According to the World Health Organization, nearly 3.2 billion people are at risk of malaria. The most deadliest form of human malaria is caused by the pathogen Plasmodium falciparum, which has claimed over 400,000 lives worldwide in 20151. Even when optimally treated with drug and donor blood therapies, severe malaria has a high mortality rate. The parasites target a patient’s red blood cells and convert them into paramagnetic units before eventually rupturing the host cell, further spreading the infection. Combination drug therapies using quinine and artemisinin derivatives are common but are either expensive or have associated toxicities from mis-dosing. Moreover, antimalarial drugs are becoming increasingly ineffective against the growing number of drug-resistant malaria strains. Combination drug and blood exchange therapies are often implemented to flush out malaria-infected red blood cells (iRBC) but consume a great quantity of donor blood, carry a high risk of transmitting other blood-borne diseases, and have no agreed upon advantage or disadvantage among clinicians. Due to the relative disadvantages of other treatment methods, small scale high gradient magnetic separation (HGMS) devices, used in a variety of biological applications, may be another treatment option to consider. mPharesis (“magnetic apheresis”) is a proposed low-cost, disposable magnetic blood filtration device which continually removes iRBCs from a patient’s whole blood by capitalizing on the iRBC’s unique magnetic properties. The proposed treatment-scale system will provide emergency care with parameters similar to continuous hemofiltration systems in terms of blood flow rates (up to approximately 500 mL min-1), vascular access, and treatment times (up to about 3 hours). A novel medium-scale high gradient magnetic separation device is detailed here. The device consists of a disposable photo-etched embedded wire array and acrylic layered housing on an external permanent magnet set. The magnetic force and flow field design were computationally optimized. In-vitro feasibility experiments were conducted at several flow rates and physiological hematocrits (Hct) using a blood mixture composed of healthy RBCs and a non-pathogenic paramagnetic blood analog called methemoglobin RBCs (metRBCs). The device was able to selectively remove paramagnetic RBCs without excessive loss of healthy RBCs. Simplified experiments were performed with 30% Hct with 20% metRBCs. At steady state, the concentration of metRBCs was reduced by 27.0±2.2% in a single pass at a flow rate of 77 μL min-1 as compared to 1.6±0.7% in control experiments without a magnet present. The experimental paramagnetic RBC removal rate was over 380 times greater than similar published HGMS devices. These successful results were applied to a theoretical transport model. The model was designed to compare the parasite removal and Hct level changes between combination drug and exchange transfusion (ET) therapy versus treatment-scale mPharesis-drug therapy. When the mPharesis flow rate was set to typical continuous dialysis rates, treatment times and donor blood volumes were reduced for all 10 cases. Calculated treatment times were all less than 60% of the reported ET-drug treatments, with times ranging from 47 to 71 minutes. The mPharesis-drug treatment was calculated to need between 4% and 53% less donor blood than the reported ET-drug treatments. Between 775 and 1772 mL of packed donor RBCs (3 to 6 units of whole blood) were estimated for the mPharesis-drug treatments, versus the average 5 to 20 units used during ET2. Treatment reference charts were generated to provide time and donor blood volume estimates for a range of patient sizes and disease severities. Based on the maximum flow rate of 500 mL min-1, a treatment-scale mPharesis system was estimated to be the size of three stacked briefcases, which is a feasible size for deployment in a clinical setting. Finally, the design, fabrication, and microscopic visualization of a simple, benchtop-fabricated continuous HGMS device was detailed. This proof-of-concept microfluidic device was implemented to test the effect of hematocrit and flow rate on the separation of mixtures of metRBCs (heat-treated and un-heated) and transparent ghost RBCs. An automated image processing protocol provided feasible cell concentration profiles for each flow and rheological condition with a 6.5 to 9.7% lower sum than manual counting for three samples. For the no magnet conditions, the average near-magnet concentration of paramagnetic RBCs at the outlet (within 10% of 130 μm channel height adjacent to the wire array) was between 1.3 and 2.4 times greater than the average of the rest of the flow field (degree of separation, DOS). The most effective separation was found to occur at the lowest flow rate 0.4 μL min-1 and with the 0.5% Hct metRBC sample with DOS=26. The addition of 30% ghost RBCs reduced the efficiency for all flow rates, with DOS=7.4 for best flow rate of 0.4 μL min-1. Heat treatment did not significantly affect separation with DOS=7.3, likely due to the low impact of the relatively low concentration of metRBCs (0.5%). The mesoscale fabrication and design process, clearance model, cell counting algorithm, and HGMS fabrication protocol and microscopy study described in this thesis provides a useful framework for future HGMS optimization and the further development of a clinical treatment system for severe malaria patients with often limited treatment options.

Platelet transfusions can be necessary during treatment of patients with thrombocytopenia or impaired platelet function. Platelet function in platelet concentrates (PC) deteriorate with storage time. Studying swirling is often used to control the quality of PC’s before transfusion but the method has some disadvantages. Therefore other methods can be useful, for example impedance aggregometry (IA, Multiplate® Analyzer) to measure platelet function.      In this study the change in platelet function over time was examined in buffy coat and apheresis platelets with IA where aggregation had been induced with adenosine diphosphate (ADP) and collagen. PC’s were tested on day 1, 4 and 7 after donation. One of the main aims of this study was to evaluate if dilution to a standardized platelet concentration (800x109 platelets/L) for IA of PC’s could be used, since platelet concentration has been shown to influence aggregation. The effect of pathogen inactivation (INTERCEPT) on platelet function and the importance of fibrinogen for aggregation were also studied.      The dilution of platelet samples reduced the range of measured values and was suitable to use with collagen but not ADP. The platelet function decreased significantly over time with both agonists. There was a significant difference between pathogen inactivated and gamma irradiated PC’s with collagen activation on day 1. Fibrinogen was shown to be of importance for platelet aggregation, but other factors in plasma seem to be necessary too.      In conclusion, IA is a suitable method for following change in aggregability over time in PC’s and sample dilution reduced variation in results.

Platelets are small fragments, but they are of crucial importance for the coagulation. The risk of spontaneous bleeding increases when the level of platelets falls below a thrombocyte particle concentration threshold value of 50 x 109/L. In those cases a platelet transfusion might be compulsory. Ongoing research tries to improve the quality of the platelets and to increase the safety of the method used. However, we still need to better understand which factors that affect how patients react upon platelet transfusion. In this study, 100 transfusions performed at Uppsala University Hospital during 2009 were examined. The platelets used had been produced with apheresis followed by pathogen inactivation by Intercept Blood SystemTM. Platelets were counted before and after transfusions and the increase was calculated in purpose to examine how well the patients responded to the platelet transfusions. These values were plotted against platelet storage time in order to examine the possible impact on the result of treatment.

O curto período de armazenamento dos concentrados de plaquetas (CP), de 5 a 7 dias, torna esse hemocomponente crítico para os serviços de hemoterapia. A criopreservação se apresenta então como uma possibilidade para a manutenção dos estoques por período mais prolongado. Essa prática exige a adição de substância crioprotetora e a adequação do volume em relação à concentração de plaquetas. O CP obtido por aférese (CPAF) é um hemocomponente proveniente de um único doador, coletado em sistema automatizado, que equivale a 6-8 unidades de CP obtidas da coleta de doadores convencional. Objetivo: Avaliar o efeito da criopreservação de CPAF no quinto dia do armazenamento, por meio de análises imunofenotípicas e funcionais das plaquetas, utilizando o crioprotetor dimetilsulfóxido (DMSO) a 5%, em freezer a -80°C e descongelamento em banho maria a 37°C, sem a remoção do crioprotetor. Material e Métodos: Foram analisadas 20 unidades de CPAF em quatro fases diferentes do processo: Fase I (pré-redução de volume), Fase II (pós-repouso, agitação e adição do DMSO), Fase III (pós-descongelamento) e Fase IV (após duas horas de descongelamento e agitação). Todas as bolsas foram avaliadas quanto à presença do \"swirling\" plaquetário, e análise microbiológica, contagem de plaquetas e leucócitos, determinação do volume plaquetário médio (VPM), análise do pH, dosagem de desidrogenase lática (DHL) e glicose, a ativação plaquetária por meio de citometria de fluxo com os marcadores CD61, CD62P e anexina V e para a avaliação funcional, as técnicas de microagregação plaquetária e retração do coágulo. Os resultados de cada fase foram analisados e comparados, considerando resultados p< 0,05 de significância estatística, avaliada pelos testes de Wilcoxon e Teste-T pareado. Resultados: Todos os CPs criopreservados apresentaram na inspeção visual presença de \"swirling\" e ausência de grumos. O pH manteve-se com mediana de 7,185 (6,076-7,528) pra fase III, análise microbiológica foi negativa em todas as unidades criopreservadas, o número mediano de plaquetas caiu de 3,04x1011/U na Fase II para 2,27x1011/U na fase III (redução de 25,32%). A ativação plaquetária na fase II foi de 23% CD62P+ para 44% CD62P+ na fase III (p=0,067). O marcador anexina V estava expresso em 13% das plaquetas na fase II e em 11% na fase III, (p=0,33). A LDH aumentou de 747 U/L (4-3079) para 1.428 U/L (662-2303) da fase II para a fase III, respectivamente (p=0,055). A glicose diminuiu em todas as fases (p<0,0001). Os testes de função plaquetária revelaram que a plaquetas descongeladas mantêm sua função. Conclusão: Os resultados obtidos mostraram que, embora tenham ocorrido ativação e redução significativa do número de plaquetas, o produto final conservou quantidade suficiente de plaquetas, cujas funções foram mantidas, o que torna viável a utilização do hemocomponente CP criopreservado. Sugere ainda que os CPAF possam ser criopreservados no quinto dia de armazenamento, com o uso do DMSO a 5%, ideal para que não se faça necessário sua remoção pós-descongelamento. / The concentration time of the platelet concentrates (CP), from 5 to 7 days, makes this blood component critical for hemotherapy services. Cryopreservation presents itself as a possibility of maintaining stocks for a longer period. This practice requires an additional cryoprotectant and a suitability of volume in relation to platelet concentration. The CP collected by the same (CPAF) is a blood component of a single dosage, being collected in an automated system, which is equivalent to 6-8 CP units from the collection of conventional donors. Objective: To evaluate the effect of CPAF cryopreservation on the fifth day of storage, by means of immunophenotypic and functional platelet analysis using the 5% DMSO cryoprotectant in a freezer at -80 ° C and thawing in a Maria bath at 37 ° C, without removal of the cryoprotectant. Material and Methods: 20 units of CPAF were analyzed in four different phases of the process: Phase I (pre-reduction of volume), Phase II (post-rest, agitation and DMSO addition), Phase III (post-thaw) and Phase IV (after two hours of thawing and shaking). All units were evaluated for platelet swirling and microbiological analysis, platelet and leukocyte counts, determination of mean platelet volume (MVP), pH analysis, lactate dehydrogenase (LDH) and glucose, platelet activation by means of flow cytometry with the markers CD61, CD62P and annexin V and for the functional evaluation, platelet microaggregation and clot retraction techniques. The results of each phase were analyzed and compared, considering results p <0.05 of statistical significance, evaluated by the Wilcoxon and Paired T-test. Results: All cryopreserved CPs presented visual presence of \"swirling\" and absence of lumps. The pH was maintained at a median of 7.185 (6.076- 7.528) for phase III, microbiological analysis was negative in all cryopreserved units, the median number of platelets fell from 3.04x1011/U in Phase II to 2.27x1011/U in phase III (reduction of 25.32%). Phase II platelet activation was 23% CD62P + to 44% CD62P + in phase III (p=0.067). The annexin V marker was expressed in 13% of platelets in phase II and 11% in phase III (p=0.33). LDH increased from 747 U/L (4-3,079) to 1,428 U / L (662-2303) from phase II to phase III, respectively (p<0,0055). Glucose decreased in all phases (p <0.0001). Platelet function tests have revealed that thawed platelets maintain their function. Conclusion: The results showed that, although activation and significant reduction of platelet count occurred, the end product preserved sufficient quantity of platelets, whose functions were maintained, which makes the use of the cryopreserved CP blood component viable. It also suggests that CPAFs can be cryopreserved on the fifth day of storage using 5% DMSO, so that their post-thaw removal is not required.

Introdução: A doação de plaquetas por aférese é um método de coleta que vem aumentando em relevância. Sabe-se que esta técnica apresenta inúmeras vantagens em comparação à doação de sangue total. Observamos que há uma preocupação na qualidade dos hemocomponentes enviados ao paciente, no entanto, não se observam muitas pesquisas em busca do cuidado com o doador. Órgãos como o Food and Drug Administration (FDA) já publicaram normas mais restritivas em relação à doação de plaquetas por aférese, pois pesquisas apontaram uma diminuição de algumas células e proteínas do sistema imunológico em doadores de repetição. Objetivos: Analisar doadores de plaquetas de repetição quanto a parâmetros hematimétricos e quantificação de subpopulações linfocitárias comparando-os com um grupo controle composto por doadores de sangue total que não doam há no mínimo um ano ou doando pela primeira vez e, ainda avaliar se a frequência de doações, o tempo de procedimento e o número de plaquetas doadas influenciam na contagem de leucócitos totais e nas subpopulações de linfócitos. Metodologia: Foram analisados 88 indivíduos em um estudo caso-controle, sendo que o grupo controle (CO) incluído foi de doadores de sangue total que haviam doado pela primeira vez ou haviam doado sangue total há mais de um ano. Os casos (CA) incluídos foram os doadores de repetição de plaquetaférese (quatro ou mais doações no último ano). O pareamento foi feito por sexo e idade. As amostras de sangue periférico foram coletadas em tubos contendo EDTA e analisadas em até 6 horas por citometria de fluxo, através da utilização de anticorpos monoclonais anti-CD3, CD4, CD8, HLADR, CD19 e CD56. Resultados: Foram avaliados 44 pares de doadores (caso vs controle). Destes, 81,8% eram homens, a média de idade dos grupos foi de 46 ±13 anos nos casos e 47 ±11 nos controles. Comparando os dois grupos, observou-se diferença estatisticamente significativa (p<0,05) na média de quantificação de leucócitos absolutos CA= 6476,6/μL vs CO=7115,4/μL (p=0,017), na média de linfócitos absolutos CA= 1862,6/μL vs CO= 2239,2/μL (p=0,007) e nos marcadores: CD3+/CD8+ (absoluto) CA= 437/μL vs CO= 597/μL (p=0,01), CD3+/CD4+(%) CA= 47,3/μL vs CO= 42,77/μL (p=0,007). Conclusões: Neste estudo foi possível observar que há uma diminuição em algumas células linfoides dos doadores de repetição em relação aos doadores convencionais, no entanto essa diferença não tem relevância clínica, demonstrando que os intervalos de doações que estes doadores estão sendo submetidos é adequado. A contagem de plaquetas dos doadores de repetição se mantiveram no decorrer do ano, este dado nos auxilia para mantermos um banco de dados de doadores de repetição com uma quantificação de plaquetas adequada, podendo ser convocado sem risco de ser bloqueado por contagem inferior ao preconizado. / Introduction: The donation of platelets by apheresis as a collection method has lately grown in relevance. This technique presents several advantages when compared to total blood donation. We understand there is a concern about the quality of the hemocomponents that are administered to the patients; however, there are not many researches concerned with caring for the donor. Entities such as the Food and Drug Administration (FDA) have published more restricting regulations regarding the donation of platelets by apheresis, since researches indicate a decrease in some cells and proteins present in the immunological systems of repeat donors. Objectives: To analyze repeat donors of platelets with regards to hematimetric parameters and quantification of lymphocyte sub-populations by comparing them with a control group consisting of total blood donors that have not donated blood for the past year at least or that are donating for the first time. Additionally, to evaluate if the frequency of donations, the duration of the procedure, and the donated platelet counts influence in the total leukocyte counts and in the sub-populations of lymphocytes. Methodology: We analyzed 88 individuals in a control case study. The control group (CG) consisted of total blood donors in their first donation or that had donated for the last time more than a year before. The cases (CA) included were the repeat donors by platelet apheresis (four or more donations in the past year). We matched the individuals by gender and age. Peripheral blood samples were collected in tubes containing EDTA and analyzed up until 6 hours later by flow cytometry, through monoclonal antibodies anti-CD3, CD4, CD8, HLADR, CD19, and CD56. Results: 44 pairs of donor were evaluated (case vs control). Among them, 81.8% were men, the average age of the groups was 46 (±13) years in the cases and 47 (±11) in the controls. When comparing the two groups, we observed a statistically significant difference (p<0,05) in the average of the quantification of absolute leukocytes CA= 6476.6/μL vs CG=7115.4/μL (p=0.017), in the average of absolute lymphocytes CA= 1862.6/μL vs CG= 2239.2/μL (p=0.007), and in the markers: CD3+/CD8+ (absolute) CA= 437/μL vs CG= 597/μL (p=0,01), CD3+/CD4+(%) CA= 47.3/μL vs CG= 42.77/μL (p=0.007). Conclusions: We were able to note in this study that there is a significant decrease in some lymphoid cells of repeat donors when compared to conventional donors. This difference, however, is not clinically relevant, which demonstrates that the donation intervals to which the donors are subject are appropriate. Platelet numbers of repeat donors remained the same throughout the year. This piece of data helps us keep a database of repeat donors with an adequate platelet number. These donors can be called for without risking of their being blocked in the screening for a number lower than the recommended.

<p>The newly established Swedish Apheresis Registry makes it possible to do national inter-center comparisons. This study was undertaken to describe and analyze the use of therapeutic apheresis and the adverse effects in such therapy. The special case of plasma exchange as rescue therapy in multi-organ failure, including renal failure, was also studied. In Sweden, plasma for transfusion is prepared and stored to ensure rapid availability. Due to new EU legislation, validation of such plasma was performed. </p><p>The analysis indicated that the use of therapeutic apheresis was in line with recommendations of other international societies. The frequency and types of adverse effects were comparable to those reported in other studies from analogous time periods. Compared with other countries, it appears that more therapeutic resources are available in Sweden and that there is a lower frequency of adverse effects in specific procedures. No fatalities were reported. The unique comparison of differences between centers regarding plasma exchange identified areas for further improvement.</p><p>The study on plasma exchange as rescue therapy in severe sepsis or septic shock is the second largest reported. The result was promising, with a survival rate of 82%. The rapid availability of plasma for transfusion appears to be of clinical importance in patients with early coagulopathy and severe trauma but the present selection and storage procedures for plasma lead to a time-dependent increase of the number of units with cold-induced activation of the contact system and C1 inhibitor consumption before day 14. Improvements of plasma quality can be attained by using plasma from male donors only and by reducing the storage time from 14 to 7 days. </p><p>Further studies are needed to define the role of plasma exchange in severe sepsis/septic shock, to evaluate the outcome of each patient’s treatment and to establish the indications for the transfusion of plasma.</p>

The newly established Swedish Apheresis Registry makes it possible to do national inter-center comparisons. This study was undertaken to describe and analyze the use of therapeutic apheresis and the adverse effects in such therapy. The special case of plasma exchange as rescue therapy in multi-organ failure, including renal failure, was also studied. In Sweden, plasma for transfusion is prepared and stored to ensure rapid availability. Due to new EU legislation, validation of such plasma was performed. The analysis indicated that the use of therapeutic apheresis was in line with recommendations of other international societies. The frequency and types of adverse effects were comparable to those reported in other studies from analogous time periods. Compared with other countries, it appears that more therapeutic resources are available in Sweden and that there is a lower frequency of adverse effects in specific procedures. No fatalities were reported. The unique comparison of differences between centers regarding plasma exchange identified areas for further improvement. The study on plasma exchange as rescue therapy in severe sepsis or septic shock is the second largest reported. The result was promising, with a survival rate of 82%. The rapid availability of plasma for transfusion appears to be of clinical importance in patients with early coagulopathy and severe trauma but the present selection and storage procedures for plasma lead to a time-dependent increase of the number of units with cold-induced activation of the contact system and C1 inhibitor consumption before day 14. Improvements of plasma quality can be attained by using plasma from male donors only and by reducing the storage time from 14 to 7 days. Further studies are needed to define the role of plasma exchange in severe sepsis/septic shock, to evaluate the outcome of each patient’s treatment and to establish the indications for the transfusion of plasma.

Der Hörsturz, seine Pathoätiogenese und Therapie werden nach wie vor kontrovers diskutiert. Basierend auf der Annahme, dass dem Hörsturz eine reduzierte kochleäre Perfusion durch eine erhöhte Plasmaviskosität zugrunde liegt, therapierten wir 85 Patienten im Zeitraum von März 2007 bis Oktober 2009 an der Klinik und Poliklinik für Hals-, Nasen-, Ohrenheilkunde der Universität Leipzig mit der HELP-Apherese. Die HELP-Apherese ist ein Verfahren, um selektiv Fibrinogen, LDL-Cholesterin und Lipoprotein(a) im Blut zu reduzieren, wodurch die Plasmaviskosität gesenkt werden kann. Verglichen haben wir die Wirksamkeit der HELP- Apherese mit einer historischen Kontrollgruppe (n=89), die im Zeitraum von September 2005 bis Oktober 2009 an der Klinik und Poliklinik für Hals-, Nasen-, Ohrenheilkunde der Universität Leipzig die Standardinfusionstherapie mit Glukokortikoiden nach dem Stennert - Schema erhalten hatte. Im Gesamtkollektiv zeigte sich hinsichtlich der Hörverbesserungen kein signifikanter Unterschied zwischen beiden Therapien, es fanden sich jedoch Hinweise für das besondere Profitieren der Apherese in bestimmten Subgruppen (Alter >60 Jahre, hochgradige Hörverluste). Außerdem wies die HELP -Gruppe trotz gleicher mittlerer Hörverbesserung signifikant mehr Vollremissionen auf als die Standardgruppe. Des Weiteren konnten wir als bedeutenden Risikofaktor für das Auftreten des akuten Hörsturzes einen hohen Fibrinogenspiegel im Blut identifizieren. Darüber hinaus erlangten wir Kenntnisse über prognosebeeinflussende Faktoren. So stellten wir fest, dass der Schweregrad des Hörverlustes, das Alter der Patienten und die Art des Hörverlustes einen Einfluss auf das therapeutische Outcome hatten. Demnach konnten wir bestätigen, was vorherige Studien bereits gezeigt hatten. Die HELP-Apherese ist eine effektive Therapiealternative zur Standardtherapie und kann höchst wahrscheinlich für besondere Subgruppen sogar bessere Ergebnisse erzielen. Vor allem, da sie hinsichtlich Lebensqualität und Nebenwirkungsprofil überlegen ist, stellt sie eine bedeutende Therapieoption dar. Auf die Frage nach der Ätiologie des Hörsturzes konnten wir Fibrinogen eine bedeutende Rolle nachweisen. Weitere multizentrische prospektive klinische Studien sind erforderlich, um die Hinweise unserer Arbeit zu belegen und eine individuelle Therapie im klinischen Alltag zu gewährleisten.

Introdução: A doação de plaquetas por aférese é um método de coleta que vem aumentando em relevância. Sabe-se que esta técnica apresenta inúmeras vantagens em comparação à doação de sangue total. Observamos que há uma preocupação na qualidade dos hemocomponentes enviados ao paciente, no entanto, não se observam muitas pesquisas em busca do cuidado com o doador. Órgãos como o Food and Drug Administration (FDA) já publicaram normas mais restritivas em relação à doação de plaquetas por aférese, pois pesquisas apontaram uma diminuição de algumas células e proteínas do sistema imunológico em doadores de repetição. Objetivos: Analisar doadores de plaquetas de repetição quanto a parâmetros hematimétricos e quantificação de subpopulações linfocitárias comparando-os com um grupo controle composto por doadores de sangue total que não doam há no mínimo um ano ou doando pela primeira vez e, ainda avaliar se a frequência de doações, o tempo de procedimento e o número de plaquetas doadas influenciam na contagem de leucócitos totais e nas subpopulações de linfócitos. Metodologia: Foram analisados 88 indivíduos em um estudo caso-controle, sendo que o grupo controle (CO) incluído foi de doadores de sangue total que haviam doado pela primeira vez ou haviam doado sangue total há mais de um ano. Os casos (CA) incluídos foram os doadores de repetição de plaquetaférese (quatro ou mais doações no último ano). O pareamento foi feito por sexo e idade. As amostras de sangue periférico foram coletadas em tubos contendo EDTA e analisadas em até 6 horas por citometria de fluxo, através da utilização de anticorpos monoclonais anti-CD3, CD4, CD8, HLADR, CD19 e CD56. Resultados: Foram avaliados 44 pares de doadores (caso vs controle). Destes, 81,8% eram homens, a média de idade dos grupos foi de 46 ±13 anos nos casos e 47 ±11 nos controles. Comparando os dois grupos, observou-se diferença estatisticamente significativa (p<0,05) na média de quantificação de leucócitos absolutos CA= 6476,6/μL vs CO=7115,4/μL (p=0,017), na média de linfócitos absolutos CA= 1862,6/μL vs CO= 2239,2/μL (p=0,007) e nos marcadores: CD3+/CD8+ (absoluto) CA= 437/μL vs CO= 597/μL (p=0,01), CD3+/CD4+(%) CA= 47,3/μL vs CO= 42,77/μL (p=0,007). Conclusões: Neste estudo foi possível observar que há uma diminuição em algumas células linfoides dos doadores de repetição em relação aos doadores convencionais, no entanto essa diferença não tem relevância clínica, demonstrando que os intervalos de doações que estes doadores estão sendo submetidos é adequado. A contagem de plaquetas dos doadores de repetição se mantiveram no decorrer do ano, este dado nos auxilia para mantermos um banco de dados de doadores de repetição com uma quantificação de plaquetas adequada, podendo ser convocado sem risco de ser bloqueado por contagem inferior ao preconizado. / Introduction: The donation of platelets by apheresis as a collection method has lately grown in relevance. This technique presents several advantages when compared to total blood donation. We understand there is a concern about the quality of the hemocomponents that are administered to the patients; however, there are not many researches concerned with caring for the donor. Entities such as the Food and Drug Administration (FDA) have published more restricting regulations regarding the donation of platelets by apheresis, since researches indicate a decrease in some cells and proteins present in the immunological systems of repeat donors. Objectives: To analyze repeat donors of platelets with regards to hematimetric parameters and quantification of lymphocyte sub-populations by comparing them with a control group consisting of total blood donors that have not donated blood for the past year at least or that are donating for the first time. Additionally, to evaluate if the frequency of donations, the duration of the procedure, and the donated platelet counts influence in the total leukocyte counts and in the sub-populations of lymphocytes. Methodology: We analyzed 88 individuals in a control case study. The control group (CG) consisted of total blood donors in their first donation or that had donated for the last time more than a year before. The cases (CA) included were the repeat donors by platelet apheresis (four or more donations in the past year). We matched the individuals by gender and age. Peripheral blood samples were collected in tubes containing EDTA and analyzed up until 6 hours later by flow cytometry, through monoclonal antibodies anti-CD3, CD4, CD8, HLADR, CD19, and CD56. Results: 44 pairs of donor were evaluated (case vs control). Among them, 81.8% were men, the average age of the groups was 46 (±13) years in the cases and 47 (±11) in the controls. When comparing the two groups, we observed a statistically significant difference (p<0,05) in the average of the quantification of absolute leukocytes CA= 6476.6/μL vs CG=7115.4/μL (p=0.017), in the average of absolute lymphocytes CA= 1862.6/μL vs CG= 2239.2/μL (p=0.007), and in the markers: CD3+/CD8+ (absolute) CA= 437/μL vs CG= 597/μL (p=0,01), CD3+/CD4+(%) CA= 47.3/μL vs CG= 42.77/μL (p=0.007). Conclusions: We were able to note in this study that there is a significant decrease in some lymphoid cells of repeat donors when compared to conventional donors. This difference, however, is not clinically relevant, which demonstrates that the donation intervals to which the donors are subject are appropriate. Platelet numbers of repeat donors remained the same throughout the year. This piece of data helps us keep a database of repeat donors with an adequate platelet number. These donors can be called for without risking of their being blocked in the screening for a number lower than the recommended.

Submitted by RAFAEL SILVA CARDOSO null (rafa.silvacardoso@gmail.com) on 2016-10-15T16:22:28Z No. of bitstreams: 1 Rafael Silva Cardoso.Dissertação Mestrado.Pesquisa e Desenvolvimento.pdf: 912570 bytes, checksum: 30f9812f4a7c979520b8ac0343efc199 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-10-20T19:53:16Z (GMT) No. of bitstreams: 1 cardoso_rs_me_bot.pdf: 912570 bytes, checksum: 30f9812f4a7c979520b8ac0343efc199 (MD5) / Made available in DSpace on 2016-10-20T19:53:16Z (GMT). No. of bitstreams: 1 cardoso_rs_me_bot.pdf: 912570 bytes, checksum: 30f9812f4a7c979520b8ac0343efc199 (MD5) Previous issue date: 2016-09-01 / Trata-se de um trabalho do tipo coorte para avaliar a espoliação dos depósitos de ferro em doadores de sangue. Sabendo-se que um percentual expressivo da população brasileira é portadora de ferro deficiência, e tendo em vista as inovações tecnológicas envolvendo os processos hemoterápicos este projeto teve como objetivo a avaliação da dinâmica de ferro com o monitoramento de parâmetros tais como: hematócrito, hemoglobina, dosagem de ferro e ferritina pré transfusionais e quatro meses após a doação, em quatro diferentes grupos de estudo:A,C,Ce D. Foi feita pelo teste ANOVA simples e para as variáveis sem distribuição normal pelo teste não paramétrico de Mann-Whitney e o teste de Kruskal Wallis, teste T-Pareado e Wilcoxon. No primeiro momento de análise (M1), com análise intra-grupos, as variações estatísticas foram presentes apenas nos parâmetros de Hb (p. 0,017), onde as variações estiveram presentes quando comparados os grupos A x D (p. 0,034) e C x D (p. 0,028) e Ht (p. <0,01) onde as variações estiveram presentes quando comparados os grupos A x D (p. 0,034) e C x D (p. 0,028). No segundo momento de análise (M2) foi identificada diferença entre os grupos, entretanto, devido à baixa significância estatística não foi possível identificar a diferença exata por grupo. Quando comparado entre os momentos um e dois- (M1 x M2) foi identificado redução da média de todos os parâmetros para os grupos A, B e C, significância estatística para o parâmetro de hemoglobina para todos os grupos e significância para o parâmetro de Ferritina exceto para o grupo B, sendo esse o único que demonstrou otimização na melhoria dos parâmetros exceto a hemoglobina. A doação de sangue diminui os índices de hemoglobina nos doadores de sangue a curto e médio prazo quando comparados em dois momentos com 4 meses de intervalo; para o indicador hematócrito e determinação de ferro sérico houve diminuição dos índices com significância estatística apenas para o grupo A (indivíduos que nunca haviam doado antes); quanto à sugestão na periodicidade das doações de CHD utilizando a modalidade de coleta aférese, o intervalo de 4 meses é insuficiente. Sugere-se 6 meses com a condição de se comprovar a viabilidade. / It is a work of the cohort to evaluate the plundering of iron deposits in blood donors. Knowing that a significant percentage of the population is disabled iron, and in view of the technological innovations involving haemotherapic processes this project aimed to evaluate the iron dynamics with the monitoring of parameters such as hematocrit, hemoglobin, iron dosing and pre transfusion ferritin months after the donation, in four different study groups: A, B, C and D. Statistical analyses was made by ANOVA and simple test for variables without normal distribution using the nonparametric Mann-Whitney and Kruskal Wallis test, Paired t-test and Wilcoxon. At first analysis (M1) and intra-group analysis, statistical variations were present only in Hb parameters (p. 0.017), where variations were present when comparing the x groups D (p. 0.034) and C x D (p. 0028) and HT (p. <0.01) in which variations were present when comparing the groups D x (p. 0.034) and C x D (p. 0028). In the second stage of analysis (M2) was identified differences between the groups, however, due to the low statistical significance was not possible to identify the exact difference per group. When compared between one and two- moments (M1 and M2) was identified reduction in the average of all parameters for groups A, B and C for the statistical significance hemoglobin parameter for all groups and significance for the parameter Ferritin except for the B group, which is the only one that showed improvement in optimization of parameters except hemoglobin. Blood donation decreases hemoglobin levels in the short and medium term blood donors when compared in two stages with 4 months apart; for the indicator hematocrit and determination of serum iron there was a decrease of the indices was statistically significant only for the group A (individuals who had never donated before); as the suggestion in the frequency of Double erythrocyte concentrate donations using the method of apheresis collection, four months range is insufficient. It is suggested that 6 months with the condition to prove the viability.