University of Macau / Institute of Chinese Medical Sciences

Project title: Optimalization of the regime of marker's examination of clinically important infections in blood donors Project author: Daniela Dušková, M.D. Project supervisor: prof. Vladimír Tesař, M.D., Dr.Sc., MBA, FASN The aim of this project is to contribute to the discussion about introducing the methods of molecular biology into the routine blood donor testing in the transfusions departments in the Czech Republic. The theoretical part includes a brief history and some turning points in transfusion medicine. The next part within the theoretical section is dedicated to the problems of infectious diseases concerning transfusion and the general examination processes used during the selection of blood donors. The end of the theoretical part concentrates on existing possibilities of markers' examination of clinically important infections in blood donors, including the list of processes performed in the Czech Republic, the European Union and other countries. The practical part describes this study, ie. the routine screening test of blood donors using the CMIA method (a routine method) and using RT-Real Time PCR method (a molecular biology method) for detecting infectious markers (HCV, HBV, HIV). Within this part, the principle of both methods and the process of actual examinations are described in...

A prevalência de anticorpos e DNA viral de HHV-8 em 400 doadores de sangue e 50 indivíduos politransfundidos da Fundação Pró-Sangue Hemocentro de São Paulo, através das técnicas de Nested PCR e IFI para antígenos lítico e latente. Foi detectada uma soroprevalência mais alta entre doadores de sangue(4%) quando comparados a indivíduos politransfundidos (14%) (p= 0,002). A prevalência também foi mais alta entre mulheres em relação a homens, 7% versus 1.8% (p= 0,012; 95% IC 1,14 - 16,6). Foram detectados DNA de HHV-8 no plasma e PBMC de um doador 0.25%. Esta amostra foi seqüenciada confirmando a presença de HHV-8. O encontro de DNA viral no plasma de um doador de sangue indica a transfusão sanguínea como um possível meio de transmissão deste agente / The prevalence of antibodies and DNA viral of HHV-8 in 400 donors of blood and 50 receiving of the Fundação Pró-Sangue Hemocentro of São Paulo, through Nested PCR'S techniques and IFI for lithic and latent antigens. A seroprevalence of 4% was detected in the donors of blood and 14% in receiving, significantly high in comparison with the prevalence in donors of blood (p = 0,002). There was a significant difference in the prevalence among men 1,8% and 7% in the women (p= 0,012; 95% CI 1,14 - 16,6). They were detected DNA of HHV-8 in the plasma and a donor's 6,3% PBMC, and it was sequenced and submitted to the Gene Bank where presented 100% of identity. This allows to define the transmission tax and the need of introducing methods for this agent's selection

A legislação brasileira obriga a realização de testes sorológicos em todas as amostras de sangue de doadores, com o objetivo de prevenir a transmissão transfusional de microorganismos causadores de doenças como hepatite B, hepatite C, síndrome da imunodeficiência adquirida (SIDA), doença de Chagas, HTLV I/II e sífilis. Este procedimento é importante, porém não é suficiente para garantir a segurança do procedimento transfusional, sendo assim durante todo o ciclo do sangue, atitudes são adotadas para minimizar sinistros no objetivo final que a transfusão. Para receptores, a mesma legislação só obriga a realização de testes imuno-hematológicos pré-transfusionais, o que não revela o perfil sorológico deles, que por vezes pode ser positivo para algum dos marcadores antes da transfusão. O objetivo do presente trabalho foi avaliar o perfil sorológico pré-transfusional para doenças infecciosas transmissíveis por transfusão sanguínea em receptores de sangue de um hospital brasileiro de referência, o Hospital Geral de Palmas (HGP), no Estado do Tocantins. Foram realizados testes sorológicos pré-transfusionais para SIDA, hepatite B, hepatite C e sífilis em 514 receptores de transfusão do HGP, no período de fevereiro de 2014 a agosto de 2014. Vinte e nove receptores (5,65%) apresentaram-se reagentes para pelo menos um dos marcadores sorológicos testados. Os receptores apresentaram sorologia positiva para hepatite B (1,57%), hepatite C (0,58%), SIDA (1,17%) e sífilis (2,33%). Dentre os receptores de sangue que apresentaram sorologia positiva para alguma das doenças infecciosas testadas, 37,93% foram atendidos no centro cirúrgico e 27,59% na unidade de tratamento intensivo do HGP. Dentre os receptores com sorologia positiva, 62% não haviam sido transfundidos anteriormente. Este estudo determinou a prevalência de doenças infecciosas em receptores de sangue de um hospital brasileiro de referência, bem como discutiu a viabilidade e importância da implementação de testes sorológicos pré-transfusionais em receptores de sangue no Brasil. A avaliação sorológica pré-transfusional tem grande relevância pois possibilita o diagnóstico precoce de infecções nos receptores de sangue, minimizando assim o risco de comorbidade. Por outro lado, confere uma maior segurança judicial para os Serviços de Hemoterapia e para o Estado, quando da necessidade de esclarecimentos sobre possíveis transmissões de doenças infecciosas por transfusão sanguínea. / Brazilian law requires serological tests in all donors of blood samples, in order to prevent transfusion transmission of disease-causing organisms such as hepatitis B, hepatitis C, acquired immunodeficiency syndrome ( AIDS), Chagas disease, HTLV I/II and syphilis. This is important, but not sufficient to ensure the safety of the transfusion procedure, so all the blood cycle, actions are taken to minimize losses in the ultimate goal that the transfusion. For receivers, the same legislation only requires the completion of pre-transfusion immuno-hematological tests, which does not reveal the serological profile of them, which can sometimes be positive for any of the markers prior to transfusion. The objective of this study was to evaluate the pre-transfusion serological profile for transfusion-transmitted infectious diseases in blood recipients from reference Brazilian hospital, Palmas General Hospital (HGP), State of Tocantins, Brazil. Pre-transfusion serological tests for HIV, HBV, HCV and syphilis were performed in 514 transfusion recipients of HGP, from February 2014 to August 2014. Twenty and nine receivers (5.65%) presented reactivity for at least one of the evaluated serological markers. The recipients were positive for hepatitis B (1.57%), hepatitis C (0.58%), AIDS (1.17%), and syphilis (2.33%). Among the blood recipients with positive serology for any of the tested infectious diseases, 37.93% were treated at the surgical center and 27.59% in the intensive care unit of the HGP, and 62% of recipients had not been previously transfused. Thus, this study determined the prevalence of infectious diseases in blood recipients from a Brazilian reference hospital, and discussed the feasibility and importance of implementing pre-transfusion serological tests for blood recipients in Brazil. The pre-transfusion serological evaluation is important because it provides both an early diagnosis of infections in blood recipients, thus minimizing the risk of comorbidity, and a greater legal certainty for Hematology Services and for the State, when clarification about possible transmissions of infectious diseases by blood transfusion are needed.

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No. of bitstreams: 1 alinedealmeidaperes.pdf: 4968992 bytes, checksum: b682ecc1e40f813d6f65a50bb5cd23a6 (MD5) Previous issue date: 2016-10-11 / Pesquisa delineada no método misto (estudos de caso com pessoas internadas para receber hemocomponentes; representações sociais do binômio usuário-profissional sobre o processo de punção venosa periférico em hemoterapia; coorte prospectiva com doadores de sangue para calcular a incidência de trauma vascular periférico; survey sobre a prática de punção desempenhada com profissionais de duas instituições e sobre o nível de satisfação de doadores de sangue sobre o processo de punção venosa para doação de sangue; e implicações da punção venosa periférica em hemoterapia para a enfermagem). Objetivou analisar as especificidades do processo de punção de vasos durante a captação e transfusão de hemocomponentes, a ocorrência de trauma vascular periférico na perspectiva do binômio usuário-profissional e as representações sociais de profissionais e doadores de sangue sobre o processo de punção realizado com fins hemoterápicos. Utilizados referenciais da Teoria das Representações Sociais (TRS) e Teoria do Cuidado Transpessoal de Jean Watson. Foram dois os cenários da investigação: hemocentro e instituição hospitalar de ensino em Minas Gerais. Delineamentos amostrais: 1) por tipicidade para estudo de caso com seis receptores de hemocomponentes; 2) seleção completa composta por 200 doadores de sangue de um hemocentro, cujas representações sociais sobre o processo de punção foram captadas nas abordagens processual e estrutural; 3) amostra de seleção completa composta de 62 profissionais oriundos de duas instituições, cujos usuários tiveram suas veias puncionadas para fins hemoterápicos e para captação de representações sociais e survey sobre a prática laboral de profissionais que puncionam veias; 4) amostra de seleção completa com 150 doadores que integraram uma coorte prospectiva e aferição de seu nível de satisfação sobre o processo de punção. Dados coletados de Junho/2015 a Agosto/2016. Estudo de caso e coortes estruturadas a partir de instrumento previamente validado, utilizando escalas mensurativas, paleta cromática e registros fotográficos de manifestações de trauma vascular. Utilizadas técnica de evocação livre a partir de termos indutores (―doar sangue‖, ―receber sangue‖ e ―punção da veia para doar sangue‖) para abordagem estrutural da TRS e gravação de áudio com entrevista semiestruturada para abordagem processual. Survey abordando práticas profissionais e aferição da satisfação de doadores quanto à punção. Atendidas recomendações éticas e legais da pesquisa com seres humanos. Integraram os estudos de caso pessoas com anemia, plaquetopenia e imunodepressão, cujos acessos venosos viabilizaram transfusões sanguíneas e infusão de terapia intravenosa. Detectada perda da punção havendo divergência entre registros profissionais e prática laboral. Complexo de fixação não atendeu aos protocolos institucionais. Foram manifestações de trauma vascular: edema e equimose. Atendidas recomendações sobre o calibre do cateter intravenoso e uso de vaso central para a infusão de drogas vesicantes. Abordagem processual das representações sociais de profissionais evidenciou três categorias temáticas que retrataram o processo de punção venosa. Técnica de evocação livre para etapas do processo hemoterápico evidenciou motivações do grupo para o ato de doação, proximidade com familiares e influência das informações compartilhadas sobre o comportamento de doadores. Identificado distanciamento do recebimento de sangue devido xviii falta de ancoragem em experiências próprias ou de terceiros. Registrado desconforto daqueles que doam sangue. Elementos simbólicos para o ato de doação e de se ter veia puncionada (―sensaçãoruim-desconforto‖, ―menor-dor‖, ―medo-ansiedade‖ e ―agulha-picada‖) retrataram representações semelhantes. Incidência de trauma vascular foi 6,7% com manifestações de dor e equimose. O survey com profissionais evidenciou sentimentos de ―sentir-se avaliado‖, ao longo das etapas da punção, ―alívio‖ ao término da punção e ―apreensão‖ no período de manutenção do acesso venoso. Foram comportamentos emergentes: ―vigilância‖ ao longo das etapas do processo e ―tagarelar‖. Aferição da satisfação dos doadores com a punção evidenciou escores ≥6/10, sendo a menor pontuação atribuída ao tempo de atendimento. Conclui-se sobre a existência de peculiaridades no processo de punção de vasos para fins hemoterápicos quando comparado à sua utilização para outras finalidades, sendo a coerência entre práticas laborais com: boas práticas, educação permanente dos profissionais e controle de eventos adversos que norteiam a reestruturação das ações terapêuticas fatores que justificam a incidência de trauma vascular de 6,7%. As implicações dos resultados para o cuidado de enfermagem, com base na Teoria do Cuidado Transpessoal, evidenciaram a necessidade de ressignificar a realização do processo de punção venosa periférico para as pessoas que o vivenciam e está em coerência com uma assistência de enfermagem que vise proporcionar um momento de cuidado autêntico aos usuários do contexto hemoterápico. / Outline research in a mixed method (case of study with hospitalized individuals to receive blood component; social representation of user-professional binomial involving the peripheral venipuncture process; prospective cohort with blood donator to calculate the peripheral venipuncture damage occurrence; survey about the puncture practice performed by professionals in two institutions and the satisfaction level of the blood donators with the venipuncture process for blood donation; and peripheral venipuncture implications in hemotherapy for nursing. The main goal is to analyze the venipuncture process specificities during withdrawal and transfusion of blood component, the peripheral venipuncture damage occurrence in a user-professional binomial approach and the social representation of professionals and blood donators in the venipuncture process performed for hemotherapy purposes. Was utilized Social Representation Theory’s references (SRT) and Jean Watson’s Human Caring Theory: Pertinent Transpersonal. Was two investigative scenarios: Hemocentro and Education Hospital Institute. Sampling Designs: 1) By types for case of study with six blood component receivers; 2) Completely selection composed by 200 blood donators of a Hemocentro, whom social representations about the venipuncture process were captured in the procedural and structural approach; 3) Sample of complete selection composed by sixty-two professionals from the two institutions. Which the users had venipuncture for the hemotherapy purposes and social representation gathering. Include professional’s venipuncture labor practice survey; 4) Sample of complete selection with a 150 donators which integrated a prospective cohort e standardization of satisfaction level in the puncture process. The data was collected from June/2015 to August/2016. Case of study and cohort were structured from an instrument previously authenticate. Was utilized measurable scale, chromatic palette, and photographic register of venous damage. Also, was utilized the free evocation technique from inductor terms: ―blood donation‖,‖ blood receiver‖, ―venipuncture for blood donation‖ for structure approach of (SRT) and audio record interview semi-structured for processual approach. Survey involving professional practice and measurement of donators satisfaction as the puncture. Was attended the ethical recommendations and statutory research with human beings. Integrate the Study’s case people with anemia, thrombocytopenia and immunosuppression people whom venous access allowed blood transfusion and venous therapy infusion. Was detected puncture loss involving divergence between professional register and labor practice. Dressing devices didn’t attempt the institute protocol. Was damage veins manifestations: edema and ecchymosis. Attended recommendations about gauge intravenous catheter and use of central vessel to infusion of vesicant drugs. The processual approach of professional’s social representation showed three thematic categories that reflects the venipuncture process. The free evocation technique for stages of the hemotherapeutic process showed group motivations for the donation act, proximity with relatives and the influence of shared information about the donor behavior. Was identified distancing from receiving blood due to lack of anchorage in own experiences or others. Was registered discomfort of those who donate blood. Symbolic elements to the act of donate and have punctured vein: ―bad feeling – uncomfortable‖, ―less pain‖, ―afraid-anxious‖, ―needle-sting‖, pictured similar representations. The vascular trauma incidence was 6,7% with manifestations of pain and ecchymosis. The professional’s survey showed feelings of ―relief‖ during the puncture steps, ―relief‖ at the puncture’s end, seizure during the maintenance of venous access. Were emerging behaviors: ―vigilance‖, along the process steps and ―chatter‖. Gauging the satisfaction of donors with the punch showed scores >= 6/10, being the lowest score assigned to the service time. Was concluded the existence of peculiarities in the vessel puncture xx process for hemotherapic purpose when compared to its use for other purposes, being the coherence between labor practices with: good practices, continuing professional’s education and control of adverse events that guide the restructuring of therapeutic actions. Factors that justify the incidence of vascular trauma by 6,7%. The implications of the results for nursing care, based on the Human Caring Theory: Pertinent Transpersonal, highlighted the need to reframe the completion of the peripheral venipuncture process for people who experienced and is coherent with a nursing care that aims to provide a moment of authentic care to the users of the hemotherapic context.

O diagnóstico da infecção pelo vírus da hepatite C (HCV) é obtido através de testes de triagem para anti-HCV pelo ELISA e para confirmar os positivos, por teste suplementar mais específico, o immunoblot (IB). A infecção ativa é determinada pelas técnicas moleculares como por exemplo a PCR. Os testes sorológicos são métodos indiretos, baseados na detecção de anticorpos específicos, estando portanto, sujeito a vários fatores que limitam a sua eficiência diagnóstica, podendo gerar resultados inespecíficos. Um dos objetivos deste trabalho foi o de avaliar a eficiência diagnóstica dos testes para o diagnóstico da infecção pelo HCV, em condições de rotina diagnóstica, em um grande número de amostras de doadores de sangue e analisar o custo benefício de três diferentes algoritmos, recentemente propostos pelo Centro de Controle de Doenças e Prevenção (EUA). Foram estudadas 692 amostras de soros provenientes de doadores de sangue, sendo 522 positivas e 170 inconclusivas pelo ELISA de triagem (ELlSA-T) realizado no momento da doação. Esses doadores retornaram à Fundação PróSangue/ Hemocentro de São Paulo para a coleta da segunda amostra de sangue para confirmação dos resultados obtidos na doação. Em todas as amostras de retorno foram realizadas ELISA (ELISA-R) e a PCR, o IB somente nas positivas ou indeterminadas no ELISA-R. A concordância global de resultados entre ELlSA-T e ELlSA-R foi de 64,5%, sendo 77,6% (405/522) entre os positivos e 24,1 % (41/170) entre os inconclusivos. Em amostras positivas nos dois ELlSAs, o IB foi positivo (aqui denominadas de sorologicamente positivas) em 69,6% (282/405) e PCR em 61,2% (248/405). Entre as 282 amostras sorologicamente positivas, viremia foi detectada em 87,6% (247/282). A ausência de viremia em 12,4% (37/282) dessas amostras pode representar amostras de indivíduos que tiveram a infecção no passado e que eliminaram espontaneamente o HCV. A avaliação de bandas individuais no IB mostrou a alta freqüência das bandas do core (C1C2 e C3C4). Entretanto sua distribuição entre as amostras de doadores virêmicos e não virêmicos foi bastante semelhante. A intensidade da reação, expressa como graduação da coloração das bandas no IB, demonstrou que C1C2 e NS3 fortemente reativas eram as mais associadas à positividade no IB e na PCR e a banda NS4 somente com a PCR. A análise dos resultados do teste de ELISA anti-HCV de doadores de sangue indicou que a relação DO/CO, aqui denominada de índice de reatividade (IR), poderia ser utilizada como preditivo de positividade no teste suplementar. O IR de todas as amostras foram estratificados em 6 grupos sendo o IR &#8805; 6 o que melhor correlacionou-se com a positividade no IB (IR de corte). Os três algoritmos para o diagnóstico da infecção pelo HCV, propostos pelo CDC foram avaliados, sendo dois novos (a e b) e o outro, o algoritmo convencional (c). a) Resultados de ELISA com IR &#8805; 6,0 é considerado positivo sem a necessidade de teste suplementar. Para amostras IR < 6,0, de preferência o IB, seria realizado; b) Teste molecular (NAT) deve ser feito para todas as amostras positivas no teste de triagem, seguido de IB naquelas negativas no NAT; c) IB realizado para todas as amostras positivas na triagem. Esses algoritmos foram aplicados em todas as amostras, com resultados muito semelhantes entre si: 287, 287 e 285 positivos, respectivamente para a, b e c. Um total de 283 amostras foi positivo nos três algoritmos e quatro, com resultados divergentes, foram analisadas separadamente. A análise do custo benefício mostrou que o algoritmo a é o mais econômico e prático podendo ser recomendado para os laboratórios com condições econômicas limitadas; o b é o mais completo para fins de decisão médica, fornecendo informações precoces sobre a presença ou a ausência da viremia; o c é importante para determinar o status imunológico e para a população de baixa prevalência da infecção pelo HCV. O custo estimado dos três algoritmos baseou-se na tabela de 2004 da Associação Médica Brasileira. Esses custos mostraram ser o algoritmo a, 40% mais econômico e o b, 18,2% mais oneroso do que o c. Os algoritmos a e c foram complementados pela realização da PCR, nas amostras indeterminadas provenientes desses algoritmos. Duas amostras resultaram em PCR positivas, as mesmas já consideradas positivas inicialmente pelo algoritmo b. Os três algoritmos estudados, puderam ser validados para o diagnóstico laboratorial da infecção pelo HCV, podendo a escolha depender do interesse clínico ou da prevalência dessa infecção na população. / The diagnosis of hepatitis C (HCV) infection is usually undertaken stepwise by screening with ELISA and confirming the positive results in screening with a more specific assay, the immunoblot (IB). Active infection by hepatitis C virus should be confirmed by molecular techniques such as PCR. Serological tests are indirect methods based on specific antibody detection. Therefore, they may be influenced by many factors which limit their diagnostic efficiency, producing false-positive results. The aim of this work was to evaluate the real diagnostic efficiency of anti-HCV screening tests, in routine condition, in a large serum sampling, and to analyze the cost-benefit of three different algorithms recently proposed by the Center for Diseases Control and Prevention. 692 serum samples were studied. The samples consisted of 522 sera from blood donors, positive in ELISA, with the sample collected by the time of donation (ELlSA-T), and of 170 inconclusive sera. Those donors returned to Fundação PróSangue Hemocentro de São Paulo to have a second sample of blood collected to confirm the former results. ELlSA-R and PCR were carried out in all second samples and all the positive and inconclusive samples were submitted to IB. The global concordance of results between ELlSA-T and ELlSA-R was 64,5%, in that 77,6%(405/522) were obtained among the positives results and 24,1% (41/170) among inconclusive. For samples positive in both ELlSAs, IB was positive (serologically positive) in 69,6% (282/405) and PCR in 61,2% (248/405). Among 282 serologically positive samples, viremia was detected in 87,6% (247/282) and it was absent in 12,4% (37/282) of the sera, which could represent samples from individuals who were infected by HCV in the past and who have spontaneously cleared the virus. Evaluation of each antigenic band of IB showed high frequency of core (C1C2 and C3C4). Their distribution among viremic and non-viremic donors was similar. The reaction intensity, expressed by the color score of the bands, demonstrated that the strongly reactive bands of C1C2 and NS3 were associated with positiveness in IB and PCR and of NS4 only with PCR positive. Analysis of anti-HCV ELISA results from blood donors indicated that the signal-to-cut-off ratio (DO/CO), named as reactive index (IR) in this study, could be used to predict supplemental test positive results for anti-HCV. IR from all samples was classified in 6 groups, being IR &#8805; 6, the index better correlated with positive on IB (cut-off IR). The CDC has recently proposed three algorithms for HCV diagnosis, in which two were new (a and c) and one was a conventional algorithm (c). a) screening-test-positive samples with IR &#8805; 6,0 can be reported as anti-HCV positive without supplemental testing. IR < 6,0 should have refiex supplemental testing performed, preferably IB; b) reflex supplemental testing in all specimens screening-test-positive by performing NAT followed by IB for specimens with NAT negative results; c) reflex supplemental testing (IB) in all specimens screening-test-positive. These algorithms were applied to all samples and resulted in very similar positive results: 287, 287 and 285, respectively for a, b and c. A total of 283 samples were positive in three algorithms. Four samples showed divergent results and were analyzed separately. Cost-benefit - The algorithm a is the most economical and practical and it could be recommended for laboratory with limited conditions and for population with a high prevalence of HCV infection; b is the most complete for medical decision providing early information about the presence or absence of viremia; c is suitable for determining immune status and for HCV infection low prevalence population. The cost of the three algorithms was estimated based on the 2004 Brazilian Medical Association Table. These costs were 40% lower than c for algorithm a, and 18,2% higher for b. The algorithm a and c were complemented by performing PCR to solve indeterminate results. This complementary test detected two samples PCR positive which were already positive by algorithm b. Therefore we could validate those algorithms for HCV infection laboratory diagnosis. Laboratories and blood banks may choose the algorithm depending on the clinical interest or on the prevalence of HCV infection in the population.

Abstract Coronary artery disease (CAD) is the leading cause of death in developed countries. In patients with complex CAD, coronary artery bypass grafting (CABG) remains the preferred treatment as it can provide long-lasting results. However, CABG carries a significant risk of excessive perioperative bleeding and other complications, which may deteriorate the prognosis. Transfusion of blood products is generally used to compensate blood loss. However, both bleeding and blood transfusions have been shown to be associated with an adverse outcome. This cohort study aimed to clarify the impact of perioperative bleeding and the use of different blood products in the development of perioperative complications in 2,764 patients undergoing isolated CABG. The universal definition of perioperative bleeding classification (UDPB) was employed to stratify the severity of bleeding. Additionally, the impact of storage time of transfused red blood cells (RBCs) on the outcome was investigated. Increased UDPB classes, particularly classes 3 and 4, were associated with significantly poorer immediate and late outcome. RBC transfusion in patients who underwent elective off-pump CABG was independently associated with increased troponin I release indicating myocardial injury. Prolonged storage duration of transfused RBCs did not affect immediate and late outcome of patients with moderate bleeding. The most remarkable risk factors for stroke after off-pump CABG were any degree of atherosclerosis of the ascending aorta as well as transfusion of platelets and/or solvent/detergent-treated plasma. The UDPB classification appears to be a promising research tool to stratify the severity of perioperative bleeding and to assess its prognostic impact after coronary surgery. Prevention of major bleeding that leads to blood transfusion may protect from myocardial injury and stroke and possibly result in better early and late outcomes. Patients with a diseased ascending aorta could be considered at high risk of stroke because of their risk of generalized atherosclerosis. In case of mildly diseased aorta, the “no-touch” aorta policy should be considered with the intention of preventing postoperative stroke. / Tiivistelmä Sepelvaltimotauti on yleisin kuolinsyy kehittyneissä maissa. Potilailla, joilla on vaikea monen suonen tai vasemman sepelvaltimon päärungon tauti, sepelvaltimoiden ohitusleikkaus on edelleen paras hoitovaihtoehto, koska sillä pystytään saavuttamaan pitkäkestoisia tuloksia. Kuitenkin ohitusleikkaukseen liittyy suuri riski leikkauksen aikaiselle tai jälkeiselle verenvuodolle ja muille haittatapahtumille, jotka osaltaan huonontavat potilaan ennustetta. Vuodon hoitona käytetään yleisesti verensiirtoa. Kuitenkin on osoitettu, että sekä verenvuoto että verituotteiden anto lisäävät riskiä komplikaatioille. Tämän kohorttitutkimuksen tavoitteena oli selvittää tarkemmin leikkauksen yhteydessä ilmenevän vuodon ja siihen liittyvän verensiirron vaikutuksia leikkauksen jälkeisten haittatapahtumien kehittymiseen 2764 ohitusleikatulla potilaalla. Universal Definition of Perioperative Bleeding (UDPB) -luokitusta käytettiin vuodon vaikeusasteen luokittelemiseen. Lisäksi tutkittiin siirrettyjen punasolujen varastointiajan vaikutusta potilaan ennusteeseen. Korkeammat UDPB-luokat, erityisesti luokat 3 ja 4, liittyivät merkittävästi huonompaan lyhyen ja pitkän aikavälin ennusteeseen. Potilailla, joille oli tehty kiireetön ohitusleikkaus ilman sydän-keuhkokoneen käyttöä, punasolujen anto oli itsenäinen riskitekijä suurentuneelle troponiini I -päästölle eli sydänlihasvauriolle. Pidentynyt punasolujen varastointiaika ei ollut yhteydessä lyhyen tai pitkän aikavälin ennusteeseen potilailla, jotka olivat vuotaneet kohtalaisesti. Merkittävimmät riskitekijät ilman sydän-keuhkokonetta tehdyn leikkauksen jälkeiselle aivoinfarktille olivat minkä tahansa asteinen nousevan aortan ateroskleroosi sekä verihiutaleiden ja/tai jääplasman anto. UDPB-luokitus vaikuttaa lupaavalta tutkimustyökalulta verenvuodon vaikeusasteen luokitteluun. Lisäksi sitä voidaan käyttää vuodon ennusteellisen vaikutuksen arvioimiseen ohitusleikkauksen jälkeen. Runsaan verenvuodon ja siihen liittyvän verensiirron ehkäiseminen saattaa suojata potilasta sydänlihasvauriolta ja aivoinfarktilta ja mahdollisesti johtaa parempaan lyhyen ja pitkän aikavälin ennusteeseen. Potilaita, joilla on nousevan aortan ateroskleroosi, voisi pitää suuressa aivoinfarktiriskissä yleisen ateroskleroosiriskin vuoksi. Potilailla, joilla on lieväkin nousevan aortan ateroskleroosi, tulisi harkita aortan jättämistä pihdittämättä aivoinfarktin ehkäisemiseksi.

Abstract Cardiovascular diseases are the leading cause of death worldwide, and coronary artery disease accounts for the majority of them. The treatment of choice for complex coronary artery disease is coronary artery bypass grafting. However, as surgery in general, cardiac surgery is associated with an increased risk of perioperative bleeding and utilization of blood products. The present study aimed to investigate the impact of preoperative anemia, perioperative bleeding and retained blood syndrome as well as blood transfusion on the outcomes after isolated coronary surgery. The severity of perioperative bleeding was assessed mainly using the E-CABG and UDPB stratification criteria. Our analyses showed that severe bleeding is associated with a significantly increased risk of stroke. Furthermore, severe bleeding increased the risk of several adverse events even in low-risk patients. Retained blood syndrome was observed to be a common complication after coronary surgery and was associated with an increased risk of postoperative complications. Preoperative anemia seems to have no significant impact on patient early and late survival. Instead, the frequent exposure to blood products may be the determinant of poorer survival observed among anemic patients. Perioperative blood loss and exposure to allogeneic blood has been shown to increase adverse events. Therefore, prevention of bleeding and measures to optimize patient blood management could improve patient outcomes after cardiac surgery. / Tiivistelmä Sydän ja verisuonitaudit ovat maailmanlaajuisesti yleisin kuoleman aiheuttaja, joista sepelvaltimotaudilla on suurin vaikutus. Sepelvaltimoiden ohitusleikkaus on käypä hoito vakavassa sepelvaltimotaudissa. Kuten kirurgiassa yleisestikin, erityisesti sydänkirurgia on yhdistetty suurentuneeseen verenevuodon ja verituotteiden saannin riskiin. Tutkimukseni tavoitteena oli selvittää preoperatiivisen anemian, perioperatiivisen verenvuodon, verituotteiden annon, sekä leikkausalueelle jääneen veren itsenäisiä vaikutuksia potilaiden lopputulemiin sepelvaltimoiden ohitusleikkauksen jälkeen. Verituotteiden ja perioperatiivisen verenvuodon määrää arvioitiin pääsääntöisesti käyttäen E-CABG ja UDPB verenvuotoluokituksia. Tuloksenamme oli, että vakava verenvuoto lisää merkitsevästi aivoinfarktin riskiä. Lisäksi vakava perioperatiivinen verenvuoto on yhteydessä useisiin komplikaatioihin myös matalan leikkausriskin potilailla. Leikkausalueelle jääneen veren huomattiin olevan yleinen ongelma sepelvaltimoiden ohitusleikkauksen jälkeen, minkä lisäksi se lisäsi riskiä useille haitta-tapahtumille. Preoperatiivisella anemialla ei ollut tilastollisesti merkitsevää vaikutusta potilaiden lyhyen ja pitkän aikavälin ennusteisiin. Sen sijaan, aneemisille potilaille annetut verensiirrot saattaisivat aiheuttaa näillä potilailla huomatun alentuneen elinajan ennusteen. Perioperatiivisen verenvuodon ja altistumisen verituotteille on osoitettu lisäävän haittatapahtumia. Siispä verenvuodon vähentäminen ja verituotteiden säästäminen voisi parantaa potilaiden ennustetta sydänkirurgiassa.

Abstract Coronary artery bypass grafting (CABG) is the treatment of choice for patients with three-vessel disease or left main stenosis. However, it is associated with considerable risk of perioperative complications such as myocardial infarction, stroke, infections, and mortality to which excessive bleeding is a contributing factor. This thesis aims to determine the factors involved in and clinical implications of bleeding after CABG. The 1st study evaluated the effects of preoperative ASA discontinuation on the patient’s outcome after CABG. The results showed that late or no discontinuation of low-dose ASA before CABG may decrease the risk of postoperative stroke without increasing the risk of postoperative bleeding. In the 2nd study the use of warfarin was found to be a safe during CABG with no excess bleeding nor other major complications. The 3rd study estimated the impact of surgeons´ performances on blood loss and need for re-exploration after CABG. With 2001 study patients, this study clearly demonstrated that an individual surgeon is a powerful determinant of postoperative bleeding and need for re-exploration after CABG. Using systematic review and meta-analysis, we estimated the risk of complications related to re-exploration for bleeding after CABG. In literature search in 2011, 8 articles with 557 923 patients fulfilled the inclusion criteria. Re-exploration for bleeding after cardiac surgery carries a significantly increased risk of postoperative mortality and morbidity, and thus has a major impact on the patient’s immediate postoperative outcome. We also studied the impact of blood transfusion on the development of post-operative stroke after CABG. Of the study population of 2 226 CABG patients, stroke occurred postoperatively in 53 patients (2.4%). The statistical analysis showed that transfusion of blood products after CABG has a strong, dose-dependent association with the risk of stroke. The use of Octaplas® and platelet transfusions seem to have an even larger impact on the development of stroke than red blood cell transfusions. The 6th study investigated the impact of transfusion of blood products on intermediate outcome after CABG in 2001 patients. The findings indicated that transfusion of any blood product is associated with a significant risk of all-cause and cardiac mortality after CABG. / Tiivistelmä Sepelvaltimotauti on yleisin kuolinsyy ja sepelvaltimoiden ohitusleikkaus hyvine pitkäaikaistuloksineen on todettu parhaaksi hoidoksi potilailla, joilla on monen suonen tai vasemman päärungon tauti. Ohitusleikkaukseen liittyy kuitenkin verenvuodon sekä näihin kytkeytyvien komplikaatioiden riski. Tämän väitöskirjan tavoitteena oli määrittää verenvuodon riskitekijöitä sekä verituotteiden siirtojen vaikutusta ohitusleikkauspotilaiden ennusteeseen. Verenhyytymistä estävien lääkkeiden tiedetään lisäävän verenvuotoja. Ensimmäinen tutkimus osoitti, että ASA:n jatkaminen keskeytyksettä ohitusleikkauksissa vähentää aivoinfarktien riskiä lisäämättä silti verenvuodon riskiä. Toisessa tutkimuksessa pitkäaikainen warfariinihoito osoittautui turvalliseksi ohitusleikkauksen aikana eikä sen käyttö lisännyt verenvuotoja eikä muita komplikaatioita. Kolmas tutkimus osoitti kirurgin taidon merkityksen verenvuotojen ja uusintaleikkausten määrään 2001 potilaalla. Verenvuotojen vuoksi tehtävien uusintaleikkausten negatiivinen vaikutus postoperatiiviseen mortaliteettiin sekä morbiditeettiin on todettu yksiselitteisesti useissa tutkimuksissa. Vuonna 2011 tehdyllä systemaattisella kirjallisuuskatsauksella ja meta-analyysillä selvitimme yhteensä 557 923 ohitusleikkauspotilaan aineistosta, että verenvuodon jälkeisiin uusintaleikkauksiin liittyy huomattava kuoleman ja komplikaatioiden riski. Verenvuotoja hoidetaan yleisesti verensiirroilla, vaikkakin useat tutkimukset ovat osoittaneet verituotteiden annon lisäävän mortaliteettia sekä komplikaatioriskiä. Viides tutkimus selvitteli sepelvaltimoleikkauksissa potilaalle annettujen verituotteiden ja leikkauksen yhteydessä sairastettujen aivoinfarktien välistä yhteyttä. Osoittautui, että verituotteiden käyttöön liittyy annosriippuvaisesti lisääntynyt riski saada aivoinfarkti leikkauksen yhteydessä. Varsinkin verihiutale- ja jääplasmasiirtoihin on todettu liittyvän vielä suurempi aivoinfarktin riski kuin punasolusiirtoihin. Kuudes tutkimus selvitteli sepelvaltimoleikkauksien yhteydessä annettujen verituotteiden vaikutusta 2001 potilaan keskipitkään ennusteeseen. Tutkimus osoitti, että minkä tahansa verituotteen antoon sepelvaltimoleikkauksissa liittyy lisääntynyt kuoleman ja sydänkuoleman riski.

Project title: Optimalization of the regime of marker's examination of clinically important infections in blood donors Project author: Daniela Dušková, M.D. Project supervisor: prof. Vladimír Tesař, M.D., Dr.Sc., MBA, FASN The aim of this project is to contribute to the discussion about introducing the methods of molecular biology into the routine blood donor testing in the transfusions departments in the Czech Republic. The theoretical part includes a brief history and some turning points in transfusion medicine. The next part within the theoretical section is dedicated to the problems of infectious diseases concerning transfusion and the general examination processes used during the selection of blood donors. The end of the theoretical part concentrates on existing possibilities of markers' examination of clinically important infections in blood donors, including the list of processes performed in the Czech Republic, the European Union and other countries. The practical part describes this study, ie. the routine screening test of blood donors using the CMIA method (a routine method) and using RT-Real Time PCR method (a molecular biology method) for detecting infectious markers (HCV, HBV, HIV). Within this part, the principle of both methods and the process of actual examinations are described in...