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1.
Int J Mol Sci ; 22(6)2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33809183

RESUMO

Packed red blood cells (pRBCs), the most commonly transfused blood product, are exposed to environmental disruptions during storage in blood banks. In this study, temporal sequence of changes in the ion exchange in pRBCs was analyzed. Standard techniques commonly used in electrolyte measurements were implemented. The relationship between ion exchange and red blood cells (RBCs) morphology was assessed with use of atomic force microscopy with reference to morphological parameters. Variations observed in the Na+, K+, Cl-, H+, HCO3-, and lactate ions concentration show a complete picture of singly-charged ion changes in pRBCs during storage. Correlation between the rate of ion changes and blood group type, regarding the limitations of our research, suggested, that group 0 is the most sensitive to the time-dependent ionic changes. Additionally, the impact of irreversible changes in ion exchange on the RBCs membrane was observed in nanoscale. Results demonstrate that the level of ion leakage that leads to destructive alterations in biochemical and morphological properties of pRBCs depend on the storage timepoint.


Assuntos
Preservação de Sangue/métodos , Eritrócitos/metabolismo , Troca Iônica , Manejo de Espécimes/métodos , Carbonatos/metabolismo , Membrana Eritrocítica , Humanos , Íons/metabolismo , Ácido Láctico/metabolismo , Microscopia de Força Atômica , Potássio/metabolismo , Sódio/metabolismo
2.
Methods Mol Biol ; 2180: 523-538, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32797432

RESUMO

Frozen blood reserves are an important component in meeting blood needs. The idea behind a frozen blood reserve is twofold: to freeze units of rare blood types for later use by patients with special transfusion needs and for managing special transfusion circumstances. The permeating additive glycerol is used as a cryoprotectant to protect red blood cells (RBCs) from freezing damage. The use of thawed RBCs has been hampered by a 24-h outdating period due to the potential bacterial contamination when a functionally open system is used for addition and removal of the glycerol. The introduction of an automated, functionally closed system for glycerolization and deglycerolization of RBCs improved the operational practice. More importantly, the closed process allowed for extended shelf life of the thawed RBCs. In the current chapter, a cryopreservation procedure for RBCs using a functionally closed processing system is described.


Assuntos
Preservação de Sangue/métodos , Criopreservação/métodos , Crioprotetores/farmacologia , Eritrócitos/citologia , Congelamento , Glicerol/farmacologia , Transfusão de Sangue , Eritrócitos/efeitos dos fármacos , Humanos
3.
Am J Clin Pathol ; 154(3): 362-368, 2020 08 05.
Artigo em Inglês | MEDLINE | ID: mdl-32445461

RESUMO

OBJECTIVES: We evaluated the impact of electronic medical record (EMR)-guided pooled cryoprecipitate dosing vs our previous practice of requiring transfusion medicine (TM) resident approval for every cryoprecipitate transfusion. METHODS: At our hospital, cryoprecipitate pooled from five donors is dosed for adult patients, while single-donor cryoprecipitate is dosed for pediatric patients (defined as patients <50 kg in weight). EMR-based dosing guidance replaced a previously required TM consultation when cryoprecipitate pools were ordered, but a consultation remained required for single-unit orders. Usage was defined as thawed cryoprecipitate; wastage was defined as cryoprecipitate that expired prior to transfusion. RESULTS: In the 6 months prior to intervention, 178 ±â€…13 doses of pooled cryoprecipitate were used per month vs 187 ±â€…15 doses after the intervention (P = .68). Wastage of pooled cryoprecipitate increased from 7.7% ±â€…1.5% to 12.7% ±â€…1.4% (P = .038). There was no change in wastage of pediatric cryoprecipitate doses during the study period. These trends remained unchanged for a full year postimplementation. CONCLUSIONS: Electronic dosing guidance resulted in similar cryoprecipitate usage as TM auditing. Increased wastage may result from reduced TM oversight. Product wastage should be balanced against the possibility that real-time audits could delay a lifesaving therapy.


Assuntos
Preservação de Sangue/métodos , Transfusão de Sangue/métodos , Registros Eletrônicos de Saúde , Humanos
4.
Rev. esp. anestesiol. reanim ; 67(5): 237-244, mayo 2020. tab
Artigo em Espanhol | IBECS | ID: ibc-199486

RESUMO

ANTECEDENTES: La implementación de los programas Patient Blood Management (PBM) es variable en Europa, incluso en centros en los que estos programas están bien establecidos, donde existe variabilidad en cuanto a prácticas transfusionales. OBJETIVOS Y MÉTODOS: Realizamos una encuesta para valorar la práctica actual sobre PBM perioperatoria en pacientes programados para artroplastia total de cadera y rodilla, entre los investigadores involucrados en el Estudio POWER.2 en España (estudio observacional prospectivo que evaluaba las vías de recuperación intensificada en cirugía ortopédica). RESULTADOS: Se obtuvo un total de 322 respuestas (37,8%). El 50% de los respondedores revisaban los niveles de hemoglobina, al menos 4 semanas antes de la cirugía; el 35% trataba a todos los pacientes anémicos, aunque el 99,7% consideraba que la detección y tratamiento de la anemia preoperatoria podrían influir en los resultados postoperatorios. La falta de infraestructuras (76%) y la falta de tiempo (51%) fueron los principales motivos para no tratar a los pacientes anémicos. El estatus del hierro es revisado antes de la cirugía por el 19% de manera rutinaria, y el 36% lo evalúa únicamente en pacientes anémicos. Hb<9,9g/dl es el valor umbral para demorar la cirugía para el 61% de los clínicos, y el 22% consideraría transfundir preoperatoriamente a los pacientes clínicamente estables sin sangrado activo. El valor umbral para transfundir a los pacientes sin enfermedad cardiovascular es 8g/dl para el 43% y 7g/dl para el 34% de los respondedores; el 75% de los facultativos considera que utiliza «umbrales restrictivos», y el 90% sigue la política transfusional uno a uno (single unit). CONCLUSIONES: Los resultados de nuestra encuesta muestran la variabilidad en la práctica clínica en PBM en cirugía ortopédica mayor, a pesar de ser el tipo de cirugía con más tradición en estos programas


BACKGROUND: Implementation of Patient Blood Management programs remain variable in Europe, and even in centres with well-established PBM programs variability exists in transfusion practices. OBJECTIBES AND METHODS: We conducted a survey in order to assess current practice in perioperative Patient Blood Management in patients undergoing total hip and knee replacement among researchers involved in POWER.2 Study in Spain (an observational prospective study evaluating enhanced recovery pathways in orthopaedic surgery). RESULTS: A total of 322 responses were obtained (37.8%). Half of responders check Haemoglobin levels in patients at least 4 weeks before surgery; 35% treat all anaemic patients, although 99.7% consider detection and treatment of preoperative anaemia could influence the postoperative outcomes. Lack of infrastructure (76%) and lack of time (51%) are the main stated reasons not to treat anaemic patients. Iron status is routinely checked by 19% before surgery, and 36% evaluate it solely in the anaemic patient. Hb<9.9 g/dl is the threshold to delay surgery for 61% of clinicians, and 22% would consider transfusing preoperatively clinically stable patients without active bleeding. The threshold to transfuse patients without cardiovascular disease is 8 g/dl for 43%, and 7 g/dl for 34% of the responders; 75% of clinicians consider they use "restrictive thresholds", and 90% follow the single unit transfusion policy. CONCLUSIONS: The results of our survey show variability in clinical practice in Patient Blood Management in major orthopaedic surgery, despite being the surgery with the greatest tradition in these programs


Assuntos
Humanos , Osteoartrite do Quadril/cirurgia , Artroplastia de Quadril/métodos , Artroplastia do Joelho/métodos , Osteoartrite do Joelho/cirurgia , Perda Sanguínea Cirúrgica/prevenção & controle , Preservação de Sangue/métodos , Implementação de Plano de Saúde/métodos , Pesquisas sobre Serviços de Saúde/estatística & dados numéricos , Transfusão de Sangue/métodos
5.
J Trauma Acute Care Surg ; 89(2): 344-350, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32301878

RESUMO

BACKGROUND: Transfusion with stored whole blood (WB) is increasingly routine practice to resuscitate bleeding trauma patients. Storage of packed red blood cells (pRBC) results in multiple biochemical, structural, and metabolic changes, referred to as to the storage lesion that may mediate adverse effects associated with transfusion of older RBC units. These include increased hemolysis, oxidative stress, and accelerated scavenging of nitric oxide (NO). Whether similar changes occur to stored WB is unclear and are characterized in this study. METHODS: Ten WB units, in citrate-phosphate-dextrose, were purchased from the American Red Cross and changes in hemolysis (increased free hemoglobin, heme, and microparticles), oxidative stress indexed by redox cycling of peroxiredoxin-2 (Prx-2) and NO-scavenging kinetics were determined at different storage times until expiration. RESULTS: Microparticle number and free hemoglobin, but not heme, increased in a storage time-dependent manner. When normalized to the initial number of RBCs in stored WB units, hemolysis rates were similar to those reported for pRBCs. Prx-2 recycling kinetics were slower at expiration compared with earlier storage times. Rates of NO dioxygenation did not change with storage, but were decreased compared with freshly isolated RBCs. CONCLUSION: Storage of WB results in changes associated with the pRBC storage lesion but not for all parameters tested. The relative rate of hemolysis (indexed by free hemoglobin and microparticles) and oxidative stress was similar to that of pRBCs. However, the absolute level of hemolysis products were lower due to lower hematocrit of stored WB units. The clinical significance of these findings requires further investigation.


Assuntos
Preservação de Sangue/efeitos adversos , Eritrócitos/patologia , Eritrócitos/fisiologia , Preservação de Sangue/métodos , Dióxido de Carbono/sangue , Micropartículas Derivadas de Células/metabolismo , Citratos , Eritrócitos/metabolismo , Glucose , Hemoglobinas/metabolismo , Hemólise , Humanos , Óxido Nítrico/metabolismo , Oxirredução , Estresse Oxidativo , Oxigênio/sangue , Peroxirredoxinas/metabolismo
6.
Transfusion ; 60(5): 1032-1041, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32237236

RESUMO

BACKGROUND: Great deformability allows red blood cells (RBCs) to flow through narrow capillaries in tissues. A number of microfluidic devices with capillary-like microchannels have been developed to monitor storage-related impairment of RBC deformability during blood banking operations. This proof-of-concept study describes a new method to standardize and improve reproducibility of the RBC deformability measurements using one of these devices. STUDY DESIGN AND METHODS: The rate of RBC flow through the microfluidic capillary network of the microvascular analyzer (MVA) device made of polydimethylsiloxane was measured to assess RBC deformability. A suspension of microbeads in a solution of glycerol in phosphate-buffered saline was developed to be used as an internal flow rate reference alongside RBC samples in the same device. RBC deformability and other in vitro quality markers were assessed weekly in six leukoreduced RBC concentrates (RCCs) dispersed in saline-adenine-glucose-mannitol additive solution and stored over 42 days at 4°C. RESULTS: The use of flow reference reduced device-to-device measurement variability from 10% to 2%. Repeated-measure analysis using the generalized estimating equation (GEE) method showed a significant monotonic decrease in relative RBC flow rate with storage from Week 0. By the end of storage, relative RBC flow rate decreased by 22 ± 6% on average. CONCLUSIONS: The suspension of microbeads was successfully used as a flow reference to increase reproducibility of RBC deformability measurements using the MVA. Deformability results suggest an early and late aging phase for stored RCCs, with significant decreases between successive weeks suggesting a highly sensitive measurement method.


Assuntos
Deformação Eritrocítica/fisiologia , Eritrócitos/citologia , Eritrócitos/fisiologia , Dispositivos Lab-On-A-Chip/normas , Técnicas Analíticas Microfluídicas , Bancos de Sangue/métodos , Bancos de Sangue/normas , Velocidade do Fluxo Sanguíneo/fisiologia , Preservação de Sangue/efeitos adversos , Preservação de Sangue/métodos , Preservação de Sangue/normas , Criopreservação , Contagem de Eritrócitos/instrumentação , Contagem de Eritrócitos/métodos , Contagem de Eritrócitos/normas , Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , Citometria de Fluxo/normas , Hemólise , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Estudo de Prova de Conceito , Reprodutibilidade dos Testes , Fatores de Tempo
7.
Vox Sang ; 115(5): 388-394, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32166752

RESUMO

BACKGROUND: Red blood cell (RBC) units accumulate morphologic and metabolic lesions during storage before transfusion. Pyruvate-inosine-phosphate-adenine (PIPA) solutions (Rejuvesol, Biomet, Warsaw, IN) can be incubated with RBC units to mitigate storage lesions. This study proposes a PIPA treatment process, termed cold 'rejuvenation', using Rejuvesol as an adjunct additive solution, to prevent biomechanical storage lesions while avoiding the 1 h PIPA incubation required with standard PIPA treatment. We compared the efficacy of cold to standard 'rejuvenation' in improving metabolic lesions that occur during cold storage of RBCs, without altering function. METHODS: Twelve leucoreduced, A-positive RBC units were obtained. Each unit was aliquoted into either control (standard storage), washed (W), standard rejuvenation (SR) or cold rejuvenation (CR) groups, the latter two requiring washing. A volume-adjusted dose of Rejuvesol was instilled into the CR group upon receipt (Day 3). After 15 days of storage, p50, RBC deformability, in-bag haemolysis and mechanical fragility were analysed. 'Any treatment' is defined as W, SR and CR, with comparisons in reference to control. RESULTS: Higher p50s were seen in rejuvenated groups (>30 mmHg vs. <19 mmHg; P < 0·0001). Any treatment significantly increased elongation index (P = 0·034) but did not significantly increase in-bag haemolysis (P = 0·062). Mechanical fragility was not significantly different between groups (P = 0·055) at baseline, but the control (CTL) group was more fragile after 2 h in a cardiac bypass simulation than any treatment (P < 0·0001). CONCLUSIONS: This study demonstrates that rejuvenation (standard or cold) prevents the leftward p50 shift of storage lesions without detrimental effect on RBC deformity, in-bag haemolysis or mechanical fragility.


Assuntos
Preservação de Sangue/métodos , Temperatura Baixa , Eritrócitos/metabolismo , Adenina , Hemoglobinas/metabolismo , Hemólise , Humanos , Inosina , Oxigênio/sangue , Ácido Pirúvico , Soluções/química
8.
Transfusion ; 60(5): 1050-1059, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32187695

RESUMO

BACKGROUND: Our previous study showed that ultraviolet C (UVC) from xenon (Xe) flash without any photoreactive compounds inactivated bacteria in platelet concentrates (PCs) with less damage to platelets (PLTs) as compared with Xe flash containing ultraviolet A, ultraviolet B, and visible light. Here, we report a UVC irradiation system for PCs under flow conditions consisting of a flow path-irradiation sheet, a peristaltic pump, and a collection bag. STUDY DESIGN AND METHODS: Platelet concentrates containing Ringer's solution (R-PCs) inoculated with bacteria were injected into a flow path sheet using a peristaltic pump, being irradiated with UVC from Xe flash. The quality of the irradiated PCs containing platelet additive solution (PAS-PCs) was assessed based on PC variables, PLT surface markers, and aggregation ability. RESULTS: Streptococcus dysgalactiae (12 tests) and Escherichia coli (11) were all negative on bacterial culture, while Staphylococcus aureus (12) and Klebsiella pneumoniae (14) grew in one and two R-PCs, respectively. Bacillus cereus spores were inactivated in 7 of 12 R-PCs. PC variables became significantly different between irradiated and nonirradiated PAS-PCs. P-selectin, first procaspase-activating compound (PAC-1) binding, and phosphatidylserine increased by irradiation. Aggregability stimulated by adenosine diphosphate, collagen, or thromboxane A2 increased in the irradiated PAS-PCs, while that by thrombin became smaller compared with nonirradiated controls. CONCLUSION: This newly developed system inactivated bacteria including spores in R-PCs. PAS-PCs irradiated by this system retained acceptable in vitro quality and aggregability. Usage of a peristaltic pump instead of agitator during irradiation may enable this system to be directly combined with an apheresis blood cell separator.


Assuntos
Plaquetas/citologia , Preservação de Sangue , Desinfecção/instrumentação , Viabilidade Microbiana , Raios Ultravioleta , Xenônio/farmacologia , Bacillus cereus/efeitos dos fármacos , Bacillus cereus/fisiologia , Bacillus cereus/efeitos da radiação , Bactérias/efeitos dos fármacos , Bactérias/efeitos da radiação , Remoção de Componentes Sanguíneos , Plaquetas/efeitos dos fármacos , Plaquetas/efeitos da radiação , Preservação de Sangue/instrumentação , Preservação de Sangue/métodos , Segurança do Sangue/instrumentação , Segurança do Sangue/métodos , Desinfecção/métodos , Contaminação de Medicamentos/prevenção & controle , Escherichia coli/efeitos dos fármacos , Escherichia coli/fisiologia , Escherichia coli/efeitos da radiação , Humanos , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/fisiologia , Klebsiella pneumoniae/efeitos da radiação , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Viabilidade Microbiana/efeitos da radiação , Soluções para Preservação de Órgãos/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Agregação Plaquetária/fisiologia , Agregação Plaquetária/efeitos da radiação , Controle de Qualidade , Solução de Ringer/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Staphylococcus aureus/efeitos da radiação , Streptococcus/efeitos dos fármacos , Streptococcus/fisiologia , Streptococcus/efeitos da radiação
9.
Transfusion ; 60(5): 1042-1049, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32187700

RESUMO

BACKGROUND: Some jurisdictions require leukoreduction of cellular blood components. The only whole blood collection set with a platelet-saving filter uses citrate-phosphate-dextrose (CPD) as storage solution. Substituting CPD with citrate-phosphate-dextrose-adenine (CPDA-1) increases shelf life from 21 to 35 days. This would simplify prehospital and rural resupply and reduce wastage. We investigated in vitro quality and hemostatic properties of CPDA-1 whole blood leukoreduced with a platelet-saving filter. STUDY DESIGN AND METHODS: CPDA-1 whole blood was leukoreduced using a platelet-saving filter and stored 35 days. EDQM requirements, hematology, metabolic parameters, thromboelastography, light transmission aggregometry, fibrinogen, factor VIII, and interleukin-6 were measured on Days 0, 1, 14, 21, and 35 and compared to non-leukoreduced blood. RESULTS: All units met EDQM requirements. Leukoreduction yielded residual white blood cell count <1 × 106 and 87% platelet recovery on Day 1. It caused reduction in thromboelastography parameters, but not aggregometry response. No hemolysis >0.8% was observed. Factor VIII was higher on Day 35 in the leukoreduced group, 37.9 (95% CI: 26.0, 49.8) versus 13.8 (9.4, 18.2) IU/dL. In both groups, aggregation was significantly reduced by Day 14. Thromboelastography showed remaining platelet activity on Day 35, MA 46.9 (42.1, 51.7) in the leukoreduced and 44.3 (39.6, 49.0) mm in the non-leukoreduced group. Fibrinogen was within reference ranges at Day 35 (>2 g/dL). Interleukin-6 was not detectable. CONCLUSION: Leukoreducing CPDA-1 whole blood with a platelet-saving filter did not compromise hemostatic properties. We encourage development of a single bag CPDA-1 whole blood collection set with in-line platelet-saving filter.


Assuntos
Adenina/química , Preservação de Sangue/métodos , Coleta de Amostras Sanguíneas/métodos , Citratos/química , Temperatura Baixa , Glucose/química , Procedimentos de Redução de Leucócitos/métodos , Fosfatos/química , Adenina/farmacologia , Sangue/efeitos dos fármacos , Plaquetas/citologia , Plaquetas/efeitos dos fármacos , Preservação de Sangue/normas , Coleta de Amostras Sanguíneas/normas , Citratos/farmacologia , Filtração/métodos , Glucose/farmacologia , Hemólise/efeitos dos fármacos , Hemostasia/efeitos dos fármacos , Humanos , Técnicas In Vitro , Procedimentos de Redução de Leucócitos/normas , Fosfatos/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Contagem de Plaquetas , Controle de Qualidade , Refrigeração/métodos
10.
Transfusion ; 60(3): 613-621, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32017135

RESUMO

BACKGROUND: Cold (4°C)-stored platelets are currently under investigation for transfusion in bleeding patients. It is currently unknown how long cold-stored platelets can be stored for clinical applications. STUDY DESIGN AND METHODS: Twenty three subjects were recruited. Twenty-one subjects were available for in vivo assessment and received indium-111 radiolabeled, cold-stored platelets. We investigated 5- (n = 5), 10- (n = 6), 15- (n = 5), and 20-day-stored (n = 5) platelets and obtained samples for in vitro testing at baseline and after the designated storage time. Twenty three units were available for in vitro testing. Five- and 7-day (n = 5 each), room temperature (RT)-stored platelets served as the current clinical standard control. RESULTS: In vivo, we found a continuous decline in platelet recovery from 5 to 20 days. Platelet survival reached a low nadir after 10 days of storage. Ex vivo, we observed the maximum platelet αIIbß3 integrin response to collagen at 5 days of cold storage, and we saw a continuous decline thereafter. However, platelet integrin activation and mitochondrial membrane integrity were better preserved after 20 days at 4°C, compared to 5 days at RT. Platelet metabolic parameters suggest comparable results between 20-day cold-stored platelets and 5- or 7-day RT-stored platelets. CONCLUSION: In summary, we performed the first studies with extended, cold-stored, apheresis platelets in plasma for up to 20 days with a fresh comparator. Storing cold-stored platelets up to 20 days yields better results in vitro, but further studies in actively bleeding patients are needed to determine the best compromise between hemostatic efficacy and storage prolongation.


Assuntos
Plaquetas/fisiologia , Preservação de Sangue/métodos , Criopreservação , Humanos , Ativação Plaquetária/fisiologia , Transfusão de Plaquetas , Plaquetoferese/métodos , Fatores de Tempo
11.
Vox Sang ; 115(4): 263-274, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32090336

RESUMO

BACKGROUND AND OBJECTIVES: Freeze-dried plasma (FDP) has logistical advantages in terms of storage and reconstitution time compared to fresh-frozen plasma. In vitro studies show FDP to be equivalent to fresh-frozen plasma regarding coagulation and clotting capacities. FDP is used in an increasing number of countries. We wanted to evaluate the clinical effects of FDP in major haemorrhage compared to standard care. METHODS: MEDLINE, Embase, Central, Biosis Previews, WHO ICTRP, Clinical Trials and Open Grey were systematically searched from inception until September 2018, without language restriction. Studies were eligible if they examined haemorrhagic adult patients transfused with FDP. Our primary outcome was mortality. Two reviewers independently assessed studies for eligibility, extracted data and assessed bias. RESULTS: Nine studies were eligible for inclusion. Three studies had a comparison group: one was a randomized controlled trial and two were before and after comparisons. Six studies were uncontrolled. A total of 606 patients received FDP, while 72 patients received non-FDP transfusion. In total, five minor adverse effects were documented. Two studies compared FDP to fresh-frozen plasma and found no difference in 30-day mortality between the groups. The included studies were heterogenous and had several methodological weaknesses, such as no control group, missing data or no protocol. CONCLUSIONS: The available research does not document the clinical effects of FDP. We cannot recommend or discourage use of FDP in major haemorrhage on base of available research.


Assuntos
Preservação de Sangue/métodos , Hemorragia/terapia , Preservação de Sangue/efeitos adversos , Transfusão de Sangue/métodos , Liofilização/métodos , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto
12.
Transfusion ; 60(4): 786-798, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32104927

RESUMO

BACKGROUND: Blood transfusion is a lifesaving intervention for millions of recipients worldwide every year. Storing blood makes this possible but also promotes a series of alterations to the metabolism of the stored erythrocyte. It is unclear whether the metabolic storage lesion is correlated with clinically relevant outcomes and whether strategies aimed at improving the metabolic quality of stored units, such as hypoxic storage, ultimately improve performance in the transfused recipient. STUDY DESIGN AND METHODS: Twelve healthy donor volunteers were recruited in a two-arm cross-sectional study, in which each subject donated 2 units to be stored under standard (normoxic) or hypoxic conditions (Hemanext technology). End-of-storage measurements of hemolysis and autologous posttransfusion recovery (PTR) were correlated to metabolomics measurements at Days 0, 21, and 42. RESULTS: Hypoxic red blood cells (RBCs) showed superior PTR and comparable hemolysis to donor-paired standard units. Hypoxic storage improved energy and redox metabolism (glycolysis and 2,3-diphosphoglycerate), improved glutathione and methionine homeostasis, decreased purine oxidation and membrane lipid remodeling (free fatty acid levels, unsaturation and hydroxylation, acyl-carnitines). Intra- and extracellular metabolites in these pathways (including some dietary purines) showed significant correlations with PTR and hemolysis, though the degree of correlation was influenced by sulfur dioxide (SO2 ) levels. CONCLUSION: Hypoxic storage improves energy and redox metabolism of stored RBCs, which results in improved posttransfusion recoveries in healthy autologous recipients-a Food and Drug Administration gold standard of stored blood quality. In addition, we identified candidate metabolic predictors of PTR for RBCs stored under standard and hypoxic conditions.


Assuntos
Preservação de Sangue/métodos , Eritrócitos/metabolismo , Hipóxia , Adulto , Doadores de Sangue , Preservação de Sangue/normas , Transfusão de Sangue/normas , Estudos Transversais , Feminino , Voluntários Saudáveis , Hemólise , Humanos , Masculino , Recuperação de Função Fisiológica , Transplante Autólogo
13.
Ann Biol Clin (Paris) ; 78(1): 27-34, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32108577

RESUMO

Unfractionated heparin (UFH) is the main anticoagulante used in intensive care unit. The anticoagulant effect is monitored by activated partial thrombin time (aPTT) and anti-Xa activity (anti-Xa) measurement. However, delayed centrifugation induces platelet factor 4 (PF4) release and anti-Xa decrease. Several studies have concluded that aPTT and anti-Xa measurement should be performed within 2 hours in citrated anticoagulant but may be delayed longer in citrate theophylline adenosine and dypiridamol (CTAD) anticoagulant. The objective of this study was to compare the stability of both aPTT and anti-Xa in citrate and CTAD samples, and to determine the effect of delayed centrifugation on both aPTT, anti-Xa results, and PF4 release in citrate samples only. METHODS: aPTT and anti-Xa were measured in citrate and CTAD anticoagulant samples from 93 patients. Delayed centrifugation was performed in citrate samples from 31 additional patients, with hourly aPTT and anti-Xa measurement from 1 to 6 hours. In 14 of these last patients, PF4 release was also evaluated with Human CXCL4/PF4 Quantikine ELISA Kit. RESULTS: We observed a significant correlation between citrate and CTAD anticoagulant for aPTT (r2=0.94) and anti-Xa (r2=0.95). With Bland-Altman correlation, a minor bias was observed for anti-Xa (-0.025±0.041). Delayed centrifugation in citrated anticoagulant showed an excellent concordance from 1 to 4 hours for aPTT (-4.0±5.3 s) and anti-Xa (1.10-9±0.058 UI/mL) measurements. Moreover, PF4 release was not different between 1 hour (31.5±14.7 ng/mL) and 4 hours (33.8±11.8 ng/mL). CONCLUSION: We have demonstrated that anti-Xa measurement for unfractionated heparin should be done 4 hours in citrated plasma and that CTAD was not better than citrate. However, these initial findings require confirmation using other aPTT and calibrated anti-Xa assays.


Assuntos
Anticoagulantes/farmacologia , Coleta de Amostras Sanguíneas/métodos , Monitoramento de Medicamentos/métodos , Heparina/uso terapêutico , Adenosina/química , Adenosina/farmacologia , Anticoagulantes/química , Anticoagulantes/uso terapêutico , Testes de Coagulação Sanguínea/métodos , Preservação de Sangue/métodos , Centrifugação/métodos , Fracionamento Químico/métodos , Ácido Cítrico/química , Ácido Cítrico/farmacologia , Dipiridamol/química , Dipiridamol/farmacologia , Fator Xa/metabolismo , Inibidores do Fator Xa/análise , Inibidores do Fator Xa/sangue , Heparina/análise , Humanos , Tempo de Tromboplastina Parcial , Teofilina/química , Teofilina/farmacologia , Tempo de Trombina , Fatores de Tempo
14.
Cryobiology ; 92: 168-179, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31935377

RESUMO

In North America, red blood cells (RBCs) are currently cryopreserved in a solution of 40% glycerol. While glycerol is not inherently toxic to humans, it must be removed prior to transfusion to prevent intravascular osmotic hemolysis. The current deglycerolization procedure requires about 45 min per RBC unit. We previously presented predictions suggesting that glycerol could be safely removed from RBCs in less than 1 min. However, experimental evaluation of these methods resulted in much higher hemolysis than expected. Here we extend our previous study by considering both concentration-dependence of permeability and variability in permeability values in the mathematical optimization algorithm. To establish a model for the concentration dependence of glycerol permeability, we combined literature data with new measurements of permeability in the presence of 40% glycerol. To account for cell-dependent variability we scaled the concentration-dependent permeability model to define a permeability range for optimization. Methods designed using a range extending to 50% of the model-predicted glycerol permeability had a duration of less than 3 min and resulted in hemolysis ranging from 34% to 83%; hemolysis values were highly dependent on the blood donor. Extending the permeability range to 5% of the model-predicted value yielded a 30 min method that resulted in an average hemolysis of 12%. Our results suggest high variability in the glycerol permeability between donors and within a population of cells from the same donor. Such variability has broad implications for design of methods for equilibration of cells with cryoprotectants.


Assuntos
Preservação de Sangue/métodos , Permeabilidade da Membrana Celular/fisiologia , Crioprotetores/metabolismo , Eritrócitos/metabolismo , Glicerol/metabolismo , Hemólise/efeitos dos fármacos , Algoritmos , Criopreservação/métodos , Humanos , Osmose/fisiologia , Permeabilidade
15.
Cytotherapy ; 22(1): 44-51, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31883947

RESUMO

BACKGROUND AIMS: In 2016, specifications for both pre-cryopreserved and post-thawed cord blood were defined in the sixth edition of NetCord Foundation for the Accreditation of Cellular Therapy (FACT) Standards for Cord Blood Banks. However, for several experts, harmonization regarding flow cytometry analysis performed on post-thawed samples is still a concern. A multicenter study led by Héma-Québec aimed to provide scientific data to support the cord blood accreditation bodies such as NetCord FACT in the revision of standards. METHODS: Twelve cord blood units were processed for plasma and red cell reduction following standard operating procedures. Cord blood unit aliquots were shipped to eight participating centers under cryogenic conditions for analysis before and after standardization of protocol. Repeatability of stem cell count, measured pre- and post-intervention with the centers, was estimated using multilevel linear regression models with a heterogeneous compound symmetry correlation structure among repeated measures. RESULTS: Excellent inter-center repeatability was reported by each participant regarding the viable CD34+ cells concentration, and a successful improvement effect of protocol standardization was also observed. However, we observed that better control over the critical parameters of the protocol did not have a significant effect on improving homogeneity in the enumeration of CD45+ cells. CONCLUSIONS: The current practice in cord blood selection should now also consider relying on post-thaw CD34+ concentration, providing that all cord blood banks or outsourcing laboratories in charge of the analysis of post-thaw CB samples take into account the consensual recommendations provided in this work and adhere to a good-quality management system.


Assuntos
Antígenos CD34/análise , Preservação de Sangue/métodos , Sangue Fetal/citologia , Antígenos Comuns de Leucócito/análise , Células-Tronco/citologia , Bioensaio , Bancos de Sangue/métodos , Contagem de Células , Ensaio de Unidades Formadoras de Colônias , Criopreservação/métodos , Citometria de Fluxo/métodos , Humanos
16.
Vox Sang ; 115(2): 167-173, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31729059

RESUMO

BACKGROUND AND OBJECTIVES: Applying pathogen reduction technologies (PRT) to platelets can extend their shelf life from 5 to 7 days, but there have been few systematic studies of the repercussions of such technologies on outdate rates. MATERIAL AND METHODS: The benefits in terms of outdate rates of applying PRT to platelets are studied via a mathematical simulation. Specifically, statistical methods are used to determine the daily production rate needed to meet demand while not exceeding a maximum amount set as a result of limitations on donations and while assuring a minimum daily stock. RESULTS: The results show that a 2-day extension in the shelf life of platelet concentrates (PC) results in reductions in outdates ranging from 88·4% to 100% at the production centres analysed. It may be the case for budgetary reasons that only part of the PCs produced can be treated. This being so, we show that if the proportion treated per annum exceeds 25% the best option is to treat part of the output every day, otherwise, it is preferable to concentrate treatment on the last two production days of the week. CONCLUSIONS: Extending the shelf life of PC from five to seven days and setting up suitable production logistics can drastically reduce outdates at production centres. If only a part of all PCs is treated, the best choices are to distribute PRT overall production days or, if the percentage of PCs treated is very low, to apply PRT on the days preceding the weekend break.


Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Preservação de Sangue/economia , Preservação de Sangue/normas , Simulação por Computador , Humanos
17.
Transfusion ; 60(2): 378-390, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31756004

RESUMO

BACKGROUND: Studies suggest that washing red cell concentrates (RCCs) to remove soluble mediators and/or inflammatory components, such as extracellular vesicles (EVs), may lead to better clinical outcomes. This study tested the hypothesis that non-red blood cell (RBC) generated vesicles in RCC are potent inflammatory mediators in vitro and washing RCCs can reduce these vesicles and subsequently decrease the inflammatory activity of RCCs. STUDY DESIGN AND METHODS: Sixteen RCCs were pooled and split into four groups based on pre-wash storage time (Day 2 or 14; n = 4/group). Each group was tested 24 hours and 7 days post-wash. Characteristics of RBCs and EVs, cytokines released by monocytes, and expression of human umbilical vein endothelial cells (HUVECs) adhesion molecules were assessed. RESULTS: All RCCs meet quality standards for hemolysis, hematocrit, and hemoglobin. Washing did not remove residual platelets from RCCs but led to a significant reduction in platelet-EV count regardless of the group. Supernatant of RCCs washed on Day 14 and stored for 24 hours had significantly lower concentrations of RBC-EVs and white blood cell EVs compared to unwashed controls. Supernatant of unwashed RCCs showed higher production of inflammatory cytokines/chemokines MCP-1, IL-8, and TNF-α, and heightened expression of HUVEC VCAM-1, which were significantly reduced by washing. Spiking washed RCC supernatants with platelet-EVs showed significant increase in IL-8, MCP-1, VCAM-1, and E-selection in groups washed on Day 14. CONCLUSIONS: Platelet-EVs in RCCs are associated with pro-inflammatory activity. As washing significantly reduced RCC immunomodulatory activity, implementation of this process may improve transfusion outcomes.


Assuntos
Plaquetas/metabolismo , Eritrócitos/metabolismo , Preservação de Sangue/métodos , Quimiocina CCL2/metabolismo , Hemólise/fisiologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Interleucina-8/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Molécula 1 de Adesão de Célula Vascular/metabolismo
18.
Transfusion ; 60(2): 367-377, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31802514

RESUMO

BACKGROUND: The risk of bacterial contamination and the deterioration of platelet (PLT) quality limit the shelf-life of platelet concentrates (PCs). The INTERCEPT pathogen inactivation system reduces the risk of pathogen transmission by inhibiting nucleic acid replication using a combination of a photo-reactive compound and UVA illumination. The goal of this study was to investigate the effects the INTERCEPT system has on the PLT metabolome and metabolic activity. STUDY DESIGN AND METHODS: Paired units of buffy coat-derived PCs were generated using a pool and split strategy (n = 8). The paired PCs were either treated with the INTERCEPT system or left untreated. Samples were collected on Days 1, 2, 4, and 7 of storage. Ultra-performance chromatography coupled with time-of-flight mass spectrometry was used to analyze the extra- and intracellular metabolomes. Constraint-based metabolic modeling was then used to predict the metabolic activity of the stored PLTs. RESULTS: A relatively large number of metabolites in the extracellular environment were depleted during the processing steps of the INTERCEPT system, in particular, metabolites with hydrophobic functional groups, including acylcarnitines and lysophosphatidylcholines. In the intracellular environment, alterations in glucose and glycerophospholipid metabolism and decreased levels of 2-hydroxyglutarate were observed following the INTERCEPT treatment. Untargeted metabolomics analysis revealed residual amotosalen dimers present in the treated PCs. Systems-level analysis of PLT metabolism indicated that the INTERCEPT system does not have a significant impact on the PLT energy metabolism and nutrient utilization. CONCLUSIONS: The INTERCEPT system significantly alters the metabolome of the stored PCs without significantly influencing PLT energy metabolism.


Assuntos
Preservação de Sangue/métodos , Furocumarinas/farmacologia , Metabolômica/métodos , Raios Ultravioleta , Metabolismo Energético
19.
Immunohematology ; 36(4): 157-165, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33544622

RESUMO

CONCLUSIONS: Storage of dithiothreitol (DTT)-treated red blood cells (RBCs) leads to hemolysis. The aim of this study was to compare 0.1 M DTT with 0.2 M DTT treatment of RBCs and to share our experience of providing components to seven patients on daratumumab (DARA). This prospective, observational study included patients who required RBC transfusion within 6 months of DARA administration. All patients underwent a baseline serologic evaluation followed by a repeat evaluation after DARA administration. In addition, use of 0.1 M DTT was compared with 0.2 M DTT in terms of concordance of results, hemolysis with storage of treated RBCs, and ease of use. A total of 22 RBC requisitions were received for seven patients. Antibody screen was positive for one patient (anti-C) at baseline; it was panreactive for all patients after DARA. Concordance of results between the two concentrations was 98.5 percent. Laboratory personnel found results obtained with use of 0.1 M DTT-treated RBCs easy to interpret. Supernatant hemoglobin was found to be significantly greater for 0.2 M DTT-treated RBCs at the sixth day of storage. In conclusion, component administration to patients on DARA can be done without delay if adequate policies and procedures are in place. Use of 0.1 M DTT-pretreated RBCs can be used to avoid delay in transfusion and reduce the burden on the laboratory of weekly preparation of 0.2 M DTT-treated RBCs.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Preservação de Sangue/métodos , Ditiotreitol , Eritrócitos , Mieloma Múltiplo/sangue , Mieloma Múltiplo/tratamento farmacológico , Humanos , Estudos Prospectivos
20.
PLoS One ; 14(12): e0225137, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31809517

RESUMO

Gene expression profiling using blood samples is a valuable tool for biomarker discovery in clinical studies. Different whole blood RNA collection and processing methods are highly variable and might confound comparisons of results across studies. The main aim of the current study is to compare how blood storage, extraction methodologies, and the blood components themselves may influence gene expression profiling. Whole blood and peripheral blood mononuclear cell (PBMC) samples were collected in triplicate from five healthy donors. Whole blood was collected in RNAgard® and PAXgene® Blood RNA Tubes, as well as in collection tubes with anticoagulants such as dipotassium ethylenediaminetetraacetic acid (K2EDTA) and Acid Citrate Dextrose Solution A (ACD-A). PBMCs were separated using sodium citrate Cell Preparation Tubes (CPT™), FICOLL™, magnetic separation, and the LeukoLOCK™ methods. After blood collection, the LeukoLOCK™, K2EDTA and ACD-A blood tubes were shipped overnight using cold conditions and samples from the rest of the collection were immediately frozen with or without pre-processing. The RNA was isolated from whole blood and PBMCs using a total of 10 different experimental conditions employing several widely utilized RNA isolation methods. The RNA quality was assessed by RNA Integrity Number (RIN), which showed that all PBMC procedures had the highest RIN values when blood was stabilized in TRIzol® Reagent before RNA extraction. Initial data analysis showed that human blood stored and shipped at 4°C overnight performed equally well when checked for quality using RNA integrity number when compared to frozen stabilized blood. Comparisons within and across donor/method replicates showed signal-to-noise patterns which were not captured by RIN value alone. Pathway analysis using the top 1000 false discovery rate (FDR) corrected differentially expressed genes (DEGs) showed frozen vs. cold shipping conditions greatly impacted gene expression patterns in whole blood. However, the top 1000 FDR corrected DEGs from PBMCs preserved after frozen vs. cold shipping conditions (LeukoLOCK™ preserved in RNAlater®) revealed no significantly affected pathways. Our results provide novel insight into how RNA isolation, various storage, handling, and processing methodologies can influence RNA quality and apparent gene expression using blood samples. Careful consideration is necessary to avoid bias resulting from downstream processing. Better characterization of the effects of collection method idiosyncrasies will facilitate further research in understanding the effect of gene expression variability in human sample types.


Assuntos
Preservação de Sangue/métodos , Coleta de Amostras Sanguíneas/métodos , Perfilação da Expressão Gênica/métodos , Leucócitos Mononucleares/metabolismo , Transcriptoma , Anticoagulantes , Humanos
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