Fresh Whole Blood Research Articles

Evaluation of Fluorescent Spot Test with Dried Blood Spots for Glucose-6-Phosphate Dehydrogenase Deficiency Screening in A Malaria-Endemic Area

The utilization of dried blood spot (DBS) samples for screening glucose-6-phosphate dehydrogenase (G6PD) deficiency remains limited in resource-constrained and malaria-endemic regions of Thailand. This study evaluated the feasibility of employing DBS for G6PD deficiency screening among 242 participants (118 males and 124 females) who provided fresh whole blood (FWB) and DBS samples for analysis. The G6PD deficiency diagnostic performances of the fluorescent spot test using DBS (FST-DBS) were compared to those using FWB (FST-FWB). The G6PD gene variant was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using DBS samples. The FST-DBS showed no significant difference in sensitivity (95.0% vs 100.0%) but superior specificity (100.0% vs 80.2%) and a positive predictive value (100.0% vs. 50.0%) compared to FST-FWB. The PCR-RFLP revealed a G6PD mutation incidence rate of 16.5% (11.1% in males and 5.4% in females). Among 40 DBS samples, 39 (97.5%) were the Mahidol (487G>A) while a sample (2.5%) which both FST-DBS and FST-FWB showed positive results had an unidentified variant. Therefore, FST-DBS is an alternative format for G6PD deficiency screening in malaria-endemic regions, particularly where resources are limited.

Abstract 79: Enabling fast and convenient immune profiling of fresh and long-term stabilized human whole blood samples with CyTOF

Abstract Accurate phenotyping of immune cells in whole blood (WB) from patients with cancer is critical for making disease prognoses and monitoring clinical efficacy of immunotherapies. Fresh WB must be analyzed within 24 hours of collection to minimize changes in cellular composition. However, WB collection and cytometric analysis are often performed at different sites, which can result in sample processing delays. Several WB preservation reagents have been developed to address this challenge, including PROT1 (Smart Tube Inc.) and Cytodelics Whole Blood Cell Stabilizer (Cytodelics). However, not all antibody panels are compatible with these reagents. A CyTOF® panel was developed to be compatible with these commercial WB stabilizers and for use in pharmaceutical and clinical research. CyTOF flow cytometry uses metal-tagged antibodies to identify cellular and functional phenotypes. Advantages and features of CyTOF technology enable rapid design and application of 50-plus-marker panels and convenient workflows in which samples can be stained and acquired in a single tube. Compensation is not required since CyTOF flow cytometry has low signal spillover and no autofluorescence. Moreover, antibody cocktails and stained samples can be frozen for later use and acquisition. Thus, CyTOF technology overcomes major hurdles of fluorescence-based cytometry and provides a streamlined and flexible workflow in clinical research. The CyTOF panel contains 20 antibodies to identify over 30 immune cell populations. For easy customization, there are more than 30 additional open channels to analyze markers of interest. The panel works with fresh and stabilized WB samples and is amenable to different staining and acquisition workflows. To show the flexibility of sample staining and stabilization, WB from three healthy donors was assessed using two stabilization workflows. First, fresh WB samples were stained with the antibody panel, followed by PROT1 or Cytodelics stabilization and storage at -80 °C. The second workflow involved immediate stabilization/fixation of WB with PROT1 or Cytodelics and storage at -80 °C. Subsequently, the samples for the second workflow were thawed, surface stained, and acquired. To reduce technical variability from staining, the antibodies were pooled together and frozen at -80 °C as single-use aliquots. Furthermore, all samples were barcoded and acquired as a single tube to reduce variability from sample acquisition. The CyTOF panel developed for broad immune profiling is compatible with WB stabilizers, which overcomes traditional and logistical challenges with WB processing and acquisition. Furthermore, freezing antibody cocktails is a unique feature of CyTOF flow cytometry, ensuring batch-to-batch consistency in clinical research. Thus, CyTOF workflows enable swift and convenient analysis of WB samples. For Research Use Only. Not for use in diagnostic procedures. Citation Format: Michael Cohen, Shakir Hasan, Stephen Li, Christina Loh. Enabling fast and convenient immune profiling of fresh and long-term stabilized human whole blood samples with CyTOF [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 79.

OMIP-101: 27-color flow cytometry panel for immunophenotyping of major leukocyte populations in fixed whole blood.

This 27-color flow cytometry antibody panel allows broad immune-profiling of major leukocyte subsets in human whole blood (WB). It includes lineage markers to identify myeloid and lymphoid cell populations including granulocytes, monocytes, myeloid dendritic cells (mDCs), natural killer (NK) cells, NKT-like cells, B cells, conventional CD4 and CD8 T cells, γδ T cells, mucosa-associated invariant T (MAIT) cells and innate lymphoid cells (ILC). To further characterize each of these populations, markers defining stages of cell differentiation (CCR7, CD27, CD45RA, CD127, CD57), cytotoxic potential (perforin, granzyme B) and cell activation/proliferation (HLA-DR, CD38, Ki-67) were included. This panel was developed for quantifying absolute counts and phenotyping major leukocyte populations in cryopreserved, fixed WB collected from participants enrolled in large multi-site tuberculosis (TB) vaccine clinical trials. This antibody panel can be applied to profile major leukocyte subsets in other sample types such as fresh WB or peripheral blood mononuclear cells (PBMCs) with only minor additional optimization.

What's new in whole blood resuscitation? In the trauma bay and beyond.

Transfusion therapy commonly supports patient care during life-threatening injury and critical illness. Herein we examine the recent resurgence of whole blood (WB) resuscitation for patients in hemorrhagic shock following trauma and other causes of severe bleeding. A growing body of literature supports the use of various forms of WB for hemostatic resuscitation in military and civilian trauma practice. Different types of WB include warm fresh whole blood (FWB) principally used in the military and low titer O cold stored whole blood (LTOWB) used in a variety of military and civilian settings. Incorporating WB initial resuscitation alongside subsequent component therapy reduces aggregate blood product utilization and improves early mortality without adversely impacting intensive care unit length of stay or infection rate. Applications outside the trauma bay include prehospital WB and use in patients with nontraumatic hemorrhagic shock. Whole blood may be transfused as FWB or LTOWB to support a hemostatic approach to hemorrhagic shock management. Although the bulk of WB resuscitation literature has appropriately focused on hemorrhagic shock following injury, extension to other etiologies of severe hemorrhage will benefit from focused inquiry to address cost, efficacy, approach, and patient-centered outcomes.

Stability Assessment of Fresh or Cryopreserved Whole Blood and Bone Marrow Samples from AML and B-ALL Patients to Monitor CD123 Receptor and Immune Cells Subsets

Introduction: Compounds targeting CD123 are currently under clinical development for the treatment of acute myeloid leukemia (AML) and high-risk myelodysplastic syndrome (MDS). As part of clinical trials, CD123 expression and immune cell subsets need be monitored by quantitative flow cytometry (qFC) in bone marrow (BM) aspirates and whole blood (WB) samples before and on-treatment. One critical parameter for analysis is the poor stability of the fresh biological BM or WB samples. No general rule can predict the stability of the expression of a given receptor on diseased or healthy cells thus the pre-analytical stability of each biomarker should be evaluated to design the sample logistics. The pre analytical stability of clinical samples is the main constraint for quantitative flow cytometry (qFC) in multicenter and worldwide clinical studies. The transportation of the samples is one of the limiting factor. The classical freezing method of peripherical whole blood (WB) or bone marrow (BM) isolated by Ficoll requires high skilled operators, is time consuming and may be difficult to set up at sampling site in hospitals. One option to make the clinical samples suitable for subsequent high dimensional analysis by qFC is to use a simple freezing method at -80°C. Our cryopreservation method could be a new option for easier preparation of clinical samples prior to centralized samples analysis. This study aims to evaluate the pre-analytical fresh or cryopreserved stability of each pharmacodynamic monitoring endpoint over time: CD123 expression and receptor density, T-cell subsets and activation profile. Methods: FreshBM and WB were collected in EDTA tubes from 5 AML/MDS patients at hospitals, in accordance with local regulations. The samples were stored at room temperature (RT) for 4, 24, 48 or 72 h before immunostaining with the standard operation procedures and analysis by qFC or mixed with the cryopreserved media (BioCytex) within 24 h post-collection before freezing at -80°C for months. After storage, the cryopreserved samples were thawed and immunostained before analysis by qFC. Using a qFC assay (Multiplex FC panel assay developed by BioCytex, Marseille, France), CD123 receptor density/cell was collected. T-cell and blast cell immunophenotypes were also assessed on fresh samples for time point 4, 24, 48, or 72 h post-collection but also for cryopreserved WB sample within the 24hs post collection. The assay profiled blasts, monocytes, plasmacytoid dendritic cells, basophils and T-cells. Results: CD123 density in blast and normal cells was stable at RT from 4 h up to 48 h after collection. The profiles of blast subsets were very similar in BM and WB samples from a particular patient, whether a single blast cell subset (CD45lowCD11b-CD34+CD38med) or several blast cell subsets (expressing CD34 and CD38 at different levels) were identified. Both T-cell distribution and activation markers were stable up to 48 h for BM and WB. A similar pattern of T-cell subsets was also observed in the BM and WB paired samples from the same donor. In addition, CD123 receptor density, immunophenotyping and percentage of blast subsets and T cells from fresh WB samples was comparable to paired cryopreserved samples with a new method and for at least 1 month. Conclusions: Based on these observations, we conclude that CD123 density, blast cells and T-cell subsets in WB and BM samples from AML patients were stable for 48 h in our experimental conditions. As a consequence, fresh WB and BM samples can be processed and analyzed by qFC within 48 h of sample collection. Furthermore, the new cryopreservation method showed sample stabilization for blasts and T cells immunophenotyping. This data opens new perspectives in clinical studies for AML This new strategy would also help to standardize preparation of clinical samples and to centralize sample analysis.

Qualification of the differential leukocyte count and immunophenotyping in cryopreserved ex vivo whole blood assay.

We developed a flow cytometry-based assay, termed Differential Leukocyte Counting and Immunophenotyping in Cryopreserved Ex vivo whole blood (DLC-ICE), that allows quantification of absolute counts and frequencies of leukocyte subsets and measures expression of activation, phenotypic and functional markers. We evaluated the performance of the DLC-ICE assay by determining inter-operator variability for processing fresh whole blood (WB) from healthy donors collected at multiple clinical sites. In addition, we assessed inter-operator variability for staining of fixed cells and robustness across different anticoagulants. Accuracy was evaluated by comparing DLC-ICE measurements to real-time cell enumeration using an accredited hematology analyzer. Finally, we developed and tested the performance of a 27-colour immunophenotyping panel on cryopreserved fixed WB and compared results to matched fresh WB. Overall, we observed <20% variability in absolute counts and frequencies of granulocytes, monocytes and lymphocytes (T, B and NK cells) when fresh WB was collected in different anti-coagulant tubes, processed or stained by independent operators. Absolute cell counts measured across operators and anti-coagulants using the DLC-ICE method exhibited excellent correlation with the reference method, complete blood count (CBC) with differential, measured using a hematology analyzer (r2 > 0.9 for majority of measurements). A comparison of leukocyte immunophenotyping on fresh WB versus DLC-ICE processed blood yielded equivalent and linear results over a wide dynamic range (r2 = 0.94 over 10-104 cells/μL). These results demonstrate low variability across trained operators, high robustness, linearity and accuracy, supporting utility of the DLC-ICE assay for large cohort studies involving multiple clinical research sites.

Effects of Freezing and Rewarming Methods on RNA Quality of Blood Samples.

Background: RNA extracted from human blood has been widely applied to biological, medical, and clinical research of numerous diseases. Previous studies have demonstrated that high-quality RNA is indispensable to guarantee the reliability of downstream assays. In this study, we investigated the effects of freezing procedures, rewarming methods, and blood components on RNA quality of blood samples. Methods: Rabbit blood samples were divided into two groups: (1) whole blood (WB) and (2) blood cell components (BCC) with plasma removed. Samples were frozen using four representative freezing procedures (snap freezing in liquid nitrogen, snap freezing at -80°C, traditional slow freezing, and programmable controlled rate freezing) and rewarmed by placing at 4°C or by vortexing. RNA was extracted using the phenol-chloroform RNA extraction method and measured by an Agilent bioanalyzer. Then, human blood was used to verify the best protocol obtained from the rabbit blood experiment. Results: For the four freezing procedures, there were no differences in RNA integrity. For different rewarming methods, RNA integrity number (RIN) values of RNA extracted from frozen WB and BCC samples in the vortex group were above 9, while RNA obtained from WB showed worse quality compared with BCC in the 4°C group. For verification using human blood, RIN values of frozen human WB rewarmed by vortexing ranged from 8.0 to 9.1. Conclusions: Blood components and rewarming methods could affect the RNA quality of blood samples. For scenarios where WB samples have already been cryopreserved, the vortex rewarming method is optimal for high-quality RNA. Otherwise, we would recommend centrifuging fresh WB and cryopreserving it in the form of BCC, which showed a tendency to obtain high-quality RNA by either of the two rewarming methods.

The U.S. Armed Services Blood Program support to U.S. Central Command 2014-2021: Transformation of combat trauma resuscitation through blood product innovation and expansion of blood availability far forward.

The United States Armed Services Blood Program (ASBP) faced complex blood supply challenges during two decades of military operations in the U.S. Central Command (CENTCOM) and through an adaptive, responsive, and agile system, gained valuable insights on blood product usage in combat casualty care. A retrospective review of blood product introduction and utilization trends was compiled from ASBP data collected during CENTCOM operations from 2014 through 2021. During the study period, several blood products were introduced to the CENTCOM area of operations including Low Titer O Whole Blood (LTOWB), Cold-Stored Platelets (CSP), Liquid Plasma (LP), and French Freeze Dried Plasma (FDP) manufactured from U.S. sourced donor plasma, all while expanding Walking Blood Bank capabilities. There was a gradual substitution of component therapy for whole blood; blood utilization peaked in 2017. Transfusion of Fresh Whole Blood (FWB) from Walking Blood Banks decreased as fully pre-tested LTOWB was supplied by the ASBP. LTOWB was initially supplied in citrate-phosphate-dextrose (CPD) anticoagulant (21-day shelf life) but was largely replaced with LTOWB in citrate-phosphate-dextrose-adenine (CPDA-1) anticoagulant (35-day shelf life) by 2019. Implementation of prehospital transfusion and expansion of surgical and resuscitation teams led to an increase in the number of sites receiving blood. ASBP introduced new products to its inventory in order to meet changing blood product demands driven by changes in the Joint Trauma System Clinical Practice Guidelines and operational demands. These products were adopted into clinical practice with a resultant evolution in transfusion strategies.

Point-of-care ketone meters may be used to estimate serum β-hydroxybutyrate concentrations in healthy African penguins (Spheniscus demersus)

To evaluate the agreement between 3 point-of-care (POC) devices and a reference laboratory for measuring β-hydroxybutyrate (β-HB) concentration in African penguin (Spheniscus demersus) whole blood (WB) and plasma samples and the precision of each POC device for measuring β-HB concentration in plasma samples. 48 healthy African penguins. Blood was obtained from the right jugular vein of each penguin, and β-HB concentration was measured on each POC device using fresh WB and heparinized plasma and at the reference laboratory using plasma. β-HB concentration was measured in plasma on each POC device. All devices overestimated serum β-HB concentrations on average by 0.46 mM relative to the reference laboratory. WB samples had less error than plasma for meters A and C. Meter A had the lowest total error observed (26.4%) and the lowest mean difference (0.19 mmol/L) relative to the reference laboratory. Controlling for other factors, the magnitude of disagreement was not affected by sex, age, packed cell volume, or serum total solids concentration. WB, not plasma, should be used for measurement of β-HB concentration on the POC meters tested. Meter A showed good correlation with the reference laboratory for WB. The use of POC devices for the measurement of β-HB concentration may be acceptable when laboratory analyzers are not available. Further research is needed for clinical application and the diagnostic value of POC meters compared with reference laboratories.

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.

Ranger O Low Titer (ROLO): Whole Blood Transfusion for Forward Deployed Units.

First-time use of Ranger O Low Titer (ROLO) blood and implementation of a forward-walking blood bank using predetermined donors proved essential in the survival of a 33-year-old active duty soldier following a complex blast injury during combat operations. The patient sustained significant bone, soft tissue, and vascular damage and continued to deteriorate despite resuscitation with cold-stored whole blood (WB). Only after utilizing the ROLO battle drill and transfusing with fresh WB was the patient able to be stabilized and evacuated. In this case report, we discuss how ROLO walking blood bank takes the next step in aiding resuscitation, providing smaller, forward-deployed units with blood resupply without the administrative burden of storage, particularly in resource-scarce environments. We provide an overview of WB and contrast its use to that of component therapy. In conjunction with the Golden Hour, ROLO can be incorporated as the standard damage control resuscitation to reduce the risks of noncompressible hemorrhage. By taking precautionary steps in the pre-deployment setting, ROLO offers an invaluable alternative to conventional resuscitation.

Platelet aggregation in whole blood is not impaired by a platelet‐sparing leukoreduction filter and instead depends upon the presence of leukocytes

Recent studies characterizing in vitro hemostatic properties of whole blood (WB) leukoreduced (LR) with a platelet-sparing filter have described subtle, if any, changes to viscoelastic clotting; however, reductions in platelet (PLT) content and impedance aggregometry (IA) responses have been noted. The effects of filtration of WB (i.e., filter-contact effects, reduction in platelet and leukocyte count) have not been rigorously investigated as to their individual impacts on platelet IA responses. WB units from healthy donors were collected and characterized to assess the effects of platelet-sparing leukoreduction (LR) upon the in vitro hemostatic measures of platelet IA and thromboelastometry. Further characterization of platelet IA responses was carried out in WB samples to delineate the effects of platelet count and leukocyte presence/absence upon the response. WB filtration reduced the platelet count and IA responses but had no impact on viscoelastic clotting measures in fresh WB. Experiments revealed that IA responses have a linear correlation with platelet count in both apheresis platelets and WB and that passage of platelets through the WB-LR filter has no impact upon the strength of this response. Further experiments in LR WB showed that addition of autologous leukocytes back to the platelets fully restored the platelet aggregation response to pre-filtration levels. WB filtration results in platelet count reduction and leukocyte removal; however, platelet IA is not degraded by passage through the filter. Apparent declines in platelet IA responses can be fully attributed to the reduction in platelet count and the removal of leukocytes.

Effect of parachute delivery on red blood cell (RBC) and plasma quality measures of blood for transfusion.

BackgroundParachute airdrop offers a rapid transfusion supply option for humanitarian aid and military support. However, its impact on longer‐term RBC survival is undocumented. This study aimed to determine post‐drop quality of RBCs in concentrates (RCC), and both RBCs and plasma in whole blood (WB) during subsequent storage.Study design and methodsTwenty‐two units of leucodepleted RCC in saline, adenine, glucose, mannitol (SAGM) and 22 units of nonclinical issue WB were randomly allocated for air transportation, parachute drop, and subsequent storage (parachute), or simply storage under identical conventional conditions (4 ± 2°C) (control). All blood products were 6–8 days post‐donation. Parachute units were packed into Credo Cubes, (Series 4, 16 L) inside a PeliCase (Peli 0350) and rigged as parachute delivery packs. Packs underwent a 4‐h tactical flight (C130 aircraft), then parachuted from 250 to 400 ft before ground recovery. The units were sampled aseptically before and after airdrop at weekly intervals. A range of assays quantified the RBC storage lesion and coagulation parameters.ResultsBlood units were maintained at 2–6°C and recovered intact after recorded ground impacts of 341–1038 m s−2. All units showed a classical RBC storage lesion and increased RBC microparticles during 42 days of storage. Fibrinogen and clotting factors decreased in WB during storage. Nevertheless, no significant difference was observed between Control and Parachute groups. Air transportation and parachute delivery onto land did not adversely affect, or shorten, the shelf life of fresh RBCs or WB.DiscussionAppropriately packaged aerial delivery by parachute can be successfully used for blood supply.

Efficacy of past, present, and future fluid strategies in an improved large animal model of non-compressible intra-abdominal hemorrhage.

Noncompressible hemorrhage is a leading cause of potentially survivable combat death, with the vast majority of such deaths occurring in the out-of-hospital environment. While large animal models of this process are important for device and therapeutic development, clinical practice has changed over time and past models must follow suit. Developed in conjunction with regulatory feedback, this study presents a modernized, out-of-hospital, noncompressible hemorrhage model, in conjunction with a randomized study of past, present, and future fluid options following a hypotensive resuscitation protocol consistent with current clinical practice. We performed a randomized controlled experiment comparing three fluid resuscitation options in Yorkshire swine. Baseline data from animals of same size from previous experiments were analyzed (n = 70), and mean systolic blood pressure was determined, with a permissive hypotension resuscitation target defined as a 25% decrease from normal (67 mm Hg). After animal preparation, a grade IV to V liver laceration was induced. Animals bled freely for a 10-minute "time-to-responder" period, after which resuscitation occurred with randomized fluid in boluses to the goal target: 6% hetastarch in lactated electrolyte injection (HEX), normal saline (NS), or fresh whole blood (FWB). Animals were monitored for a total simulated "delay to definitive care" period of 2 hours postinjury. At the end of the 2-hour study period, 8.3% (1 of 12 swine) of the HEX group, 50% (6 of 12 swine) of the NS group, and 75% (9 of 12 swine) of the FWB had survived (p = 0.006), with Holm-Sidak pairwise comparisons showing a significant difference between HEX and FWB and (p = 0.005). Fresh whole blood had significantly higher systemic vascular resistance and hemoglobin levels compared with other groups (p = 0.003 and p = 0.001, respectively). Survival data support the movement away from HEX toward NS and, preferably, FWB in clinical practice and translational animal modeling. The presented model allows for future research including basic science, as well as translational studies of novel diagnostics, therapeutics, and devices.

Is fresh, leucodepleted, whole blood transfusion superior to blood component transfusion in pediatric patients undergoing spinal deformity surgeries? A prospective, randomized study analyzing postoperative serological parameters and clinical recovery.

PurposeTo compare the effectiveness of fresh whole blood (FWB) and blood component transfusion in improving clinical outcome and serological parameters in the early postoperative period following spinal deformity surgery.MethodsPatients undergoing major spinal deformity surgeries involving ≥ 6 levels of fusion and expected blood loss ≥ 750 ml between September 2017 and August 2018 were included in the study. The patients were randomized into two groups: FWBG and CG, receiving fresh whole blood and component transfusions, respectively.ResultsA total of 65 patients with spinal deformities of different etiologies were included. The mean age was 14.0 and 14.9 years in FWB and CG, respectively. All other preoperative parameters were comparable. The mean fusion levels and surgical time were 11.1 and 221.20 min in FWB, as compared with 10.70 and 208.74minutes in CG, respectively. Intraoperative blood losses were 929 ml (FWBG) and 847 ml(CG), and the mean volumes of transfusion were 1.90 (FWBG) and 1.65 units (CG). FWBG was significantly superior to CG in the following clinical and laboratory parameters: duration of oxygen dependence [36.43 (FWBG) vs. 43.45 h (CG); P = 0.0256], mean arterial pH [7.442 (FWBG) vs. 7.394 (CG); p < 0.001], interleukin-6 [30.04 (FWBG) vs. 35.10 (CG); p < 0.019], mean duration of HDU stay [40.6 hours (FWBG) vs 46.51 hours (CG); p = 0.0234] and postoperative facial puffiness [7/30 in FWBG vs. 18/35 (CG) (P < 0.02)].ConclusionFWB transfusion can potentially improve the immediate postoperative outcome in patients undergoing major spinal deformity surgeries by reducing the duration of intensive care unit stay and oxygen dependence. The other potential benefits of this practice, based on our study, include a reduced inflammatory response (reduced lactate and IL-6) and postoperative facial puffiness. However, further large-scale validation studies in future are necessary to precisely determine the role of FWB in spine surgeries.Level of evidence IIDiagnostic: individual cross-sectional studies with the consistently applied reference standard and blinding.

Random laser emission from whole blood as the active medium

We introduce a proof of concept for multimode random laser (RL) emission with fresh whole blood (WB) as the active medium. The experimental principle is adapted from RL emission using rhodamine 6G (R6G). We achieved conditions for fresh WB to fluoresce with stochastic amplification of stimulated emission of radiation. The random conditions are placed with SiO2 particles, suspended in isotonic solvent. The results we report are for: (1) R6G-RL, with 2 nm bandwidth centered at 567 nm, and (2) RL emission for WB at 969 nm and 437 nm, of sub-3 nm bandwidth. For validation purposes, we show that the pumping energy threshold for RL emission from blood is consistent with R6G-RL.

Operation Blood Rain: The Effect of Airdrop on Fresh Whole Blood.

Administration of fresh whole blood (FWB) is a life-saving treatment that prolongs life until definitive surgical intervention can be performed; however, collecting FWB is a time-consuming and resource-intensive process. Furthermore, it may be difficult to collect sufficient FWB to treat critically wounded patients or multiple-hemorrhaging casualties. This study describes the effect of airdrop on FWB and explores the possibility of using airdrop to deliver FWB to combat medics treating casualties in the prehospital setting when FDA-approved, cold-stored blood products are not readily available and timely casualty evacuation (CASEVAC) is not feasible. Four units of FWB were collected from volunteer donors and loaded into a blood cooler that was dropped from a fixed-wing aircraft under a standard airdrop training bundle (SATB) parachute. A control group of 4 units of FWB was stored in a blood cooler that was not dropped. Baseline and postintervention laboratory samples were measured in both airdropped and control units, including full blood counts, prothrombin time/partial thromboplastin time/international normalized ratio (PT/PTT/INR), pH, lactate, potassium, indirect bilirubin, glucose, fibrinogen, lactate dehydrogenase, and peripheral blood smears. The blood cooler, cooling bags, and all 4 FWB units did not sustain any damage from the airdrop. There was no evidence of hemolysis. All airdropped blood met parameters for transfusion per the Whole Blood Transfusion Clinical Practice Guideline of the Joint Trauma System (JTS). Airdrop of FWB in a blood cooler with a SATB parachute may be a viable way of delivering blood products to combat medics treating hemorrhaging patients in the prehospital setting, although further research is needed to fully validate the safety of this method of FWB delivery.

Improved survival in critically injured combat casualties treated with fresh whole blood by forward surgical teams in Afghanistan.

The objective of this study was to assess transfusion strategies and outcomes, stratified by the combat mortality index, of casualties treated by small surgical teams in Afghanistan. Resuscitation that included warm fresh whole blood (FWB) was compared to blood component resuscitation. Casualties treated by a Role 2 surgical team in Afghanistan from 2008 to 2014 who received 1 or more units of red blood cells (RBCs) or FWB were included. Patients were excluded if they had incomplete data or length of stay less than 30 minutes. Patients were separated into two groups: 1) received FWB and 2) did not receive FWB; moreover, both groups potentially received plasma, RBCs, and platelets. The analysis was stratified by critically versus noncritically injured patients using the prehospital combat mortality index. Kaplan-Meier plot, log-rank test, and multivariable Cox regression were performed to compare survival. In FWB patients, median units of FWB and total blood product were 4.0 (interquartile range [IQR], 2.0-7.0) and 16.0 (IQR, 10.0-28.0), respectively. The Kaplan-Meier plot demonstrated that survival was similar between FWB (79.1%) and no-FWB (74.5%) groups (p = 0.46); after stratifying patients by the combat mortality index, the risk of mortality was increased in the no-FWB group (hazard ratio, 2.8; 95% confidence interval, 1.2-6.4) compared to the FWB cohort. In forward-deployed environments, where component products are limited, FWB has logistical advantages and was associated with reduced mortality in casualties with a critical combat mortality index. Additional analysis is needed to determine if these effects of FWB are appreciable in all trauma patients or just in those with severe physiologic derangement.

Implementation of a low titer group O whole blood program for a law enforcement tactical team.

The Texas Ranger Special Operations Group (SOG) performs high-risk warrant service and responds to callouts for evolving kinetic situations and special missions as required. These operations may occur many hours from a trauma center. Fresh whole blood (FWB) transfusions may offer a stopgap for those who are critically injured. To make FWB transfusions a viable option, several steps must be implemented. The following lays out how the Texas Ranger SOG will implement and conduct FWB transfusions using low titer group O whole blood. The techniques outlined may be useful for communities that may face critical blood shortage in disasters.

Practical Considerations for a Military Whole Blood Program.

Prehospital care in the combat environment has always been of great importance to the U.S. military, and trauma resuscitation has remained a cornerstone. More evidence continues to demonstrate the advantages of intervention with early transfusion of blood products at the point of injury. The military has recognized these benefits; as such, the Department of Defense Joint Trauma System and the Committee on Tactical Combat Casualty Care have developed new advanced resuscitation guidelines, which now encourage the use of whole blood (WB) in the prehospital setting. This general review of peer-reviewed journal articles was performed through an extensive electronic search from the databases of PubMed Central (MEDLINE) and the Cochrane Library. Based on this literature search, the current evidence suggests that transfusion with WB is safe and efficacious. Additionally, soldier function is preserved after donating fresh WB in the field. Currently, the collection and implementation of WB is accomplished through several different protocol-driven techniques. WB has become the favored transfusion product as it provides all of the components of blood in a convenient package that is easy to store and transport. Specifically, group O WB containing low titers of anti-A and -B antibodies has become the transfusion product of choice, offering the ability to universally fluid resuscitate patients despite not knowing their blood group. This new ability to obtain low titer group O WB has transformed the approach to the management of hemorrhagic shock in the prehospital combat environment.