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1.
Vox Sang ; 114(8): 783-794, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31637738

RESUMO

INTRODUCTION: In vitro qualitative differences exist in red cell concentrates (RCCs) units processed from whole blood (WB) depending on the method of processing. Minimal literature exists on differences in processing and variability in quality data. Therefore, we collected information from blood manufacturers worldwide regarding (1) details of WB collection and processing used to produce RCCs and (2) quality parameters and testing as part of routine quality programmes. METHODS: A secure web-based survey was developed, refined after pilot data collection and distributed to blood centres. Descriptive analyses were performed. RESULTS: Data from ten blood centres in nine countries were collected. Six blood centres (60%) processed RCCs using the top-and-top (TAT) method which produces RCCs and plasma, and eight centres (80%) used the bottom-and-top (BAT) which additionally produces buffy coat platelets. Five of the centres used both processing methods; however, four favoured BAT processing. One centre utilized the Reveos automated system exclusively. All centres performed pre-storage leucoreduction. Other parameters demonstrated variability, including active cooling at collection, length of hold before processing, donor haemoglobin limits, acceptable collection weights, collection sets, time to leucoreduction, centrifugation speeds, extraction devices and maximum RCC shelf life. Quality marker testing also differed amongst blood centres. Trends towards higher RCC unit volume, haemolysis and residual leucoctyes were seen in the TAT compared with BAT processing across centres. CONCLUSION: Methods and parameters of WB processing and quality testing of RCCs differ amongst surveyed blood manufacturers. Further studies are needed to assess variations and to potentially improve methods and product quality.


Assuntos
Bancos de Sangue/normas , Preservação de Sangue/normas , Eritrócitos/citologia , Bancos de Sangue/métodos , Preservação de Sangue/métodos , Humanos , Inquéritos e Questionários
2.
Vox Sang ; 114(7): 701-710, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31392743

RESUMO

BACKGROUND AND OBJECTIVES: During the in vitro storage of red blood cells (RBCs), unfavourable changes (storage lesions) cause a rapid consumption of intracellular diphosphoglycerate. The latter deregulates the oxygen-haemoglobin binding potential, subsequently increasing oxygen saturation (SO2 ) and membrane degradation, transforming RBCs from biconcave discs to rigid spherical bodies (spheroechinocytes). Current laboratory techniques invasively extract RBC samples to assess the quality of red cell concentrate (RCC) units. Optical technologies could provide a means of assessing quality non-invasively. MATERIALS AND METHODS: A photoacoustic (PA) imaging technique was developed for acquiring the SO2 of blood bags non-invasively. Seven RCC units were monitored every 3-5 days until expiry (6 weeks). Measurements were validated against a conventional blood gas analyzer (BGA). Using an image flow cytometry assay, morphological profile trends were compared against the SO2 trends during blood bag storage. RESULTS: A strong correlation (r2  ≥ 0·95) was found when comparing temporal data between PA and BGA SO2 measurements. Inter-sample PA variability was found to be similar to that produced by BGA (±0·8%). A strong correlation was found to exist between the temporal changes in SO2 and relative spheroechinocyte population (0·79 ≤ r2  ≤ 0·97). CONCLUSION: This study suggests that PA imaging can non-invasively track the SO2 of stored RBCs non-invasively. By longitudinally monitoring the change in SO2 , it is possible to infer the effects of the storage lesion on RBC morphology. This non-invasive monitoring technique allows for the assessment of blood bags, without compromising sterility pre-transfusion.


Assuntos
Preservação de Sangue/normas , Técnicas Fotoacústicas/métodos , Preservação de Sangue/métodos , Eritrócitos/citologia , Estudos de Viabilidade , Citometria de Fluxo/métodos , Humanos
3.
Transfus Clin Biol ; 26(3): 164-170, 2019 Sep.
Artigo em Francês | MEDLINE | ID: mdl-31400933

RESUMO

BACKGROUND: The collection of granulocytes by apheresis requires volunteer donor stimulation by corticoids and the use of HES, a compound which is currently challenged by potential safety issues. Preparation of pooled granulocytes concentrates from whole blood buffy coats (PGC) represent an alternative to apheresis with a better benefit/risk for the donors. METHOD: Whole blood is collected in a bottom and top blood bag for buffy coat preparation. After centrifugation and separation, buffy coat are obtained. Twenty ABO matched buffy coats are selected for processing into one PGC. Four pools of five buffy coats were made, platelet additive solution is added to each pool, mixed gently and centrifuged. The red cell residue, supernatant and granulocyte rich layer are separated. Two granulocyte rich layers are pooled and added with 70mL of ABO matched plasma from the initial donations (=PGC10). The final PGC (=PGC20) is obtained by pooling two PGC10 into a platelet storage bag. Neutrophil content and in-vitro functionality are assessed at day of preparation (D1) and at expiry hour, 48 hours after collection (D2). RESULTS: On N=18, mean: Volume=408±4mL, 2.2*1010±0.24 neutrophils, Hematocrit=18%±3%, 4.7*1011platelets. Viability is well preserved: 95%±6% day of PGC preparation, 85%±7% after 24h of storage (D2). Functionality (ROS production measurement) is well preserved: 1.36±0.25 at D1 and 1.38±0.18 at D2. Expression and modulation of adhesion molecules after stimulation are normal at D1 and slightly decreased at D2 but still normal. CONCLUSIONS: PGC20 in vitro characteristics are in conformance with the EDQM guide (V19) and similar to apheresis for granulocytes content and hematocrit. The viability and two mean indicators which explore neutrophil function are well maintained during PGC preparation and after 24 hours of storage.


Assuntos
Buffy Coat/citologia , Separação Celular/métodos , Granulócitos , Sistema do Grupo Sanguíneo ABO/análise , Remoção de Componentes Sanguíneos , Doadores de Sangue , Plaquetas , Preservação de Sangue/instrumentação , Preservação de Sangue/métodos , Separação Celular/instrumentação , Centrifugação , Granulócitos/imunologia , Humanos , Masculino
5.
Blood Transfus ; 17(4): 263-273, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31385799

RESUMO

BACKGROUND: Sex hormone intake in blood donors may affect the quality of red blood cell (RBC) products via modulation of RBC function and predisposition to haemolysis during cold storage. The aims of this study were to evaluate the association between female sex hormone intake and RBC storage outcomes, and to examine possible mechanisms by which sex hormones interact with RBCs. MATERIALS AND METHODS: Sex hormone intake by race/ethnicity and menopausal status, and association analyses between hormone intake and donor scores of storage, osmotic or oxidative haemolysis, were evaluated in 6,636 female donors who participated in the National Heart, Lung and Blood Institute's RBC-Omics study. A calcium fluorophore, Fluo-3AM, was used to define RBC calcium influx in response to exogenous sex hormones or transient receptor potential cation (TRPC) channel drugs. RESULTS: Sex hormone intake was more prevalent in premenopausal women from all racial groups (18-31%) than in postmenopausal women (4-8%). Hormone intake was significantly (p<0.0001) associated with reduced storage haemolysis in all females, reduced osmotic haemolysis in postmenopausal donors (23.1±10.2% vs 26.8±12.0% in controls, p<0.001), and enhanced susceptibility to oxidative haemolysis in premenopausal women. In vitro, supraphysiological levels of progesterone (10 µmol/L), but not 17ß-oestradiol or testosterone, inhibited calcium influx into RBC and was associated with lower spontaneous haemolysis after 30 days of cold storage (0.95±0.18% vs 1.85±0.35% in controls, p<0.0001) or in response to a TRPC6 activator. CONCLUSIONS: Sex hormone intake in female donors is associated with changes in RBC predisposition to haemolysis. Menstrual status and the type of hormone preparation may contribute to differences in haemolytic responses of female RBCs to osmotic and oxidative stress. Progesterone modulates calcium influx into RBC via a mechanism that may involve interactions with membrane TRPC6 channels.


Assuntos
Doadores de Sangue , Eritrócitos/metabolismo , Hormônios Esteroides Gonadais/farmacologia , Hemólise , Progesterona/farmacologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Preservação de Sangue/métodos , Cálcio/metabolismo , Criopreservação/métodos , Eritrócitos/citologia , Eritrócitos/efeitos dos fármacos , Feminino , Hormônios Esteroides Gonadais/administração & dosagem , Hemólise/efeitos dos fármacos , Humanos , Menopausa , Pessoa de Meia-Idade , Progesterona/administração & dosagem , Adulto Jovem
6.
Blood Transfus ; 17(4): 289-295, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31385801

RESUMO

There are inter-individual differences in the quality of refrigerator-stored red blood cells (RBCs). Possible sources of these variations include nutritional and genetic factors. Glucose-6-phosphate dehydrogenase (G6PD) deficiency, the most common enzyme deficiency worldwide that affects the ability of RBCs to respond to oxidative stress, has been implicated as a genetic factor that affects the quality of stored RBCs. This review considers the literature concerning G6PD-deficient RBCs. It discusses RBC unit variables such as in vitro storage, 24-hour post-transfusion recovery (PTR), post-transfusion survival, and post-transfusion clinical outcomes.There are several differences in the in vitro storage characteristics between G6PD-deficient and G6PD-normal RBCs. Recent studies identified differences in the pathways related to glycolysis, purine metabolism, glutathione homeostasis, and fatty acid metabolism. In vitro experiments modelling the transfusion of G6PD-deficient RBCs, as well as autologous PTR studies in vivo, demonstrate increased haemolysis and decreased PTR, respectively, both indicators of a decrease in quality as compared to G6PD-normal RBCs. Finally, studies transfusing G6PD-deficient and G6PD-normal RBCs show that, in certain clinical settings, G6PD-deficient RBCs are associated with increased haemolysis.In summary, G6PD deficiency is associated with a decrease in the quality of RBCs after storage and its impact is often under-estimated. Understanding the underlying mechanisms by which G6PD deficiency affects RBC storage and transfusion outcomes may provide important clues to help optimise the future efficacy and safety of transfusions.


Assuntos
Doadores de Sangue , Preservação de Sangue/métodos , Transfusão de Eritrócitos/métodos , Eritrócitos/metabolismo , Glucosefosfato Desidrogenase/metabolismo , Animais , Seleção do Doador/métodos , Eritrócitos/citologia , Deficiência de Glucosefosfato Desidrogenase/metabolismo , Hemólise , Humanos , Resultado do Tratamento
7.
Blood Transfus ; 17(4): 321-330, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31385802

RESUMO

Reports from both adult and paediatric populations indicate that approximately two-thirds of platelet transfusions are used prophylactically to prevent bleeding, while the remaining one-third are used therapeutically to manage active bleeding. These two indications, prophylactic and therapeutic, serve two very distinct purposes and therefore will have two different functional requirements. In addition, disease aetiology in a given patient may require platelets with different functional characteristics. These characteristics can be derived from the various manufacturing methods used in platelet product production, including collection methods, processing methods, and storage options. The iterative combinations of manufacturing methods can result in a number of unique platelet products with different efficacy and safety profiles, which could potentially be used to benefit patient populations by meeting diverse clinical needs. In particular, cold storage of platelet products causes many biochemical and functional changes, of which the most notable characterised to date include increased haemostatic activity and altered expression of molecules inherent to platelet:leucocyte interactions. The in vivo consequences, both short- and long-term, of these molecular and cellular cold-storage-induced changes have yet to be clearly defined. Elucidation of these mechanisms would potentially reveal unique biologies that could be harnessed to provide more targeted therapies. To this end, in this new era of personalised medicine, perhaps there is an opportunity to provide individual patients with platelet products that are tailored to their clinical condition and the specific indication for transfusion.


Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Hemostasia , Animais , Plaquetas/imunologia , Plaquetas/metabolismo , Comunicação Celular , Temperatura Baixa , Células Endoteliais/citologia , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Humanos , Leucócitos/citologia , Leucócitos/imunologia , Leucócitos/metabolismo , Transfusão de Plaquetas/métodos , Medicina de Precisão/métodos
8.
Vox Sang ; 114(7): 711-720, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31373012

RESUMO

BACKGROUND AND OBJECTIVES: There is a growing concern for shortage in future blood supply, caused by a predicted decrease in eligible blood donors and simultaneous increase in recipients. Cryopreservation of split red blood cell units could increase stock supply by reducing waste of rare blood. This would be particularly useful in paediatric care where very small volumes often are transfused. The aim of this study was to develop a cryopreservation protocol for split units using the closed-system automated cell processor ACP215, as such protocols are currently missing. MATERIALS AND METHODS: Using a pool-and-split design, red blood cell units (N = 8) were glycerolized and frozen, either as standard volume reference units, or further divided into three smaller split units each. After thawing/deglycerolization, the supernatant of the smaller splits was reduced by additional centrifugation, and new SAGM was added to 60% haematocrit. The units were analysed for storage lesion effects up to ten days post-thawing. RESULTS: Haemolysis and extracellular potassium ion levels were lower in the split units than in the whole units from day three onwards. The metabolic parameters pH, ATP, glucose and lactate were also lower, though likely caused by lower additive solution pH rather than storage. CONCLUSION: Split units of red blood cells can be successfully cryopreserved using the ACP215. In addition to favourably low haemolysis and potassium, they also have higher haematocrit than corresponding whole units and enable involvement of fewer donors at repeated transfusions. These characteristics are all desirable features in paediatric care.


Assuntos
Preservação de Sangue/métodos , Criopreservação/métodos , Eritrócitos/metabolismo , Antígenos de Grupos Sanguíneos , Criança , Eritrócitos/efeitos dos fármacos , Congelamento/efeitos adversos , Glucose/metabolismo , Glicerol/farmacologia , Hemólise , Humanos , Ácido Láctico/metabolismo , Potássio/metabolismo
9.
Vox Sang ; 114(7): 694-700, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31286533

RESUMO

OBJECTIVE: Fresh whole blood (WB) has been used in military applications and cardiac surgery. We undertook a study of the coagulation properties of refrigerated WB stored for 21 days and compared them with the properties of reconstituted WB. STUDY DESIGN AND METHODS: Ten WB units were obtained from healthy volunteer donors and stored at 4 ± 2°C. Samples were obtained on Days 1, 2, 4, 6, 8, 10, 14 and 21 from the WB units. Ten units of reconstituted WB were prepared with a ratio of red cells, platelets and plasma of 1:1:1. Tests included complete blood count, electrolyte, routine coagulation, blood coagulation factor and thromboelastography. RESULTS: There was a progressive decline in Hb, WBC, PLT, sodium and coagulation factors but a progressive increase in APTT, PT and potassium in WB. The concentrations of factor (F)V and FVIII as well as FII and FX of WB were higher before Days 4, 2, 8 and 14, respectively, compared with the concentrations of reconstituted WB. The concentrations of FVII, FIX, FXI and FXII in WB were found to be equal to or higher than those in reconstituted WB throughout the course of 21 days. TEG variables in all WB units were normal throughout the course of 10 days. The mean PT and APTT of WB were lower than those of reconstituted WB before Days 14 and 10, respectively. CONCLUSION: This study suggests that the coagulation properties of refrigerated WB were equal to or superior to those of reconstituted WB for a minimum of 10 days.


Assuntos
Coagulação Sanguínea , Preservação de Sangue/métodos , Criopreservação/métodos , Preservação de Sangue/efeitos adversos , Humanos , Tromboelastografia/métodos , Tempo de Coagulação do Sangue Total/métodos
10.
Semin Thromb Hemost ; 45(5): 433-448, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31291676

RESUMO

Many preanalytical variables may affect the results of routine coagulation assays. While advances in laboratory instrumentation have partially addressed the laboratory's ability to recognize some of these variables, there remains an increased reliance on laboratory personnel to recognize the three potential areas where coagulation testing preanalytical issues may arise: (1) specimen collection (including patient selection), (2) specimen transportation and stability, and (3) specimen processing and storage. The purpose of this article is to identify the preanalytical variables associated with coagulation-related testing and provide laboratory practice recommendations in an effort to improve the quality of coagulation testing and accuracy of result reporting.


Assuntos
Testes de Coagulação Sanguínea/métodos , Preservação de Sangue/métodos , Humanos
11.
Transfus Clin Biol ; 26(3): 174-179, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31262629

RESUMO

Whole blood, that is blood that is not manufactured into its component red blood cells (RBC) plasma, and platelets (PLT) units, was the mainstay of transfusion for many years until it was discovered that the component parts of a blood donation could be stored under different conditions thereby optimizing the storage length of each product. The use of low anti-A and -B titer group O whole blood (LTOWB) has recently been rediscovered for use in massively bleeding trauma patients. Whole blood has several advantages over conventional component therapy for these patients, including simplifying the logistics of the resuscitation, being more concentrated than whole blood that is reconstituted from conventional components, and providing cold-stored PLTs, amongst other benefits. While randomized controlled trials to determine the efficacy of using LTOWB in the resuscitation of massively bleeding trauma patients are currently underway, retrospective data has shown that massively bleeding recipients of LTOWB with traumatic injury do not have worse outcomes compared to patients who received conventional components and, in some cases, recipients of LTOWB have more favourable outcomes. This paper will describe some of the advantages of using LTOWB and will discuss the emerging evidence for its use in massively bleeding patients.


Assuntos
Transfusão de Sangue/métodos , Hemorragia/terapia , Doença Aguda , Anticoagulantes/efeitos adversos , Tipagem e Reações Cruzadas Sanguíneas/métodos , Preservação de Sangue/métodos , Substitutos Sanguíneos/efeitos adversos , Substitutos Sanguíneos/uso terapêutico , Citratos/efeitos adversos , Soluções Cristaloides/efeitos adversos , Soluções Cristaloides/uso terapêutico , Serviços Médicos de Emergência , Glucose/efeitos adversos , Hemorragia/etiologia , Humanos , Procedimentos de Redução de Leucócitos , Ressuscitação , Choque Hemorrágico/etiologia , Choque Hemorrágico/mortalidade , Choque Hemorrágico/terapia , Reação Transfusional/prevenção & controle , Resultado do Tratamento , Ferimentos e Lesões/complicações
12.
J Thromb Thrombolysis ; 48(3): 430-438, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31292828

RESUMO

Upon platelet activation, inside-out signals synergistically induced by a variety of agonists and adhesion molecules can enhance the affinity of platelet main integrin, αIIbß3 to its ligands. Integrin ligation with fibrinogen induces potent signals which develop platelet function including aggregation, release and spreading of which platelet spreading is considered as a major early consequence of αIIbß3 outside-in signaling. Study presented here evaluated platelet spreading on fibrinogen matrix as a marker of platelet activation during storage. PRP-platelet concentrates were subjected to flowcytometry analysis and the expression levels of P-selectin, CD61, GPIbα and active conformation of the αIIbß3 (PAC-1 binding) were examined on day 0, 1, 3 and 5 post-storage. Concurrently platelet adhesion and spreading on fibrinogen matrix, glucose concentration and LDH activity were also determined at the same intervals. Results showed significant decreases in platelet spreading on fibrinogen matrix during storage. Spreading was dominant pattern of adhesion till the first day of storage. In 3 day-stored platelets, filopodial or lamellipodial formation was dominant event whereas 5 day-stored platelets simply adhered to fibrinogen with less protrusion formation and partially failed to spread. Compared to simple adhesion, reduction of platelet spreading was also more significantly correlated with the usual markers of platelet storage lesion including P-selectin (r = - 0.88; p < 0.0001) and GPIbα expression (r = 0.76; p = 0.0001), PAC-1 binding (r = 0.66; p = 0.001), glucose concentration and LDH activity. Taken together, we introduced platelet spreading on fibrinogen matrix as a reliable and sensitive marker of platelets functional activity during storage. As a valid marker which is directly and obviously relevant to the platelet functional capacities, the rapid reduction of platelet spreading during storage overshadows other markers of platelet storage lesion while raising serious question about the quality of 5 day-stored platelets.


Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Fibrinogênio/metabolismo , Ativação Plaquetária , Biomarcadores/metabolismo , Preservação de Sangue/normas , Adesão Celular , Humanos , Testes de Função Plaquetária
13.
Blood Transfus ; 17(4): 281-288, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31184577

RESUMO

Red blood cells (RBCs) have been historically regarded as a critical model to investigate cellular and oxidant stress biology. First of all, they are constantly exposed to oxidant stress, as their main function is to transport and deliver oxygen to tissues. Second, they are devoid of de novo protein synthesis capacity, which prevents RBCs from replacing irreversibly oxidised proteins with newly synthesised ones. As such, RBCs have evolved to (i) protect themselves from oxidant stress, in order to prevent oxidant damage from reactive species; (ii) repair oxidatively damaged proteins, through mechanisms that involve glutathione and one-carbon metabolism; (iii) destroy irreversibly oxidised proteins through proteasomal or protease-dependent degradation; and (iv) sacrifice membrane portions through mechanism of vesiculation. In this brief review we will summarize these processes and their relevance to RBC redox biology (within the context of blood storage), with a focus on how polymorphisms in RBC antioxidant responses could contribute to explaining the heterogeneity in the progression and severity of the RBC storage lesion that can be observed across the healthy donor population.


Assuntos
Preservação de Sangue/métodos , Eritrócitos/metabolismo , Estresse Oxidativo , Animais , Doadores de Sangue , Eritrócitos/citologia , Glucosefosfato Desidrogenase/metabolismo , Humanos , Metabolômica/métodos , Modelos Moleculares
15.
Clin Lab ; 65(5)2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31115238

RESUMO

BACKGROUND: Determination and quantification of serum free light chains (FLC) is essential for the diagnosis and monitoring of patients with monoclonal gammopathies. Currently, the Freelite assay (The Binding Site, United Kingdom) and the N Latex FLC assay (Siemens Healthineers, Germany) are available for FLC determination. Data concerning stability of FLC following long-term storage are limited. Therefore, the aim of the present study is to investigate the stability of FLC in frozen samples determined by the N latex FLC assay. METHODS: One hundred eighty-eight serum samples from 47 patients with monoclonal gammopathies were analyzed at the beginning and after long-term storage (-20°C, 193 - 568 days). Serum free light chain kappa (κFLC) and lambda (λFLC) concentrations were measured, and the corresponding kappa/lambda (κ/λ FLC) ratios were calculated. All samples were analyzed with the N latex FLC assay on a single BNII System. RESULTS: Comparison analyses between fresh and stored samples revealed very strong correlations for the determination of κFLC (r = 0.993), λFLC (r = 0.998) and the calculated κ/λ FLC ratio (r = 0.997). Median percentage changes of κFLC (-0.5%) and λFLC (-0.8%) measurements and the calculated κ/λ FLC ratios (-3.7%) were within the calculated analytical imprecision of the FLC assay. Changes of κFLC (r = 0.17, p = 0.02) and λFLC (r = 0.28, p < 0.01) concentrations and of the κ/λ FLC ratio (r = 0.18, p = 0.01) over time were very weak, but statistically significant. CONCLUSIONS: Serum free light chains in serum samples are sufficiently stable following long-term frozen storage and are therefore suitable to be measured with the N Latex FLC assay.


Assuntos
Preservação de Sangue/métodos , Criopreservação/métodos , Cadeias Leves de Imunoglobulina/sangue , Cadeias kappa de Imunoglobulina/sangue , Cadeias lambda de Imunoglobulina/sangue , Paraproteinemias/sangue , Humanos , Cadeias Leves de Imunoglobulina/química , Cadeias kappa de Imunoglobulina/química , Cadeias lambda de Imunoglobulina/química , Látex/química , Paraproteinemias/diagnóstico , Estabilidade Proteica , Reprodutibilidade dos Testes , Fatores de Tempo
16.
Vox Sang ; 114(5): 478-486, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31045253

RESUMO

BACKGROUND AND OBJECTIVES: Bacterial contamination of red blood cells (RBC) remains a rare but serious clinical concern. Despite the low temperature storage of RBC, some bacteria can proliferate. The impact of RBC additive solutions (AS), manufacturing method or donor sex on bacterial growth/survival in RBC was addressed in this pilot study. MATERIALS AND METHODS: Using a partial pool-and-split design, bacterial growth/survival was assessed in intentionally inoculated RBC, manufactured separately from male and female donors using three different manufacturing methods (two whole blood [WB] filtration methods; one RBC filtration method), and resuspended in one of four AS: SAGM, PAGGSM, AS-1 or AS-3. At the beginning of storage, RBC were inoculated with 10 CFU/ml of either Klebsiella pneumoniae, Staphylococcus epidermidis, Yersinia enterocolitica or Propionibacterium acnes. Manufacturing, inoculation, storage (until day 42) and monitoring of bacterial growth were conducted at two sites: Canadian Blood Services and Héma-Québec. RESULTS: Yersinia enterocolitica was the only bacterium that proliferated during storage at both sites. RBC tested at Canadian Blood Services had higher bacterial concentrations than those at Héma-Québec (P = 0·0044). At Héma-Québec, where two different manufacturing methods were used, Y. enterocolitica reached significantly higher bacterial concentrations in AS-3 RBC (WB filtration method) compared to units prepared in the other three AS (RBC filtration method; P < 0·05). Bacterial survival/growth dependent on donor sex was not uniformly noted. CONCLUSION: Only one of four bacteria grew under RBC storage conditions. The results indicate that RBC manufacturing variables, rather than AS or donor sex, affect bacterial growth in RBC.


Assuntos
Preservação de Sangue/métodos , Eritrócitos/microbiologia , Filtração/métodos , Canadá , Feminino , Humanos , Klebsiella pneumoniae , Masculino , Projetos Piloto , Propionibacterium acnes , Staphylococcus epidermidis , Yersinia enterocolitica
17.
J Thromb Thrombolysis ; 48(2): 277-283, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31098816

RESUMO

Unfractionated heparin (UFH) is the main anticoagulant 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 h 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. 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 h. In 14 of these last patients, PF4 release was also evaluated with Human CXCL4/PF4 Quantikine ELISA Kit. 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 h 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 h (31.5 ± 14.7 ng/ml) and 4 h (33.8 ± 11.8 ng/ml). We have demonstrated that anti-Xa measurement for unfractionated heparin should be done 4 h 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
Preservação de Sangue/métodos , Citratos/farmacologia , Dipiridamol/farmacologia , Monitoramento de Medicamentos/métodos , Heparina/farmacocinética , Anticoagulantes/farmacologia , Testes de Coagulação Sanguínea , Centrifugação , Heparina/uso terapêutico , Humanos , Inibidores de Fosfodiesterase/farmacologia , Teofilina , Fatores de Tempo
18.
Vox Sang ; 114(5): 487-494, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31056755

RESUMO

BACKGROUND AND OBJECTIVES: The 30-min rule has been used to maintain a core temperature (CT) of red-blood-cell (RBC) units below 10°C during transportation. We evaluated the utility of temperature-sensitive indicators (TIs) to monitor the surface temperature (ST) of RBC units and to explore whether TIs can help with compliance with the 30-min rule by extrapolating or correlating temperature change with time. MATERIALS AND METHODS: Two US FDA-approved TIs, Safe-T-Vue 10 (STV10; Temptime Corporation, Morris Plains, NJ, USA) and Timestrip Blood Temp 10 (BT10; Timestrip UK Ltd, Cambridge, UK), were attached to 50 RBC units. After issue, their colour change indicating 10°C was monitored, and temperature excursions were measured by standard reading. In additional 18 RBC units, both ST and CT were monitored simultaneously. RESULTS: In 50 RBC units, 94% of STV10 and 100% of BT10 showed colour change indicating 10°C within 30 min; 4% of STV10 and 18% of BT10 showed it during transportation. The time for colour change indicating 10°C differed significantly between STV10 and BT10 (19·0 vs. 5·6 min, P < 0·001). In additional 18 RBC units, 83·3% of STV10, 100% of BT10 and 88·9% of CT reached 10°C within 30 min, and the time for colour change indicating 10°C was 24·4 min in STV10, 14·6 min in BT 10 and 24·2 min in CT (P < 0·001). CONCLUSION: In two TIs, the time for colour change indicating 10°C varied considerably. To enhance the utility of TIs, further improvement and standardization would be needed.


Assuntos
Preservação de Sangue/normas , Eritrócitos , Temperatura Ambiente , Preservação de Sangue/métodos , Humanos , Indicadores e Reagentes
19.
J Trauma Acute Care Surg ; 87(1): 49-53, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31033893

RESUMO

BACKGROUND: Recent data demonstrate the safety of uncrossmatched cold-stored whole blood (WB) transfusion in pediatric trauma patients. The hemostatic capabilities of platelets within the cold-stored WB unit have been demonstrated via in vitro studies and animal models. However, platelet function has not been evaluated in pediatric recipients of cold-stored WB transfusions. METHODS: Injured children, 2 years or older and 10 kg or greater with hemorrhagic shock received up to 30 mL/kg of cold-stored, low titer (<50) anti-A and -B, leukoreduced, group O- WB during their initial resuscitation. Patients were included if (1) they received WB and no conventional platelets, and (2) platelet count and thromboelastography maximum amplitude were measured both before and after transfusion. These data and relevant clinical outcomes (mortality, intensive care unit length of stay [LOS], hospital LOS and ventilator days) were compared to a historical cohort of pediatric trauma patients who received uncrossmatched red blood cells (RBC) and conventional room temperature platelets. RESULTS: Twenty-two children were included in the study; 14 in the component cohort versus 8 in the WB cohort. Neither posttransfusion platelet count (129 × 109/L vs. 135 × 109/L) nor function (thromboelastography maximum amplitude, 59.5 mm vs. 60.2 mm) differed significantly between children receiving cold-stored platelets within the WB unit versus children who received conventional warm platelets. Median (interquartile range) weight-adjusted platelet transfusion volume in the historical cohort was 4.6 (2.5-7.7) mL/kg vs. 2.4 (1.3-4.0) mL/kg in the WB cohort (p = 0.03). There was no difference between groups in age, race, mechanism of injury, Injury Severity Score, vital signs, and severe traumatic brain injury (TBI). Outcomes, including mortality, intensive care unit LOS, hospital LOS, and ventilator days, were not significantly different between groups. CONCLUSION: No difference was seen in posttransfusion platelet number or function in severely injured children receiving cold-stored WB platelets as compared to those receiving conventional room temperature-stored platelets. Larger cohorts are required to confirm these findings. LEVEL OF EVIDENCE: Therapeutic, level IV.


Assuntos
Plaquetas/fisiologia , Preservação de Sangue , Transfusão de Sangue , Choque Hemorrágico/terapia , Ferimentos e Lesões/terapia , Adolescente , Preservação de Sangue/métodos , Transfusão de Sangue/métodos , Criança , Pré-Escolar , Feminino , Humanos , Tempo de Internação/estatística & dados numéricos , Masculino , Contagem de Plaquetas , Refrigeração , Choque Hemorrágico/sangue , Ferimentos e Lesões/sangue
20.
Drug Test Anal ; 11(8): 1231-1237, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30950199

RESUMO

The erythrocyte membrane is composed of a phospholipid bilayer, which is known to undergo physicochemical changes during storage at low temperatures. This study was conducted to identify marker phospholipids that indicate alteration during deep-frozen storage and to determine the amount of marker phospholipids. Our research suggested a method to detect phospholipids by profiling analysis of thermally injured red blood cells (RBCs) without protecting agents. Human blood was stored at -80°C for 72 days. The RBC membrane phospholipids were extracted through a modified Bligh and Dyer method. Six selected phospholipids were analyzed and quantified using liquid chromatography-tandem mass spectrometry, and an in vitro model system was developed. The intracellular level of N-nervonoyl-D-erythro-sphingosylphosphorylcholine significantly increased in the thermally injured RBCs, and multiple biomarker candidates were evaluated by profiling analysis and mass spectrometry technology for targeted metabolomics.


Assuntos
Preservação de Sangue/métodos , Criopreservação/métodos , Eritrócitos/química , Fosfolipídeos/análise , Membrana Eritrocítica/química , Eritrócitos/citologia , Humanos , Fosforilcolina/análogos & derivados , Fosforilcolina/análise , Esfingosina/análogos & derivados , Esfingosina/análise
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