Storage of canine packed red blood cells (pRBCs) can increase erythrocyte phosphatidylserine (PS) expression and eicosanoid concentrations. The objective of this study was to determine the effects of leukoreduction on erythrocyte PS expression and eicosanoid concentrations in stored units of canine pRBCs. Units of whole blood were leukoreduced (LR) or non-leukoreduced (non-LR), and stored (10 and 21 days) as pRBCs. Samples were collected at donation, and before and after a simulated transfusion. PS expression was measured by flow cytometry, and concentrations of arachidonic acid (AA), prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), prostaglandin D2 (PGD2), thromboxane B2 (TXB2), 6-keto-prostaglandin F1α (6-keto-PGF1α) and leukotriene B4 (LTB4) were quantified by liquid chromatography-mass spectrometry. Our study demonstrated that PS expression on the surface of erythrocytes was not affected by leukoreduction or storage duration. Additionally, the passage of blood through a leukoreduction filter causes an immediate and dramatic increase in TXB2 and PGF2α concentrations, but these concentrations then decrease during subsequent storage. Despite leukoreduction, the concentration of 6-keto-PGF1α continued to increase during storage and simulated transfusion. Overall, when compared to non-LR units, the addition of a leukoreduction step prior to storage had a minimal impact on the accumulation of eicosanoids in canine units of pRBCs. While leukoreduction may be beneficial in other aspects of transfusion medicine, based on the results in this study, using leukoreduction to decrease PS expression and eicosanoid concentrations does not appear to be effective.

Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-05-03T14:28:20Z No. of bitstreams: 1 NayannaBSF_TESE.pdf: 3573082 bytes, checksum: ddfdfc7465913dd1c55ac398eab942c2 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-05-04T16:04:27Z (GMT) No. of bitstreams: 1 NayannaBSF_TESE.pdf: 3573082 bytes, checksum: ddfdfc7465913dd1c55ac398eab942c2 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-05-04T16:04:57Z (GMT) No. of bitstreams: 1 NayannaBSF_TESE.pdf: 3573082 bytes, checksum: ddfdfc7465913dd1c55ac398eab942c2 (MD5) / Made available in DSpace on 2017-05-04T16:05:14Z (GMT). No. of bitstreams: 1 NayannaBSF_TESE.pdf: 3573082 bytes, checksum: ddfdfc7465913dd1c55ac398eab942c2 (MD5) Previous issue date: 2016-10-31 / Fundação de Apoio à Pesquisa do RN / This work aimed to evaluate the clinical, hematological, biochemical, hemogasometric, oxidative stress, lipid peroxidation and transfusion reactions of goats submitted to homologous transfusion of fresh whole blood or stored for 15 and 35 days. Eighteen healthy adult male goats were submitted to the induction of anemia and six hours later were transfused with whole blood stored in CPDA-1 bags (Citrate, phosphate, dextrose and adenine) according to the composite experimental group For 6 animals each: G0 (received fresh blood); G15 and G35 (received blood stored for 15 and 35 days, respectively). The transfusion reactions were recorded during the course of the transfusion, whereas clinical, hematological, biochemical, hemogasometric, oxidative stress and lipid peroxidation parameters were assessed by blood sampling at the following moments: before induction of anemia (TC0); 6 hours after phlebotomy and before transfusion (CT scan); 1, 6, 12, 24 and 96 hours after transfusion (T1, T6, T12, T24 and T96, respectively); 8, 16 and 32 days after transfusion (T8d, T16d and T32d, respectively). Samples of blood were also withdrawn from the pockets, prior to transfusion, for the same analyzes. The pockets of blood stored for 15 and 35 days presented more biochemical changes, hemogasometric, oxidative stress and lipid peroxidation than fresh blood bags. However, the hematological parameters were similar to each other. All groups presented transfusion reactions, among which fasciculations and vocalization occurred more frequently. In the hematological analysis, the animals that received fresh blood presented higher globular volume, number of red blood cells and hemoglobin concentration when compared to the animals that received stored blood. As for the biochemical analysis, after the transfusion it was observed an increase of the proteins in the 3 groups; Transient increase of serum glucose in G0 and G35; Elevation of total bilirubin, direct bilirubin, and urea in G15 and G35; In addition, there was elevation of creatine kinase in the 3 groups. The changes observed in the hemogasometric analysis had no clinical significance, as they were within the reference values for the species. As for oxidative stress, the animals that received stored blood presented disorder in their antioxidant system demonstrated by the alteration of the activity of SOD until the T12. In the analysis of lipid peroxidation there was no difference between the groups for the concentration of malondialdehyde. Thus, it can be concluded that transfusion of fresh total homologous blood was more efficient in restoring blood volume, number of erythrocytes and hemoglobin concentration than the transfusion of stored whole blood. However, transfusion of fresh or stored whole blood did not compromise the blood gas, lipid peroxidation, hepatic and renal functions of goats and proved to be efficient in restoring, among other components, the total proteins. Thus, blood transfusion, as performed in this study, was shown to be safe for use in the clinical practice of this species / Este trabalho teve como objetivo avaliar as respostas clínica, hematológica, bioquímica, hemogasométrica, estresse oxidativo, peroxidação lipídica e as reações transfusionais de caprinos submetidos à transfusão homóloga de sangue total fresco ou armazenado por 15 e 35 dias. Foram utilizados 18 caprinos machos, adultos, hígidos, os quais foram submetidos à indução de anemia e, seis horas após, transfundidos com sangue total armazenado em bolsas CPDA-1 (Citrato, fosfato, dextrose e adenina) de acordo com o grupo experimental composto por 6 animais cada: G0 (recebeu sangue fresco); G15 e G35 (receberam sangue armazenado por 15 e 35 dias, respectivamente). As reações transfusionais foram registradas durante o decorrer da transfusão, enquanto que parâmetros clínicos, hematológicos, bioquímicos, hemogasométricos, estresse oxidativo e peroxidação lipídica foram avaliados através da coleta de amostras de sangue nos seguintes momentos: antes da indução da anemia (TC0); 6 horas após a flebotomia e antes de transfusão (TC1); 1, 6, 12, 24 e 96 horas após a transfusão (T1, T6, T12, T24 e T96, respectivamente); 8, 16 e 32 dias após a transfusão (T8d, T16d e T32d, respectivamente). Amostras de sangue também foram retiradas das bolsas, antes da transfusão, para realização das mesmas análises. As bolsas de sangue armazenado por 15 e 35 dias apresentaram mais alterações bioquímicas, hemogasométricas, estresse oxidativo e peroxidação lipídica do que as bolsas de sangue fresco, porém quanto aos parâmetros hematológicos elas foram semelhantes entre si. Todos os grupos apresentaram reações transfusionais, entre as quais fasciculações e vocalização ocorreram com maior frequência. Na análise hematológica, os animais que receberam sangue fresco apresentaram maior volume globular, número de hemácias e concentração de hemoglobina quando comparados aos animais que receberam sangue armazenado. Quanto à análise bioquímica, após a transfusão observou-se aumento das proteínas nos 3 grupos; aumento transitório da glicose sérica no G0 e G35; elevação da bilirrubina total, bilirrubina direta e uréia no G15 e G35; além disso, houve elevação da creatina quinase nos 3 grupos. As alterações observadas na análise hemogasométrica não tiveram significado clínico, pois estavam dentro dos valores de referência para a espécie. Quanto ao estresse oxidativo, os animais que receberam sangue armazenado apresentaram desordem no seu sistema antioxidante demonstrada pela alteração da atividade da SOD até o T12. Na análise da peroxidação lipídica não houve diferença entre os grupos para a concentração de malondialdéido. Sendo assim, pode-se concluir que a transfusão de sangue homólogo total fresco foi mais eficiente em repor a volemia, número de hemácias e concentração de hemoglobina do que a transfusão de sangue total armazenado. Porém, a transfusão seja de sangue total fresco ou armazenado não comprometeu a gasometria, peroxidação lipídica, funções hepática e renal dos caprinos e mostrou-se eficiente em repor entre outros componentes, as proteínas totais. Sendo assim, a transfusão sanguínea, conforme realizada neste estudo, mostrou-se segura para ser utilizada na prática clínica desta espécie / 2017-05-03

Résumé 1 Hypotension et réactions hypotensives aigues isolées associées aux transfusions de concentrés de plaquettes Objectifs: Les transfusions de concentrés de plaquettes sont associées à plusieurs types de réactions transfusionnelles impliquant de l’hypotension, notamment des réactions hypotensives aigues isolées médiées par la relâche de bradykinines. L’objectif de cette étude est de déterminer l’incidence d’épisodes d’hypotension et plus particulièrement de réactions hypotensives aigues isolées associées aux transfusions de concentrés de plaquettes. Nous avons aussi tenté de déterminer si ces évènements sont associés à un niveau élevé de bradykinines. Matériels et Méthodes: Il s’agit d’une étude prospective descriptive portant sur les transfusions de concentrés de plaquettes au Centre Hospitalier Universitaire Sainte- Justine. L’étude s’est déroulée sur 28 mois. Durant cette période, tous les rapports d’incidents/accidents associés aux transfusions (RIATs) de concentrés de plaquettes transmis à la banque de sang du CHU Sainte-Justine et impliquant un épisode d’hypotension ont été identifiés. Ces RIATs furent revus par un comité d’adjudicateurs qui ont évalué et déterminé l’imputabilité de chaque réaction à la transfusion de plaquettes. Tous les sacs et tubulures des concentrés de plaquettes transfusés ont été retournés à la banque de sang après chaque transfusion. La concentration de bradykinines fut mesurée dans les 168 premiers sacs de concentrés de plaquettes retournés à la banque de sang. Les niveaux de bradykinines des concentrés de plaquettes associés à un épisode d’hypotension et des concentrés de plaquettes non associés à de l’hypotension ont par la suite été comparés. Résultats: 3672 sacs de concentrés de plaquettes furent retournés à la banque de sang parmi lesquels 25 furent associés avec un épisode d’hypotension. Les adjudicateurs ont identifiés 5 épisodes hypotensifs attribuables aux concentrés de plaquettes dont une réaction hypotensive aigue isolée (incidence par transfusion: 0.03%). Le niveau de bradykinines dans cette dernière réaction était de 10 pg/ml, alors qu’il était de 226.2 ±1252 pg/ml (95%CI : 20.0-432.4 pg/ml) dans les 143 concentrés de plaquettes contrôles. Conclusion: L’incidence d’hypotension suivant l’administration de concentrés de plaquettes est faible. Nous n’avons identifié qu’une seule réaction hypotensive aigue isolée. Nous n’avons pas été en mesure d’identifier de corrélation entre le niveau de bradykinines et l’incidence de ces réactions. Résumé 2 Transfusions de plaquettes aux soins intensifs pédiatriques Objectifs: Caractériser l’épidémiologie et les déterminants des transfusions de plaquettes dans une unité de soins intensifs pédiatriques et vérifier s’il existe une association entre ces transfusions et une augmentation de la morbidité et de la mortalité. Méthode: Étude prospective observationnelle unicentrique, combinée à un questionnaire. Lieu: Unité de soins intensifs pédiatriques du Centre Hospitalier Universitaire Sainte- Justine. Patients: Tous les enfants admis à l’unité de soins intensifs d’avril 2009 à avril 2010. Intervention: Aucune. Résultats: Parmi les 842 patients admis consécutivement aux soins intensifs, 60 patients (7.1%) ont reçu au moins une transfusion de plaquettes durant leur séjour. Les déterminants de transfusions de plaquettes identifiés à l’analyse univariée sont un score de PRISM à l’admission >10 (rapport de cotes (RC): 6.80; 95%CI: 2.5-18.3, p <0.01), un score de PELOD >20 (RC: 26.9; 95%CI: 8.88-81.5, p<0.01), un antécédent de néoplasie (RC: 5.08; 95%CI: 2.43-10.68, p <0.01), une thrombocytopénie (compte plaquettaire <50 x 109/L ou< 50,000/mm3) (RC: 141; 95%CI: 50.4-394.5, p <0.01), l’utilisation d’héparine (RC:3.03; 95%CI 1.40-6.37, p <0.01), un état de choc (RC: 5.73; 95%CI: 2.85-11.5, p<0.01) et un syndrome de défaillance multiviscérale (RC: 10.41; 95%CI: 5.89-10.40, p <0.01). À l’analyse multivariée, seuls le compte de plaquettes de<50 x109/L (RC: 138; 95%CI: 42.6-449, p<0.01) et un âge <12 mois (RC: 3.06; 95%CI: 1.03-9.10, p=0.02) demeurent des déterminants statistiquement significatifs. Les intensivistes ont répondu qu’ils avaient prescrits une transfusion de plaquettes principalement à cause d’une 6 thrombocytopénie (compte moyen de plaquettes pré-transfusion de 32 ±27 x109/L (médiane 21)), ou d’un saignement actif (compte moyen de plaquettes pré-transfusion de 76 ±39 x109/L (médiane 72)). Les transfusions de plaquettes sont associées avec le développement d’un syndrome de défaillance multiviscérale (RC: 2.53; 95%CI: 1.18- 5.43, p=0.03) et la mortalité (RC: 10.1; 95%CI: 4.48-22.7, p<0.01). Conclusions: 7.1% des enfants admis aux soins intensifs reçoivent au moins une transfusion de plaquettes durant leur séjour. La thrombocytopénie et un saignement actif sont des déterminants significatifs de transfusion de plaquettes. Les patients recevant une transfusion de plaquettes ont un risque plus élevé de développer un syndrome de défaillance multi-viscérale et ont un risque plus élevé de mortalité. / Abstract 1 Incidence of hypotension and acute isolated hypotensive transfusion reactions following platelet concentrate transfusions Background and objectives: Platelet concentrates (PCs) are associated with transfusion reactions involving hypotension, particularly bradykinin-mediated acute isolated hypotensive transfusion reactions. This study aims to determine the incidence of hypotensive events and more specifically acute isolated hypotensive transfusion reactions associated with PC transfusions. We also sought to ascertain whether these reactions are associated with high bradykinin levels. Materials and Methods: This is a prospective descriptive study of PCs administered at Sainte-Justine Hospital over 28 months. All PCs administered during this period were screened for hypotension through review of all transfusion-associated reaction reports (TARRs) sent to the blood bank. All residual PC bags were returned to the blood bank. TARRs associated with hypotension were reviewed by adjudicators who established the imputability of the PC transfusion to the reaction. Bradykinin levels were measured in the first 168 PC bags returned to the blood bank. Levels were compared between PCs associated with hypotension and control PCs not associated with hypotension. Results: A total of 3672 PC bags were returned to the blood bank; 25 PCs were associated with hypotension. Adjudicators ascertained that five hypotensive events were 8 imputable to PCs of which one was an acute isolated hypotensive transfusion reaction (incidence per transfusion: 0.03%). Bradykinin level in the latter PC was 10 pg/ml, whereas levels were 226.2 pg/ml (95%CI : 20.0-432.4 pg/ml) in the 143 control PCs. Conclusion: Our results show a low incidence of hypotension after PC transfusion. We identified only one acute isolated hypotensive transfusion reaction. No correlation between bradykinin level and the occurrence of acute isolated hypotensive reactions could be observed given that only one event was identified. Abstract 2 Platelet transfusions in pediatric intensive care Objectives: To characterize the epidemiology and the determinants of platelet transfusion (PT) in a pediatric intensive care unit (PICU) and determine whether there exists an association between PT and adverse outcomes. Design: Prospective observational single center study, combined with a self-administered survey. Setting: PICU of Sainte-Justine Hospital, a university-affiliated tertiary care institution. Patients: All children admitted to the PICU from April 2009 to April 2010. Intervention: None. Measurements and Main Results: Among 842 consecutive PICU admissions, 60 patients (7.1%) received at least one PT while in PICU. In the univariate analysis, significant determinants for PT transfusion were admission PRISM >10 (odds ratio (OR): 6.80; 95%CI: 2.5-18.3, p <0.01) and PELOD scores >20 (OR: 26.9; 95%CI: 8.88- 81.5, p<0.01), history of malignancy (OR: 5.08; 95%CI: 2.43-10.68, p <0.01), thrombocytopenia (platelet count <50 x 109/L or < 50,000/mm3) (OR: 141; 95%CI: 50.4- 394.5, p <0.01), use of heparin (OR:3.03; 95%CI 1.40-6.37, p <0.01), shock (OR: 5.73; 95%CI: 2.85-11.5, p<0.01) and multiple organ dysfunction syndrome (MODS) (OR: 10.41; 95%CI: 5.89-10.40, p <0.01). In the multivariate analysis, platelet count <50 x109/L (OR: 138; 95%CI: 42.6-449, p<0.01) and age less than 12 months (OR: 3.06; 95%CI: 1.03-9.10, p=0.02) remained statistically significant determinants. The attending physicians were asked why they gave a PT; the most frequent justification was prophylactic platelet transfusion in presence of thrombocytopenia with an average pre10 transfusion platelet count of 32 ±27 x109/L(median 21), followed by active bleeding with an average pre-transfusion platelet count of 76 ±39 x109/L(median 72). PTs were associated with the subsequent development of MODS (OR: 2.53; 95%CI: 1.18-5.43, p=0.03) and mortality (OR: 10.1; 95%CI: 4.48-22.7, p<0.01). Conclusions: 7.1% of children received at least one PT while in PICU. Thrombocytopenia and active bleeding are significant determinants of PT. Patients that received PTs had a higher risk of developing MODS and had a higher risk of mortality.

La médecine transfusionnelle féline a connu une croissance exponentielle au cours des dix dernières années. La découverte du système de groupes sanguins AB et la meilleure compréhension des mécanismes d’incompatibilité donneur-receveur, ainsi que le développement de systèmes de collecte adaptés au chat et de techniques de typage sanguin au chevet du patient ont permis d’améliorer la sécurité des transfusions. Notre travail s’est intégré dans cette volonté d’optimiser la sécurité des transfusions sanguines chez le chat en se concentrant sur deux aspects différents : le prélèvement de sang et les antigènes érythrocytaires à l’origine d’incompatibilités donneur-receveur non expliquées par le système AB. Dans un premier temps, un système de collecte de sang adapté au chat a été fabriqué afin de permettre un prélèvement fermé et autoriser le stockage des produits sanguins. Ce système a été comparé à un système ouvert composé de seringues. Le système fermé apparaît adapté au prélèvement de sang car aucune différence significative n’a été enregistrée dans les paramètres vitaux des donneurs après le prélèvement, le succès du prélèvement, ou la qualité du produit sanguin en termes de contamination bactérienne et d’hémolyse, entre les deux systèmes. Le net avantage du système fermé est qu’il assure un temps de prélèvement plus rapide que le système ouvert. Dans un second temps, en réalisant des tests de compatibilité chez 258 chats de type A, nous avons montré que la probabilité de détecter des incompatibilités entre deux chats de groupe A sélectionnés aléatoirement était de 3.9 %, et que 7 % des chats de groupe A présentaient des allo-anticorps naturels extérieurs au système AB. Sept des 18 allo-anticorps naturels détectés ont été utilisés comme réactifs lors d’un typage sanguin extensif. Les analyses sur l’accord des résultats du typage obtenus avec les différents réactifs ont permis d’identifier cinq nouveaux antigènes érythrocytaires félins différents, nommés FEA 1 à FEA 5 (pour feline erythrocyte antigen), dont l’hérédité, la prévalence, la distribution géographique et par race, ainsi que l’immunogénicité restent encore à déterminer. / Feline transfusion medicine has grown exponentially over the past decade. The discovery of the AB blood group system and the better understanding of the mechanisms of donor-receiver incompatibility, as well as the development of cat-friendly collection systems and bedside blood typing techniques have improved transfusion safety. Our work has been part of this effort to optimize the safety of blood transfusions in cats by focusing on two different aspects: blood collection and red blood cell antigens that cause donor-recipient incompatibilities unexplained by the AB system. First, a blood collection system adapted to the cat was manufactured to allow collection in a closed-system and storage of blood products. This system was compared to an open system consisting of syringes. The closed system appeared well-adapted for feline blood collection because no significant difference was found in the vital parameters of the donors after collection, the success of the collection, or the quality of the blood product in terms of bacterial contamination and hemolysis. The distinct advantage of the closed system was that it provided a shorter duration of collection than the open system. Secondly, based on the systematic crossmatches of 258 cats, we showed that the probability of detecting incompatibilities by randomly crossmatching two type A cats was 3.9 %, which resulted in at least 7 % of type A cats having naturally occurring alloantibodies outside the AB blood group system. Seven of the 18 detected naturally occurring alloantibodies were used as reagents in an extensive blood typing. Comparison of the results obtained from this extensive blood typing supports the existence of five, presumably different, new feline erythrocyte antigens, named FEA 1 to FEA 5, whose mode of inheritance, geographical and breed distribution, frequency and immunogenicity have yet to be determined.