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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.
Anesthesiology ; 134(3): 395-404, 2021 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-33503656

RESUMO

BACKGROUND: Removal of cytokines, chemokines, and microvesicles from the supernatant of allogeneic erythrocytes may help mitigate adverse transfusion reactions. Blood bank-based washing procedures present logistical difficulties; therefore, we tested the hypothesis that on-demand bedside washing of allogeneic erythrocyte units is capable of removing soluble factors and is feasible in a clinical setting. METHODS: There were in vitro and prospective, observation cohort components to this a priori planned substudy evaluating bedside allogeneic erythrocyte washing, with a cell saver, during cardiac surgery. Laboratory data were collected from the first 75 washed units given to a subset of patients nested in the intervention arm of a parent clinical trial. Paired pre- and postwash samples from the blood unit bags were centrifuged. The supernatant was aspirated and frozen at -70°C, then batch-tested for cell-derived microvesicles, soluble CD40 ligand, chemokine ligand 5, and neutral lipids (all previously associated with transfusion reactions) and cell-free hemoglobin (possibly increased by washing). From the entire cohort randomized to the intervention arm of the trial, bedside washing was defined as feasible if at least 75% of prescribed units were washed per protocol. RESULTS: Paired data were available for 74 units. Washing reduced soluble CD40 ligand (median [interquartile range]; from 143 [1 to 338] ng/ml to zero), chemokine ligand 5 (from 1,314 [715 to 2,551] to 305 [179 to 488] ng/ml), and microvesicle numbers (from 6.90 [4.10 to 20.0] to 0.83 [0.33 to 2.80] × 106), while cell-free hemoglobin concentration increased from 72.6 (53.6 to 171.6) mg/dl to 210.5 (126.6 to 479.6) mg/dl (P < 0.0001 for each). There was no effect on neutral lipids. Bedside washing was determined as feasible for 80 of 81 patients (99%); overall, 293 of 314 (93%) units were washed per protocol. CONCLUSIONS: Bedside erythrocyte washing was clinically feasible and greatly reduced concentrations of soluble factors thought to be associated with transfusion-related adverse reactions, increasing concentrations of cell-free hemoglobin while maintaining acceptable (less than 0.8%) hemolysis.


Assuntos
Remoção de Componentes Sanguíneos/métodos , Quimiocinas , Citocinas , Transfusão de Eritrócitos/métodos , Eritrócitos/química , Reação Transfusional/prevenção & controle , Preservação de Sangue , Estudos de Coortes , Eritrócitos/citologia , Humanos , Sistemas Automatizados de Assistência Junto ao Leito , Estudos Prospectivos
3.
Cryobiology ; 98: 73-79, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33359645

RESUMO

Despite recent advances in biostabilization, clinical blood supplies still experience shortages and storage limitations for red blood cells (RBCs) have not yet been sufficiently addressed. Storing RBCs in a frozen or dried state is an appealing solution to address storage limitations, but many promising cryoprotectants, including the non-reducing sugar trehalose, are impermeant to mammalian cell membranes and cannot be utilized effectively using currently available compound-loading methods. We found that transient pore formation induced by ultrasound and microbubbles (sonoporation) offers an effective means of loading trehalose into RBCs to facilitate long-term storage in a frozen or desiccated state. The protective potential of trehalose loading was demonstrated by freezing processed RBCs at -1 °C/min to -80 °C, then either storing the cells at -80 °C or lyophilizing them. RBCs were either thawed or rehydrated after 42 days of storage and evaluated for membrane integrity and esterase activity to estimate recovery and cell viability. The intracellular concentration of trehalose reached 40 mM after sonoporation and over 95% of treated RBCs were recovered after loading. Loading of trehalose was sufficient to maintain RBC morphology and esterase activity in most cells during freezing (>90% RBC recovery) and to a lower degree after lyophilization and rehydration (>20% recovery). Combining sonoporation with an integrated fluidics device allowed for rapid loading of up to 70 mM trehalose into RBCs. These results demonstrate the potential of sonoporation-mediated trehalose loading to increase recovery of viable RBCs, which could lead to effective methods for long-term stabilization of RBCs.


Assuntos
Preservação de Sangue , Criopreservação , Eritrócitos , Trealose , Criopreservação/métodos , Crioprotetores , Humanos
4.
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
5.
Lancet Haematol ; 7(10): e756-e764, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32628911

RESUMO

The COVID-19 pandemic has major implications for blood transfusion. There are uncertain patterns of demand, and transfusion institutions need to plan for reductions in donations and loss of crucial staff because of sickness and public health restrictions. We systematically searched for relevant studies addressing the transfusion chain-from donor, through collection and processing, to patients-to provide a synthesis of the published literature and guidance during times of potential or actual shortage. A reduction in donor numbers has largely been matched by reductions in demand for transfusion. Contingency planning includes prioritisation policies for patients in the event of predicted shortage. A range of strategies maintain ongoing equitable access to blood for transfusion during the pandemic, in addition to providing new therapies such as convalescent plasma. Sharing experience and developing expert consensus on the basis of evolving publications will help transfusion services and hospitals in countries at different stages in the pandemic.


Assuntos
Betacoronavirus , Bancos de Sangue/estatística & dados numéricos , Doadores de Sangue/provisão & distribução , Transfusão de Sangue , Infecções por Coronavirus , Pandemias , Pneumonia Viral , Anticorpos Antivirais/uso terapêutico , Transtornos da Coagulação Sanguínea/etiologia , Transtornos da Coagulação Sanguínea/terapia , Preservação de Sangue , Segurança do Sangue , Transfusão de Sangue/estatística & dados numéricos , Infecções por Coronavirus/sangue , Infecções por Coronavirus/complicações , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/terapia , Seleção do Doador , Procedimentos Cirúrgicos Eletivos , Alocação de Recursos para a Atenção à Saúde , Política de Saúde , Necessidades e Demandas de Serviços de Saúde , Hemoglobinopatias/complicações , Hemoglobinopatias/terapia , Humanos , Imunização Passiva , Pandemias/prevenção & controle , Pneumonia Viral/sangue , Pneumonia Viral/complicações , Pneumonia Viral/prevenção & controle , Pneumonia Viral/terapia , Ensaios Clínicos Controlados Aleatórios como Assunto
8.
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
9.
Transfusion ; 60(8): 1778-1784, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32442333

RESUMO

BACKGROUND: The aim of this study was to estimate the number of blood donors during the COVID-19 incubation period across China. STUDY DESIGN AND METHODS: In this study, we developed a predictive model to estimate the number of blood donors during the COVID-19 incubation period among 34 provincial regions in China. Our main assumption was that blood donors of all ages in different regions have a stable blood donation intention and the same infection risk. RESULTS: First, we estimated the number of blood donors during the COVID-19 incubation period in Wuhan city, Hubei Province, and China, from December 31, 2019 to March 17, 2020. Second, we compared the number of blood donors during the COVID-19 incubation period in all provinces across China. In addition, we found that if all RBCs, plasma, and cryoprecipitation were stored in isolation until the 14th day, the potential risk of SARS-CoV-2 transmission through blood transfusion was reduced by at least 65.77% after the blood donor safely passed the COVID-19 incubation period. Moreover, if the detection of SARS-CoV-2 RNA was carried out on all platelets, the potential risk would be reduced by 77.48%. CONCLUSIONS: Although the risk is low, with the rapid spread of the COVID-19 and the appearance of alarmingly high infectivity and a high fatality rate, appropriate measures should be taken by health departments to ensure the safety of clinical blood.


Assuntos
Doadores de Sangue/estatística & dados numéricos , Segurança do Sangue/métodos , Transfusão de Sangue/normas , Infecções por Coronavirus/transmissão , Pneumonia Viral/transmissão , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , Doadores de Sangue/provisão & distribução , Preservação de Sangue , China , Infecções por Coronavirus/prevenção & controle , Humanos , Período de Incubação de Doenças Infecciosas , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Quarentena , RNA Viral/sangue
10.
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
11.
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
12.
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
13.
Transfusion ; 60(4): 806-814, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32159862

RESUMO

BACKGROUND: During platelet storage, there are extensive changes in cytoskeleton and phosphatidylserine exposure. The intrinsic mitochondrial pathway of apoptosis, activated in stored platelets, is a major mediator these changes. Cofilin-1 is an effector of actin reorganization. We examined the effect of cofilin-1 deficiency on cytoskeleton and phosphatidylserine exposure during storage and following activation of apoptosis. METHODS AND RESULTS: We assessed actin filaments by Alexa-647-phalloidin and phosphatidylserine exposure by fluorescein isothiocyanate-lactadherin by fluorescence microscopy. In fresh platelets, actin filaments are distributed in the subcortical region, and they do not express phosphatidylserine in the outer surface. In stored platelets, there is retraction of actin filaments from the subcortical region with increased phosphatidylserine expression. These changes are seen in 20% of platelets of 6 days old and increases further with storage. Treatment with ABT-737, which activates the mitochondrial apoptosis, induces similar cytoskeletal changes in actin filaments with increased phosphatidylserine. Cofilin-1 is activated in stored platelets as well as in ABT-737 treated platelets by dephosphorylation. In cofilin-1 deficient murine platelets actin filaments are abnormal and ABT-737 induces less phosphatidylserine. Despite these changes in vitro, platelet survival of cofilin-1 deficient platelets in mice was not significantly different from their wild-type controls. CONCLUSION: These results show that cofilin-1 plays a role in apoptosis-induced actin rearrangement and phosphatidylserine exposure during storage. Despite the defects in platelet cytoskeleton and phosphatidylserine exposure in cofilin-1-deficient platelets, the in vivo life span of platelets is similar to littermate controls, indicating multiple redundant pathways for the clearance of platelets in vivo.


Assuntos
Actinas/metabolismo , Plaquetas/citologia , Cofilina 1/fisiologia , Actinas/efeitos dos fármacos , Animais , Apoptose , Compostos de Bifenilo/farmacologia , Preservação de Sangue , Cofilina 1/deficiência , Citoesqueleto/metabolismo , Humanos , Camundongos , Nitrofenóis/farmacologia , Fosfatidilserinas/metabolismo , Piperazinas/farmacologia , Sulfonamidas/farmacologia
14.
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
15.
Vox Sang ; 115(5): 395-404, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32166810

RESUMO

BACKGROUND AND OBJECTIVES: Red-blood-cell (RBC) transfusion is associated with lung injury, which is further exacerbated by mechanical ventilation. Manufacturing methods of blood products differ globally and may play a role in the induction of pulmonary cell activation through alteration of the immunomodulatory property of the products. Here, the effect of different manufacturing methods on pulmonary cell activation was investigated in an in vitro model of mechanical ventilation. MATERIALS AND METHODS: Pulmonary type II cells were incubated with supernatant from fresh and old RBC products obtained via whole blood filtration (WBF), red cell filtration (RCF), apheresis-derived (AD) or whole blood-derived (WBD) methods. Lung cells were subjected to 25% stretch for 24 h. Controls were non-stretched or non-incubated cells. RESULTS: Fresh but not old AD products and WBF products induce lung cell production of pro-inflammatory cytokines and chemokines, which was not observed with WBD or RCF products. Effects were associated with an increased amount of platelet-derived vesicles and an increased thrombin-generating capacity. Mechanical stretching of lung cells induced more severe cell injury compared to un-stretched controls, including alterations in the cytoskeleton, which was further augmented by incubation with AD products. In all read-out parameters, RCF products seemed to induce less injury compared to the other products. CONCLUSIONS: Our findings show that manufacturing methods of RBC products impact pulmonary cell activation, which may be mediated by the generation of vesicles in the product. We suggest RBC manufacturing method may be an important factor in understanding the association between RBC transfusion and lung injury.


Assuntos
Preservação de Sangue , Transfusão de Eritrócitos/efeitos adversos , Inflamação , Pulmão/patologia , Citocinas , Eritrócitos , Humanos , Respiração Artificial , Trombina
16.
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
17.
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
18.
Transfusion ; 60(3): 513-523, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32064619

RESUMO

BACKGROUND: Red blood cell (RBC) transfusions result in the sequestration and metabolism of storage-damaged RBCs within the spleen and liver. These events are followed by increased plasma iron concentrations that can contribute to oxidant stress and cellular injury. We hypothesized that administration of a ferroportin inhibitor (FPN-INH) immediately after acute RBC exchange transfusion could attenuate posttransfusion circulatory compartment iron exposure, by retaining iron in spleen and hepatic macrophages. STUDY DESIGN AND METHODS: Donor guinea pig blood was leukoreduced, and RBCs were preserved at 4°C. Recipient guinea pigs (n = 5/group) were exchange transfused with donor RBCs after refrigerator preservation and dosed intravenously with a small-molecule FPN-INH. Groups included transfusion with vehicle (saline), 5 mg/kg or 25 mg/kg FPN-INH. A time course of RBC morphology, plasma non-transferrin-bound iron (NTBI) and plasma hemoglobin (Hb) were evaluated. End-study spleen, liver, and kidney organ iron levels, as well as renal tissue oxidation and injury, were measured acutely (24-hr after transfusion). RESULTS: RBC transfusion increased plasma NTBI, with maximal concentrations occurring 8 hours after transfusion. Posttransfusion iron accumulation resulted in tubule oxidation and acute kidney injury. FPN inhibition increased spleen and liver parenchymal/macrophage iron accumulation, but attenuated plasma NTBI, and subsequent renal tissue oxidation/injury. CONCLUSION: In situations of acute RBC transfusion, minimizing circulatory NTBI exposure by FPN inhibition may attenuate organ-specific adverse consequences of iron exposure.


Assuntos
Proteínas de Transporte de Cátions/metabolismo , Ferro/sangue , Animais , Preservação de Sangue , Proteínas de Transporte de Cátions/antagonistas & inibidores , Transfusão de Eritrócitos/métodos , Cobaias , Humanos , Masculino , Estresse Oxidativo/fisiologia
19.
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
20.
BMC Vet Res ; 16(1): 25, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000762

RESUMO

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.


Assuntos
Bancos de Sangue/normas , Plaquetas/fisiologia , Preservação de Sangue/veterinária , Cães/sangue , Animais , Plaquetas/metabolismo , Ativação Plaquetária , Agregação Plaquetária , Testes de Função Plaquetária/veterinária , Controle de Qualidade
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