Effect of post-storage filters vs. pre-storage filters for leukoreduction of blood components on clinical outcomes: a systematic review and meta-analysis.

BACKGROUND: Leukoreduction has been used to limit the risk of adverse events. The most commonly used methodology is filtration (pre- or post-storage). However, whether pre-storage filtration is better than post-storage filtration needs to be clearly defined, particularly for countries that still use post-storage filtration. This study aimed to synthesize the best available evidence on the effectiveness of pre-storage filters compared with post-storage filters for transfusion reactions, for the occurrence of infections, for the length of hospital stay, and for the death of patients undergoing leukoreduced transfusion. METHODS: We searched the MEDLINE (PubMed), CINAHL (EBSCO), PsycINFO (APA), Scopus (Elsevier), The Cochrane Library (J. Wiley), Web of Science Core Collection (Clarivate Analytics), Embase (Elsevier), and LILACS (VHL) databases and gray literature for eligible studies in August 2020 and updated the search in October 2023. The Joanna Briggs Institute critical assessment tools were applied to analyze the quality appraisal of the studies. GRADE was used to determine the certainty of the evidence. RESULTS: The meta-analysis showed that pre-storage filtration was a protective factor for the occurrence of febrile non-hemolytic transfusion reaction in red blood cells (RR 0.49, 95% CI 0.41-0.59) and platelet concentrate transfusions (RR 0.16, 95% CI 0.12-0.22). The same did not occur for post-surgical infection after platelet concentrate transfusions (RR 0.82, 95% CI 0.65-1.04). Only one study analyzed the length of hospital stay and showed no significant difference between patients who received leukoreduced transfusions according to the type of filter used. According to the GRADE criteria, the certainty of the evidence for febrile non-hemolytic transfusion reactions was low for red blood cells and very low for platelet concentrate due to the high risk of bias. Infection was a low risk due to imprecision. CONCLUSIONS: The results of this review showed that the certainty of recommending the best type of filter (pre- or post-storage) for the benefit of the outcomes analyzed is still fragile; therefore, more robust evidence is needed. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42020192202.

Early Cold Stored Platelet Transfusion Following Severe Injury: A Randomized Clinical Trial.

OBJECTIVE: To determine the feasibility, efficacy, and safety of early cold stored platelet transfusion compared with standard care resuscitation in patients with hemorrhagic shock. BACKGROUND: Data demonstrating the safety and efficacy of early cold stored platelet transfusion are lacking following severe injury. METHODS: A phase 2, multicenter, randomized, open label, clinical trial was performed at 5 US trauma centers. Injured patients at risk of large volume blood transfusion and the need for hemorrhage control procedures were enrolled and randomized. The intervention was the early transfusion of a single apheresis cold stored platelet unit, stored for up to 14 days versus standard care resuscitation. The primary outcome was feasibility and the principal clinical outcome for efficacy and safety was 24-hour mortality. RESULTS: Mortality at 24 hours was 5.9% in patients who were randomized to early cold stored platelet transfusion compared with 10.2% in the standard care arm (difference, -4.3%; 95% CI, -12.8% to 3.5%; P =0.26). No significant differences were found for any of the prespecified ancillary outcomes. Rates of arterial and/or venous thromboembolism and adverse events did not differ across treatment groups. CONCLUSIONS AND RELEVANCE: In severely injured patients, early cold stored platelet transfusion is feasible, safe and did not result in a significant lower rate of 24-hour mortality. Early cold stored platelet transfusion did not result in a higher incidence of arterial and/or venous thrombotic complications or adverse events. The storage age of the cold stored platelet product was not associated with significant outcome differences. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT04667468.

Genetic regulation of carnitine metabolism controls lipid damage repair and aging RBC hemolysis in vivo and in vitro.

ABSTRACT: Recent large-scale multiomics studies suggest that genetic factors influence the chemical individuality of donated blood. To examine this concept, we performed metabolomics analyses of 643 blood units from volunteers who donated units of packed red blood cells (RBCs) on 2 separate occasions. These analyses identified carnitine metabolism as the most reproducible pathway across multiple donations from the same donor. We also measured l-carnitine and acyl-carnitines in 13 091 packed RBC units from donors in the Recipient Epidemiology and Donor Evaluation study. Genome-wide association studies against 879 000 polymorphisms identified critical genetic factors contributing to interdonor heterogeneity in end-of-storage carnitine levels, including common nonsynonymous polymorphisms in genes encoding carnitine transporters (SLC22A16, SLC22A5, and SLC16A9); carnitine synthesis (FLVCR1 and MTDH) and metabolism (CPT1A, CPT2, CRAT, and ACSS2), and carnitine-dependent repair of lipids oxidized by ALOX5. Significant associations between genetic polymorphisms on SLC22 transporters and carnitine pools in stored RBCs were validated in 525 Diversity Outbred mice. Donors carrying 2 alleles of the rs12210538 SLC22A16 single-nucleotide polymorphism exhibited the lowest l-carnitine levels, significant elevations of in vitro hemolysis, and the highest degree of vesiculation, accompanied by increases in lipid peroxidation markers. Separation of RBCs by age, via in vivo biotinylation in mice, and Percoll density gradients of human RBCs, showed age-dependent depletions of l-carnitine and acyl-carnitine pools, accompanied by progressive failure of the reacylation process after chemically induced membrane lipid damage. Supplementation of stored murine RBCs with l-carnitine boosted posttransfusion recovery, suggesting this could represent a viable strategy to improve RBC storage quality.

Regulation of kynurenine metabolism by blood donor genetics and biology impacts red cell hemolysis in vitro and in vivo.

ABSTRACT: In the field of transfusion medicine, the clinical relevance of the metabolic markers of the red blood cell (RBC) storage lesion is incompletely understood. Here, we performed metabolomics of RBC units from 643 donors enrolled in the Recipient Epidemiology and Donor Evaluation Study, REDS RBC Omics. These units were tested on storage days 10, 23, and 42 for a total of 1929 samples and also characterized for end-of-storage hemolytic propensity after oxidative and osmotic insults. Our results indicate that the metabolic markers of the storage lesion poorly correlated with hemolytic propensity. In contrast, kynurenine was not affected by storage duration and was identified as the top predictor of osmotic fragility. RBC kynurenine levels were affected by donor age and body mass index and were reproducible within the same donor across multiple donations from 2 to 12 months apart. To delve into the genetic underpinnings of kynurenine levels in stored RBCs, we thus tested kynurenine levels in stored RBCs on day 42 from 13 091 donors from the REDS RBC Omics study, a population that was also genotyped for 879 000 single nucleotide polymorphisms. Through a metabolite quantitative trait loci analysis, we identified polymorphisms in SLC7A5, ATXN2, and a series of rate-limiting enzymes (eg, kynurenine monooxygenase, indoleamine 2,3-dioxygenase, and tryptophan dioxygenase) in the kynurenine pathway as critical factors affecting RBC kynurenine levels. By interrogating a donor-recipient linkage vein-to-vein database, we then report that SLC7A5 polymorphisms are also associated with changes in hemoglobin and bilirubin levels, suggestive of in vivo hemolysis in 4470 individuals who were critically ill and receiving single-unit transfusions.

Using umbilical cord blood as a source of paediatric packed red blood cells: Processing and quality control.

BACKGROUND AND OBJECTIVES: Umbilical cord blood (UCB) has been used as a source of red blood cells (RBCs) for neonatal/paediatric transfusion purposes. This study adopted two different procedures to obtain umbilical RBC (U-RBC) to compare its quality control parameters to those of fractionated adult RBC (A-RBC), for paediatric purposes. MATERIALS AND METHODS: UCB units (24) were filtered and processed based on two different methods, namely, conventional/manual (P1;n12) and automatic (P2;n12). They were compared to five fractionated A-RBCs. U-RBC and A-RBC were stored for 14 days and had their haematological, biochemical, haemolytic and microbiological parameters analysed at D1, D7 and D14. Cytokines and growth factors (GFs) in residual U-RBC plasma were measured. RESULTS: Mean volume of processed U-RBC units was 45 mL for P1 and 39 mL for P2; the mean haematocrit level reached 57% for P1 and 59% for P2. A-RBC recorded a mean volume of 44 mL. Haematologic and biochemical parameters analysed in U-RBC and A-RBC presented similar behaviours during storage time, except for parameter values, which differed between them. Pro-inflammatory and immunomodulatory cytokines, as well as GFs, were higher in U-RBC residual plasma than in that A-RBC. CONCLUSION: UCB can be processed into RBC based on either manual or automated protocols. U-RBC units met the referenced quality parameters defined for A-RBC. Some features, mainly the biochemical ones, should be further investigated to help improve quality parameters, with emphasis on differences found in, and particularities of, this material and on recipients of this new transfusion practice.

Defining the Healthy Infant Metabolome: Liquid Chromatography Tandem-Mass Spectrometry Analysis of Dried Blood Spot Extracts from the Prospective Research on Early Determinants of Illness and Children's Health Trajectories Birth Cohort Study.

Newborn screening using dried plasma spots offers preanalytical advantages over conventional cards for plasma-associated targets of interest. Herein we present dried plasma spot-based methods for measuring metabolites using a 250+ compound liquid chromatography tandem mass spectrometry library. Quality assurance reduced this library to 134, and from these, 30 compounds determined the normal newborn reference ranges.

Sangre total leucorreducida y filtro ahorrador de plaquetas preserva su función hemostática por 21 días: ¿La resucitación hemostática podría ser una realidad en Colombia? / Leukoreduced whole blood and platelet-sparing filter preserves its hemostatic function for 21 days: Could hemostatic resuscitation become a reality in Colombia?

Introducción. La resucitación hemostática es una estrategia para compensar la pérdida sanguínea y disminuir el impacto de la coagulación inducida por trauma. Debido a que la disponibilidad de transfundir una razón equilibrada de hemocomponentes es difícil de lograr en el entorno clínico, la sangre total ha reaparecido como una estrategia fisiológica, con ventajas logísticas, que le permiten ser accesible para iniciar tempranamente la resucitación hemostática. El objetivo de este estudio fue evaluar las propiedades celulares, coagulantes y viscoelásticas de la sangre total almacenada por 21 días. Métodos. Las unidades de sangre total fueron obtenidas de 20 donantes voluntarios sanos. Se procesaron mediante un sistema de leucorreducción ahorrador de plaquetas y fueron almacenadas en refrigeración (1-6°C) sin agitación. Se analizaron los días 0, 6, 11 y 21. Las bolsas fueron analizadas para evaluar las líneas celulares, niveles de factores de coagulación y propiedades viscoelásticas mediante tromboelastografía. Resultados. El conteo eritrocitario y la hemoglobina se mantuvieron estables. El conteo de plaquetas tuvo una reducción del 50 % al sexto día, pero se mantuvo estable el resto del seguimiento. Los factores de coagulación II-V-VII-X, fibrinógeno y proteína C se mantuvieron dentro del rango normal. La tromboelastografía mostró una prolongación en el tiempo del inicio de la formación del coágulo, pero sin alterar la formación final de un coágulo estable. Conclusiones. La sangre total leucorreducida y con filtro ahorrador de plaquetas conserva sus propiedades hemostáticas por 21 días. Este es el primer paso en Colombia para la evaluación clínica de esta opción, que permita hacer una realidad universal la resucitación hemostática del paciente con trauma severo.

Background. Hemostatic resuscitation is a strategy to compensate blood loss and reduce the impact of trauma-induced coagulopathy. However, balanced resuscitation presents challenges in its application in the clinical setting. Whole blood has re-emerged as a physiologic strategy with logistical advantages that offer the opportunity for early initiation of hemostatic resuscitation. The study aims to evaluate the cellular, coagulation, and viscoelastic properties of whole blood preserved for 21 days. Methods. Whole blood units were donated by 20 healthy volunteers. These units were processed using a platelet-sparing leukoreduction filtration system. Units were stored under refrigeration (1-6°C) without agitation and were sampled on days 0, 6, 11, 16, and 21. The units were tested to assess its cellular properties and coagulation factors levels. In addition, viscoelastic features were tested using tromboelastography.Results. Red blood cells count and hemoglobin levels remained stables. Platelet count had a 50% reduction on day 6, and then remained stable for 21 days. Factors II-V-VII-X, fibrinogen, and protein C remained within normal range. Tromboelastrography test showed that the reaction time of clot formation is prolonged, but the final clot formation is not altered. Conclusion. Whole blood retains its hemostatic properties for 21 days. This is the first step to evaluate the use of whole blood in the resuscitation protocols for Colombia allowing hemostatic resuscitation become a universal reality.

Biochemical and hematologic changes in whole blood from Brazilian horses stored in citrate-phosphate-dextrose-adenine pouches for up to 28 days.

BACKGROUND: Anaerobic cellular metabolism causes a series of structural and physiologic changes during storage that could compromise post-transfusion viability, reducing the safety of using blood stored for an extended period. OBJECTIVE: We aimed to follow the biochemical and hematologic alterations of equine blood stored in plastic bags containing citrate-phosphate-dextrose-adenine (CPDA-1) for up to 28 days. METHODS: Whole blood samples (450 mL) were collected from 20 Brazilian Saddle horses into CPDA-1 pouches and stored between 2°C and 6°C in a blood bank. On days 0, 7, 14, 21, and 28 of storage, blood samples were taken and submitted for biochemical (sodium [Na+ ], potassium [K+ ], glucose, and lactate) and hematologic (hemoglobin [Hb], hematocrit [HCT], mean corpuscular volume [MCV], percent hemolysis [% hemolysis]) analyses. RESULTS: The only time the blood pH levels dipped below 7 was after D21 of storage, and the levels were significantly lower than those on the first storage day (D0). Potassium concentrations showed significant increases from D7 and then remained increased throughout the experimental period. Chloride and lactate concentrations revealed a significantly increased trend from D7 that was maintained over time. Mean corpuscular volumes increased significantly on D7 and D14 and, thereafter, remained stable. The mean % hemolysis increased on D28, which was significantly higher than D0. No bacterial growth was found in any pouch after 28 days of storage. CONCLUSIONS: Significant and gradual biochemical changes were observed in equine whole blood during prolonged storage. These changes could compromise the clinical conditions of patients requiring transfusion. In vivo studies are needed to evaluate the effects as well as survival rates and efficacy of transfused red blood cells in recipients.

Biochemical and blood gas alterations in buffalo (Bubalus bubalis) whole blood stored in CPDA-1 and CPD/SAG-M bags.

OBJECTIVE: The objective of this study was to evaluate the biochemical and blood gas alterations of whole blood of buffaloes that was stored in citrate-phosphate-dextrose with adenine (CPDA-1) and CPD/SAG-M blood bags for 42 days. DESIGN: Prospective study. INTERVENTIONS: Ten male buffaloes were used in this study. A total volume of 900 mL of blood was collected from each buffalo so that 450 mL was stored in CPDA-1 and 450 mL was stored in CPD/SAG-M bags at 2-6°C for 42 days. The stored blood was evaluated at 7 time points (D): D0 (immediately after blood collection) and 7 (D7), 14 (D14), 21 (D21), 28 (D28), 35 (D35), and 42 (D42) days after collection. Blood gas, biochemical, and microbiological parameters were monitored. MEASUREMENTS AND MAIN RESULTS: The overall blood pH decreased from 6.997 ± 0.05 at D0 to 6.784 ± 0.09 at D42, differing from baseline from D14 onward (P < 0.05). There were increases in partial pressure of oxygen (pO2 ), partial pressure of carbon dioxide (pCO2 ), lactate, and potassium (K) and decreases in the concentrations of sodium, bicarbonate, glucose, and pH (P < 0.05) during storage in both bags but no alterations in total protein concentration. Most of the variables were consistently similar between the 2 types of blood bags (P > 0.05) evaluated, with the exception of pCO2 , HCO3, cholesterol, and total protein, which had higher values in the CPDA-1 bag (P < 0.05). The K, pO2 , and lactate had the highest alterations during storage, with increases from baseline to D42 of 563%, 317%, and 169%, respectively. CONCLUSION: In general, no significant changes of clinical importance were observed after storage of whole blood samples from buffaloes for 42 days in the 2 types of blood bags that are indicated for use with this species.

Blood serum stability limit and maximum storage time of bovine samples

Measuring metabolic parameters in the blood has been an indispensable tool for assessing the productive and health status of dairy cows for more than 100 years. The values of laboratory parameters depend on various preanalytical, analytical and postanalytical factors. The most important preanalytical factors are sample transport time and temperature, hemolysis, anticoagulant type, and sample volume. Preanalytical factors can lead to reduced stability of the analyte in the sample, which changes their concentration. Loss of stability changes the time of storage and manipulation of the sample, which determines the criteria for its acceptance or rejection. The two stability indicators are stability limit and maximum permissible instability. A stability limit (SL) is defined as the period of time in which a property variation does not exceed a maximum permissible instability (MPI). The aim of this study was to determine the SL and MPI for each analyte in the blood serum of cows and to determine whether SL differs in the function of the presence of preanalytical errors in the blood sample. Three hundred samples of dairy cow origin in different periods of lactation participated in this research. They were classified into 6 groups of 50 samples: according to the time from sampling to processing in the laboratory (0-4 h, 4-8 h and over 8 h; all transported on dry ice, protected from environmental factors, without preanalytical errors) and according to the presence of preanalytical errors (group with hemolysis, a group transported at ambient temperature and a group with a small sample volume). Each sample was aliquoted in two portions. One portion was left at +4°C and tested once a day for 6 days of sample storage, and the second portion, placed at -20°C, was tested once a month for 6 months. The MPI had a value ranging from 1.51 to 8.4. Metabolic profile analytes with lower MPI values (1.51-3.22) were albumin (ALB), total protein (TPROT), UREA, glucose (GLU), calcium (Ca), and phosphorus (P). Higher MPI values (5.1-8.4) were found for nonesterified fatty acids (NEFA), beta-hydroxybutirate (BHB), cholesterol (CHOL), triglycerides (TGC), total bilirubin (TBIL) and aspartat aminotransferase (AST). For most parameters, we can conclude that their PD% changed faster in storage conditions at +4°C compared to the regime of -20°C. The largest number of biochemical analytes in bovine blood serum shows preserved stability in the first 6 days at +4°C or 6 months at -20°C if transported to the laboratory within 8 h after sampling in ideal conditions and without the action of preanalytical errors. Prolonged transport under ideal conditions or the existence of preanalytical errors such as transport at room temperature, hemolysis or small sample volume shorten the stability of the ALB, NEFA, GLU, UREA and P. Concentration of all analytes decreases during the stability test except for UREA, NEFA, BHB and for CHOL and TGC in some groups. Variations in parameters such as BHB, NEFA, TBIL, AST, and Ca have shown potential clinical significance. At storage conditions at +4°C, clinically significant variations at at least one measurement point were found for AST (7.5% of samples), BHB (6.1% of samples), NEFA (9.9% of samples) and for TBIL (in 7% of samples). This study can help define acceptable delay times and storage conditions for bovine blood samples, which is of great importance because in working with farm animals it is often not possible to take samples in a short time and deliver them to the laboratory, and samples are often burdened with certain preanalytical errors with limited possibilities of re-sampling.(AU)

Impact of blood storage duration on hematologic, blood gas, biochemical, and oxidative stress variables in sheep undergoing allogeneic blood transfusions.

BACKGROUND: Hemotherapy in ruminants is limited to whole blood transfusions, sometimes with stored blood for up to 42 days, but little attention has been given to the effect of blood storage times and recipient responses after transfusions. OBJECTIVES: We aimed to evaluate the hematologic and serum biochemical effects after allogeneic blood transfusion with either fresh or stored blood in sheep. We also sought to examine hematologic and biochemical analyte changes in the store blood. METHODS: Eighteen sheep underwent a single phlebotomy to remove 40% of their blood volume. The sheep were divided into three experimental groups, G0, G15, and G35, which included six animals, each receiving 20 mL/kg of either fresh blood or blood stored in citrate, phosphate, dextrose, and adenine (CPDA-1) bags for 15 and 35 days, respectively. Biochemical, hematologic, coagulation, blood gas, lipid peroxidation, and oxidative stress test evaluations were performed using the blood samples gathered at T0 (before transfusion), 30 minutes (T30m), 6, 12, 24, 48, 72, and 96 hours (T6h-T96h), 8 days (T8d), and 16 days (T16d) after transfusions. RESULTS: Sheep exhibited increases in packed cell volumes, red blood cell counts, and total hemoglobin concentrations at T30m (P < .05). G35 animals had greater plasma hemoglobin concentrations at T12h and decreased blood pH values at T6h, characterized by slight metabolic acidemia. Regarding oxidative stress, G35 animals had decreased catalase activities from T0 at T30m, T6h, T12h, and T24h, indicating that hemolysis had occurred, which was supported by concomitant increases in bilirubin. CONCLUSIONS: Sheep transfused with 35-day stored blood exhibited greater hematologic, blood gas, biochemical, and oxidative alterations; however, anemic animals without comorbidities effectively reversed those alterations.

Evaluation of hematologic, biochemical, and blood gas variables in stored canine packed red blood cells, and the impact of storage time on blood recipients.

BACKGROUND: Canine packed red blood cells (pRBCs) can be stored under refrigeration for several days; however, cellular metabolism remains active inside the units, thus producing substances that affect their quality. OBJECTIVES: We aimed to evaluate hematologic, biochemical, and blood gas variable alterations that occur in canine pRBCs during storage, and their effects on recipient clinicopathologic parameters. METHODS: The study was conducted in two phases. In phase I, 15 pRBC units containing CPDA-1 were stored for 28 days; samples were collected weekly from the units of days 0 to 28 to measure the packed cell volume (PCV), pH, partial pressure carbon dioxide (PCO2 ), partial pressure oxygen (PO2 ), concentrations of lactate and potassium, and the percent hemolysis. In phase II, another 22 canine pRBC units stored for different time periods (maximum of 21 days) were transfused, and the recipients were evaluated before and after transfusion for changes in clinical parameters (heart rate, respiratory rate, systolic arterial pressure, and rectal temperature) and hematologic variables (PCV, lactate and potassium concentrations, pH, PCO2 , the ratio of arterial oxygen partial pressure to fractional inspired oxygen [PO2 /FiO2 ] ratio, oxygen saturation [SaO2 ], base excess, and bicarbonate [HCO3 ]). RESULTS: In the pRBC units, the PCV increased from 70% to 78.33%, the lactate concentration increased 627%, the potassium concentration increased 183%, the percent hemolysis reached 0.69%, and the pH decreased 9% after 28 days. However, the dogs who received transfusions were not negatively affected. There was a significant increase in PCVs, and a significant decrease in heart rates. CONCLUSION: Canine pRBCs undergo hematologic, blood gas, and biochemical alterations during storage; however, the transfusion of pRBCs stored for up to 21 days increased PCVs without causing harm to the dogs.

Serum biochemistry of the Arrau turtle (Podocnemis expansa): Frozen analyte stability and the effects of long-term storage.

BACKGROUND: Turtles are a major source of protein for riverside human populations in Brazil. The encouragement of commercial breeding meets conservation efforts for these animals, and it is, therefore, crucial to understand the physiologic and behavioral aspects of semi-aquatic species in captive conditions. Serum biochemical tests are ancillary diagnostic tools, and sample storage is a main problem since clinical laboratories are not always available near the habitats of these species. OBJECTIVES: The aim of this study was to provide information about the stability of albumin, aspartate aminotransferase (AST), calcium, creatinine kinase (CK), total cholesterol (Chol), alkaline phosphatase (ALP), gamma glutamyltransferase (GGT), total protein (TP), and urea at different storage times. METHODS: In all, 17 Arrau turtles (Podocnemis expansa) were used, and the serum obtained was separated into aliquots and analyzed at 0, 4, 8, 16, and 32 days after being stored at -20°C. RESULTS: The results showed that albumin, AST, CK, GGT, and TP suffered interference due to the long storage times. CONCLUSION: Analytes such as ALP, calcium, Chol, and urea can be evaluated for up to 1 month after freezing. Albumin, AST, and TP can be analyzed up to 1 week after freezing without alterations, and CK GGT are best evaluated on fresh samples.

Quality control in veterinary blood banks: evaluation of canine platelet concentrates stored for five days.

BACKGROUND: Platelets undergo structural, biochemical and functional alterations when stored, and platelet storage lesions reduce platelet function and half-life after transfusion. The objective of this study was to evaluate stored canine platelet concentrates with platelet aggregation, flow cytometry and biochemistry assays. Twenty-two bags of canine platelet concentrates were obtained by the platelet-rich plasma method and were assessed on days 1, 3 and 5 after collection. Parameters such as platelet counts, residual leukocytes, platelet swirling, glucose, lactate, pH, CD62P expression (platelet activation), JC-1 (mitochondrial function) and annexin V (apoptosis and cell death) were assessed. RESULTS: Over the five days of storage there was a significant decrease in glucose, HCO3, pCO2, ATP, pH, swirling and mitochondrial function, associated with a significant increase in lactate levels and pO2. At the end of storage pH was 5.9 ± 0.6 and lactate levels were 2.8 ± 1.2 mmol/L. Results of the quality parameters evaluated were similar to those reported in human platelets studies. The deleterious effects of storage were more pronounced in bags with higher platelet counts (> 7.49 × 1010/unit), suggesting that canine platelet concentrates should not contain an excessive number of platelets. CONCLUSIONS: Quality parameters of canine platelets under standard storage conditions were similar to those observed in human platelets. Our results have potential to be used for the routine evaluation and quality control in veterinary blood banks.

Red blood cell metabolism in Rhesus macaques and humans: comparative biology of blood storage.

Macaques are emerging as a critical animal model in transfusion medicine, because of their evolutionary similarity to humans and perceived utility in discovery and translational science. However, little is known about the metabolism of Rhesus macaque red blood cells (RBC) and how this compares to human RBC metabolism under standard blood banking conditions. Metabolomic and lipidomic analyses, and tracing experiments with [1,2,3-13C3]glucose, were performed using fresh and stored RBC (sampled weekly until storage day 42) obtained from Rhesus macaques (n=20) and healthy human volunteers (n=21). These results were further validated with targeted quantification against stable isotope-labeled internal standards. Metabolomic analyses demonstrated inter-species differences in RBC metabolism independent of refrigerated storage. Although similar trends were observed throughout storage for several metabolic pathways, species- and sex-specific differences were also observed. The most notable differences were in glutathione and sulfur metabolites, purine and lipid oxidation metabolites, acylcarnitines, fatty acyl composition of several classes of lipids (including phosphatidylserines), glyoxylate pathway intermediates, and arginine and carboxylic acid metabolites. Species-specific dietary and environmental compounds were also detected. Overall, the results suggest an increased basal and refrigerator-storage-induced propensity for oxidant stress and lipid remodeling in Rhesus macaque RBC cells, as compared to human red cells. The overlap between Rhesus macaque and human RBC metabolic phenotypes suggests the potential utility of a translational model for simple RBC transfusions, although inter-species storage-dependent differences need to be considered when modeling complex disease states, such as transfusion in trauma/hemorrhagic shock models.

Freeze-dried versus fresh platelet-rich plasma in acute wound healing of an animal model.

Aim: To compare freeze-dried and fresh platelet-rich plasma (PRP) preparations, in a pre-clinical study. Materials & methods: 30 Wistar male rats were used to compare and characterize human PRP which was applied at the perilesional area in an acute wound model, evaluated by macroscopical and histological analysis. Results: Despite the increased growth factor concentration after the freeze-drying process, no change in the healing kinetics was observed in vivo. Nevertheless, a significant increased number of myofibroblasts was demonstrated in comparison with the fresh PRP group. We also demonstrated a significant increased percentage of blood vessels in comparison with controls in both the superficial and deep epidermis. Conclusion: These results encourage randomized clinical trials to evaluate the effectiveness of freeze-dried PRP for skin ulcer treatment.

Effects of aged stored autologous red blood cells on human plasma metabolome.

Cold storage of blood for 5 to 6 weeks has been shown to impair endothelial function after transfusion and has been associated with measures of end-organ dysfunction. Although the products of hemolysis, such as cell-free plasma hemoglobin, arginase, heme, and iron, in part mediate these effects, a complete analysis of transfused metabolites that may affect organ function has not been evaluated to date. Blood stored for either 5 or 42 days was collected from 18 healthy autologous volunteers, prior to and after autologous transfusion into the forearm circulation, followed by metabolomics analyses. Significant metabolic changes were observed in the plasma levels of hemolytic markers, oxidized purines, plasticizers, and oxidized lipids in recipients of blood stored for 42 days, compared with 5 days. Notably, transfusion of day 42 red blood cells (RBCs) increased circulating levels of plasticizers (diethylhexyl phthalate and derivatives) by up to 18-fold. Similarly, transfusion of day 42 blood significantly increased circulating levels of proinflammatory oxylipins, including prostaglandins, hydroxyeicosatrienoic acids (HETEs), and dihydroxyoctadecenoic acids. Oxylipins were the most significantly increasing metabolites (for 9-HETE: up to ∼41-fold, P = 3.7e-06) in day 42 supernatants. Measurements of arginine metabolism confirmed an increase in arginase activity at the expense of nitric oxide synthesis capacity in the bloodstream of recipients of day 42 blood, which correlated with measurements of hemodynamics. Metabolic changes in stored RBC supernatants impact the plasma metabolome of healthy transfusion recipients, with observed increases in plasticizers, as well as vasoactive, pro-oxidative, proinflammatory, and immunomodulatory metabolites after 42 days of storage.