Storage Of Whole Blood Research Articles

Hemostatic Evaluation of Refrigerated Whole Blood Stored 7 Days Post-Expiration.

The United States Army has shifted doctrine to focus on large-scale combat operations against peer to near-peer adversaries. Future conflicts could result in a limited supply chain, leaving medical providers with only expired blood products for treatment of hemorrhagic shock. This study evaluated quality, function, and safety metrics of whole blood stored for 1 week past regulated expiration (i.e., 35 days, in CPDA-1). Whole blood units (n = 6) were collected in citrate phosphate dextrose adenine-1 (CPDA-1) anticoagulant and stored refrigerated for up to 42 days. Units were sampled on days 35, 37, 39, and 42 of storage and evaluated for the following: complete blood count, blood metabolism and chemistries, clotting dynamics, and presence of bacteria. The majority of evaluated parameters fell outside of normal clinical ranges beginning at day 35 of storage. At 42 days, blood pH was 6.58 ± 0.038, hemolysis was significantly increased (P = .037 vs day 35), and bacterial contamination was not evident. Glucose levels continuously dropped during extended storage. K+ was significantly increased at day 42 compared to day 35 (P = .010). A significant reduction in clot strength, factor V activity, and factor VIII activity was evident beginning at day 39 of storage. Storage of whole blood out to 42 days results in a continuous decline in function, but further in vivo safety studies should be performed to determine if the benefits of expired blood outweigh the risks. Other methods to safely extend storage of whole blood that maintain hemostatic function and preserve safety should be investigated, with emphasis placed on methods that reduce potassium leak and/or hemolysis.

Emergency transfusion of different rhesus blood type: Will there be complications?

Objective: Antepartum hemorrhage (APH) is one of the leading causes of maternal and fetal mortality worldwide, complicating 2–4% of pregnancies. Considering the probability of blood transfusion, blood testing and cross-matching are essential for the anesthesiologist during perioperative management. Rhesus (Rh) is the second most significant blood group system after ABO. Hence, this blood type system needs to be taken into consideration. Furthermore, in Indonesia, Rh-negative blood types are rare. Case Presentation: A 39-year-old woman with Gravida IV, three term pregnancies, no preterm pregnancies, no abortions, and three living children (GIVP3003) and gestational age 26 to 27 weeks arrived at the hospital with massive bleeding from the birth canal three hours before admission. The bleeding was fresh red, and the patient had changed diapers twice before arriving at the hospital. She presented hypovolemic shock, a blood pressure of 71/39 mm Hg, and anemic conjunctivas, with a history of B Rh (-) blood type, ongoing HIV therapy, and completed pulmonary tuberculosis (TB) treatment. The hospital’s and PMI’s (Indonesian Red Cross) blood bank had no Rh (-) bloodstock. An emergency cesarean section under general anesthesia was performed, and a 900 g neonate was delivered. The patient received a total of four stored whole blood (SWB) bags of type B Rh (+) blood products, one SWB bag during surgery, and three SWB bags during recovery in the intensive care unit (ICU). Conclusion: Rh (+) transfusion in patients with Rh (-) should be the last option in an emergency. The transfusion reaction did not occur right away in the initial transfusion. Under prompt management, the transfusion response and the life-threatening condition were then successfully managed.

Abstract 995: Preservation of patient whole blood preserves cancer immune cell biology at single cell resolution

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.

Evaluating the effects of hypoxic storage on platelet function and health using a novel storage system.

Thousands of units of whole blood (WB) and blood components are transfused daily to treat trauma patients. Improved methods for blood storage are critical to support trauma-related care. The Hemanext ONE® system offers a unique method for hypoxic storage of WB, with successfully demonstrated storage of clinically viable RBCs. This work evaluated the system for the storage of WB, focusing on platelet health and function. WB was collected from healthy donors and processed through the Hemanext ONE® system. Hemoglobin oxygen saturation (HbSO2) levels of WB were depleted to 10%, 20%, or 30% of total HbSO2 and then stored in PVC bags sealed in oxygen-impermeable bags (except for normoxic control) with samples collected on days 1, 7, and 14 post-processing. Flow cytometry assessed the activation and apoptosis of platelets. Clot dynamics were assessed based on aggregometry and thromboelastography assays, as well as thrombin generation using a calibrated-automated thrombogram method. Hypoxic storage conditions were maintained throughout the storage period. Hypoxia triggered increased lactate production, but pH changes were negligible compared to normoxic control. Storage at 10% HbSO2 had a significant impact on platelet function, resulting in increased activation and reduced clot formation and aggregation. These effects were less significant at 20% and 30% HbSO2. This study indicates that platelets are sensitive to hypoxic storage and suffer significant metabolic and functional deterioration when stored at or below 10% HbSO2.

Blood storage impacts on the hematological indices of healthy subjects and patients with iron-deficiency anemia and beta-thalassemia – A comparative study

Background: There are scientific evidence confirmed specific changes in blood cell counts, reducing the efficacy and feasibly the safeness of blood transmission when storing blood at 4°C for 5 weeks or more.Objectives: The study aimed to investigate the effects of stored blood obtained from healthy subjects and patients with anemia due to iron deficiency and beta-thalassemia, on hematological indices.Materials and Methods: A total of 37 participants, consisting of 14 healthy subjects, 13 patients with iron-deficiency anemia, and 10 patients with beta-thalassemia minor, were recruited from Hiwa Hospital between November 2021 and July 2022. Blood samples were obtained from the participants and stored at 4°C for 5 weeks. Hematological indices, including red cell distribution, platelet distribution width, and mean platelet volume, were determined using a hematology analyzer at weekly intervals.Results: Blood storage caused significantly increased mean values of hematological indices among healthy subjects as well as among patients with iron-deficiency anemia and beta-thalassemia, although the pattern of changes was differed.Conclusions: The storage of whole blood significantly increased hematological indices, showing variations in both healthy subjects and patients with iron-deficiency anemia and beta-thalassemia. The pattern of raise in these hematological indices is specific to iron-deficiency anemia and thalassemia when compared with healthy subjects.

The stability of 65 biochemistry analytes in plasma, serum, and whole blood.

The pre-analytical stability of various biochemical analytes requires careful consideration, as it can lead to the release of erroneous laboratory results. There is currently significant variability in the literature regarding the pre-analytical stability of various analytes. The aim of this study was to determine the pre-analytical stability of 65 analytes in whole blood, serum and plasma using a standardized approach. Blood samples were collected from 30 healthy volunteers (10 volunteers per analyte) into five vacutainers; either SST, Li-heparin, K2-EDTA, or Na-fluoride/K-oxalate. Several conditions were tested, including delayed centrifugation with storage of whole blood at room temperature (RT) for 8 h, delayed centrifugation with storage of whole blood at RT or 4 °C for 24 h, and immediate centrifugation with storage of plasma or serum at RT for 24 h. Percent deviation (% PD) from baseline was calculated for each analyte and compared to the maximum permissible instability (MPI) derived from intra- and inter-individual biological variation. The majority of the analytes evaluated remained stable across all vacutainer types, temperatures, and timepoints tested. Glucose, potassium, and aspartate aminotransferase, among others, were significantly impacted by delayed centrifugation, having been found to be unstable in whole blood specimens stored at room temperature for 8 h. The data presented provides insight into the pre-analytical variables that impact the stability of routine biochemical analytes. This study may help to reduce the frequency of erroneous laboratory results released due to exceeded stability and reduce unnecessary repeat phlebotomy for analytes that remain stable despite delayed processing.

The effect of whole blood logistics on neutrophil non-specific activation and kinetics ex vivo

While the exquisite sensitivity of neutrophils enables their rapid response to infection in vivo; this same sensitivity complicates the ex vivo study of neutrophils. Handling of neutrophils ex vivo is fraught with unwanted heterogeneity and alterations that can diminish the reproducibility of assays and limit what biological conclusions can be drawn. There is a need to better understand the influence of ex vivo procedures on neutrophil behavior to guide improved protocols for ex vivo neutrophil assessment to improve inter/intra-experimental variability. Here, we investigate how whole blood logistics (i.e., the procedure taken from whole blood collection to delivery of the samples to analytical labs and storage before neutrophil interrogation) affects neutrophil non-specific activation (i.e., baseline apoptosis and NETosis) and kinetics (i.e., activation over time). All the experiments (60+ whole blood neutrophil isolations across 36 blood donors) are performed by a single operator with optimized isolation and culture conditions, and automated image analysis, which together increase rigor and consistency. Our results reveal: (i) Short-term storage (< 8 h) of whole blood does not significantly affect neutrophil kinetics in subsequent two-dimensional (2D) cell culture; (ii) Neutrophils from long-term storage (> 24 h) in whole blood show significantly higher stability (i.e., less non-specific activation) compared to the control group with the isolated cells in 2D culture. (iii) Neutrophils have greater non-specific activation and accelerated kinetic profiles when stored in whole blood beyond 48 h.

Direct Analysis of Whole Blood by a Disposable Monolithic Column Mass Spectrometry Analysis Kit†

Comprehensive SummaryA monolithic column‐based mass spectrometry (MS) analysis kit was prepared for whole blood analysis with MS. The kit is disposable and can be used for purification, storage, transportation and direct analysis of whole blood. The kit mainly consists of a capillary for quantitative microsampling, a cation exchange monolithic column for purification and storage, and a syringe for loading sample. This kit is very friendly to various users that one can easily siphon the blood in the kit followed by rapid clean‐up. We established a quantitative method using the kit with a limit detection as low as 0.33 nmol/L, and achieved more than five orders of magnitude enhancement in sensitivity compared to direct nanoelectrospray ionization MS analysis. The column can avoid analyte exposure to environment, which helps the storage of the sample for laboratory analysis. The relative standard deviation of immediate blood analysis and storage blood analysis within 10 d was less than 10%. This method has been successfully applied to the quantitative analysis of procainamide hydrochloride in 2 μL rat blood. These results indicate that this disposable kit does have the potential to achieve highly sensitive quantitative MS analysis in biological samples, which is expected to become a cost‐effective and powerful tool for in vitro diagnostics.

Growth and Distribution of Bacteria in Contaminated Whole Blood and Derived Blood Components

Introduction: Bacterial contamination of blood products presumably occurs mainly during blood collection, starting from low initial concentrations of 10–100 colony-forming units (CFUs) per bag. As little is known about bacterial growth behavior and distribution in stored whole blood (WB) and WB-derived blood products, this study aims to provide data on this subject. Methods: WB units were inoculated with transfusion-relevant bacterial species (Acinetobacter baumannii, Bacillus cereus, Escherichia coli, Klebsiella pneumoniae, Listeria monocytogenes, Pseudomonas fluorescens, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus dysgalactiae, Streptococcus pyogenes, Yersinia enterocolitica; n = 12 for each species), stored for 22–24 h at room temperature, and then centrifuged for separation into plasma, red blood cells (RBCs), and buffy coats (BCs). The latter were pooled with 3 random donor BCs and one unit of PAS-E each to yield plasma-reduced platelet concentrates (PCs). Samples for bacterial colony counting were collected after WB storage and immediately after blood component production. Sterility testing in PCs (n = 12 for each species) was performed by bacterial culture after 7 days of storage. Results: Bacterial growth in WB varied remarkably between donations and species. Streptococcus species produced the highest titers in WB, whereas Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas fluorescens did not multiply. Centrifugation resulted in preferential accumulation of bacteria in BCs, with titers of up to 3.5 × 103 CFU/mL in BCs and up to ≤0.9 × 103 CFU/mL in BC-derived PCs. Overall, 72/144 PCs (50%) tested positive for bacteria after storage. Sterility test results were species-dependent, ranging from 12 of 12 PCs tested positive for Streptococcus pyogenes to 1 of 12 PCs positive for Escherichia coli. Bacterial contamination of RBC and plasma units was much less common and was associated with higher initial bacterial counts in the parent WB units. Conclusions: Bacterial growth in WB is species-dependent and varies greatly between donations. Preferential accumulation of bacteria in BCs during manufacturing is a critical determinant of the contamination risk of BC-derived pooled PCs.

Development of blood collection tubes for glucose measurement using adenosine 3-phosphate and sodium fluoride as glycolytic inhibitors.

Blood collection tubes with sodium fluoride (NaF) added as a glycolytic inhibitor are widely used for glucose measurement. However, the glycolytic inhibitory effects of NaF are insufficient, and decreases in glucose levels over time after blood collection have become a problem. Blood from a volunteer collected using an NaF tube was used to compare the glycolysis inhibitory abilities of ATP and ADP. Blood samples from 10 volunteers were collected in NaF tubes and NaF tubes with added ATP (NaF-ATP tubes). The stability of glucose and haemoglobin (Hb)A1c after whole-blood storage from immediately after blood collection to 24 h later was compared. ATP and ADP had similar inhibitory effects on glycolysis, but ATP was selected as an additive for blood collection tubes because ADP was more haemolytic than ATP. We verified the ability of NaF blood collection tubes supplemented with ATP to inhibit glycolysis. Mean (± standard deviation) glucose levels (n=10) after storage for 24 h after blood collection decreased to -9.0 ± 2.7 mg/dL (-0.50 ± 0.15 mmol/L) in conventional NaF tubes. NaF-ATP(20) tubes with 20 mg (0.036 mmol) ATP added showed a reduced decrease, with a mean of -5.8 ± 2.9 mg/dL (-0.32 ± 0.16 mmol/L). NaF-ATP tubes also had no effect on HbA1c measurement. This study reports on a blood collection tube that enables the measurement of glucose and HbA1c. Based on the results of validation, we conclude that NaF-ATP tubes can reduce decreases in glucose over time in stored whole blood compared to conventional NaF tubes.

Pyrophosphate as a novel anticoagulant for storage of whole blood: A proof-of-concept study.

Citrate is the only anticoagulant currently Food and Drug Administration (FDA)-approved for the long-term storage of blood for transfusion. Citrate inhibits phosphofructokinase and may play a pro-inflammatory role, suggesting that there may be an advantage to using alternative anticoagulants. Here, we examine the use of pyrophosphate as an anticoagulant. Whole blood samples from healthy donors were anticoagulated either with citrate-phosphate-adenine-dextrose (CPDA-1) or our novel anticoagulant mixture pyrophosphate-phosphate-adenine-dextrose (PPDA-1). Samples were assessed for coagulation capacity by thromboelastography immediately after anticoagulation (T0) with and without recalcification, as well as 5 hours after anticoagulation (T1) with recalcification. Complete blood counts were taken at both timepoints. Flow cytometry to evaluate platelet activation as well as blood smears to evaluate cellular morphology were performed at T1. No clotting was detected in samples anticoagulated with either solution without recalcification. After recalcification, clotting function was restored in both groups. R-Time in recalcified PPDA-1 samples was shorter than in CPDA-1 samples. A reduction in platelet count at T1 compared to T0 was observed in both groups. No significant platelet activation was observed in either group at T1. Blood smear indicated platelet clumping in PPDA-1. We have shown initial proof of concept that pyrophosphate functions as an anticoagulant at the dose used in this study, though there is an associated loss of platelets over time that may limit its usefulness for blood storage. Further dose optimization of pyrophosphate may limit or reduce the loss of platelets.

Cold storage of whole blood in an additive solution containing apoptotic and necrotic inhibitors.

Whole blood (WB) reigns superior to component therapy for the treatment of hemorrhagic shock on the battlefield. Though cold storage of WB offers a shelf life of 21 to 35 days, storage lesions and the potential for blood wastage remain. Storing WB in an additive solution (AS) containing apoptotic inhibitors may help preserve blood cell viability and improve blood quality over extended cold storage. Non-leukoreduced WB was obtained from healthy individuals and dosed with: AS, AS+Necrostatin-1 (AS+N1), AS+Boc-D-fmk (AS+B; apoptosis inhibitor), AS+Q-VD-OPh (AS+Q; apoptosis inhibitor), and Control (0.9% saline). Blood bags were kept refrigerated (1°-6°C) for 21 days. Bags were tested on days 0, 7, 14, and 21 for complete blood count, metabolism, clot formation, aggregation function, platelet activation, and red blood cell quality. Platelet count was better preserved in all AS-containing samples. All groups displayed increased glucose consumption and lactate production with storage. Furthermore, all groups displayed a similar decline in clot strength (max amplitude) over the 21-day storage period. Bags that received AS displayed greater preservation of GPIIb expression and lower phosphatidylserine exposure. P-selectin expression was increased in all AS groups. Treatment of hemorrhagic shock with WB transfusion is logistically simpler than component therapy. Results from our study suggest that refrigerated WB stored with an AS containing apoptotic and necrotic inhibitors helps better preserve platelet count but does not improve platelet function. The future development of WB ASs is warranted to optimize both platelet quality and hemostatic function.

Establishment of preanalytical conditions for microRNA profile analysis of clinical plasma samples.

The relationship between the expression of microRNAs (miRNAs) in blood and a variety of diseases has been investigated. MiRNA-based liquid biopsy has attracted much attention, and cancer-specific miRNAs have been reported. However, the results of analyses of the expression of these miRNAs vary among studies. The reproduction of results regarding miRNA expression levels could be difficult if there are differences in the data acquisition process. Previous studies have shown that the anticoagulant type used during plasma preparation and sample storage conditions could contribute to differences in measured miRNA levels. Thus, the impact of these preanalytical conditions on comprehensive miRNA expression profiles was examined. First, the miRNA expression profiles of samples obtained from healthy volunteers were analyzed using next-generation sequencing. Based on an analysis of the library concentration, human genome identification rate, ratio of unique sequences and expression profiles, the optimal preanalytical conditions for obtaining highly reproducible miRNA expression profiles were established. The optimal preanalytical conditions were as follows: ethylenediaminetetraacetic acid (EDTA) as the anticoagulant, whole-blood storage at room temperature within 6 hours, and plasma storage at 4°C or -20°C within 30 days. Next, plasma samples were collected from 60 cancer patients (3 facilities × 20 patients/facility), and miRNA expression profiles were analyzed. There were no significant differences in measurements except in the expression of erythrocyte-derived hsa-miR-451a. However, the variation in hsa-miR-451a levels was smaller among facilities than among individuals. This finding suggests that samples obtained from the same facility could show significantly different degrees of hemolysis across individuals. We found that the standardization of anticoagulant use and storage conditions contributed to reducing the variation in sample quality across facilities. The findings from this study could be useful in developing protocols for collecting samples from multiple facilities for cancer screening tests.

Laboratory Medicine and Research in Pathology

The laboratory is where medicine and science meet. This supplementary issue on ‘Laboratory Medicine and Research in Pathology’ celebrates that partnership. The laboratory study of pathology informs not only diagnosis, but also prognosis and disease pathogenesis. Crucially, a diagnostic test is only as good as its sample and the rigour of its protocol. In this issue, readers can review -20°C and -80°C sample storage of whole blood for HbA1c measurements, analyse the haematological investigations in a rare case of lymphoplasmacytic lymphoma with paraprotein IgG, examine CD14 expression in urothelial bladder carcinoma tissue to correlate inflammation with prognosis, and more. Continuous and looping dialogue between medicine and science, physician and laboratory specialist betters the care of the patient. We hope this series of articles demonstrates that. Sincere thanks to our reviewers and the MJMHS Editorial Board and production team for bringing this special issue to fruition and we hope you enjoy the read.

High-dose radiation exposure of cold-stored whole blood does not affect hemostatic function.

Whole blood (WB) transfusion is routinely used to resuscitate severely injured military trauma patients. Blood can be stored refrigerated while still maintaining reasonable function but is susceptible to environmental influences, including radiation exposure. Immune-compromised patients are transfused with irradiated blood to inactivate donor lymphocyte function (25 Gy per Association for the Advancement of Blood and Biotherapies [AARB] standard 5.7.3.2). However, there is limited information on function of WB exposed to high radiation doses. This study aimed to determine if stored irradiated WB still retains function. This will be important if the stored blood supply is exposed to radiation in a combat situation or mass casualty incident when the need for blood will be high. Whole blood collected from healthy donors was irradiated at 0, 25, or 75 Gy and stored at 4°C. Blood cell count, blood gas chemistry, thromboelastometry, platelet aggregation, and reactive oxygen species were measured before irradiation and at 1, 7, and 14 days of storage. Irradiated WB was compared with nonirradiated WB controls. Irradiated WB stored for up to 14 days was not significantly different than nonirradiated WB in most of the parameters measured. Stored blood showed expected changes associated with functional decline at longer storage times, but irradiation did not hasten the decline. There was a significant change in potassium and sodium ion concentrations after irradiation, but the functional relevance is not clear. High-dose irradiation had little effect on stored WB. Although there were changes in plasma sodium and potassium levels, there was little to no effect on hemostasis and blood cell viability. This suggests that stored blood subjected to a radiation event generating at least a dose of 75 Gy is still suitable for transfusion, which could be particularly important in the event of a mass casualty event where a large amount of blood is needed.

US Central Command military blood utilization practices 2011 to 2020.

Military involvement in Afghanistan ended in 2021, and while low-intensity troop engagements continue globally, casualty numbers are dwindling. To understand the clinical and operational connections between blood utilization and clinical paradigm shifts in resuscitation strategies, a review of blood product utilization and the changes in the last decade was conducted within the US Central Command area of responsibility. The intent of this review was to assess patterns of blood use during the last decade of the United States' involvement in the most recent major conflicts to potentially inform future blood requirements. Blood product and types transfused between January 1, 2011, and December 31, 2020, were acquired from the Medical Situational Awareness in Theater blood reports. All reported blood usage data in the US Central Command area of responsibility were queried. Packed red blood cells and fresh frozen plasma (FFP) usage showed no statistically significant change over time ( τb = 0.24, p = 0.3252; τb = -0.47, p = 0.0603). Fresh and stored whole blood (SWB) use increased overtime ( τb = 0.69, p = 0.0056; τb = 0.83, p = 0.0015). A strong inverse relationship was found between SWB and FFP usage ( r = -0.68, p = 0.0309) and liquid plasma and FFP usage ( r = -0.65, p = 0.0407) over time. Whole blood usage increased significantly over time with a preference for SWB. Component therapy is anticipated to remain a critical element of resuscitation in the event of large-scale combat operations secondary to supply chain and longer storage times. Therapeutic/care management; Level III.

Stored whole blood transfusion initiates serum amyloid A activation monitored by real-time dynamic imaging.

Transfusion of stored whole blood (SWB) to resuscitate severe traumatic haemorrhage patients in military operations and civilian emergency centres is being increasingly used in routine practice. It has been well established that transfusion of red blood cells (RBCs) after prolonged storage has harmful effects, mainly mediated by inflammation. Whether the side effects of inflammation are brought about by SWB transfusion remains unclear. A hepatocyte SAA (serum amyloid A) specific reporter mouse that facilitated non-invasive imaging of hepatocyte SAA expression was used to evaluate acute inflammation and acute-phase reaction after the transfusion of SWB or components separated from end-storage whole blood. The whole blood of C57BL/6 donor mouse was used to model an allogeneic transfusion to BALB/c recipient mouse. End-storage whole blood (14 days of storage) transfusion induced the most significant SAA expression, while 10-day storage resulted in a much weaker signal compared to their fresh and 5-day storage counterparts. RBCs rather than white blood cells and plasma-containing platelets are thought to be responsible for the systemic inflammatory and SAA activation during end-storage whole blood transfusion. Circulatory and hepatic pro-inflammatory cytokines secreted by M1-polarised macrophage initiated the SAA expression in hepatocytes through nuclear transcription factor NF-κB. Storage lesions will also occur during the storage of whole blood, which is related to the change in RBCs with prolonged storage. The side effect induced by systemic inflammation and acute-phase reaction should be considered before resuscitation with long-term storage whole blood transfusion.

Whole Blood Storage Temperature Investigation in Austere Environments.

Military medical research has affirmed that early administration of blood products and timely treatment save lives. The US Navy's Expeditionary Resuscitative Surgical System (ERSS) is a Role 2 Light Maneuver team that functions close to the point of injury, administering blood products and providing damage-control resuscitation and surgery. However, information is lacking on the logistical constraints regarding provisions for and the stability of blood products in austere environments. ERSS conducted a study on the United States Central Command (USCENTCOM) area of responsibility. Expired but properly stored units of stored whole blood (SWB) were subjected to five different storage conditions, including combinations of passive and active refrigeration. The SWB was monitored continuously, including for external ambient temperatures. The time for the SWB to rise above the threshold temperature was recorded. The main outcome of the study was the time for the SWB to rise above the recommended storage temperature. Average ambient temperature during the experiment involving conditions 1 through 4 was 25.6°C (78.08°F). Average ambient temperature during the experiment involving condition 5 was 34.8°C (94.64°F). Blood temperature reached the 6°C (42.8°F) threshold within 90 minutes in conditions 1 and 2, which included control and chemically activated ice packs in the thermal insulated chamber (TIC). Condition 2 included prechilling the TIC in a standard refrigerator to 4°C (39.2°F), which kept the units of SWB below the threshold temperature for 490 minutes (approximately 8 hours). Condition 4 entailed prechilling the TIC in a standard freezer to 0.4°C (32.72°F), thus keeping the units of SWB below threshold for 2,160 minutes (i.e., 36 hours). Condition 5 consisted of prechilling the TIC to 3.9°C (39.02°F) in the combat blood refrigerator, which kept the SWB units below the threshold for 780 minutes (i.e., 13 hours), despite a higher average ambient temperature of almost +10°C (50°F). Combining active and passive refrigeration methods will increase the time before SWB rises above the threshold temperature. We demonstrate an adaptable approach of preserving blood product temperature despite refrigeration power failure in austere settings, thereby maintaining mission readiness to increase the survival of potential casualties.

Operation Blood Rain Phase 2: Evaluating the Effect of Airdrop on Fresh and Stored Whole Blood.

Transfusion of whole blood (WB) is a lifesaving treatment that prolongs life until definitive surgical intervention can be performed; however, collecting WB is a time-consuming and resource-intensive process. Furthermore, it may be difficult to collect sufficient WB at the point of injury to treat critically wounded patients or multiple hemorrhaging casualties. This study is a follow-up to the proof-of-concept study on the effect of airdrop on WB. In addition, this study confirms the statistical significance for the plausibility of using airdrop to deliver WB to combat medics treating casualties in the pre-hospital setting when Food and Drug Administration (FDA)-approved cold-stored blood products are not available. Forty-eight units of WB were collected and loaded into a blood cooler that was dropped from a fixed-wing aircraft under a Standard Airdrop Training Bundle (SATB) parachute or 68-in pilot chute. Twenty-four of these units were dropped from a C-145 aircraft, and 24 were dropped from a C-130 aircraft. A control group of 15 units of WB was storedin a blood cooler that was not dropped. Baseline and post-intervention laboratory tests were measured in both airdroppedand control units, including complete blood count; prothrombin time/partial thromboplastin time (PT/PTT); pH, lactate,potassium, bilirubin, glucose, fibrinogen, and lactate dehydrogenase (LDH) levels; and peripheral blood smears. The blood cooler, cooling packs, and all 48 WB units did notsustain any major damage from the airdrop. There was noevidence of hemolysis. Except for the one slightly damagedbag that was not sampled, all airdropped blood met parameters for transfusion per the Joint Trauma System Whole BloodTransfusion Clinical Practice Guideline and the Associationfor the Advancement of Blood and Biotherapies (AABB) Circular of Information for the Use of Human Blood and BloodComponents. Airdrop of fresh or stored WB in ablood cooler with a chute is a viable way of delivering bloodproducts to combat medics treating hemorrhaging patientsin the pre-hospital setting. This study also demonstrated theportability of this technique for multiple aircraft. The techniques evaluated in this study have the potential for utilizationin other austere settings such as wilderness medicine or humanitarian disasters where an acute need for WB delivery by airdrop is the only option.

A comparison of platelet function in cold-stored whole blood and platelet concentrates.

There is renewed interest in the use of whole blood (WB) for the resuscitation of trauma patients. Platelet function in stored WB compared to platelet concentrates is not well established and was assessed in vitro in this study. Leucocyte-depleted cold-stored WB (CS-WB) was prepared using a Terumo WB-SP Imuflex kit and held at 2-6°C alongside: (A) UK standard pooled platelets stored at 20-24°C (RT-PLTS), (B) pooled platelets stored at 2-6°C (CS-PLTS), and (C) platelet-rich plasma produced using the Terumo kit (CS-PRP), for 21 days. A series of in vitro assays were assessed platelet function. Platelet count was retained to 57 ± 14% of starting number at day 21 in CS-WB. Over time, CS-WB platelets become more activated, with increased CD62P expression (day 1: 7± 3.7% vs. day 21: 59 ± 17.1%) and annexin V binding (day 1: 2± 0.2% vs. day 21: 21 ± 15.1%). For comparison, 18.6± 6% of platelets in RT-PLTS demonstrated CD62P expression at day 7, whereas annexin V binding in RT-PLTS at day 7 was 2.6± 0.5%. Over storage, aggregatory response to agonists decreased in all arms. Functional platelet microparticles increased steadily in CS-WB throughout storage. During storage, platelet count reduced in CS-WB, whereas CD62P expression and annexin V binding increased. This was accompanied by a reduced aggregatory response, although compared to 7-day-old RT-PLTS, CS-WB maintained a maximal response to agonists for longer, suggesting that the shelf life for CS-WB can be considered for up to 21 days.