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1.
Transfusion ; 61 Suppl 1: S22-S31, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34269432

RESUMO

BACKGROUND: Civilian and military guidelines recommend early balanced transfusion to patients with life-threatening bleeding. Low titer group O whole blood was introduced as the primary blood product for resuscitation of massive hemorrhage at Haukeland University Hospital, Bergen, Norway, in December 2017. In this report, we describe the whole blood program and present results from the first years of routine use. STUDY DESIGN AND METHODS: Patients who received whole blood from December 2017 to April 2020 were included in our quality registry for massive transfusions. Post-transfusion blood samples were collected to analyze isohemagglutinin (anti-A/-B) and hemolysis markers. Administration of other blood products, transfusion reactions, and patient survival (days 1 and 30) were recorded. User experiences were surveyed for both clinical and laboratory staff. RESULTS: Two hundred and five patients (64% male and 36% female) received 836 units in 226 transfusion episodes. Patients received a mean of 3.7 units (range 1-35) in each transfusion episode. The main indications for transfusion were trauma (26%), gastrointestinal (22%), cardiothoracic/vascular (18%), surgical (18%), obstetric (11%), and medical (5%) bleeding. There was no difference in survival between patients with blood type O when compared with non-group O. Haptoglobin level was lower in the transfusion episodes for non-O group patients, however no clinical hemolysis was reported. No patients had conclusive transfusion-associated adverse events. Both clinical and laboratory staff preferred whole blood to component therapy for massive transfusion. DISCUSSION: The experience from Haukeland University Hospital indicates that whole blood is feasible, safe, and effective for in-hospital treatment of bleeding.


Assuntos
Transfusão de Sangue , Ressuscitação , Reação Transfusional/etiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Transfusão de Sangue/métodos , Criança , Pré-Escolar , Feminino , Hemólise , Hospitais , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Noruega/epidemiologia , Ressuscitação/métodos , Reação Transfusional/sangue , Reação Transfusional/patologia , Adulto Jovem
2.
Transfusion ; 61 Suppl 1: S80-S89, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34269444

RESUMO

BACKGROUND: Collection of non-leukoreduced citrate-phosphate-dextrose-adenine (CPDA-1) whole blood is performed in walking blood banks. Blood collected under field conditions may have increased risk of bacterial contamination. This study was conducted to examine the effects of WBC reduction and storage temperature on growth of Escherichia coli (ATCC® 25922™) in CPDA-1 whole blood. METHODS: CPDA-1 whole blood of 450 ml from 10 group O donors was inoculated with E. coli. Two hours after inoculation, the test bags were leukoreduced with a platelet-sparing filter. The control bags remained unfiltered. Each whole blood bag was then split into three smaller bags for further storage at 2-6°C, 20-24°C, or 33-37°C. Bacterial growth was quantified immediately, 2 and 3 h after inoculation, on days 1, 3, 7, and 14 for all storage temperatures, and on days 21 and 35 for storage at 2-6°C. RESULTS: Whole blood was inoculated with a median of 19.5 (range 12.0-32.0) colony-forming units per ml (CFU/ml) E. coli. After leukoreduction, a median of 3.3 CFU/ml (range 0.0-33.3) E. coli remained. In the control arm, the WBCs phagocytized E. coli within 24 h at 20-24°C and 33-37°C in 9 of 10 bags. During storage at 2-6°C, a slow self-sterilization occurred over time with and without leukoreduction. CONCLUSIONS: Storage at 20-24°C and 33-37°C for up to 24 h before leukoreduction reduces the risk of E. coli-contamination in CPDA-1 whole blood. Subsequent storage at 2-6°C will further reduce the growth of E. coli.


Assuntos
Preservação de Sangue , Segurança do Sangue , Infecções por Escherichia coli/microbiologia , Escherichia coli/crescimento & desenvolvimento , Procedimentos de Redução de Leucócitos , Adenina/química , Preservação de Sangue/métodos , Citratos/química , Escherichia coli/isolamento & purificação , Glucose/química , Humanos , Temperatura
3.
Transfusion ; 61 Suppl 1: S101-S110, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34269459

RESUMO

BACKGROUND: There is a global increase in whole blood usage and at the same time, emerging pathogens give cause for pathogen reduction technology (PRT). The Mirasol PRT has shown promising results for plasma and platelet concentrate products. Treatment of whole blood with subsequent platelet survival and recovery analysis would be of global value. STUDY DESIGN AND METHODS: A two-arm, open-label laboratory study was performed with 40 whole blood collections in four groups: non-leukoreduced non-PRT-treated, non-leukoreduced PRT-treated, leukoreduced non-PRT-treated, and leukoreduced PRT-treated. Leukoreduction and/or PRT-treatment was performed on the day of collection, then all WB units were stored at room temperature for 24 h. Sampling was performed after hold-time and after 24-h storage in RT. If PRT-treatment or leukoreduction, samples were also taken subsequently after treatment. Thirteen healthy volunteer blood donors completed the in vivo study per protocol. All WB units were non-leukoreduced and PRT-treated. Radioactive labeling of platelets from RT-stored, PRT-treated whole blood, sampling of subjects, recovery, and survival calculations were performed according to the Biomedical Excellence for Safer Transfusion Collaborative protocol. RESULTS: In vitro characteristics show that PRT-treatment leads to increased levels of hemolysis, potassium, and lactate, while there are decreased levels of glucose, FVIII, and fibrinogen after 24 h of storage. All values are within requirements for WB. In vivo recovery and survival of platelets were 85.4% and 81.3% of untreated fresh control, respectively. CONCLUSIONS: PRT-treatment moderately reduces whole blood quality but is well within the limits of international guidelines. Recovery and survival of platelets are satisfactory after Mirasol treatment.


Assuntos
Plaquetas/efeitos dos fármacos , Plaquetas/efeitos da radiação , Segurança do Sangue , Fármacos Fotossensibilizantes/farmacologia , Riboflavina/farmacologia , Raios Ultravioleta , Plaquetas/citologia , Preservação de Sangue , Humanos , Testes de Função Plaquetária , Fatores de Tempo
4.
Stem Cell Res Ther ; 11(1): 351, 2020 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-32962723

RESUMO

BACKGROUND: Human platelet lysate (HPL) is emerging as the preferred xeno-free supplement for the expansion of mesenchymal stromal cells (MSCs) for bone tissue engineering (BTE) applications. Due to a growing demand, the need for standardization and scaling-up of HPL has been highlighted. However, the optimal storage time of the source material, i.e., outdated platelet concentrates (PCs), remains to be determined. The present study aimed to determine the optimal storage time of PCs in terms of the cytokine content and biological efficacy of HPL. METHODS: Donor-matched bone marrow (BMSCs) and adipose-derived MSCs (ASCs) expanded in HPL or fetal bovine serum (FBS) were characterized based on in vitro proliferation, immunophenotype, and multi-lineage differentiation. Osteogenic differentiation was assessed at early (gene expression), intermediate [alkaline phosphatase (ALP) activity], and terminal stages (mineralization). Using a multiplex immunoassay, the cytokine contents of HPLs produced from PCs stored for 1-9 months were screened and a preliminary threshold of 4 months was identified. Next, HPLs were produced from PCs stored for controlled durations of 0, 1, 2, 3, and 4 months, and their efficacy was compared in terms of cytokine content and BMSCs' proliferation and osteogenic differentiation. RESULTS: BMSCs and ASCs in both HPL and FBS demonstrated a characteristic immunophenotype and multi-lineage differentiation; osteogenic differentiation of BMSCs and ASCs was significantly enhanced in HPL vs. FBS. Multiplex network analysis of HPL revealed several interacting growth factors, chemokines, and inflammatory cytokines. Notably, stem cell growth factor (SCGF) was detected in high concentrations. A majority of cytokines were elevated in HPLs produced from PCs stored for ≤ 4 months vs. > 4 months. However, no further differences in PC storage times between 0 and 4 months were identified in terms of HPLs' cytokine content or their effects on the proliferation, ALP activity, and mineralization of BMSCs from multiple donors. CONCLUSIONS: MSCs expanded in HPL demonstrate enhanced osteogenic differentiation, albeit with considerable donor variation. HPLs produced from outdated PCs stored for up to 4 months efficiently supported the proliferation and osteogenic differentiation of MSCs. These findings may facilitate the standardization and scaling-up of HPL from outdated PCs for BTE applications.


Assuntos
Plaquetas , Células-Tronco Mesenquimais , Osteogênese , Engenharia Tecidual , Técnicas de Cultura de Células , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , Manejo de Espécimes , Fatores de Tempo
5.
Anesthesiology ; 133(6): 1173-1183, 2020 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-32902572

RESUMO

BACKGROUND: This pilot trial focused on feasibility and safety to provide preliminary data to evaluate the hemostatic potential of cold-stored platelets (2° to 6°C) compared with standard room temperature-stored platelets (20° to 24°C) in adult patients undergoing complex cardiothoracic surgery. This study aimed to assess feasibility and to provide information for future pivotal trials. METHODS: A single center two-stage exploratory pilot study was performed on adult patients undergoing elective or semiurgent complex cardiothoracic surgery. In stage I, a two-armed randomized trial, platelets stored up to 7 days in the cold were compared with those stored at room temperature. In the subsequent single-arm stage II, cold storage time was extended to 8 to 14 days. The primary outcome was clinical effect measured by chest drain output. Secondary outcomes were platelet function measured by multiple electrode impedance aggregometry, total blood usage, immediate and long-term (28 days) adverse events, length of stay in intensive care, and mortality. RESULTS: In stage I, the median chest drain output was 720 ml (quartiles 485 to 1,170, n = 25) in patients transfused with room temperature-stored platelets and 645 ml (quartiles 460 to 800, n = 25) in patients transfused with cold-stored platelets. No significant difference was observed. The difference in medians between the room temperature- and cold-stored up to 7 days arm was 75 ml (95% CI, -220, 425). In stage II, the median chest drain output was 690 ml (500 to 1,880, n = 15). The difference in medians between the room temperature arm and the nonconcurrent cold-stored 8 to 14 days arm was 30 ml (95% CI, -1,040, 355). Platelet aggregation in vitro increased after transfusion in both the room temperature- and cold-stored platelet study arms. Total blood usage, number of adverse events, length of stay in intensive care, and mortality were comparable among patients receiving cold-stored and room temperature-stored platelets. CONCLUSIONS: This pilot trial supports the feasibility of platelets stored cold for up to 14 days and provides critical guidance for future pivotal trials in high-risk cardiothoracic bleeding patients.


Assuntos
Plaquetas/fisiologia , Preservação de Sangue/métodos , Procedimentos Cirúrgicos Cardíacos , Criopreservação/métodos , Transfusão de Plaquetas , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Viabilidade , Feminino , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Agregação Plaquetária/fisiologia , Temperatura , Fatores de Tempo
6.
Transfusion ; 60(7): 1544-1551, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32319702

RESUMO

BACKGROUND: Increasing numbers of emergency medical service agencies and hospitals are developing the capability to administer blood products to patients with hemorrhagic shock. Cold-stored whole blood (WB) is the only single product available to prehospital providers who aim to deliver a balanced resuscitation strategy. However, there are no data on the safety and in vitro characteristics of prehospital stored WB. This study aimed to describe the effects on in vitro quality of storing WB at remote helicopter bases in thermal insulating containers. STUDY DESIGN AND METHODS: We conducted a two-armed single-center study. Twenty units (test) were stored in airtight thermal insulating containers, and 20 units (controls) were stored according to routine procedures in the Haukeland University Hospital Blood Bank. Storage conditions were continuously monitored during emergency medical services missions and throughout remote and blood bank storage. Hematologic and metabolic variables, viscoelastic properties, and platelet (PLT) aggregation were measured on Days 1, 8, 14, and 21. RESULTS: Storage conditions complied with the EU guidelines throughout remote and in-hospital storage for 21 days. There were no significant differences in PLT aggregation, viscoelastic properties, and hematology variables between the two groups. Minor significantly lower pH, glucose, and base excess and higher lactate were observed after storage in airtight containers. CONCLUSION: Forward cold storage of WB is safe and complies with EU standards. No difference is observed in hemostatic properties. Minor differences in metabolic variables may be related to the anaerobic conditions within the thermal box.


Assuntos
Resgate Aéreo , Glicemia/metabolismo , Plaquetas/metabolismo , Preservação de Sangue , Agregação Plaquetária , Plaquetas/citologia , Feminino , Humanos , Concentração de Íons de Hidrogênio , Masculino , Estudos Prospectivos , Fatores de Tempo
7.
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
8.
Transfusion ; 59(8): 2652-2661, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31150571

RESUMO

BACKGROUND: Cold storage of platelets may extend shelf life compared to room temperature storage. This study aimed to investigate in vitro platelet quality and function in cold-stored and delayed-cold-stored nonagitated apheresis platelets in platelet additive solution during storage for 21 days. STUDY DESIGN AND METHODS: Ten double apheresis platelet concentrates in 37% plasma/63% PAS-IIIM were split into two groups; nonagitated 2 to 6°C storage (CSPs) and delayed cold storage (DCSPs) with 7 days agitated storage at 20-24°C followed by nonagitated cold storage for 14 additional days. Platelet count, metabolism, viscoelastic properties, and aggregation ability were measured on Days 1, 7, 14, and 21. RESULTS: All platelet units, both CSPs and DCSPs, complied with the EU guidelines throughout storage for 21 days. Swirling was not detectable after cold storage. Cold storage improved platelet function; however, DCSP on Day 7 showed poorer results compared to CSP. Cold storage slowed down metabolism, with lower lactate and higher glucose concentrations in the CSP compared to the DCSP throughout storage for 21 days. CONCLUSION: Cold storage of platelets improved platelet function in in vitro assays, even though delayed cold storage on Day 7 showed poorer results compared to continuous cold storage. This difference could be explained by accelerated metabolism and higher glucose consumption during the period of room temperature storage. Cold storage and delayed cold storage could ease inventory management. Further studies investigating the in vitro and clinical effects of cold-stored and delayed-cold-stored platelets are encouraged.


Assuntos
Plaquetas/metabolismo , Preservação de Sangue , Temperatura Baixa , Plaquetoferese , Plaquetas/citologia , Feminino , Humanos , Masculino , Testes de Função Plaquetária , Estudos Prospectivos , Fatores de Tempo
9.
Vaccine ; 36(16): 2213-2219, 2018 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-29548607

RESUMO

The 2009 pandemic H1N1 (A(H1N1)pdm09) virus had a highly divergent hemagglutinin (HA) compared to pre-2009 seasonal H1N1 strains. Most peoples were immunologically naïve to the A(H1N1)pdm09, although hospital workers were exposed early in the pandemic before pandemic vaccines became available. Here, we evaluated how pre-existing antibodies influence the induction of cross-functional HA stalk antibodies following A(H1N1)pdm09 vaccination. Fifty-six healthcare workers vaccinated with AS03 adjuvanted A(H1N1)pdm09 vaccine were chosen by their pre-vaccination priming status (primed HI titers ≥ 40 or unprimed < 40). We analyzed the HA head- and stalk-specific serum IgG subclasses at pre- and 21 days post-vaccination. We also assessed the functionality of the HA stalk-specific antibodies to neutralize virus and mediate antibody dependent cellular cytotoxicity (ADCC). Primed individuals had higher pre-existing HA head- and stalk-specific IgG1, as well as higher ADCC functionality of stalk antibodies. However, following vaccination with the adjuvanted pandemic vaccine, only the quantity of HA head specific IgG1 antibodies were significantly higher than in unprimed individuals. The priming status did not impact upon the cross-reactive HA stalk specific IgG antibodies or their ability to neutralize virus or induce ADCC post-vaccination. In conclusion, a single dose of AS03 adjuvanted pandemic vaccine elicited similar levels of functional antibodies in naïve and primed individuals. These findings are important for understanding the immunological factors that impact or modulate pandemic vaccine responses in humans.


Assuntos
Reações Cruzadas/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Memória Imunológica , Vírus da Influenza A Subtipo H1N1/imunologia , Vacinas contra Influenza/imunologia , Influenza Humana/imunologia , Influenza Humana/prevenção & controle , Adjuvantes Imunológicos , Adulto , Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Citotoxicidade Celular Dependente de Anticorpos/imunologia , Feminino , Pessoal de Saúde , Testes de Inibição da Hemaglutinação , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Vacinas contra Influenza/administração & dosagem , Masculino , Pessoa de Meia-Idade , Vigilância em Saúde Pública , Vacinação
10.
Sci Rep ; 7(1): 8725, 2017 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-28821815

RESUMO

Epac1 (Exchange protein directly activated by cAMP 1) limits fluid loss from the circulation by tightening the endothelial barrier. We show here that Epac1-/- mice, but not Epac2-/- mice, have prolonged bleeding time, suggesting that Epac1 may limit fluid loss also by restraining bleeding. The Epac1-/- mice had deficient in vitro secondary hemostasis. Quantitative comprehensive proteomics analysis revealed that Epac1-/- mouse platelets (thrombocytes) had unbalanced expression of key components of the glycoprotein Ib-IX-V (GPIb-IX-V) complex, with decrease of GP1bß and no change of GP1bα. This complex is critical for platelet adhesion under arterial shear conditions. Furthermore, Epac1-/- mice have reduced levels of plasma coagulation factors and fibrinogen, increased size of circulating platelets, increased megakaryocytes (the GP1bß level was decreased also in Epac1-/- bone marrow) and higher abundance of reticulated platelets. Viscoelastic measurement of clotting function revealed Epac1-/- mice with a dysfunction in the clotting process, which corresponds to reduced plasma levels of coagulation factors like factor XIII and fibrinogen. We propose that the observed platelet phenotype is due to deficient Epac1 activity during megakaryopoiesis and thrombopoiesis, and that the defects in blood clotting for Epac1-/- is connected to secondary hemostasis.


Assuntos
Plaquetas/metabolismo , Fatores de Troca do Nucleotídeo Guanina/deficiência , Hemorragia/sangue , Hemorragia/metabolismo , Complexo Glicoproteico GPIb-IX de Plaquetas/metabolismo , Difosfato de Adenosina/farmacologia , Animais , Fatores de Coagulação Sanguínea/metabolismo , Plaquetas/ultraestrutura , Tamanho Celular , Colágeno/farmacologia , Exocitose , Feto/metabolismo , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Fígado/embriologia , Megacariócitos/efeitos dos fármacos , Megacariócitos/metabolismo , Camundongos Endogâmicos C57BL , Selectina-P/metabolismo , Fenótipo , Trombina/farmacologia
11.
Transfusion ; 53 Suppl 1: 124S-130S, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23301964

RESUMO

The Blood Far Forward (BFF) research program was established to conduct blood product efficacy and safety studies, donor performance studies, and research on optimal training methods to improve the safety of blood collection and transfusion performed by Norwegian Naval Special Operation Commando soldiers. The use of intravenous fluids for volume replacement during hemorrhagic shock is controversial, but it is currently the standard of care. In the far-forward environment, large volume resuscitation for massive bleeding is a great challenge. Crystalloid and colloid solutions add weight and bulk to the medic's kit, require temperature sensitive storage, and should be warmed before infusion to prevent hypothermia. Excessive use of these solutions causes a dilutional coagulopathy, acidosis, and potentially increased inflammatory injury compared with blood products. Type-specific whole blood from an uninjured combat companion on the other hand is almost always available. It is warm, replaces intravascular volume, and provides oxygen delivery and hemostatic capacity to prevent or treat shock and coagulopathy. Whole blood may be ideal for the resuscitation of combat casualties with hemorrhagic shock. BFF program pilot studies on use of platelet-sparing leukoreduction filters, whole blood transport tolerance, donor performance, and autologous reinfusion of 24-hour ambient temperature stored whole blood have been performed and suggest the feasibility of expanding whole blood use in resuscitation. If successful, the BFF program will change tactics, techniques, and procedures with a new lifesaving capability.


Assuntos
Transfusão de Sangue/métodos , Hemorragia/terapia , Infusões Intravenosas/métodos , Medicina Militar/métodos , Militares/educação , Ressuscitação/métodos , Transfusão de Sangue/normas , Meio Ambiente , Hemorragia/etiologia , Humanos , Infusões Intravenosas/normas , Medicina Militar/educação , Medicina Militar/normas , Avaliação de Programas e Projetos de Saúde , Ressuscitação/educação , Ferimentos e Lesões/complicações
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