This study monitored the storage lesion of 15 units of canine platelet concentrates harvested by differential centrifugation. Canine platelet concentrates were stored at 20-24°C in a platelet rotator for a total of 9 days; the storage lesion of three second generation platelet storage containers was compared. The battery of in vitro tests used to monitor the storage lesion were selected from previous studies performed with human platelet concentrates separated by differential centrifugation. Based on these tests, canine platelet concentrates exhibited a storage lesion similar to human platelet concentrates. Metabolic analytes demonstrated decreasing pH, carbon dioxide, bicarbonate and glucose concentrations concurrent with increasing oxygen and lactate dehydrogenase activity over the 9-day period. Platelet structural changes were monitored by mean platelet volume, which began to increase on Day-5. Platelet function appeared to be compromised, as indicated by aggregation studies using collagen and adenosine diphosphate as agonists. Product sterility was maintained. There was no consensus of data supporting superior performance of one platelet storage container. This study indicates that canine platelet concentrates may be harvested by differential centrifugation of whole blood. In vitro studies utilizing three second-generation platelet storage bags support a previous study and concurs that canine platelet concentrates stored at 20-24°C using continuous agitation are viable for at least 5 days. / Master of Science

Platelet products have a short shelf life (5 to 7 days) owing in part to the deterioration of the quality of platelets stored at 22°C. This creates significant inventory challenges, and blood banks may suffer shortages and high wastage as a result. Proteomics offers a global quantitative approach to investigate changes occurring in stored blood products. These data sets can identify processes leading to storage-associated losses of blood component quality such as the platelet storage lesion (PSL). Changes to the platelet proteome between days 1 and 7 of storage were analysed with 3 complementary proteomic approaches with final mass spectrometric (MS) analysis: 2-dimensional (2D) gel electrophoresis/differential gel electrophoresis (DIGE), isobaric tagging for relative and absolute quantification (iTRAQ), and isotope-coded affinity tagging (ICAT). Although proteomics analyses identified many storage-associated protein changes, these varied significantly by method suggesting that a combination of protein-centric (2D gel or DIGE) and peptide-centric (iTRAQ or ICAT) approaches is necessary to acquire the most informative data. Validation of the proteomics results by western blotting, flow cytometry, quantitative real-time polymerase chain reaction (qRT PCR) and ³ٰ⁵S-methionine incorporation confirmed that platelets are capable of synthesising biologically relevant proteins ex vivo throughout a 10-day storage period with particularly long-lived mRNA (half-life of approximately 2.4 days), and has provided the first evidence for one of the mechanisms of the PSL. The development of an ³ٰ⁵Smethionine assay has since shown that stored human blood platelets incorporate ³ٰ⁵S-methionine at a rate that is proportional to time and substrate concentration, and is slower for freshly drawn platelets than those stored in pooled buffy coat derived units for 10 days. More interesting still are the observations that the overall ³ٰ⁵S-methionine incorporation rate was higher in pooled buffy coat platelet units versus freshly drawn platelets, that this rate increased upon agonist exposure in both, and that day 8 platelets showed significantly greater total protein translation than on days 2,3,7 and 10 of storage. This may be indicative of translational regulation of the platelet proteome during storage and upon activation. Translational control is a consequence of remarkable cellular specialisation and precise biochemical pathways which, in the case of platelets, may lead to storage-associated losses of blood component quality and must be understood if platelet storage times are to be extended.

Platelet products have a short shelf life (5 to 7 days) owing in part to the deterioration of the quality of platelets stored at 22°C. This creates significant inventory challenges, and blood banks may suffer shortages and high wastage as a result. Proteomics offers a global quantitative approach to investigate changes occurring in stored blood products. These data sets can identify processes leading to storage-associated losses of blood component quality such as the platelet storage lesion (PSL). Changes to the platelet proteome between days 1 and 7 of storage were analysed with 3 complementary proteomic approaches with final mass spectrometric (MS) analysis: 2-dimensional (2D) gel electrophoresis/differential gel electrophoresis (DIGE), isobaric tagging for relative and absolute quantification (iTRAQ), and isotope-coded affinity tagging (ICAT). Although proteomics analyses identified many storage-associated protein changes, these varied significantly by method suggesting that a combination of protein-centric (2D gel or DIGE) and peptide-centric (iTRAQ or ICAT) approaches is necessary to acquire the most informative data. Validation of the proteomics results by western blotting, flow cytometry, quantitative real-time polymerase chain reaction (qRT PCR) and ³ٰ⁵S-methionine incorporation confirmed that platelets are capable of synthesising biologically relevant proteins ex vivo throughout a 10-day storage period with particularly long-lived mRNA (half-life of approximately 2.4 days), and has provided the first evidence for one of the mechanisms of the PSL. The development of an ³ٰ⁵Smethionine assay has since shown that stored human blood platelets incorporate ³ٰ⁵S-methionine at a rate that is proportional to time and substrate concentration, and is slower for freshly drawn platelets than those stored in pooled buffy coat derived units for 10 days. More interesting still are the observations that the overall ³ٰ⁵S-methionine incorporation rate was higher in pooled buffy coat platelet units versus freshly drawn platelets, that this rate increased upon agonist exposure in both, and that day 8 platelets showed significantly greater total protein translation than on days 2,3,7 and 10 of storage. This may be indicative of translational regulation of the platelet proteome during storage and upon activation. Translational control is a consequence of remarkable cellular specialisation and precise biochemical pathways which, in the case of platelets, may lead to storage-associated losses of blood component quality and must be understood if platelet storage times are to be extended.

Platelet products have a short shelf life (5 to 7 days) owing in part to the deterioration of the quality of platelets stored at 22°C. This creates significant inventory challenges, and blood banks may suffer shortages and high wastage as a result. Proteomics offers a global quantitative approach to investigate changes occurring in stored blood products. These data sets can identify processes leading to storage-associated losses of blood component quality such as the platelet storage lesion (PSL). Changes to the platelet proteome between days 1 and 7 of storage were analysed with 3 complementary proteomic approaches with final mass spectrometric (MS) analysis: 2-dimensional (2D) gel electrophoresis/differential gel electrophoresis (DIGE), isobaric tagging for relative and absolute quantification (iTRAQ), and isotope-coded affinity tagging (ICAT). Although proteomics analyses identified many storage-associated protein changes, these varied significantly by method suggesting that a combination of protein-centric (2D gel or DIGE) and peptide-centric (iTRAQ or ICAT) approaches is necessary to acquire the most informative data. Validation of the proteomics results by western blotting, flow cytometry, quantitative real-time polymerase chain reaction (qRT PCR) and ³ٰ⁵S-methionine incorporation confirmed that platelets are capable of synthesising biologically relevant proteins ex vivo throughout a 10-day storage period with particularly long-lived mRNA (half-life of approximately 2.4 days), and has provided the first evidence for one of the mechanisms of the PSL. The development of an ³ٰ⁵Smethionine assay has since shown that stored human blood platelets incorporate ³ٰ⁵S-methionine at a rate that is proportional to time and substrate concentration, and is slower for freshly drawn platelets than those stored in pooled buffy coat derived units for 10 days. More interesting still are the observations that the overall ³ٰ⁵S-methionine incorporation rate was higher in pooled buffy coat platelet units versus freshly drawn platelets, that this rate increased upon agonist exposure in both, and that day 8 platelets showed significantly greater total protein translation than on days 2,3,7 and 10 of storage. This may be indicative of translational regulation of the platelet proteome during storage and upon activation. Translational control is a consequence of remarkable cellular specialisation and precise biochemical pathways which, in the case of platelets, may lead to storage-associated losses of blood component quality and must be understood if platelet storage times are to be extended. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate

Red blood cells (RBCs) are vital for oxygen delivery to tissues and constitute the vast majority of all cells in blood. After leaving the red bone marrow as mature cells, RBCs have a lifespan of approximately 120 days before they are removed from the circulation by macrophages, mainly in the spleen and liver. RBC transfusion is a common therapy in modern healthcare. Major surgery, numerous cancer treatments and other, often lifesaving, interventions would be unthinkable without available blood supply. For this reason, hospitals store donated RBCs in blood banks. The metabolic and structural changes that occur during prolonged storage of RBCs (the storage lesion) have been studied in detail in vitro and include oxidative stress, a reduction in glycolysis, increased membrane rigidity and shedding of microparticles from the RBC membrane. Stored RBCs share several features of senescent RBCs, but also with RBCs undergoing an apoptotic-like process called eryptosis. A consequence of the storage lesion is the fact that as much as 25% of stored RBCs could be rapidly removed from the circulation within 24 hours after transfusion. The mechanisms behind this rapid macrophage-mediated recognition and removal of stored RBCs, and its immunological consequences, remain largely unknown. Therefore, the aims of this thesis were to investigate if cryopreserved human RBCs induced an inflammatory response following autologous transfusion into healthy volunteers, and to further understand the mechanisms behind macrophage recognition of stored RBCs in vitro and in vivo. Autologous transfusion of two units of cryopreserved RBCs into healthy human recipients was found to be associated with an increased extravascular RBC elimination already at 2 hours after transfusion. However, there were no signs of an increased production of any of the investigated pro-inflammatory cytokines, indicating that an increase in the destruction of RBCs per se did not induce an inflammatory response. Eryptosis is a form of induced RBC death associated with an increased cytoplasmic Ca2+ uptake. We found that a subset of human RBCs increased their Ca2+ permeability during prolonged storage at +4°C. Using a murine model, to further understand how RBCs with an increased Ca2+ permeability were eliminated by phagocytic cells in the spleen, it was found that such RBCs were taken up by marginal zone macrophages and dendritic cells (DCs) in a manner distinct from that of naturally senescent RBCs. The DC population particularly efficient in this process expressed CD207 and are known for their ability to promote immunological tolerance. Eryptotic cell uptake was not regulated by the phagocytosis-inhibitory protein CD47 on the RBCs. To investigate how RBCs damaged during liquid storage are recognized and taken up by macrophages, a model to store and transfuse murine RBCs was developed. This storage model generated murine RBCs with several characteristics similar to that of stored human RBCs (i.e. loss of ATP, formation of RBC microparticles and rapid clearance of up to 35% of the RBCs during the first 24 h after transfusion). In vitro phagocytosis of human as well as murine stored RBCs was serum dependent and could be inhibited by blocking class A scavenger receptors using fucoidan or dextran sulphate. In conclusion, the findings of this thesis contribute to further understanding how changes inflicted to RBCs during storage direct the fate of these cells in their interaction with cells of the immune system after transfusion. The observation of an increased Ca2+ permeability of stored RBCs, and the possible recognition of such cells by tolerance-promoting DCs, in combination with the findings that class A scavenger receptors and serum factors may mediate recognition of stored RBCs, may result in novel new directions of research within the field of transfusion medicine.

The purpose of this dissertation was to evaluate outcomes associated with blood component (BC) transfusion in adult trauma patients. Specific aims were to: 1) explore the relationship between traumatic injury, hemorrhage, and BC transfusion, focusing on consequences of the component storage lesion through presentation of a conceptual model; 2) systematically review research literature comparing outcomes of massively transfused major trauma patients based on ratios of BCs received; 3) evaluating the relationship between type of blood transfusion trauma patients received (whole blood versus BCs) and mortality likelihood after controlling for demographic and clinical variables; 4) evaluating the relationship between volume and ratio of BCs transfused to trauma patients and development of inflammatory complications (ICs) after controlling for demographic and clinical variables. Specific aim one was addressed through the development of a conceptual model, depicting the current state of knowledge regarding the storage lesion, and short-/long-term outcomes of traumatic injury, hemorrhage, and blood transfusion. The second specific aim was addressed through a systematic review of studies that grouped critically injured, massively transfused patients based on ratios of BCs they received, and compared clinical outcomes among groups. Findings from this analysis revealed increased survival likelihood with massive transfusion of BCs in a 1:1:1 (packed red blood cells [PRBCs], fresh frozen plasma [FFP], platelets [PLTs]) fashion. The third specific aim involved a secondary analysis of the National Trauma Data Bank to evaluate the relationship between type of transfusion trauma patients received (whole blood versus BCs) and mortality. Patients who received BCs experienced a higher mortality likelihood compared with those who received whole blood. The fourth specific aim was addressed through a secondary analysis of the Inflammation and Host Response to Injury Trauma-Related Data Base, to evaluate the relationship between volume and ratio of BCs transfused and development of ICs in patients with major trauma. Findings revealed that total transfused volume of PRBCs, injury severity, and comorbidities were associated with development of ICs. There were no differences in time to complication between PRBCs:FFP or PRBCs:PLTs ratio groups.

O sangue total passa por processo de centrifugação a fim de que o paciente receba apenas o componente sanguíneo do qual necessita. O controle de qualidade, inicia com inspeção visual, onde são avaliados: coloração, lipemia, presença de coágulos e vazamentos. No Hemocentro Coordenador do Paraná, a principal causa de descarte de plasma fresco (PF) é a lipemia. Os PF turvos são descartados e os respectivos concentrados de hemácias (CH) permanecem em estoque. Durante o armazenamento, os CH passam por processo de alterações bioquímicas e morfológicas conhecido como lesão de armazenamento, cuja última etapa é a hemólise. É comprometida a função terapêutica e segurança transfusional. Alguns autores relatam a relação entre lipemia e hemólise. Lipemia é o aspecto turvo do plasma, devido à presença de lipoproteínas e tem relação principalmente com a dieta do doador. O objetivo deste estudo foi comparar os parâmetros de qualidade dos CH que tiveram os plasmas descartados por lipemia com os CH de plasmas límpidos. Os PF lipêmicos foram separados e foi realizado registro por fotografia e dosagem dos triglicerídeos. Conforme o grau de turbidez os plasmas foram classificados como: ligeiramente turvos, moderadamente turvos, intensamente turvos ou leitosos. Conforme a concentração de triglicerídeos foram classificados como normal (<175 mg/dl), limítrofe (175 a 199 mg/dl), elevado (200 a 499 mg/dl) ou muito elevado (>500 mg/dl). Os respectivos CH foram avaliados durante o período de validade. Os experimentos nos CH foram realizados entre 1º e 10º, entre 11º e 22º, entre 23º e 34º e entre 35º e 42º dias de armazenamento. Foram avaliados: esterilidade, índices hematimétricos, ERO, TBARS, metemoglobina. Nos sobrenadantes dos CH foram avaliados: Na+, K+, Cl-, lactato, glicose, pH e grau de hemólise. Os experimentos também foram realizados com controles, CH de plasmas límpidos do mesmo dia de doação do grupo teste. Houve aumento da hemólise mais expressivo nos CH teste, demonstrando que a lipemia impacta negativamente na qualidade do CH durante o armazenamento. A partir dos resultados do estudo foram elaboradas estratégias para processamento, controle de qualidade, modificação e distribuição dos CH de doações com lipemia, garantindo a distribuição de um hemocomponente seguro e eficaz, minimizando descarte por hemólise e efeitos adversos à transfusão. / The blood processing allows the patient receives only the blood component that he needs. The quality control starts with visual inspection and are evaluated: coloration, lipemia, clots and leaks. At the HEMEPAR, the main loss of fresh plasma (FP) is caused by lipemia. The lipemic F are discarded and the related red blood cell (RBC) remain in stock. During storage, RBCs go through the process of biochemical and morphological changes known as storage lesions, and the last one hemolysis. Therapeutic function and transfusion safety are compromised. Some authors report the relationship between lipemia and hemolysis. Lipemia is the turbid aspect of plasma, due to the presence of lipoproteins and is mainly related to the diet of the donor. The objective of this study was to compare the quality parameters of the RBCs from lipemic donations with RBC from clear donations. Lipemic FP were photographed and quantificated to triglycerides. According to the degree of turbidity the plasmas were classified as: slightly cloudy, moderately cloudy, intensely turbid or milky. Triglyceride concentrations were classified as normal (<175 mg / dl), borderline (175 to 199 mg / dl), high (200 to 499 mg / dl) or very high (> 500 mg / dl). The respective RBCs were evaluated during the period of validity. The CH experiments were performed between 1 and 10º, 11º and 22º, 23º and 34º and 35º and 42º days of storage. The following: sterility, hematimetric indexes, ROS, TBARS and methemoglobin were evaluated. In the supernatants Na +, K +, Cl-, lactate, glucose, pH and degree of hemolysis were evaluated. The experiments were also carried out with controls, RBC from clear donations of the same day of donation of the test group. There was an increase in hemolysis in the RBC test. It shows that lipemia negatively impacts the quality of RBC during storage. From the results of the study, were established strategies for processing, quality control, modification and distribution of RBC of donations with lipemic plasma to ensure the use of safe and effective blood product for transfusion.

O sangue total passa por processo de centrifugação a fim de que o paciente receba apenas o componente sanguíneo do qual necessita. O controle de qualidade, inicia com inspeção visual, onde são avaliados: coloração, lipemia, presença de coágulos e vazamentos. No Hemocentro Coordenador do Paraná, a principal causa de descarte de plasma fresco (PF) é a lipemia. Os PF turvos são descartados e os respectivos concentrados de hemácias (CH) permanecem em estoque. Durante o armazenamento, os CH passam por processo de alterações bioquímicas e morfológicas conhecido como lesão de armazenamento, cuja última etapa é a hemólise. É comprometida a função terapêutica e segurança transfusional. Alguns autores relatam a relação entre lipemia e hemólise. Lipemia é o aspecto turvo do plasma, devido à presença de lipoproteínas e tem relação principalmente com a dieta do doador. O objetivo deste estudo foi comparar os parâmetros de qualidade dos CH que tiveram os plasmas descartados por lipemia com os CH de plasmas límpidos. Os PF lipêmicos foram separados e foi realizado registro por fotografia e dosagem dos triglicerídeos. Conforme o grau de turbidez os plasmas foram classificados como: ligeiramente turvos, moderadamente turvos, intensamente turvos ou leitosos. Conforme a concentração de triglicerídeos foram classificados como normal (<175 mg/dl), limítrofe (175 a 199 mg/dl), elevado (200 a 499 mg/dl) ou muito elevado (>500 mg/dl). Os respectivos CH foram avaliados durante o período de validade. Os experimentos nos CH foram realizados entre 1º e 10º, entre 11º e 22º, entre 23º e 34º e entre 35º e 42º dias de armazenamento. Foram avaliados: esterilidade, índices hematimétricos, ERO, TBARS, metemoglobina. Nos sobrenadantes dos CH foram avaliados: Na+, K+, Cl-, lactato, glicose, pH e grau de hemólise. Os experimentos também foram realizados com controles, CH de plasmas límpidos do mesmo dia de doação do grupo teste. Houve aumento da hemólise mais expressivo nos CH teste, demonstrando que a lipemia impacta negativamente na qualidade do CH durante o armazenamento. A partir dos resultados do estudo foram elaboradas estratégias para processamento, controle de qualidade, modificação e distribuição dos CH de doações com lipemia, garantindo a distribuição de um hemocomponente seguro e eficaz, minimizando descarte por hemólise e efeitos adversos à transfusão. / The blood processing allows the patient receives only the blood component that he needs. The quality control starts with visual inspection and are evaluated: coloration, lipemia, clots and leaks. At the HEMEPAR, the main loss of fresh plasma (FP) is caused by lipemia. The lipemic F are discarded and the related red blood cell (RBC) remain in stock. During storage, RBCs go through the process of biochemical and morphological changes known as storage lesions, and the last one hemolysis. Therapeutic function and transfusion safety are compromised. Some authors report the relationship between lipemia and hemolysis. Lipemia is the turbid aspect of plasma, due to the presence of lipoproteins and is mainly related to the diet of the donor. The objective of this study was to compare the quality parameters of the RBCs from lipemic donations with RBC from clear donations. Lipemic FP were photographed and quantificated to triglycerides. According to the degree of turbidity the plasmas were classified as: slightly cloudy, moderately cloudy, intensely turbid or milky. Triglyceride concentrations were classified as normal (<175 mg / dl), borderline (175 to 199 mg / dl), high (200 to 499 mg / dl) or very high (> 500 mg / dl). The respective RBCs were evaluated during the period of validity. The CH experiments were performed between 1 and 10º, 11º and 22º, 23º and 34º and 35º and 42º days of storage. The following: sterility, hematimetric indexes, ROS, TBARS and methemoglobin were evaluated. In the supernatants Na +, K +, Cl-, lactate, glucose, pH and degree of hemolysis were evaluated. The experiments were also carried out with controls, RBC from clear donations of the same day of donation of the test group. There was an increase in hemolysis in the RBC test. It shows that lipemia negatively impacts the quality of RBC during storage. From the results of the study, were established strategies for processing, quality control, modification and distribution of RBC of donations with lipemic plasma to ensure the use of safe and effective blood product for transfusion.

Introdução: As transfusões de sangue e as Reações transfusionais (RT) têm tido grande destaque nas discussões e estudos da hemoterapia atual, devido a necessidade e relevância para a prática transfusional e na busca em qualificar as transfusões e refinar a classificação das RT. As reações transfusionais febris não hemolíticas (RTFNH) apresentam um crescente no número de notificações e despertam a necessidade de mais estudos. Durante a estocagem dos hemocomponentes, ocorrem uma série de alterações morfológicas, aumento de potássio (K+) extracelular, hemólise e aumento de hemoglobina (Hb) sobrenadante. Analisar a qualidade e viabilidade do hemocomponente pode nos levar a verificar os fatores preditores de uma RT, procurando minimizar os riscos e selecionar um hemocomponente de melhor qualidade ao paciente. Objetivos: Avaliar potenciais fatores etiológicos na precipitação das RTFNH por meio da mensuração na concentração de sódio (Na+) e K+ no sobrenadante, a contagem leucocitária por mcL, o cultural e o Hematócrito (Ht) e Hb da bolsa de concentrado de hemácias (CH) envolvidas, comparando estes parâmetros em relação a um grupo controle de bolsas de CH. Analisar e comparar o perfil dos pacientes envolvidos com a RTFNH e do grupo controle e, estimar a frequência de culturais coletados positivos e os germes envolvidos. Metodologia: Estudo de caso-controle com seleção de amostras a partir de notificações de suspeita de RTFNH ao Serviço de Hemoterapia de um Hospital Universitário de Porto Alegre - RS, no período de setembro de 2015 a setembro de 2016. O grupo controle foi selecionado a partir da mesma população de bolsas, sendo pareadas por tipagem sanguínea e data de vencimento do hemocomponente, numa proporção de 1:2,1. Resultados: o total incluído foi de 124 bolsas, sendo 39(30,5%) do grupo RT e 85(69,5%) do grupo controle, onde uma série de variáveis foram avaliadas. A média de dias de estocagem das bolsas foi de 10,7(DP=6,7) dias, sendo que no grupo RT 12,1(DP=8,1), foi significativamente maior que no grupo controle 10(DP=5,8) com (P=0,037). Também quando avaliamos as dosagens de Ht as médias verificadas foram de 68,3(DP=7,27), sendo no grupo RT 71(DP=81) e 67(DP=6,5) no grupo controle e, na comparação dos grupos, observamos um P<0,001. Dessa forma, a cada dia a mais de estocagem e, a cada ponto a mais no HT da bolsa, há um aumento na chance de aparecimento de RTFNH. Conclusões: a lesão de estocagem é uma temática importante no momento da oferta de hemocomponentes ao paciente, principalmente aos pacientes em tratamento oncológico de tumores sólidos. A avaliação do HT e do tempo de estocagem da bolsa demonstraram ter relevância estatística e clínica na predição de aparecimento de RTFNH. O manejo de estoque adequado para poder haver essa oferta se faz necessário. Novos estudos serão necessários para verificarmos os mecanismos desencadeantes da RTFNH comparado com o Ht da bolsa e, também estudos relacionados à utilização de pré medicação nas transfusões. / Introduction: Blood transfusions and the transfusion reactions (TR) have had great emphasis in current hemotherapy discussions and studies, due to its importance in transfusion practice and with the aim of qualifying the transfusions and refining TR classifications. The non-hemolytic febrile transfusion reaction (NHFTR) show an increasing number of notifications and arouse the necessity for further studies. During the storage of blood products a series of morphologic alterations occur, such as extracellular potassium (K+) increase, hemolysis and supernatant Hemoglobin (Hb) increase. Analyzing the blood product quality and availability may lead us to verifying predictive factors of a TR, seeking to minimize the risks and select a blood product of a superior quality for the patient. Objective: Evaluate potential etiological factors in the NHFTR precipitation through sodium (Na+) concentration measurement and K+ in the supernatant, the leukocyte count by mcL, the cultural and the Hematocrit (Ht),and Hb of erythrocyte concentrate bag (EC) involved, comparing those parameters in relation to a control group of EC blood bags. Analyze and compare the profile of the patients involved with a NHFTR to the control group and estimate the frequency of positive cultures collected and the germs involved. Methodology: Case-control study with sampling selections from a notification of NHFTR suspicion at a Hemotherapy Service in a College Hospital in Porto Alegre, RS, during the period from September 2015 to September 2016, where the control-group was selected from the same blood bag population, being grouped by blood type and blood product expiry date, in proportion 1:2.1. Results: Were studied 124 blood bags, being 39(39,5%) from the TR group and 85(69,5%) from the control group, where a series of invariables were evaluated. The mean of blood bag storage was 8.5 days, 10,7(PD=6,7) in the TR group and 10(DP=5,8) in the control group, and when compared they showed a P=0.037. Moreover, when we analyzed the Ht dosage, it was verified an mean of 68,3(DP=7,27), in the TR group and 71(DP=81), 67(DP=6,5) in the control group and, comparing both groups, we observed a P=<0.001. Therefore, with each additional storage day and, with each additional point in the Ht bool bag, the chance of NHFTR appearance increases. Conclusions: Storage injury is an important topic at the moment of the offer of blood components to the patient, especially to the ones with ongoing oncological treatments for solid tumors. The HT evaluation and the storage time of the blood bag demonstrate clinical and statistical relevance in the prediction of NHFTR appearance. The management of adequate storage is fundamental for the offer’s availability. Further studies are needed to verify the triggering mechanisms of NHFTR compared to the Ht of the bag, as well as studies associated with the use of premedication in transfusions.

Introdução: As transfusões de sangue e as Reações transfusionais (RT) têm tido grande destaque nas discussões e estudos da hemoterapia atual, devido a necessidade e relevância para a prática transfusional e na busca em qualificar as transfusões e refinar a classificação das RT. As reações transfusionais febris não hemolíticas (RTFNH) apresentam um crescente no número de notificações e despertam a necessidade de mais estudos. Durante a estocagem dos hemocomponentes, ocorrem uma série de alterações morfológicas, aumento de potássio (K+) extracelular, hemólise e aumento de hemoglobina (Hb) sobrenadante. Analisar a qualidade e viabilidade do hemocomponente pode nos levar a verificar os fatores preditores de uma RT, procurando minimizar os riscos e selecionar um hemocomponente de melhor qualidade ao paciente. Objetivos: Avaliar potenciais fatores etiológicos na precipitação das RTFNH por meio da mensuração na concentração de sódio (Na+) e K+ no sobrenadante, a contagem leucocitária por mcL, o cultural e o Hematócrito (Ht) e Hb da bolsa de concentrado de hemácias (CH) envolvidas, comparando estes parâmetros em relação a um grupo controle de bolsas de CH. Analisar e comparar o perfil dos pacientes envolvidos com a RTFNH e do grupo controle e, estimar a frequência de culturais coletados positivos e os germes envolvidos. Metodologia: Estudo de caso-controle com seleção de amostras a partir de notificações de suspeita de RTFNH ao Serviço de Hemoterapia de um Hospital Universitário de Porto Alegre - RS, no período de setembro de 2015 a setembro de 2016. O grupo controle foi selecionado a partir da mesma população de bolsas, sendo pareadas por tipagem sanguínea e data de vencimento do hemocomponente, numa proporção de 1:2,1. Resultados: o total incluído foi de 124 bolsas, sendo 39(30,5%) do grupo RT e 85(69,5%) do grupo controle, onde uma série de variáveis foram avaliadas. A média de dias de estocagem das bolsas foi de 10,7(DP=6,7) dias, sendo que no grupo RT 12,1(DP=8,1), foi significativamente maior que no grupo controle 10(DP=5,8) com (P=0,037). Também quando avaliamos as dosagens de Ht as médias verificadas foram de 68,3(DP=7,27), sendo no grupo RT 71(DP=81) e 67(DP=6,5) no grupo controle e, na comparação dos grupos, observamos um P<0,001. Dessa forma, a cada dia a mais de estocagem e, a cada ponto a mais no HT da bolsa, há um aumento na chance de aparecimento de RTFNH. Conclusões: a lesão de estocagem é uma temática importante no momento da oferta de hemocomponentes ao paciente, principalmente aos pacientes em tratamento oncológico de tumores sólidos. A avaliação do HT e do tempo de estocagem da bolsa demonstraram ter relevância estatística e clínica na predição de aparecimento de RTFNH. O manejo de estoque adequado para poder haver essa oferta se faz necessário. Novos estudos serão necessários para verificarmos os mecanismos desencadeantes da RTFNH comparado com o Ht da bolsa e, também estudos relacionados à utilização de pré medicação nas transfusões. / Introduction: Blood transfusions and the transfusion reactions (TR) have had great emphasis in current hemotherapy discussions and studies, due to its importance in transfusion practice and with the aim of qualifying the transfusions and refining TR classifications. The non-hemolytic febrile transfusion reaction (NHFTR) show an increasing number of notifications and arouse the necessity for further studies. During the storage of blood products a series of morphologic alterations occur, such as extracellular potassium (K+) increase, hemolysis and supernatant Hemoglobin (Hb) increase. Analyzing the blood product quality and availability may lead us to verifying predictive factors of a TR, seeking to minimize the risks and select a blood product of a superior quality for the patient. Objective: Evaluate potential etiological factors in the NHFTR precipitation through sodium (Na+) concentration measurement and K+ in the supernatant, the leukocyte count by mcL, the cultural and the Hematocrit (Ht),and Hb of erythrocyte concentrate bag (EC) involved, comparing those parameters in relation to a control group of EC blood bags. Analyze and compare the profile of the patients involved with a NHFTR to the control group and estimate the frequency of positive cultures collected and the germs involved. Methodology: Case-control study with sampling selections from a notification of NHFTR suspicion at a Hemotherapy Service in a College Hospital in Porto Alegre, RS, during the period from September 2015 to September 2016, where the control-group was selected from the same blood bag population, being grouped by blood type and blood product expiry date, in proportion 1:2.1. Results: Were studied 124 blood bags, being 39(39,5%) from the TR group and 85(69,5%) from the control group, where a series of invariables were evaluated. The mean of blood bag storage was 8.5 days, 10,7(PD=6,7) in the TR group and 10(DP=5,8) in the control group, and when compared they showed a P=0.037. Moreover, when we analyzed the Ht dosage, it was verified an mean of 68,3(DP=7,27), in the TR group and 71(DP=81), 67(DP=6,5) in the control group and, comparing both groups, we observed a P=<0.001. Therefore, with each additional storage day and, with each additional point in the Ht bool bag, the chance of NHFTR appearance increases. Conclusions: Storage injury is an important topic at the moment of the offer of blood components to the patient, especially to the ones with ongoing oncological treatments for solid tumors. The HT evaluation and the storage time of the blood bag demonstrate clinical and statistical relevance in the prediction of NHFTR appearance. The management of adequate storage is fundamental for the offer’s availability. Further studies are needed to verify the triggering mechanisms of NHFTR compared to the Ht of the bag, as well as studies associated with the use of premedication in transfusions.