RESUMO
BACKGROUND AND OBJECTIVES: According to European guidelines, the temperature of whole blood (WB) has to be maintained at 20-24°C until processing within 24 h, but in blood bank practice, WB is frequently held at temperatures between 18-25°C. We aimed to assess the impact of these small temperature deviations on the quality of the blood components. MATERIALS AND METHODS: After rapid cooling, 7 WB units were held overnight at 18°C and 8 units at 25°C, reflecting worst case holding conditions, and separated into a red cell concentrate (RCC), plasma and buffy coat (BC). RCCs were filtered at test temperature and stored for 42 days at 2-6°C. BCs were processed to single-BC platelet concentrates (sPC) and stored up to Day 8 at 20-24°C. RESULTS: After overnight hold at 18°C, 2,3-DPG in WB decreased by 34 ± 9%, while at 25°C the decrease was 82 ± 6%. Accordingly, the 2,3-DPG levels in the RCCs in the 25°C group were significantly lower than in the 18°C group (2·2 ± 1·4 vs. 10·4 ± 2·9 µmol/g Hb). RCCs and sPCs in the 25°C group showed higher initial lactate levels and lower pH compared to the 18°C group, but these differences levelled off at the end of storage. RCCs showed small differences in ATP levels and haemolysis. Plasma in both groups showed comparable Factor VIII:C levels. CONCLUSION: The temperature of WB during overnight hold strongly affects initial 2,3-DPG levels of RCCs and supports the maintenance of temperature limits between 20 and 24°C. Other in vitro effects of the temperature deviations were small and of no practical relevance.
Assuntos
Plaquetas/citologia , Preservação de Sangue , Eritrócitos/citologia , 2,3-Difosfoglicerato/análise , Trifosfato de Adenosina/metabolismo , Plaquetas/metabolismo , Eritrócitos/metabolismo , Hemólise , Humanos , Concentração de Íons de Hidrogênio , Ácido Láctico/análise , Temperatura , Fatores de TempoRESUMO
BACKGROUND AND OBJECTIVES: Blood operators routinely monitor the pH of apheresis platelets as a marker of the so-called storage lesion, which can result from manufacturing problems. It is also suspected that some donor characteristics can increase the risk of poor platelet storage. To explore this hypothesis, we analysed a large, multinational data set of quality control (QC) pH test results on apheresis platelets. MATERIALS AND METHODS: For the period between September 2011 and August 2014, seven blood operators in Canada, the USA, the Netherlands, the United Kingdom, France and Australia provided pH QC test results and donor characteristics on a total of 21,671 apheresis platelets. RESULTS: Some variations in pH distribution between blood operators were in part explained by differences in collection, processing and testing methods. Younger age and female gender were significantly associated with a pH value below the 10th percentile. Among donors who had two or more pH measurements (n = 3672), there was a strong correlation between pH results (r = 0·726; P < 0·0001). CONCLUSION: The strong intradonor correlation of pH measurements and the association between donor characteristics and pH results suggest that donor factors play a role in the quality of platelets.
Assuntos
Seleção do Doador , Plaquetoferese/normas , Controle de Qualidade , Adolescente , Adulto , Fatores Etários , Idoso , Feminino , Humanos , Concentração de Íons de Hidrogênio , Masculino , Pessoa de Meia-Idade , Plaquetoferese/métodos , Fatores Sexuais , Preservação de Tecido/normas , Adulto JovemRESUMO
BACKGROUND: For a clinical platelet (PLT) transfusion trial conducted in three countries, the production of PLT concentrates (PCs) that were pathogen inactivated with the Mirasol technology was set up and validated. While the Mirasol procedure is applied to an established PLT product, the PLT processing procedure still had to be modified to ensure a treated PC was of sufficient quality. Further, the effect of simulated transport conditions and the effect of ambient light on Mirasol-treated PCs was determined. STUDY DESIGN AND METHODS: Platelet concentrates in plasma were made from pooled buffy coats followed by Mirasol treatment. To mimic transport conditions, units were left unagitated for 6 h at room temperature. To mimic ambient light exposure, units were held unagitated for 4 h in direct fluorescent tube light. RESULTS: Measures had to be taken to allow 7-day storage of treated concentrates. In one site, PCs made from five buffy coats with >450 × 109 PLTs were removed from inventory. Another site went from five to four buffy coats per pool. Interruption of agitation for 6 h on day 3 did not induce meaningful changes in in vitro measures, even when stored up to 7 days. Exposure to ambient light for 4 h, either on day 3 or 6, had no effect on in vitro measures. CONCLUSION: The Mirasol pathogen inactivation process can be implemented in routine, but changes to current PLT processing methods might be needed. Transport conditions and 4-h-long ambient light exposure have no negative effect on the in vitro quality of Mirasol-treated PCs.
Assuntos
Plaquetas/efeitos dos fármacos , Riboflavina/farmacologia , Raios Ultravioleta , Plaquetas/efeitos da radiação , Preservação de Sangue/métodos , Humanos , Contagem de Plaquetas , Temperatura , Inativação de Vírus/efeitos dos fármacos , Inativação de Vírus/efeitos da radiaçãoRESUMO
BACKGROUND AND OBJECTIVES: The accumulation of non-polar lipids arachidonic acid, 5-hydroxyeicosatetraenoic acid (HETE), 12-HETE and 15-HETE during storage of transfusion products may play a role in the onset of transfusion-related acute lung injury (TRALI), a syndrome of respiratory distress after transfusion. MATERIALS AND METHODS: We investigated non-polar lipid accumulation in red blood cells (RBCs) stored for 42 days, plasma stored for 7 days at either 4 or 20°C and platelet (PLT) transfusion products stored for 7 days. Furthermore, we investigated whether transfusion of RBCs with increased levels of non-polar lipids induces TRALI in a 'two-hit' human volunteer model. All products were produced following Dutch Blood Bank protocols and are according to European standards. Non-polar lipids were measured with high-performance liquid chromotography followed by mass spectrometry. RESULTS: All non-polar lipids increased in RBCs after 21 days of storage compared to baseline. The non-polar lipid concentration in plasma increased significantly, and the increase was even more pronounced in products stored at 20°C. In platelets, baseline levels of 5-HETE and 15-HETE were higher than in RBCs or plasma. However, the non-polar lipids did not change significantly during storage of PLT products. Infusion of RBCs with increased levels of non-polar lipids did not induce TRALI in LPS-primed human volunteers. CONCLUSION: We conclude that non-polar lipids accumulate in RBC and plasma transfusion products and that accumulation is temperature dependent. Accumulation of non-polar lipids does not appear to explain the onset of TRALI (Dutch Trial Register - NTR4455).
Assuntos
Lesão Pulmonar Aguda/etiologia , Lipídeos/sangue , Reação Transfusional , Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico/sangue , Adolescente , Adulto , Ácido Araquidônico/sangue , Plaquetas/citologia , Plaquetas/metabolismo , Preservação de Sangue , Transfusão de Sangue Autóloga , Cromatografia Líquida de Alta Pressão , Eritrócitos/citologia , Eritrócitos/metabolismo , Humanos , Ácidos Hidroxieicosatetraenoicos/sangue , Lipopolissacarídeos/toxicidade , Masculino , Modelos Teóricos , Transfusão de Plaquetas/efeitos adversos , Sistema de Registros , Espectrometria de Massas em Tandem , Temperatura , Fatores de Tempo , Adulto JovemRESUMO
BACKGROUND AND OBJECTIVES: There are concerns about the haemostatic function of platelets stored in platelet additive solution (PAS). Aim of this study was to compare the haemostatic function of PAS-C-platelets to plasma-platelets in reconstituted whole blood. MATERIALS AND METHODS: In our experiment, whole blood was reconstituted with red blood cells, solvent-detergent (SD) plasma and either PAS-C-platelets or plasma-platelets (n = 7) in a physiological ratio. On storage days 2, 5, 8 and 13, the agonist-induced aggregation (multiple electrode aggregometry), clot formation (thromboelastography) and agonist-induced CD62P responsiveness (flow cytometry) were measured. RESULTS: Samples with PAS-C-platelets showed significantly lower aggregation than plasma-platelets when induced with adenosine diphosphate, -6 U (95% confidence interval: -8; -4) or thrombin receptor-activating protein, -15 U (-19; -10). Also when activated with collagen and ristocetin, the PAS-C-platelets showed less aggregation, although not statistically significant. All samples with PAS-C-platelets showed significantly lower agonist-induced CD62P responsiveness than samples with plasma-platelets. However, there was no difference regarding all TEG parameters. CONCLUSION: Our findings demonstrate that the function - aggregation and CD62P responsiveness - of PAS-C-platelets in reconstituted whole blood is inferior to that of plasma-platelets, which may have implications in the setting of massive transfusions.
Assuntos
Plaquetas/fisiologia , Preservação de Sangue , Hemostasia/fisiologia , Difosfato de Adenosina/farmacologia , Plaquetas/efeitos dos fármacos , Colágeno/farmacologia , Impedância Elétrica , Eritrócitos , Humanos , Selectina-P/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária , Ristocetina/farmacologia , TromboelastografiaRESUMO
BACKGROUND AND OBJECTIVES: Buffy coat-derived granulocytes have been described as an alternative to the apheresis product from donors pretreated with dexamethasone and granulocyte colony-stimulating factor (G-CSF). The latter is - dependent on the local and national settings - obtained following a demanding and time-consuming procedure, which is undesirable in critically ill septic patients. In contrast, buffy coat-derived products have a large volume and are often heavily contaminated with red cells and platelets. We developed a new pooled buffy coat-derived product with high purity and small volume, and performed a comprehensive functional characterization of these granulocytes. MATERIALS AND METHODS: We pooled ten buffy coats following the production of platelet concentrates. Saline 0·9% was added to decrease the viscosity and the product was split into plasma, red cells and a 'super' buffy coat. Functional data of the granulocytes were compared to those obtained with granulocytes from healthy controls and G-CSF/dexamethasone-pretreated donors. RESULTS: Buffy coat-derived granulocytes showed adhesion, chemotaxis, reactive oxygen species production, degranulation, NETosis and in vitro killing of Staphylococcus aureus, Escherichia coli and Aspergillus species comparable to control and G-CSF/dexamethasone-derived granulocytes. Candida killing was superior compared to G-CSF/dexamethasone-derived granulocytes. Immunophenotyping was normal; especially no signs of activation in the buffy coat-derived granulocytes were seen. Viability was reduced. Buffy coats are readily available in the regular blood production process and would take away the concerns around the apheresis product. CONCLUSION: The product described appears a promising alternative for transfusion purposes.
Assuntos
Buffy Coat/citologia , Dexametasona/farmacologia , Fator Estimulador de Colônias de Granulócitos/farmacologia , Granulócitos/efeitos dos fármacos , Adulto , Antígenos de Superfície/metabolismo , Remoção de Componentes Sanguíneos , Doadores de Sangue , Plaquetas/citologia , Adesão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Quimiotaxia/efeitos dos fármacos , Granulócitos/citologia , Granulócitos/imunologia , Granulócitos/metabolismo , Humanos , Imunofenotipagem , Contagem de Leucócitos , Masculino , NADPH Oxidases/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
BACKGROUND AND OBJECTIVES: Interventions to prevent and detect bacterial contamination of platelet concentrates (PCs) have reduced, but not eliminated the sepsis risk. Standardized bacterial strains are needed to validate detection and pathogen reduction technologies in PCs. Following the establishment of the First International Reference Repository of Platelet Transfusion-Relevant Bacterial Reference Strains (the 'repository'), the World Health Organization (WHO) Expert Committee on Biological Standardisation (ECBS) endorsed further repository expansion. MATERIALS AND METHODS: Sixteen bacterial strains, including the four repository strains, were distributed from the Paul-Ehrlich-Institut (PEI) to 14 laboratories in 10 countries for enumeration, identification and growth measurement on days 2, 4 and 7 after low spiking levels [10-25 colony-forming units (CFU)/PC bag]. Spore-forming (Bacillus cereusPEI-B-P-07-S, Bacillus thuringiensisPEI-B-P-57-S), Gram-negative (Enterobacter cloacaePEI-B-P-43, Morganella morganiiPEI-B-P-74, PEI-B-P-91, Proteus mirabilisPEI-B-P-55, Pseudomonas fluorescensPEI-B-P-77, Salmonella choleraesuisPEI-B-P-78, Serratia marcescensPEI-B-P-56) and Gram-positive (Staphylococcus aureusPEI-B-P-63, Streptococcus dysgalactiaePEI-B-P-71, Streptococcus bovisPEI-B-P-61) strains were evaluated. RESULTS: Bacterial viability was conserved after transport to the participating laboratories with one exception (M. morganiiPEI-B-P-74). All other strains showed moderate-to-excellent growth. Bacillus cereus, B. thuringiensis, E. coli, K. pneumoniae, P. fluorescens, S. marcescens, S. aureus and S. dysgalactiae grew to >106 CFU/ml by day 2. Enterobacter cloacae, P. mirabilis, S. epidermidis, S. bovis and S. pyogenes achieved >106 CFU/ml at day 4. Growth of S. choleraesuis was lower and highly variable. CONCLUSION: The WHO ECBS approved all bacterial strains (except M. morganiiPEI-B-P-74 and S. choleraesuisPEI-B-P-78) for repository enlargement. The strains were stable, suitable for spiking with low CFU numbers, and proliferation was independent of the PC donor.
Assuntos
Plaquetas/microbiologia , Segurança do Sangue/normas , Transfusão de Plaquetas , Bancos de Espécimes Biológicos , Escherichia coli/crescimento & desenvolvimento , Humanos , Klebsiella pneumoniae/crescimento & desenvolvimento , Padrões de Referência , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento , Organização Mundial da SaúdeRESUMO
BACKGROUND: Platelet transfusion therapy poses many challenges in veterinary clinical practice. Lack of readily available blood donors, short shelf-life, and inability to administer a sufficient number of platelets to meet a dog's transfusion need are the major difficulties encountered. Platelet additive solutions are already in use at American and European human blood banks, showing to be a realistic alternative. This study compares the in vitro platelet function in plasma, Composol, or SSP+ during storage for 13 days. Platelet rich plasma-platelet concentrate with 35% plasma and 65% platelet additive solutions (Composol or SSP+) and a control group (100% plasma) were prepared. Swirling, platelet count, blood gases, metabolic variables, platelet activation markers, and apoptosis markers were analyzed on days 1, 5, 9 and 13. RESULTS: Swirling was well preserved and pH was acceptable (> 6.2) during storage for all platelet additive solutions units until day 9. SSP + units showed more stable pH and metabolic variables until day 13. Platelets in plasma showed higher glucose consumption than in Composol or in SSP+. The platelet additive solutions units showed better platelet metabolism maintenance, reduced glucose consumption and lactate production. The apoptotic markers were still low for 9 days in platelet concentrates with platelet additive solutions, suggesting the possibility to extend the shelf life with the use of SSP+ or Composol. CONCLUSIONS: Our findings suggest that the uses of Composol and SSP+ in canine platelet concentrates are potential alternatives in veterinary blood banks.
Assuntos
Plaquetas , Preservação de Sangue/veterinária , Meios de Cultura , Cães/sangue , Animais , Plaquetas/fisiologia , Meios de Cultura/química , Estudos de Viabilidade , Contagem de Plaquetas , Soluções/químicaRESUMO
BACKGROUND AND OBJECTIVES: Pathogen inactivation technologies require continuous development for adjustment to different blood components and products. With Theraflex UV-Platelets, a system using shortwave ultraviolet C (UVC) light (254 nm), efficient mixing of platelet concentrates (PCs) during UVC treatment is essential to ensure homogeneous illumination of the blood components. In this study, we investigated the impact of increasing the agitation speed during UVC treatment on pathogen inactivation capacity and platelet quality. MATERIAL AND METHODS: The pathogen inactivation efficacy of UVC treatment was evaluated at two agitation speeds (110 vs. 180 rpm) using four different transfusion-relevant bacteria strains and three model viruses. Using a pool-and-split design, the in vitro quality of buffy coat-derived PCs stored in SSP+ additive solution for up to 7 days was assessed in UVC-treated PCs agitated at either 110 rpm (standard speed) or 180 rpm (increased speed) and in untreated controls. RESULTS: The higher agitation speed improved bacterial inactivation but did not influence viral inactivation. Metabolic activity (glucose consumption and lactate accumulation) in UVC-treated platelets was slightly higher than in untreated controls. Increases in parameters such as CD62P expression and annexin A5 binding indicated moderate activation of UVC-treated platelets. Quality variables for UVC-treated platelets agitated at standard vs. increased agitation speed were comparable. CONCLUSION: The mixing rate during illumination may be a process parameter for further development of UVC-based pathogen inactivation procedures for PLT concentrates.
Assuntos
Raios Ultravioleta , Anexina A5/metabolismo , Bactérias/efeitos da radiação , Plaquetas/metabolismo , Plaquetas/efeitos da radiação , Vírus da Diarreia Viral Bovina/fisiologia , Vírus da Diarreia Viral Bovina/efeitos da radiação , Herpesvirus Suídeo 1/fisiologia , Herpesvirus Suídeo 1/efeitos da radiação , Humanos , Selectina-P/metabolismo , Inativação de Vírus/efeitos da radiaçãoRESUMO
BACKGROUND AND OBJECTIVES: In mice, loss of sialic acid resulting in shedding of glycoprotein (GP) Ibα and GPV has been linked to platelet survival. The aim of this study was to determine whether loss of sialic acid and the GPIb-IX-V complex contributes to development of the platelet storage lesion (PSL) in human platelet concentrates (PCs). MATERIALS AND METHODS: PCs (stored in plasma (with or without Mirasol treatment); PAS-C or PAS-E) were stored at room temperature. Flow cytometry was used to monitor membrane expression of the GPIb-IX-V complex, CD62P, surface glycans and PS exposure. The functionality of stored platelets was determined employing aggregometry and ristocetin-induced VWF binding. RESULTS: Storage time of PCs in blood banks is limited to 7 days. During this time period, a minor but gradually increasing subpopulation of GPIbα-negative platelets was observed. Also, ristocetin-induced VWF binding was impaired in a small population of platelets. Mean surface expression of GPIbα and GPV remained stable until day 9, whereas CD62P expression increased; also a rapid decrease in ADP-induced aggregation was observed for PAS-C, PAS-E and Mirasol-treated PCs. Upon prolonged storage (>9 days), a slow decline in surface expression of GPIbα and GPV was observed; no major changes were observed in surface sialylation with the exception of Mirasol-treated platelets. CONCLUSION: In a small population of stored platelets, changes in GPIbα occur from day 2 onwards. Loss of sialic acid and subsequent shedding of GPIbα and GPV is not an early event during the development of the PSL.
Assuntos
Plaquetas/metabolismo , Complexo Glicoproteico GPIb-IX de Plaquetas/metabolismo , Plaquetas/citologia , Plaquetas/efeitos dos fármacos , Preservação de Sangue , Crioprotetores/farmacologia , Citometria de Fluxo , Humanos , Ácido N-Acetilneuramínico/metabolismo , Selectina-P/metabolismo , Ligação Proteica , Ristocetina/farmacologia , Fator de von Willebrand/química , Fator de von Willebrand/metabolismoRESUMO
BACKGROUND AND OBJECTIVES: The use of a functionally closed system (ACP215, Haemonetics) for (de)glycerolization of RBCs allows for prolonged post-thaw storage. Currently, glycerolization is followed by supernatant glycerol reduction before freezing. The aim of this study was to investigate the influence of supernatant glycerol reduction before freezing on the stability of thawed, deglycerolized RBCs during subsequent cold storage. MATERIALS AND METHODS: Leucoreduced RBCs were stored for 6 days at 2-6°C before glycerolization. The RBCs were pooled and split, and glycerol was added using the ACP215 to a final concentration of 40%. Units were either frozen as such (n = 4) or supernatant reduced before freezing (n = 4). After storage at -80°C, the units were thawed, deglycerolized and resuspended in SAGM. An additional sixteen units, frozen without supernatant reduction, were resuspended in either AS-3 (n = 8) or SAGM (n = 8) after deglycerolization. During cold storage (2-6°C), the red cells were analysed for their stability and in vitro quality. RESULTS: The freeze-thaw-wash recovery was comparable for both volume reduced and non-reduced units. During post-thaw storage in SAGM, non-glycerol reduced units showed significantly less potassium leakage and haemolysis and higher ATP levels. AS-3 strongly reduced haemolysis during post-thaw storage of non-glycerol reduced units: haemolysis remained below 0.8% for up to 28 days of storage. CONCLUSION: Omitting glycerol supernatant reduction before freezing simplifies the cryopreservation procedure and increases the stability and therefore the outdating period of thawed RBCs. This increases the practical applicability of cryopreserved RBCs in both civil (rare blood) and military blood transfusion practice.
Assuntos
Preservação de Sangue/métodos , Criopreservação/métodos , Crioprotetores/farmacologia , Eritrócitos/efeitos dos fármacos , Glicerol/farmacologia , HumanosRESUMO
Plastic blood bags improve the safety and effectiveness of blood component separation and storage. Progress towards optimal storage systems is driven by medical, scientific, business and environmental concerns and is limited by available materials, consumer acceptance and manufacturing and regulatory concerns. Blood bag manufacturers were invited to submit lists of the bags they manufacture. The lists were combined and sorted by planned use. The lists were analysed by experts to assess the degree to which the products attend to scientific problems. Specific issues addressed included the use of di-ethylhexyl phthalate (DEHP) as plasticizer for polyvinyl chloride (PVC) blood bags, the size, material and thickness of platelet bags, and the fracture resistance of plasma bags. Alternatives to DEHP for red blood cell (RBC) storage exist, but are mostly in a developmental stage. Plastic bags (DEHP-free, PVC-free) for platelet storage with better gas diffusion capabilities are widely available. Alternatives for plasma storage with better fracture resistance at low temperatures exist. Most RBC products are stored in DEHP-plasticized PVC as no fully satisfactory alternative exists that ensures adequate storage with low haemolysis. A variety of alternative platelet storage systems are available, but their significance - other than improved oxygen transport - is poorly understood. The necessity to remove DEHP from blood bags still needs to be determined.
Assuntos
Preservação de Sangue , Embalagem de Produtos , Plaquetas/química , Dietilexilftalato , Eritrócitos/química , Hemólise , Humanos , Ácidos Ftálicos/química , Plasma/química , Plastificantes/análise , Cloreto de Polivinila/químicaRESUMO
BACKGROUND AND OBJECTIVES: Semi-automatic separation devices can be used for the separation of centrifuged whole blood into leucoreduced red cell concentrate (LR-RCC), plasma and buffy coat (BC) and to make platelet concentrates (PC) from pooled BCs. To improve and to obtain a more uniform and standardized process, the CompoMat G5 (Fresenius) was implemented, a new-generation semi-automated device. MATERIALS AND METHODS: Uniform programs for WB separation and preparation of PCs were validated, using collection and pooling systems with CompoFlow (CF) closures, which can be automatically opened by the G5. Cell counts were performed and compared with historic data of blood components obtained with the formerly used Compomat G4 and Optipress II. After implementation, different adjustments were made to improve product quality. RESULTS: Leucoreduced red cell concentrates (280 ± 15 ml, 53 ± 5 g haemoglobin) and plasma (317 ± 16 ml) met European guidelines. BCs (48 ± 2 ml, 0·42 ± 0·05 l/l, 93 ± 25 × 10(9) platelets) contained a similar platelet (PLT) content as BC prepared before with the Compomat G4. A relatively high percentage (4-6%) of PCs (330 ± 17 ml, 330 ± 50 × 10(9) PLT, 0·12 ± 0·21 × 10(6) leucocytes) contained <250 × 10(9) PLT which was the subject of improvement studies. After implementation, RCC and BC discard decreased and workload was less. Operator complaints were also less frequent. CONCLUSION: The same high-quality blood components can be prepared by using the CompoMat G5 as previously with other semi-automated devices. Improvement was realized by automation of the opening process by the use of collection systems with CF closures, which led to a decrease in discarded units and workload.
Assuntos
Buffy Coat/citologia , Separação Celular/instrumentação , Transfusão de Componentes Sanguíneos , Plaquetas/citologia , Centrifugação , Eritrócitos/citologia , Humanos , Leucócitos/citologia , Plasma/citologiaRESUMO
Anaerobic Propionibacterium acnes and Staphylococcus saccharolyticus are frequently isolated during platelet screening with anaerobic culture methods. Although neither P. acnes nor S. saccharolyticus proliferates during platelet storage, both species survive well in this environment. This study was aimed at determining whether strains of P. acnes and/or S. saccharolyticus form surface-attached bacterial cell aggregates, known as biofilms, under platelet storage conditions. We report that these organisms are able to adhere to the inner surface of platelet containers in tight interaction with activated platelets.
Assuntos
Biofilmes , Plaquetas/microbiologia , Propionibacterium acnes/fisiologia , Staphylococcus/fisiologia , Aderência Bacteriana , Segurança do Sangue , HumanosRESUMO
BACKGROUND: Bacterial contamination of platelet concentrates (PCs) still remains a significant problem in transfusion with potential important clinical consequences, including death. The International Society of Blood Transfusion Working Party on Transfusion-Transmitted Infectious Diseases, Subgroup on Bacteria, organised an international study on Transfusion-Relevant Bacteria References to be used as a tool for development, validation and comparison of both bacterial screening and pathogen reduction methods. MATERIAL AND METHODS: Four Bacteria References (Staphylococcus epidermidis PEI-B-06, Streptococcus pyogenes PEI-B-20, Klebsiella pneumoniae PEI-B-08 and Escherichia coli PEI-B-19) were selected regarding their ability to proliferate to high counts in PCs and distributed anonymised to 14 laboratories in 10 countries for identification, enumeration and bacterial proliferation in PCs after low spiking (0·3 and 0·03 CFU/ml), to simulate contamination occurring during blood donation. RESULTS: Bacteria References were correctly identified in 98% of all 52 identifications. S. pyogenes and E. coli grew in PCs in 11 out of 12 laboratories, and K. pneumoniae and S. epidermidis replicated in all participating laboratories. The results of bacterial counts were very consistent between laboratories: the 95% confidence intervals were for S. epidermidis: 1·19-1·32 × 10(7) CFU/ml, S. pyogenes: 0·58-0·69 × 10(7) CFU/ml, K. pneumoniae: 18·71-20·26 × 10(7) CFU/ml and E. coli: 1·78-2·10 × 10(7) CFU/ml. CONCLUSION: The study was undertaken as a proof of principle with the aim to demonstrate (i) the quality, stability and suitability of the bacterial strains for low-titre spiking of blood components, (ii) the property of donor-independent proliferation in PCs, and (iii) their suitability for worldwide shipping of deep frozen, blinded pathogenic bacteria. These aims were successfully fulfilled. The WHO Expert Committee Biological Standardisation has approved the adoption of these four bacteria strains as the first Repository for Transfusion-Relevant Bacteria Reference Strains and, additionally, endorsed as a project the addition of six further bacteria strain preparations suitable for control of platelet contamination as the next step of enlargement of the repository.
Assuntos
Plaquetas/microbiologia , Transfusão de Sangue , Infecções Bacterianas/prevenção & controle , Técnicas de Tipagem Bacteriana/métodos , Técnicas Bacteriológicas , Bancos de Espécimes Biológicos , Transfusão de Componentes Sanguíneos/métodos , Plaquetas/citologia , Escherichia coli/metabolismo , Humanos , Cooperação Internacional , Klebsiella pneumoniae/metabolismo , Garantia da Qualidade dos Cuidados de Saúde/métodos , Reprodutibilidade dos Testes , Staphylococcus epidermidis/metabolismo , Streptococcus pyogenes/metabolismoRESUMO
BACKGROUND: Whole blood is stored at room temperature (RT) until processing into components. After separation and filtration, the RCC has to be cooled from RT to +2 - 6°C. Different start times of the cooling process and different cooling rates can be encountered in daily routine. The effect of these parameters of the initial cooling of leucoreduced red cell concentrates (LR-RCC) on in vitro quality is not known. METHODS: In paired experiments (n=12), LR-RCCs in SAGM were cooled immediately after preparation from RT to +2 to +6°C either 'fast' (within 2½ h) or 'slow' (within 10-24 h) or 'slow' after a holding period of 6, 12, 18 or 24 h. Units were then stored at +2-6°C for 42 days and sampled at regular intervals for in vitro analysis. RESULTS: Irrespective of the start time and cooling rate during the initial cooling process, all units maintained good in vitro quality up to Day 42 with haemolysis <0·8%. Adenosine triphosphate (ATP) levels remained >2·7 µmol/g Hb in 99% of all units up to Day 35. Differences in pH, ATP content and 2,3-DPG content between the groups were largest at Day 2 or 3 but generally disappeared during storage. CONCLUSION: Start time and cooling rate of the initial cooling process had minor effects on in vitro quality of red cells. LR-RCCs can be stored up to 24 h before cooling down to +2-6°C without deleterious effects on in vitro parameters during 42-day storage.
Assuntos
2,3-Difosfoglicerato/sangue , Trifosfato de Adenosina/sangue , Preservação de Sangue , Eritrócitos/metabolismo , Hemoglobinas/metabolismo , Hemólise , Temperatura Baixa , Eritrócitos/citologia , HumanosRESUMO
Three EBA specified blood bag configurations ('Eurobloodpack') are described which are capable of meeting >80% of its member's requirements. These include a 'top-and-top' and two 'bottom-and-top' packs enabling aseptic, pre-donation collection of up to 40 ml of samples, 427.5-522.5 ml of whole blood and the preparation of an extensive range of blood components. Features currently beyond the scope of ISO standardisation have been controlled including: anticoagulant and additive volumes; collection needle and sampling system; transfer tubing; cross-match line; base label; leucodepletion filter performance; compatibility of access ports and transfusion sets. Eurobloodpack has significant advantages for blood services and blood bag manufacturers.
Assuntos
Transfusão de Componentes Sanguíneos/instrumentação , Transfusão de Componentes Sanguíneos/normas , Doadores de Sangue , Procedimentos de Redução de Leucócitos/instrumentação , Procedimentos de Redução de Leucócitos/normas , União Europeia , HumanosRESUMO
Phthalates are the most widely used plasticisers in the world and have been in medical use since 1955. Di(2-ethylhexyl)phthalate (DEHP) is present in many industrial and domestic products, and is a common plasticiser used in blood bags and tubing sets. Concerns have been raised by environmental groups that DEHP exposure may result in gender birth defects and the feminisation of boys although there were no studies performed of intravenous exposure to DEHP on higher mammals or humans which report similar effects. This article reports on the toxicology of DEHP and considers concerns regarding its use in the context of transfusion medicine. The benefits of using DEHP are evaluated and the use of alternative plasticisers to DEHP is explored. The article reports on international and European regulatory recommendations for DEHP use in transfusion medicine and examines the impact of legislation on the medical devices and blood industries. The article also reviews labelling considerations for blood components and offers some guidance for blood establishments in respect of DEHP use.