples are routine yet challenging pieces of evidence that provide investigators with information that helps them solve crimes. However, this type of evidence can be easily lost if the correct collection method is not used. This problem could be overcome with an optimal method of collection that increases the amount of touch DNA collected from different types of surfaces. Better-quality touch DNA can increase the chances of getting a full genetic profile. This study was divided into two parts which aimed to assess whether the type of swab used on different surfaces will significantly increase DNA recovery, concentrations, and the DNA preservation during three different timeframes (24h, 1 month and 3 months). Two different cotton swabs and Nylon swabs were used to lift touch DNA on three different surfaces (glass, plastic and wood) to identify the most suitable method of collection across all three surfaces. A total of 72 samples were lifted (3 replicates from each swab on 3 different surfaces) from two different participants (Male and Female) which were left to dry for 14 days in room temperature prior to DNA extraction. DNA preservation of the swabs was observed while using three dilutions of blood sample which was prepared from one of the volunteers (1:1 – 1:10 – 1:20) where 10 uL of each dilution was pipetted onto the four types of swabs in three replicates (n=36) to observe the preservation over three different timeframes 24h storage, 1 Month and 3 Months with a total of 108 samples. The COPAN CLASSIQSwabsTM Dry swab showed an overall average result during the storage periods of 24h with (1:1) dilution by (2.694ng/μL), (1:10) dilution with (0.548ng/μL) and (1:20) dilution with (0.143ng/μL). Results for the period of 1 Month also showed an average of (1:1) dilution with (2.825ng/μL), (1:10) dilution with (0.361ng/μL) and (1:20) dilution with (0.156ng/μL). These findings can be helpful for laboratories and crime scene investigators to optimize DNA sample collection and preservation based on their workflow.
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Davis, Thomas A., and Fred Gage. "Maximal Cord Blood Recovery and CD34+ Progenitor Cell Collection Using Machine Pulsatile Perfusion of Placentas." Blood 108, no. 11 (November 16, 2006): 3643. http://dx.doi.org/10.1182/blood.v108.11.3643.3643.
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Abstract Umbilical cord blood (UCB) is an attractive source of hematopoietic stem cells (HSC) because of greater availability, less stringent HLA matching requirements, and lower incidence and severity of GVHD. Currently, UCB transplant procedures in adults are limited by low collection volumes of total nucleated cells and CD34+ HSC. Approaches to ex vivo expand long-term engraftable HSC have been widely unsuccessful. Recent studies have clearly demonstrated that infusion of a greater number of cells UCB cells enhances the rate of engraftment and lowers the risk of transplantation-related mortality. Machine pulsatile perfusion (MPP) has been successfully used to select cadaveric renal allografts for transplantation, to isolate human islets from pancreata and shown to be a useful cardiac preservation technique in canine heart transplant studies. In this study, the feasibility of using machine pulsatile perfusion to collect human UCB total nucleated cells and CD34+ HSCs was evaluated using placentas designated for research purposes. Immediately following delivery UCB (65 ± 15 mL, n=5) was first collected by needle aspiration from the umbilical cord vein in accordance with standard procedures then followed by MPP (~500 ml) of the placental arteries within 2–3 hours of delivery. Clinically total nucleated cells count (TNC), CD34+ cell numbers and myeloid, erythroid and multipotent CFU progenitor cell content of UCB units are used as predictive measurements of hematopoietic/engraftment potential. Low-density cells (<1.077 g/mL) were isolated by density centrifugation. The median number of viable low density cells obtained was 488 × 106 (range, 240–652 × 106), and 1541 × 106 (range, 888–1800 × 106) for UCB and MPP collections, respectively. MMP low density cell preparations contained significantly more mature segmented neutrophils with a low percentage (<0.1%) of sheared-off vessel wall endothelial cells. Both UCB and MPP low density cells collections showed similar number of assayable CFU-GEMM, CFU-GM, CFU-M, and CFU-G progenitor cells. In contrast, MMP collected cells contained 2–3 times more erythroid BFU-E colonies than UCB collections. Equivalent numbers of CD34+ HSC were enumerated by FACS analysis and subsequently isolated by positive immunomagnetic MACS selection from MPP and UCB collections. Likewise, the progenitor cell content (CFU-GEMM, CFU-GM, CFU-M, CFU-G and BFU-e) of the isolated CD34+ cell populations derived from each cell collection were very similar. These results demonstrate that pulsatile perfusion can be performed easily after traditional UCB collection procedures. This technique effectively recovers on average twice as many TNC and multilineage CD34+ HSC cells when compared to traditional UCB collection procedures. Altogether these results are particular promising since increased numbers of UCB HSC available for infusion should result in accelerated hematopoietic recovery. Moreover, the demonstrated enhanced HSC cell yield together with the simplicity of collection could potentially widen the clinical applicability of UCB transplants in adults.
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Conrad, Kelvin F., R. J. Robertson, and P. T. Boag. "Difficulties Storing and Preserving Tyrant Flycatcher Blood Samples used for Genetic Analyses." Condor 102, no. 1 (February 1, 2000): 191–93. http://dx.doi.org/10.1093/condor/102.1.191.
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Abstract We stored blood samples of Eastern Phoebes (Sayornis phoebe) in a lysis buffer (“QLB”) that has been used successfully to preserve blood samples of many other species. We found that although samples from adults were not affected greatly, samples of nestling blood stored for more than a few days did not reliably produce the quantity and quality of DNA useful for multi-locus DNA fingerprinting. We also were unable to extract usable DNA from blood samples collected from Eastern Kingbird (Tyrannus tyrannus) nestlings, but obtained usable DNA from blood of Least Flycatcher (Empidonax minimus) nestlings stored for more than a year. We recommend that anyone planning DNA research with tyrant flycatchers should conduct their DNA extractions as soon as possible after collection. A pilot study to test methods of storage, preservation, and extraction may be necessary before beginning a large-scale project.
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Palmirotta, Raffaele, Maria Laura De Marchis, Giorgia Ludovici, Barbara Leone, Annalisa Savonarola, Cristiano Ialongo, Antonella Spila, et al. "Impact of Preanalytical Handling and Timing for Peripheral Blood Mononuclear Cells Isolation and RNA Studies: The Experience of the Interinstitutional Multidisciplinary BioBank (BioBIM)." International Journal of Biological Markers 27, no. 2 (April 2012): 90–98. http://dx.doi.org/10.5301/jbm.2012.9235.
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Multicenter studies and biobanking projects require blood transportation from the participating center to a central collection or diagnostic laboratory. The impact of time delays between venous blood collection and peripheral blood mononuclear cells (PBMC) isolation prior to RNA extraction may affect the quality and quantity of isolated nucleic acids for genomic applications. Thus, standard operating procedure (SOP) optimization for the treatment of biological samples before RNA extraction is crucial in a biological repository. In order to define SOPs for whole blood preservation prior to RNA extraction, we sought to determine whether different blood storage times (0, 3, 6, 10, 24, and 30 hours) prior to PBMCs isolation and storage at –80°C, could affect the quality and quantity of extracted RNA. After spectrophotometric quantification, the quality and integrity of RNA were assessed by agarose gel electrophoresis, RNA integrity number and real time-PCR (RT-PCR). Across the different time points we did not observe significant differences within the first 24 hours of blood storage at room temperature, while a significant loss in RNA yield and integrity was detected between 24 and 30 hours. We conclude that time delays before PBMCs isolation prior to RNA extraction may have a significant impact on downstream molecular biological applications.
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Illarionov, R. A., O. V. Kosyakova, E. S. Vashukova, N. O. Yurkina, M. O. Bakleicheva, Yu S. Dolgova, T. A. Sushko, et al. "Collection of samples from women at different stages of pregnancy to search for early biomarkers of preterm birth." Cardiovascular Therapy and Prevention 19, no. 6 (December 31, 2020): 2708. http://dx.doi.org/10.15829/1728-8800-2020-2708.
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Aim. To create a collection of samples from women at different stages of pregnancy to search for early biomarkers of preterm birth.Material and methods. In order to standardize the sample collection, standard operation procedures have been developed with a step-by-step protocol for each research member at the clinical (collection of medical data and biological material) and laboratory (transportation, sample preparation, storage, quality control) stages.Results. As of October 1, 2020, the collection includes peripheral blood samples from 182 women. Whole blood, serum, plasma, buffy coat and urine were collected during pregnancy, and placenta and umbilical cord blood samples — during labor. Clinical and medical history data was obtained about each pregnant woman, which includes data on the woman’s health status, the course and outcome of pregnancy. An electronic catalog has been created with information on samples (data on clinical characteristics and the number of aliquots of each sample type). The quality control (assessment of DNA and microRNA) was carried out, which showed the compliance of the obtained samples with the quality criteria and the preservation of initial characteristics during long-term storage. On the basis of collection, a study has begun to assess the level of microRNA expression in various types of biomaterial, in order to search for early biomarkers of premature birth.Conclusion. The creation of a collection of samples from pregnant women is a significant groundwork for future fundamental and applied research in various fields of biomedicine. This collection may provide an in-depth study of the pathogenesis of various pregnancy complications and the development of new methods for their diagnosis and treatment.-
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Lal, Renu B., Subhash K. Hira, Rita R. Dhawan, and Peter L. Perine. "Long-Term Preservation of Whole Blood Samples for Flow Cytometry Analysis in Normal and HIV-Infected Individuals from Africa." International Journal of STD & AIDS 1, no. 1 (January 1990): 38–45. http://dx.doi.org/10.1177/095646249000100110.
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A whole blood method requiring less than 4 ml of heparinized blood was developed to assess the practicality of preparing whole blood samples that could be easily stored, transported and readily used to determine the lymphocyte phenotypes and proliferation responses of individuals from remote areas who are infected with the human immunodeficiency virus. Minor modifications in standard whole blood procedure for lymphocyte phenotyping have significantly increased the stability of light scatter and fluorescence intensity of the cells for subsequent flow cytometry (FC) analysis. These changes include removal of lysis solution prior to fixation, fixation of monoclonal antibody-stained cells in 1% paraformaldehyde for 30 minutes and storage of fixed samples in medium containing 1% bovine serum albumin. Lymphocyte subsets and their functional subsets could reliably be determined on samples stored for up to 4 weeks. Further, blood samples could be kept at room temperature for up to 96 hours or at ambient temperature during transportation from Africa before staining for FC without affecting their quantitation. While samples could be processed for FC analysis under field-laboratory conditions, proliferation assays could only be performed on samples that were transported within 48 hours of their collection. The whole blood method saves time and expense and decreases the volumes of blood required to perform phenotypic analysis and functional assays on specimens collected in remote areas.
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De Schauwer, C., S. Piepers, M. K. Hoogewijs, J. L. J. Govaere, T. Rijsselaere, K. Demeyere, E. Meyer, and A. Van Soom. "280 ISOLATION, PRESERVATION, AND CHARACTERIZATION OF EQUINE UMBILICAL CORD BLOOD STEM CELLS." Reproduction, Fertility and Development 20, no. 1 (2008): 220. http://dx.doi.org/10.1071/rdv20n1ab280.
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The isolation, preservation, and identification of hematopoietic and mesenchymal stem cells from fresh umbilical cord blood (UCB) has been extensively reported in humans. Although both types of stem cells may be of therapeutic interest in horses, data on equine UCB cells are scarce. In the present study, two separation methods to isolate stem and progenitor cells from equine UCB and two cryoprotectant solutions for their subsequent freezing were compared. Characterization of the isolated cells was evaluated flow cytometrically, based on the presence of the cytosolic enzyme aldehyde dehydrogenase (ALDH), which has been shown to be highly expressed in primitive hematopoietic cells in a number of species. Cord blood was collected from 15 foals immediately after birth. While the placenta was still in utero, the umbilical cord was clamped and disinfected. A sterile blood bag collection system containing citrate-phosphate-dextrose-adenine anticoagulant was used to collect the UCB by gravity. The UCB units were stored at 4�C and processed within 36 h. Percoll density gradient separation and rouleaux formation induced by hydroxyethyl starch (HES) were tested in parallel on equal volumes of each UCB unit. The enriched progenitor cell fraction was cryopreserved at 10 � 106 nucleated cells mL–1 using two cryoprotectant solutions based on plasma or RPMI 1640, and both containing 10% DMSO and DNase I (20 IU mL–1). Before and after thawing, cells were labeled using a fluorescent ALDH substrate (Aldefluor�, StemCell Technologies SARL, Grenoble, France) including a negative control. Cell viability was simultaneously evaluated by means of exclusion of propidium iodide. Cryopreservation was performed using a programmable freezer (–1�C/min–1 until –70�C, then –10�C/min–1 until –140�C) prior to storage in liquid nitrogen. Results were analyzed statistically with a nonparametric Mann-Whitney test. The concentration of the isolated UCB cells ranged from 0.3 to 4 � 106 cells mL–1 for Percoll and from 0.4 to 7.3 � 106 cells mL–1 for HES. The average viability before and after freezing was 94% and 93% for Percoll-, and 93% and 94% for HES-separated cells, respectively. No significant differences in concentration or in viability were observed between both isolation procedures and both cryoprotectant solutions. Before freezing, the proportion of Aldefluor�-positive cells after Percoll and HES isolation ranged between 0.5 and 38% and between 1 and 60.5%, respectively. No significant differences were found. In conclusion, the percentage of ALDH-positive cells as determined by flow cytometry was highly variable between foals, but was independent of the isolation procedures used. Whether the isolated cells represent true progenitor cells remains to be confirmed. Ongoing, flow cytometrical experiments showed that the isolated cells are CD29+ and CD44+, which may be indicative for their mesenchymal origin.
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Parenrengi, Andi. "NON-CRYOGENIC PRESERVATION ON GROUPER MUSCLE TISSUE FOR DNA ANALYSIS." Indonesian Fisheries Research Journal 8, no. 1 (June 5, 2017): 41. http://dx.doi.org/10.15578/ifrj.8.1.2002.41-44.
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Sample preservation is one of the problems frequently faced in collecting materials such as muscle tissue and blood samples in the field and during transportation to the laboratory prior to DNA extraction.
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Devi, Seeta, Anupkumar M. Bongale, Minyechil Alehegn Tefera, Prashant Dixit, and Prasad Bhanap. "Fresh Umbilical Cord Blood—A Source of Multipotent Stem Cells, Collection, Banking, Cryopreservation, and Ethical Concerns." Life 13, no. 9 (August 23, 2023): 1794. http://dx.doi.org/10.3390/life13091794.
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Umbilical cord blood (UCB) is a rich source of hematopoietic cells that can be used to replace bone marrow components. Many blood disorders and systemic illnesses are increasingly being treated with stem cells as regenerative medical therapy. Presently, collected blood has been stored in either public or private banks for allogenic or autologous transplantation. Using a specific keyword, we used the English language to search for relevant articles in SCOPUS and PubMed databases over time frame. According to our review, Asian countries are increasingly using UCB preservation for future use as regenerative medicine, and existing studies indicate that this trend will continue. This recent literature review explains the methodology of UCB collection, banking, and cryopreservation for future clinical use. Between 2010 and 2022, 10,054 UCB stem cell samples were effectively cryopreserved. Furthermore, we have discussed using Mesenchymal Stem Cells (MSCs) as transplant medicine, and its clinical applications. It is essential for healthcare personnel, particularly those working in labor rooms, to comprehend the protocols for collecting, transporting, and storing UCB. This review aims to provide a glimpse of the details about the UCB collection and banking processes, its benefits, and the use of UCB-derived stem cells in clinical practice, as well as the ethical concerns associated with UCB, all of which are important for healthcare professionals, particularly those working in maternity wards; namely, the obstetrician, neonatologist, and anyone involved in perinatal care. This article also highlights the practical and ethical concerns associated with private UCB banks, and the existence of public banks. UCB may continue to grow to assist healthcare teams worldwide in treating various metabolic, hematological, and immunodeficiency disorders.
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Md Shohag, Mohammad Raguib Munif, Mst Nargis Jahan, Md Mizanur Rahman, and Md Rafiqul Alam. "Haematobiochemical changes of ovine (Ovis aries) blood during storage for transfusion." Research in Agriculture Livestock and Fisheries 7, no. 1 (April 26, 2020): 113–20. http://dx.doi.org/10.3329/ralf.v7i1.46838.
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Haematobiochemical changes of ovine (sheep) blood were investigated during preservation and storage with Citrate Phosphate Dextrose Adenine-1 (CPDA-1) and Acid Citrate Dextrose (ACD) for transfusion. Twelve healthy sheep were selected and divided into two equal groups: group X (n=6) and group Y (n=6). Thirty-five ml of blood was collected from each animal and preserved with CPDA-1 in group X and ACD in group Y under 4°C in refrigerator for 28 days. Haematological changes viz., total erythrocyte count (TEC), total leukocyte count (TLC), haemoglobin (Hb), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC); and biochemical changes viz., total protein (TP) and pH were evaluated immediately after blood collection and thereafter on day-1, day-3, day-7, day-14, day-21 and day-28 for both groups. In ACD preserved blood; TEC, TLC, Hb and PCV decreased significantly (P<0.01) from day-14 onward, whereas in CPDA-1 preserved blood, these parameters decreased significantly (P<0.01) from day-21 onward. Blood preserved in ACD showed significant changes (P<0.01) in MCV, MCH and MCHC respectively from day- 7, day-14 and day-21 onward, whereas blood preserved in CPDA-1 showed no significant changes in the same parameters throughout the experiment. In both groups, no significant changes were noticed in TP but significant changes (P<0.01) were observed in pH with the progression of storage period. These findings elicited that both ACD and CPDA-1 exert certain haematobiochemical changes in stored sheep blood, however, CPDA-1 was more efficient than ACD in terms of maintaining proper levels of TEC, TLC, Hb., PCV, MCV, MCH and MCHC during preservation and storage of sheep blood for transfusion.
Res. Agric., Livest. Fish.7(1): 113-120, April 2020
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Seutin, Gilles, Bradley N. White, and Peter T. Boag. "Preservation of avian blood and tissue samples for DNA analyses." Canadian Journal of Zoology 69, no. 1 (January 1, 1991): 82–90. http://dx.doi.org/10.1139/z91-013.
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A problem frequently faced by researchers involved in collecting tissues for DNA isolation is the preservation of samples in the field prior to and during their transportation to the laboratory. Prevention of DNA degradation is usually achieved through freezing. As this is not always practical, we have tested the efficiency of chemical solutions containing high concentrations of salts (e.g., NaCl, EDTA, and diaminocyclohexanetetraacetate) and detergent at preserving DNA in bird tissue and blood samples stored at ambient temperature for extended periods of time. For blood samples, we recommend the use of a buffer that lyses the cells and nuclei and contains 0.01 M Tris, 0.01 M NaCl, 0.01 M EDTA, and 1% n-lauroylsarcosine, Ph 7.5. Tissue samples are best preserved as small pieces in a saline solution made of 20% dimethyl sulfoxyde, 0.25 M EDTA, saturated with NaCl, pH 8.0. DNA extracted from samples preserved in these solutions for up to 24 weeks was compared with DNA recovered from tissue samples stored at −70 °C and blood samples stored at −70 and −20 °C. Yields were similar, averaging 300 μg/0.2 g of tissue and 500 μg/50 μL of blood. Quality of DNA in terms of fragment size, ability to be cut by restriction enzymes, and ability to hybridize to radioactive probes was also similar between cryopreserved and chemically preserved samples. Yields of DNA recovered from tissue samples preserved in 70% ethanol for 6 or 11 weeks was very low and significant degradation was observed. We have also examined how DNA contained in crude avian blood samples withstands freeze–thaw cycles. We found normal yields and no significant degradation of DNA in samples that experienced up to six cycles. We encourage field researchers who refrain from preserving tissue samples because of logistical problems, such as transporting liquid nitrogen containers in the field, to consider using these solutions. Both solutions can also facilitate exchanges of samples between laboratories, and they form an alternative to storage of samples at −70 °C for laboratories and museums with limited access to deep-freezing facilities.
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Gambarino, Stefano, Ilaria Galliano, Anna Clemente, Cristina Calvi, Paola Montanari, Anna Pau, Maddalena Dini, and Massimiliano Bergallo. "Characteristics of RNA Stabilizer RNApro for Peripheral Blood Collection." Diagnostics 14, no. 10 (May 7, 2024): 971. http://dx.doi.org/10.3390/diagnostics14100971.
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Peripheral blood is the most practical tissue for human immune system gene expression profiling because it is easily accessible, whereas the site of primary infection in certain diseases may not be easily accessible. Due to the ex vivo instability of RNA transcripts, a key challenge in the gene expression analysis of blood samples is the rapid sample handling and stabilization of the mRNA by adding an RNA preservative (PAXgeneTM Blood RNA Tubes, TempusTM Blood RNA tubes, RNAlater Stabilization Reagent, RNAgard® Blood Tubes). BioMole (Turin, Italy) has developed a novel blood stabilizer, called RNApro, in which RNA is stabilized during phlebotomy and sample storage. In this study, RNApro performance intended as RNA yield, integrity, and stability was evaluated. Our results show that blood samples stored at −80 °C and re-extracted after 7 years show no differences in terms of quantity, quality, and amplificability. The samples in the RNAlater stabilization solution can be stored at room temperature for up to one week or at 4 °C for up to one month. Similar results can also be observed for PAXgene tubes, Tempus tubes, and RNAgard tubes. In agreement with these data, the RNApro stabilization solution preserves the RNA from degradation for up to 1 month at 4 °C and 1 week at room temperature. RNApro can be stored indifferently at −80, −20, 4 °C, or room temperature for up to 2 months after, and then could be stored at −80 °C for up to seven years. In summary, our study is the first to analyze the performance of an RNA stabilizer called RNApro. We can conclude that several studies have shown significant differences in gene expression analysis when the sample was preserved in different RNA stabilizers. Therefore, it is desirable to standardize the method of nucleic acid conservation when comparing data from transcriptomic analyses.
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Bao, Hua, Xiaoxi Chen, Xuxiaochen Wu, Wanxiangfu Tang, Min Wu, Shiting Tang, Xue Wu, and Yang Shao. "Abstract 6093: Evaluation of preanalytical and physiological variables affecting cfDNA-based multi-cancer early detection test." Cancer Research 84, no. 6_Supplement (March 22, 2024): 6093. http://dx.doi.org/10.1158/1538-7445.am2024-6093.
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Abstract Background: The multi-cancer early detection (MCED) tests utilizing blood-based circulating cell-free DNA (cfDNA) have gained significant attention in the market as a promising approach for early cancer detection. However, potential effects of preanalytical and physiological variables, including extraction methods, preservation techniques, and physiological factors such as diet, on the accuracy and reliability of MCED results remains poorly investigated. In this study, we aimed to evaluate the impact of physiological variables during blood collection and preanalytical procedures on the outcomes of a MCED test. Methods: A total of 105 plasma samples from 19 healthy donors was collected and analyzed using a MCED test (MERCURY) which leverages the low-coverage whole-genome sequencing and a set of genome-wide features based on cfDNA fragmentomics. Samples were controlled for preanalytical procedures, including transportation condition and preservation time or physiological conditions (before/after meal or exercise) at blood collection. Results: Repeat blood collection (N=4) from the 5 healthy participants consistently yielded negative results for cancer (PPA: 100.0%; 95% CI [56.6%,100.0%]). Among the 5 healthy participants, plasma samples frozen within 1 year (7 days, 3 months, 9 months) showed the same agreement with the reference condition as non-frozen samples (PPA: 100.0%; 95% CI [56.6%,100.0%]), except for one sample frozen for 1 year, which showed a higher risk score and became a positive signal (PPA:80.0%; 95% CI [37.6%,96.4%]). Transportation conditions within 2 hours, 24 hours, 48 hours, and 96 hours at room temperature or 4℃ did not affect the test outcomes, as all samples remained in agreement with the reference condition as of 2 hours at 4℃ (PPA:100.0%; 95% CI [61.0%,100.0%]). Physiological conditions, including pre- and post-meal as well as pre- and post-exercise states, did not exert any influence on the test results, as all samples exhibited agreement with the reference condition (PPA:100.0%; 95% CI [43.9%,100.0%]). Conclusions: The results of this study suggest that variables such as time of blood collection and plasma freezing time may not significantly affect the accuracy of the MCED test in healthy participants. Transportation conditions and physiological conditions evaluated in this study did not have a notable influence on the test outcomes. Nevertheless, it is advised that the freezing duration of plasma samples should not exceed one year. Citation Format: Hua Bao, Xiaoxi Chen, Xuxiaochen Wu, Wanxiangfu Tang, Min Wu, Shiting Tang, Xue Wu, Yang Shao. Evaluation of preanalytical and physiological variables affecting cfDNA-based multi-cancer early detection test [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6093.
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ANDREWS, N. J., and K. BLOOR. "Autologous blood collection in abdominal vascular surgery. Assessment of a low pressure blood salvage system with particular reference to the preservation of cellular elements, triglyceride, complement and bacterial content in the collected blood." Clinical & Laboratory Haematology 5, no. 4 (June 28, 2008): 361–70. http://dx.doi.org/10.1111/j.1365-2257.1983.tb00509.x.
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Kiyaga, Charles, Caroline Makoha, Ivan Nkugwa, Christopher Okiira, Richard Okwir, Sirak Zenebe Gebreab, Patricia Rodriguez-Ventosa Suarez, Benjamin LaBrot, and Ana Carrasco Durán. "The plasma separation card as a novel solution for enhancing central laboratory capability for HIV-1 viral load monitoring in limited-access settings." PLOS Global Public Health 3, no. 6 (June 28, 2023): e0002099. http://dx.doi.org/10.1371/journal.pgph.0002099.
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Measurement of HIV-1 viral load (VL) is essential for monitoring antiretroviral treatment (ART) efficacy. The preferred specimen type for VL is plasma, but in remote settings where collection and preservation of plasma many not be possible, dried blood spots (DBS) are often used instead. A new specimen collection matrix, the cobas plasma separation card (PSC, Roche Diagnostics Solutions), enables specimen preparation from a finger prick or venous blood, using a multi-layer absorption and filtration design that results in a specimen similar to dried plasma. We sought to confirm the correlation between VL results obtained using PSC prepared from venous blood to those from plasma or DBS, as well as PSC prepared with capillary blood from a finger prick. PSC, DBS and plasma were prepared with blood from HIV-1 infected persons attending a primary care clinic in Kampala, Uganda. VL in PSC and plasma was measured using cobas HIV-1 (Roche Diagnostics), while VL in DBS was measured with RealTime HIV-1 (Abbott Diagnostics). The correlation between VL from plasma and PSC made from capillary or venous blood was high (regression coefficient of determination r2 between 0.87 and 0.91), and there was good agreement based on mean bias (-0.14 to 0.24 log10 copies/mL) and classification of VL above or below 1000 copies/mL (91.4% agreement). In contrast, VL from DBS was lower than plasma or PSC (mean bias 0.51 to 0.63 log10 copies/mL) and not as well correlated (r2 0.78 to 0.81, 75.1–80.5% agreement). These results confirm the utility of PSC as an alternative specimen type for HIV-1 viral load measurement in areas where preparation and optimal storage or shipment of plasma is an obstacle to provision of treatment and care of HIV-1 infected people.
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Kit, O. I., N. V. Gnennaya, S. Yu Filippova, T. V. Chembarova, I. B. Lysenko, I. A. Novikova, L. Ya Rozenko, S. N. Dimitriadi, E. V. Shalashnaya, and O. G. Ishonina. "Cryostorage of peripheral blood hematopoietic stem cells in transplantology: current status and prospects." Cardiovascular Therapy and Prevention 22, no. 11 (December 10, 2023): 3691. http://dx.doi.org/10.15829/1728-8800-2023-3691.
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Peripheral blood hematopoietic stem cell (HSC) transplantation is a well-established procedure for the treatment of hematological, cancer and autoimmune diseases. In cancer patients, HSC transplantation allows the use of high-dose cytotoxic drugs in combination with radiation therapy during treatment, which provides a pronounced antitumor effect. The hematological toxicity of such treatment is eliminated by the sequential introduction of stem cells, which contribute to hematopoiesis restoration. Before transplantation, peripheral blood HSCs are subjected to collection and cryopreservation for further storage. An important requirement for cryopreservation is viable HSCs responsible for hematopoietic restoration. The aim of the review was to analyze the literature devoted to the influence of various methods of cryopreservation of human peripheral blood HSCs on the preservation of cell viability after thawing, as well as the development of adverse events in patients. Issues related to the use of various cryoprotectants, as well as methods for storing HSC grafts, are considered. The presented data indicate the need for further study of the effect of cryoprotectants on the human body and the cellular composition of the graft and improvement of protocols for HSC cryopreservation.
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D'Alesandro, Michele M., Dale F. Gruber, H. Lester Reed, Kevin P. O'Halloran, and Robert Robertson. "Effects of collection methods and storage on the in vitro stability of canine plasma catecholamines." American Journal of Veterinary Research 51, no. 2 (February 1, 1990): 257–59. http://dx.doi.org/10.2460/ajvr.1990.51.02.257.
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SUMMARY Norepinephrine (ne) and epinephrine (epi) collected from dogs were sequentially and temporally measured in blood and plasma at 24 C. Heparin and edta anticoagulants, in combination with reduced glutathione and edta as a preservative, were also compared. Norepinephrine and epi concentrations were measured by high-pressure liquid chromatography with electrochemical detection. In heparinized plasma, ne and epi concentrations were relatively stable in the absence or presence of preservative after 24 hours at 24 C. In edta plasma, ne and epi values were less stable when compared with those in heparinized samples. Norepinephrine concentrations in EDTA plasma without preservative decreased by 163.2 ± 8.88 pg over 24 hours, compared with an 86.6 ± 7.92 pg loss of NE in heparinized plasma. The degradation of epi in edta plasma without preservative was also twofold greater, compared with that in heparinized plasma. Addition of preservative had no stabilizing effect on ne or epi in heparinized or edta plasma. During long-term storage at −70 C, plasma ne and epi values decreased < 0.6 and <0.1 pg/d, respectively. Norepinephrine and epi values were stable in heparinized blood for 6 hours but decreased to < 25% and < 6% of initial base line values, respectively, when plasma separation was delayed 24 hours.
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Qin, Jianbing, Jodi R. Alt, Bradford A. Hunsley, Thomas L. Williams, and M. Fernando. "Stabilization of circulating tumor cells in blood using a collection device with a preservative reagent." Cancer Cell International 14, no. 1 (2014): 23. http://dx.doi.org/10.1186/1475-2867-14-23.
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Hobson, Keith A., Mark L. Gloutney, and H. Lisle Gibbs. "Preservation of blood and tissue samples for stable-carbon and stable-nitrogen isotope analysis." Canadian Journal of Zoology 75, no. 10 (October 1, 1997): 1720–23. http://dx.doi.org/10.1139/z97-799.
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Researchers engaged in collecting animal material for stable-carbon and stable-nitrogen isotope analysis are frequently faced with the need to preserve tissues prior to transportation to the laboratory. In many cases, freezing is not possible in the field, so we investigated the potential of several techniques for preserving tissues for this purpose. We also included preservation techniques used for DNA analyses in order to evaluate how they might alter δ13C and δ15N values in tissues and, ultimately, whether archived DNA samples could be used for stable-isotope assay. Tissues included blood and pectoral muscle from quail (Coturnix coturnix japonica) and blood from sheep (Ovis aries). Preservation techniques for blood included freeze-drying (control), drying on precombusted glass-fibre filter paper, and storing in 70% ethanol, 10% buffered formalin, ABI lysis buffer, and Queen's lysis buffer. After 8 weeks, the use of both lysis buffers and formalin resulted in significant depletion of 13C and 15N in blood. Values for samples dried on glass-fibre filter paper or stored in 70% ethanol did not differ significantly from those for the control. Muscle tissue was freeze-dried (control) or stored in 70% ethanol, 10% buffered formalin, or DMSO solution. Both the DMSO and formalin treatments resulted in significant depletion of 13C and 15N compared with the control. Only the 70% ethanol treatment did not result in changes to either isotope ratio in muscle. Where freezing is not possible, we recommend that blood samples be dried or stored in 70% ethanol. Our study provides an estimate of isotopic correction factors that may be applied to tissues archived for DNA analysis or stored in formalin.
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Weikart, Christopher M., Adam P. Breeland, Matt S. Wills, and Martin E. Baltazar-Lopez. "Hybrid Blood Collection Tubes: Combining the Best Attributes of Glass and Plastic for Safety and Shelf life." SLAS TECHNOLOGY: Translating Life Sciences Innovation 25, no. 5 (May 19, 2020): 484–93. http://dx.doi.org/10.1177/2472630320915842.
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SiO2 Medical Products, Inc. developed hybrid blood collection tubes (BCTs) that combine the breakage resistance of plastic and a shelf life approaching that of glass. These blended attributes provide improved BCT safety and reliability for patients and clinical workers. A shelf life of at least 2 y with less than 10% draw volume variation was demonstrated on evacuated hybrid BCTs, which is approximately 7 times longer than standard polyethylene terephthalate (PET) BCTs. This translates into more consistent and reliable blood draw volumes over a longer shelf life. The moisture vapor barrier of hybrid BCTs is 5 times lower than that of PET BCTs, which significantly reduces preservative evaporation over their shelf life. As a result, the risk of preservative gelation and alteration to the blood-to-preservative ratio mix is practically eliminated. Cyclic olefin polymer (COP) exhibits superior impact resistance to breakage because of its high ductility and impact strength and is not influenced by defects and flaws as is glass. Although COP has a mechanical toughness comparable with that of PET, it maintains this over a wider range of temperatures (–70 to 121 °C). As a result, COP can tolerate steam sterilization and cold storage temperatures without mechanical fatigue, deformation, or breakage. Lastly, extreme centrifugation of water-filled BCTs did not impose breakage of any kind.
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Castellini, Michael A., J. Margaret Castellini, and Vicky L. Kirby. "Effects of standard anticoagulants and storage procedures on plasma glucose values in seals." Journal of the American Veterinary Medical Association 201, no. 1 (July 1, 1992): 145–48. http://dx.doi.org/10.2460/javma.1992.201.01.145.
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Summary Standard methods for the preservation of vertebrate blood samples for glucose analysis include collecting and storing the blood in evacuated tubes containing sodium fluoride (glycolytic inhibitor) and potassium oxalate (anticoagulant). We found that blood collected from 5 seals by venipuncture and transferred into these tubes had a significantly (P < 0.05) lower plasma glucose value than blood transferred into tubes containing heparin. In species in which rbc glucose concentration is significantly less than that in the plasma, fluoride and oxalate-induced hemolysis dilutes the plasma with cytoplasm and lowers the measured concentration of glucose in plasma. Therefore, although plasma glucose is used extensively in experimental and clinical analyses, standard techniques for handling the blood may create errors that could confuse comparisons between individuals or species.
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Bock, JL, B. Wenz, and RK Gupta. "Changes in intracellular Mg adenosine triphosphate and ionized Mg2+ during blood storage: detection by 31P nuclear magnetic resonance spectroscopy." Blood 65, no. 6 (June 1, 1985): 1526–30. http://dx.doi.org/10.1182/blood.v65.6.1526.bloodjournal6561526.
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Abstract 31P nuclear magnetic resonance (NMR) spectroscopy was used to measure changes in intra-erythrocyte Mg adenosine triphosphate (MgATP) and free Mg2+ during blood storage at 4 degrees C in standard citrate preservation media. The extent of Mg2+ complexation of ATP and the concentration of free Mg2+ were measured from the Mg2+-dependent chemical shift differences, at 22 degrees C, between the P beta and P alpha resonances of intracellular ATP. This difference changed from 721.0 +/- 1.4 Hz (mean +/- SE) on the day of collection to 741.0 +/- 3.4 Hz after three to seven days and 774.0 +/- 2.8 Hz after 11 to 40 days storage in either acid-citrate-dextrose (ACD) or citrate-phosphate- dextrose-adenine (CPDA-1). Changes in intracellular pH, detected from shifts in the intracellular Pi resonance, averaged 0.27 units after 11 to 40 days of storage. These data indicate a sizable decrease in the extent of Mg2+ complexation of ATP, and a decrease by a factor of 2.6 in free Mg2+, during the shelf-life of blood stored in ACD or CPDA-1.
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Zhang, Nan, Jiayan Lei, Qing Liu, Wei Huang, Hua Xiao, and Han Lei. "The Effectiveness of Preoperative Trimetazidine on Myocardial Preservation in Coronary Artery Bypass Graft Patients: A Systematic Review and Meta-Analysis." Cardiology 131, no. 2 (2015): 86–96. http://dx.doi.org/10.1159/000375289.
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Background: Coronary artery bypass grafting (CABG) is a key and effective surgical treatment modality for coronary artery disease. Unfortunately, ischemia-reperfusion injury during and after CABG can lead to reversible and irreversible myocardial damage. Trimetazidine [1-(2,3,4-trimethoxybenzyl)piperazine dihydrochloride] is a metabolic anti-ischemic agent with demonstrated cardioprotective effects; however, its effects with respect to myocardial preservation in CABG patients remain unclear. Methods: We conducted a systematic review and meta-analysis of randomized controlled trials (RCTs) to investigate the effectiveness of myocardial preservation of preoperative trimetazidine therapy in CABG patients by assessing the postoperative levels of several blood-based biochemical markers of myocardial injury, including creatine kinase (CK), creatine kinase-muscle and brain (CK-MB), creatine phosphokinase (CPK), troponin T (TnT) and troponin I (TnI). The RCTs were classified into two subgroup analyses by the timing of sample collection (either ≤12 or >12 h after CABG). Results: Six RCTs were finally included in the meta-analysis. The pooled effect sizes showed significantly lower postoperative levels of CK, CK-MB, TnT and TnI in the trimetazidine-treated CABG patients relative to control CABG patients. However, there were no significant differences in the postoperative CPK levels between trimetazidine-treated CABG patients relative to control CABG patients. In both the ≤12 and >12 h post-CABG subgroup analyses, significant differences in CK, CK-MB, TnT and TnI were detected between the trimetazidine-treated CABG patients relative to control CABG patients. Conclusions: Preoperative trimetazidine therapy appears to have a positive effect on myocardial preservation in CABG patients.
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Satpathy, Sarthak, Beena Thomas, William Pilcher, Mojtaba Bakhtiari, Lori A. Ponder, Rafal Pacholczyk, Sampath Prahalad, Swati Bhasin, David H. Munn, and Manoj Bhasin. "Simple Preservation of Single Cells (SENSE): One-Step Robust Whole Blood Cryopreservation Method Enables the Generation of High-Quality Single-Cell Immune Profiles." Blood 142, Supplement 1 (November 28, 2023): 2289. http://dx.doi.org/10.1182/blood-2023-190127.
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Introduction: Single-cell profiling (SCP) has revolutionized our understanding of cellular and molecular states in health and disease. Current multistep methods for cryopreserving single-cell suspensions from blood for single-cell RNA sequencing (scRNA-seq) are time-consuming, require trained personnel and special equipment, limiting their clinical adoption and sample collection/availability. It is critical to develop protocols for efficient capture of precious clinical samples/implementation of SCP in clinics. We developed and validated the Simple prEservatioN of Single cElls (SENSE) method for single-step cryopreservation of whole blood (WB), which, along with granulocyte depletion during single-cell assay, generates high-quality SCPs. Methods: Blood samples (n=6) were split into two parts and cryopreserved using the SENSE and peripheral blood mononuclear cells (PBMC) methods. In the SENSE method, WB was cryopreserved by mixing 1:1 blood and freezing solution (80% FBS, 20% DMSO) and granulocytes removed using EasySep CD15 positive selection kit. For PBMC method, density-gradient method was employed for isolating PBMCs followed by cryopreservation. Single cell assay was performed using 10x Genomics kits. The raw sequencing reads were aligned using 10x Genomics Cell Ranger (Zheng et al., Nat. Comm., 2017) to the reference human genome, and the gene-expression matrices were analyzed using the Seurat package (Hao et al., Cell, 2021). Cell markers were identified by comparing target cell types with others using the Wilcoxon Rank Sum test (adjusted P<.10, average log2FC>0.25, percent cell expression>25%). Potential doublets were marked using the doubletFinder (McGinnis et al., Cell Syst., 2019) algorithm and batch-effect was quantified using Shannon entropy calculated by the CellMixS (Lutge et al., Life Sci Alliance. 2021) package for quality assessment of two methods. Cellular communications were assessed using CellChat, which measures cell-to-cell interactions based on the ligands and receptors expression (Jin et al., Nature Comm., 2021). Results: Highly viable (86.3±1.51%) single-cell suspensions (22,353 cells) were obtained from the SENSE method's WB cryopreserved samples. The captured median gene counts, unique molecular identifiers (UMIs)/cell, and mitochondrial content were comparable between the two methods. Similar proportions of membrane, extracellular, and ribosomal ontology-related genes demonstrated the robustness of the SENSE method in capturing high-quality cells without introducing cellular damage artifacts. Lower doublets (2.4%) were observed with the SENSE method compared to the PBMC method (4.8%). These rigorous quality assessments demonstrated the effectiveness of the SENSE method for obtaining high-quality cells for SCP. Both methods yielded similar transcriptomic profiles with split UMAPs of the integrated scRNA-seq data revealing similar clustering patterns except for some variations in the myeloid cells and T-cells clusters. The SENSE method cryopreserved samples exhibited significantly higher T-cell enrichment, enabling deeper characterization of T-cell subtypes. The functional landscape of T/NK cells was assessed using the CellChat tool. Most pathways showed a similar information flow pattern, indicating the capture of similar signaling networks between the cells processed using either of the two methods. However, the SENSE method captured lower number of myeloid cells, attributable to filtering out of sticky CD15+ myeloid/granulocytes that resulted in the loss of single-cells captured. Nevertheless, the SENSE method recapitulated the myeloid compartment associated with the disease as we observed similar patient-wise differences in cell types from both methods. Finally, we compared the transcriptome profiles of both methods to a publicly available PBMC dataset (Zheng et al., Nat. Comm., 2017). Concordance between the three datasets was observed, with all cell types being consistently identified using key marker gene expression. Conclusions: Comparative analysis of scRNA-seq datasets obtained with the two cryopreservation methods, i.e., SENSE and PBMC methods, yielded similar cellular and molecular profiles, confirming the suitability of the former method's incorporation in clinics/labs for cryopreserving and obtaining high-quality single-cells for conducting critical translational research.
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Shalabaev, B. A., and S. Berdiakhmetkyzy. "Exploration of ways to preserve the collection strain of Тrypanosoma equiperdum in an out-of-body." BULLETIN of the L.N. Gumilyov Eurasian National University. BIOSCIENCE Series 138, no. 1 (2021): 29–37. http://dx.doi.org/10.32523/2616-7034-2022-138-1-29-37.
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The article discusses results of the research work carried out at the Kazakh Veterinary Research Institute to consider the methods of storage and maintenance of the stored collection strain of Тrypanosoma equiperdum in artificial nutrient media or at low temperatures (liquid nitrogen -196 0C). During storage, it is important that the strain fully preserves their growth, morphological, biological, toxic, antigenic and pathogenic properties. A diagnostic drug of high sensitivity and specificity is prepared from a strain that has completely preserved its properties.Breeding stallions and mares that have had trypanosomiasis are not amenable to treatment. Of great importance is the diagnosticum used for the examination of animals to prevent the disease. The collection strain of Тrypanosoma equiperdum used in practice is stored only in a living organism, that is, in the body of laboratory animals (white mouse, rat and guinea pig). In this regard, the search for alternative ways of its development and storage is an urgent task. The conducted scientific studies on the preservation of viruses and bacteria in various nutrient media and at low temperatures (-196 oC) are found in literary sources.For the first time, we conducted a study of ways to preserve the Тrypanosoma equiperdum strain outside of a living organism (laboratory animals). Blood parasites of Тrypanosoma equiperdum were first isolated from the genital tract of a sick mare, which were initially transmitted to a rabbit by vaccination in several stages in order to adapt to them a white mouse, rat, guinea pig. Currently, this strain of trypanosomes is stored in the laboratory of parasitology of Kazakh Scientific Research Veterinary Institute. The strain of Тrypanosoma equiperdum is very intolerant to the external environment, it does not form spore capsules. A sample of blood parasites from an infected mouse is inactivated within 1.5-2 hours, that is, it dies, so it is scientifically important to find ways to preserve it outside the body.
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Abendschein, D. R., H. L. Fontanet, and R. Nohara. "Optimized preservation of isoforms of creatine kinase MM isoenzyme in plasma specimens and their rapid quantification by semi-automated chromatofocusing." Clinical Chemistry 36, no. 5 (May 1, 1990): 723–27. http://dx.doi.org/10.1093/clinchem/36.5.723.
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Abstract We report a convenient chromatofocusing procedure for rapid and sensitive quantification of isoforms of the MM isoenzyme of creatine kinase (EC 2.7.3.2) in plasma and efficient methods for preserving isoform profiles during handling of specimens. The assay involves use of prepacked, re-usable Mono P chromatofocusing columns and a "Fast Protein Liquid Chromatograph" (FPLC) system with on-line detection of isoform enzymatic activity in column effluent. Profiles of isoforms are analyzed within 25 min with the use of a 1-mL column; the lower limit of sensitivity for CK activity is 5 mU, and recovery of each isoform is within 1% of the amount added to plasma. Collection of blood specimens in Vacutainer Tubes containing 28.5 mumol of EDTA (final concentration in plasma, 7 to 10 mmol/L) inhibited carboxypeptidase activity in plasma by 76%, sufficient to essentially abolish isoform conversion in vitro at room temperature. These methods should facilitate applications of isoform analysis for diagnosis of myocardial infarction and coronary artery recanalization.
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Chan, A. Y., C. S. Ho, T. Y. Chan, and R. Swaminathan. "D-Mannose as a Preservative of Glucose in Blood Samples." Clinical Chemistry 38, no. 3 (March 1, 1992): 411–13. http://dx.doi.org/10.1093/clinchem/38.3.411.
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Abstract We studied the changes in blood glucose concentration in blood samples collected in heparinized specimen tubes containing no other preservative, or containing NaF, D-mannose, or a combination of NaF and D-mannose. Blood concentration in samples taken into NaF decreased by 0.40 mmol/L in the first 2 h; thereafter, there was no change. In samples collected into mannose there was a small but significant decrease in blood glucose concentration with time. When samples containing mannose were analyzed immediately after collection, the concentration of glucose was higher than in later analyses, probably because of an exchange of intracellular glucose for extracellular mannose. When a combination of NaF and mannose was used, the blood glucose concentration was relatively stable but slightly higher than nonpreserved samples for the next 24 h. However, samples containing mannose were unsuitable for electrolyte analysis. We conclude that a combination of D-mannose and NaF may be a better preservative for blood glucose than either compound alone.
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Gudkov, A. G., V. Yu Leushin, S. V. Agasieva, A. V. Chechetkin, A. D. Kasyanov, E. A. Kiseleva, I. A. Sidorov, V. N. Lemondzhava, G. A. Gudkov, and D. A. Gorbachev. "Devices for Metered Collection of Donor Blood into Polymer Containers and Mixing It with a Preservative." Biomedical Engineering 55, no. 3 (September 2021): 161–63. http://dx.doi.org/10.1007/s10527-021-10093-z.
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Mursalin, Supardi, Siti Nurjanah, Abraham Ethan Martupa Sahat Marune, Muhamad Hasan Sebyar, and Hina Al Kindiya. "Pecoah Kohon: The Restriction on Inter-Cousins Marriage in Indigenous the Rejang Society." JURIS (Jurnal Ilmiah Syariah) 22, no. 1 (June 13, 2023): 69. http://dx.doi.org/10.31958/juris.v22i1.9025.
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This study aims to examine the prohibition of Pecoah Kohon in the indigenous marriage Rejang tribe. Pecoah Kohon is a union between a man and woman related by blood, namely one grandmother. In this study, the problem emphasized a customary prohibition on the union, regardless of its non-prohibition by Islam. This qualitative-field analysis used a normative-sociological approach, with the implemented data collection techniques prioritizing interviews and documentation. A purposive sampling technique was also used to determine the informants. The results showed that the survival of Pecoah Kohon tradition was due to the socialization carried out by traditional officials in a systematic, structured, and hierarchical pattern toward the Rejang community. Sanctions were also considered quite strict against customary violations. Moreover, the good communication and cooperation between traditional officials and the community was a strong foundation for the preservation of Pecoah Kohon tradition and the Islamic religious insights of the Rejang community were increased. From this context, the debate about the tradition had various meeting points and solutions. This indicated that Pecoah Kohon supporters believed the tradition did not include prohibitions and cancellations of marriage, with its performance considered a tribute and cultural preservation. Religious experts also understood that Pecoah Kohonwas solely a custom and not a belief exceeding or equalling religion, indicating needless argumentative efforts.
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Ciesielski, Wojciech Jakub, Alicja Majos, Konrad Kosztowny, Mirosław Stelągowski, Mateusz Tomaszewski, Janusz Strzelczyk, Adam Durczyński, and Piotr Hogendorf. "A case of anastomotic leak due to <i>Candida albicans</i> infection in a 64-year-old female renal transplant patient treated with an emergency suprapubic iliofemoral bypass graft." Lekarz Wojskowy 102, no. 1 (May 13, 2024): 68–72. http://dx.doi.org/10.53301/lw/175143.
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IntroductionVascular complications are a rare but important risk factor for failure and loss of kidney graft, increasing the mortality of renal graft recipients.Case reportWe present the case of massive haemorrhage due to the rupture of the arterial anastomosis caused by <i>Candida albicans</i> infection in a 64-year old female kidney graft recipient managed with suprapubic iliofemoral bypass during emergency life-saving graft nephrectomy performed in our unit. The postoperative period was complicated by femoral vein thrombosis and intraabdominal fluid collection. After 33 days, with a well-functioning iliofemoral bypass, the patient was discharged to the nephrology unit. At the 12-month follow-up, the patient is functioning well, receiving nephrological care and haemodialysis treatment.ConclusionsRoutine fungal cultures of graft preservation fluid and radiological follow-up in high-risk patients after transplantation may be helpful in the prevention of fatal complications in the high-risk patient group – especially if digestive tract injury occurred during organ harvesting. Nevertheless, histological examination remains the gold standard for the detection of fungal arteritis. Negative culture samples from the preservation fluid, blood and urine before the transplantation do not exclude the risk of fungal infection. Urgent allograft nephrectomy with resection and reconstruction of changed vessels seems to be the safest approach in ruptured anastomotic pseudoaneurysm, as shown in our case.
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Bedrov, A. Ya, A. A. Moiseev, A. V. Belozertseva, A. N. Morozov, G. G. Khubulava, Yu A. Pugachenko, and A. V. Baykova. "The patency of internal iliac arteries and its role in the development of buttock claudication syndrome in the remote period after open infrarenal aortic aneurysm repair." Grekov's Bulletin of Surgery 178, no. 4 (September 9, 2019): 34–41. http://dx.doi.org/10.24884/0042-4625-2019-178-4-34-41.
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The OBJECTIVE was to study the patency of the internal iliac artery and its effect to gluteus muscles blood supply and frequency of buttock claudication occurrence in the remote period after open infrarenal aortic aneurysm repair. MATERIAL AND METHODS. Examination of 37 patients after open infrarenal aortic aneurysm repair included collection of complaints, anamnesis, making CT scan with contrast and pelvic perfusion tomography. These methods allowed to assess the patency of the prosthesis and iliac arteries, calculate average blood flow rate in buttock muscles and frequency of buttock claudication occurrence depending on the lesion of the internal iliac arteries. RESULTS. Five-year patency of the internal iliac artery was 93 %. In case of passable internal iliac artery, the average blood flow rate in the ipsilateral buttock muscles was authentically higher than the same indicator in groups with stenotic or occlusive lesion of the internal iliac artery and its branches. In case of the disturbed internal iliac artery patency, the frequency of occurrence of the buttock claudication in the same side reached 50 %. CONCLUSION. High five-year internal iliac artery patency after open infrarenal aortic aneurysm repair attested the necessity of preservation the main blood flow in these arteries during the open infrarenal aortic aneurysm repair for the purpose of buttock claudication prevention. The CT scan allowed to evaluate the internal iliac artery patency and the average blood flow rate in the buttock muscles through perfusion tomography method which was necessary for differential diagnosis of the buttock claudication syndrome.
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Faraj, Kharman, Samera Abdullah, and Sameen Muhammad. "EFFECTS OF DIFFERENT DOSES OF GAMMA RAYS AND ASCORBIC ACID CONCENTRATION ON HUMAN RBCS FOR CONSERVATION PURPOSE." Iraqi Journal of Medical Sciences 17, no. 1 (March 31, 2019): 50–56. http://dx.doi.org/10.22578/ijms.17.1.8.
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Background: Blood preservation and the development of sterile collection sets made possible the developments in blood components preparation, storage and transfusion that we have in today's blood banks and transfusion services. Objective: To investigate the effects of gamma irradiation, ascorbic acid and the combined effect of both on the lifespan of erythrocytes through determining red blood cells hemolysis for conserving it as long as possible without any change in erythrocytes biophysical properties. Methods: The blood was drawn from 10 healthy (5 males and 5 females) volunteers. Sample has been irradiated using 137Cs source. Different concentrations of ascorbic acid were used as an anti-oxidative agent for erythrocytes in blood suspension samples. A spectrometer was used for recording the data. Results: The results showed that 25% of RBCs hemolysis occurred after irradiation with 5Gy of gamma ray during 5th week of storage time while in un-irradiated sample 33.8% of RBCs hemolysis occurred during the 5th week. 25% of RBCs hemolysis for 10 μM of ascorbic acid concentration started after 7th week while for control started after 4th week. The minimum rates of RBCs hemolysis observed in the samples which pre-treated with (7 and 10) μM concentrations of ascorbic acid then irradiated with 1 Gy. Conclusion: The results indicated that irradiation of human blood with a certain doses of gamma ray, treated with small concentration of ascorbic acid or both, the two factors together can protect the blood from hemolysis for a longer time and the minimum rate of red blood cells hemolysis was observed for 10 μM ascorbic acid concentration then irradiation to 1 Gy of gamma ray. Keywords: Gamma ray, ascorbic acid, blood storage, red blood cells, oxidative damage Citation: Faraj KA, Abdullah SH, Muhammad SF. Effects of different doses of gamma rays and ascorbic acid concentration on human RBCs for conservation purpose. Iraqi JMS. 2019; 17(1): 50-56. doi: 10.22578/IJMS.17.1.8
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Zhulkov, M. O., A. R. Tarkova, I. S. Zykov, A. G. Makaev, A. V. Protopopov, M. N. Murtazaliev, F. Yu Kosimov, et al. "Long-term normothermic autoperfusion of the cardiopulmonary complex ex vivo as a method of effective graft conditioning: an experimental study." Patologiya krovoobrashcheniya i kardiokhirurgiya 27, no. 4 (December 26, 2023): 33–42. http://dx.doi.org/10.21688/1681-3472-2023-4-33-42.
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Objective: To compare the effectiveness of 6-hour normothermic autoperfusion of a heart graft ex vivo with pharmaco-cold preservation using Bretschneider's solution (Custodiol, Dr Franz Köhler Chemie GmbH, Bensheim, Germany).Methods: Landrace pigs weighing 50 ± 5 kg and aged 4–5 months (n = 10) were selected as a model for a series of acute experiments. Cardiopulmonary conditioning using autoperfusion was conducted for 6 hours on the experimental group (n = 5). On the other hand, the control group underwent a 6-hour pharmaco-cold preservation with Bretschneider solution to recover the heart's pumping function. Graft preservation effectiveness was evaluated by measuring hemodynamic parameters, heartbeat, and myocardial ischemia marker concentrations.Results: After reperfusion and isolation of the working cardiopulmonary complex, cardiac output was 0.63 [0.37; 0.8] L/min and 0.37 [0.23; 0.37] L/min in the experimental and control groups, respectively (P < .05). The levels of CPK-MB, LDH, troponin-I, and lactate in the coronary sinus blood was significantly higher in the control group.Conclusion: The study demonstrated significant benefits of normothermic autoperfusion in maintaining the morphofunctional status of the donor heart compared to pharmaco-cold preservation using Bretschneider's solution for 6 hours of ex vivo graft conditioning. Received 16 July 2023. Revised 8 September 2023. Accepted 11 September 2023. Funding: The study was carried out within the framework of project No. 23-25-10013 (agreement No. 23-25-10013 dated April 20, 2023 with the Russian Science Foundation, agreement No. р-52 dated April 3, 2023 with the Ministry of Science and Innovation Policy of the Novosibirsk Region). Conflict of interest: The authors declare no conflict of interest. Contribution of the authorsConception and study design: M.O. Zhulkov, D.A. Sirota, I.S. ZykovData collection and analysis: M.O. Zhulkov, A.R. Tarkova, I.S. Zykov, A.G. Makaev, A.V. Protopopov, M.N. Murtazaliev, F.Yu. Kosimov, N.A. Karmadonova, Ya.M. Smirnov, E.E. Kliver, A.M. Volkov, H.A. Agaeva, D.A. SirotaStatistical analysis: M.O. ZhulkovDrafting the article: M.O. ZhulkovCritical revision of the article: M.O. Zhulkov, D.A. Sirota, I.S. ZykovFinal approval of the version to be published: M.O. Zhulkov, A.R. Tarkova, I.S. Zykov, A.G. Makaev, A.V. Protopopov, M.N. Murtazaliev, F.Yu. Kosimov, N.A. Karmadonova, Ya.M. Smirnov, E.E. Kliver, A.M. Volkov, H.A. Agaeva, D.A. Sirota
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Farrugia, A., S. Douglas, J. James, G. Whyte, and R. Herrington. "Use of Plasma with High Levels of lonised Calcium in the Production of Model Scale Goagulation Factor Concentrates." Thrombosis and Haemostasis 64, no. 03 (1990): 374–78. http://dx.doi.org/10.1055/s-0038-1647322.
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SummaryWe have attempted to exploit the Ca2+ -dependent stability of factor VIII in producing factor VIII concentrates of higher yield. Plasma levels of ionised calcium were increased in two ways: (a) whole blood collection into half-strength citrate CPD anticoagulant, leading to free Ca2+ levels of ca 120 µM and (b) apheresis collection of plasma which was then recalcified to free Ca2+ levels of ca 300 µM under heparin cover. Coagulation factor concentrates were prepared using model versions of our industrial scale manufacturing methods. Factor VIII yield was increased through low citrate collection. This did not compromise factor IX yield or thrombogenic potential. Use of recalcified heparinised plasma did not lead to any improvement in factor VIII yield and resulted in a marked drop in factor IX recovery, possibly from interference by heparin of factor IX binding in ion-exchange chromatography. The benefits accruable through the use of half-strength citrate CPD anticoagulant support the continued evaluation of this preservative in large scale blood collection and fractionation. The deleterious effects of heparin in charge-mediated plasma fractionations may pose serious difficulties in harvesting vitamin K dependent factors.
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Auma, Erick, Tom Hall, Simran Chopra, Sam Bilton, Laxmee Ramkhelawon, Fahimah Amini, Anna Calvert, et al. "Using Dried Blood Spots for a Sero-Surveillance Study of Maternally Derived Antibody against Group B Streptococcus." Vaccines 11, no. 2 (February 4, 2023): 357. http://dx.doi.org/10.3390/vaccines11020357.
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Vaccination during pregnancy could protect women and their infants from invasive Group B Streptococcus (GBS) disease. To understand if neonatal dried blood spots (DBS) can be used to determine the amount of maternally derived antibody that protects infants against invasive GBS disease, a retrospective case-control study was conducted in England between 1 April 2014 and 30 April 2015. The DBS of cases with invasive GBS disease (n = 61) were matched with healthy controls (n = 125). The haematocrit, DBS storage temperature, freeze-thaw cycle, and paired serum/DBS studies were set up to optimise the antibody assessment. The samples were analysed using a multiplex immunoassay, and the results were assessed using parametric and nonparametric tests. Antibody concentrations were stable at haematocrits of up to 50% but declined at 75%. DBS storage at room temperature was stable for three months compared with storage from collection at −20 °C and rapidly degraded thereafter. Total IgG levels measured in DBS and paired serum showed a good correlation (r2 = 0.99). However, due to suboptimal storage conditions, no difference was found in the GBS IgG levels between DBS samples from cases and controls. We have demonstrated a proof of concept that assays utilising DBS for assessing GBS serotype-specific antibodies in infants is viable. This method could be used to facilitate future large sero-correlate studies, but DBS samples must be stored at −20 °C for long term preservation of antibody.
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Lebert, Brittany, Maxine Gonzalez-Vega, Samer Elbabaa, Julia Hegert, Avery Wright, Ana Aguilar-Bonilla, and Amy Smith. "OTHR-10. COLLABORATIVE EFFORTS FOR HOSPITAL-BASED BIOBANK DEVELOPMENT AND OPTIMIZATION TO ENSURE QUALITY PEDIATRIC SPECIMENS FOR TRANSLATIONAL RESEARCH." Neuro-Oncology 25, Supplement_1 (June 1, 2023): i76. http://dx.doi.org/10.1093/neuonc/noad073.292.
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Abstract In recent years, drastic improvements in sequencing technologies have led to a better understanding of the specific genetic drivers of cancer. However, one of the greatest barriers to these discoveries is access to primary patient tumor tissue, especially in cancers of the Central Nervous System (CNS). In many cases tissue from surgery is processed and stored in Formalin-Fixed Paraffin-Embedded (FFPE) blocks, which sets limitations on downstream applications. In 2015, our team launched an initiative to collect and efficiently store samples from pediatric CNS tumor patients during surgeries and autopsies. Through extensive collaboration between Neuro-Oncology, Neurosurgery, Pathology, and our Translational Laboratory, we have optimized the collection process to effectively preserve sample integrity and therefore maximize their potential usage. To date, we have collected over 4,509 samples from 293 patients (636 flash-frozen tissue, 1,026 plasma & buffy coat, 748 serum, 996 whole blood, 316 Cerebrospinal Fluid (CSF), and 787 parental blood samples). In addition, we have cultured 31 primary cell lines and collected over 242 dissociated tumor specimens for implantation into patient-derived xenograft (PDX) mouse models. Through our partnership with neurosurgery and pathology, we are permitted to attend surgeries and autopsies to immediately flash-freeze tumor tissue in liquid nitrogen, typically within minutes of the tissue being removed from the body. This immediate flash-freezing process aims to ensure specimen integrity and preserve molecular profiles. From there, the tissue is either stored in our -140oC freezer for future use or sent to partnered consortiums, including the Children’s Brain Tumor Network (CBTN) and Gift from a Child. Since the optimization of the collection process, the focus has now shifted to investigating the efficacy of our storage methods by quantifying DNA/RNA integrity over time. By ensuring the successful preservation of the samples, we can maximize their impact in the pediatric neuro-oncology research field.