With the emergence of the concept of crowdfunding, a new alternative has arisen for non-profit organizations to procure funding for their cause. But several of the issues plaguing non-profits still remain. Attracting people to give in the first place is one major challenge, but retaining the relationship and keeping the donors giving is an even bigger one. This thesis delves into the issue of getting donors to sustain their giving, more commonly known as donor retention. More specifically, this thesis aims to identify what factors are important when aiming to improve donor retention in both charity and crowdfunding contexts. Additionally, the thesis explores what is important in the general structure and content of a non-profit crowdfunding platform, and what possible measures can be taken in order to improve donor retention through the platform. This was achieved by conducting a literature study in the field of donor retention in both a charity as well as crowdfunding context to find important factors as well as measures to be considered. In parallel, 15 interviews were conducted to gather insights on the attitudes, donation habits and motivations of people who had previous experience of giving to charity. In sequence with the interviews, 15 user tests on the crowdfunding platform of the non-profit organization Help to Help were performed. The user tests aimed to evaluate and collect donors' opinions and conceptions of the contents and layout of an existing non-profit crowdfunding platform. From the literature study several factors found to improve donor retention were identified. These were: satisfaction, trust, commitment and involvement. The literature study, interviews and user tests resulted in finding a large number of possible actions to take when constructing a non-profit crowdfunding platform. The measures were summarized and condensed into a collection of recommendations and considerations to take when designing such a platform to increase donor retention. The thesis also identified that crowdfunding components can make a charity website appear more transparent and innovative. Additionally, the thesis found that many donors continue to donate based on their initial conviction. Where the ability to provide proof of where the donations end up and ability to appear transparent were the most cited reasons for deciding to support an organization.

The thesis presents current understanding of the concept of death and criteria for diagno- sis of death in the context of organ donation. We will argue that 1) the dead donor rule should not be the necessary condition for retrieving organs for transplantation and 2) it should be permissible to retrieve organs from patients that are imminently dying (not dead yet), with respect to the principle of autonomy and non-maleficence. We will first present the impossibility and current inconsistencies in determining the exact "moment of death" and we will then demonstrate that current organ donors do not fulfill biological criteria for death and that the dead donor rule is not respected in clinical practice. We suggest that in the context of recent major technological advances in the field of critical care medicine the dead donor rule becomes irrelevant and does not contribute to the transplantation ethics. The legal concept of death and the biological phenomenon of death become more and more distant. We argue that declaring death is not necessary for ethically justified policy in transplantation. Both the societal trust and the protection of vulnerable individuals can be ensured by different ethical principles (i.e. the principle of autonomy and the principle of non- maleficence). The sound ethical...

My degree work focused on the problem of involuntary childlessness, which is a situation that a number of couples are facing nowadays and this issue has been growing. The work specifically deals with the donor programme in assisted reproduction. I described the method of treatment with the use of donor gametes and ethical and legislative issues related to it. The empiric part of the work is divided into two research projects each of which corresponds with one objective of the work. The first one monitors the situation when receiving couples are making a decision whether to join the assisted reproduction programme with donor gametes and the mental and social consequences of this situation. This research part is based on partly structured and non-structured interviews with 10 couples and on observations. The data was transcribed and subsequently qualitatively analysed. The second research comprises data obtained from non-structured interviews with female donors and from on-site observations. This part relates to the second objective of the degree work. In the discussion section of the work the interviews and observations are assessed and compared with literature on this topic. I focused mainly on the issue of anonymous donation, on motivation of oocyte donors and on subsequent comparison with receivers´ expectations. Based on the research I determined a hypothesis that donation of gametes is a difficult problem which our society should deal with in detail. This degree work could be used by people from centres of assisted reproduction. It can also be used as a study material by students of the Faculty of Health and Social Studies, especially those who attend the midwife course.

abstract: Natural photosynthesis dedicates specific proteins to achieve the modular division of the essential roles of solar energy harvesting, charge separation and carrier transport within natural photosynthesis. The modern understanding of the fundamental photochemistry by which natural photosynthesis operates is well advanced and solution state mimics of the key photochemical processes have been reported previously. All of the early events in natural photosynthesis responsible for the conversion of solar energy to electric potential energy occur within proteins and phospholipid membranes that act as scaffolds for arranging the active chromophores. Accordingly, for creating artificial photovoltaic (PV) systems, scaffolds are required to imbue structure to the systems. An approach to incorporating modular design into solid-state organic mimics of the natural system is presented together with how conductive scaffolds can be utilized in organic PV systems. To support the chromophore arrays present within this design and to extract separated charges from within the structure, linear pyrazine-containing molecular ribbons were chosen as candidates for forming conductive linear scaffolds that could be functionalized orthogonally to the linear axis. A series of donor-wire-acceptor (D-W-A) compounds employing porphyrins as the donors and a C60 fullerene adduct as the acceptors have been synthesized for studying the ability of the pyrazine-containing hetero-aromatic wires to mediate photoinduced electron transfer between the porphyrin donor and fullerene acceptor. Appropriate substitutions were made and the necessary model compounds useful for dissecting the complex photochemistry that the series is expected to display were also synthesized. A dye was synthesized using a pyrazine-containing heteroaromatic spacer that features two porphyrin chromophores. The dye dramatically outperforms the control dye featuring the same porphyrin and a simple benzoic acid linker. A novel, highly soluble 6+kDa extended phthalocyanine was also synthesized and exhibits absorption out to 900nm. The extensive functionalization of the extended phthalocyanine core with dodecyl groups enabled purification and characterization of an otherwise insoluble entity. Finally, in the interest of incorporating modular design into plastic solar cells, a series of porphyrin-containing monomers have been synthesized that are intended to form dyadic and triadic molecular-heterojunction polymers with dedicated hole and electron transport pathways during electrochemical polymerization. / Dissertation/Thesis / Ph.D. Chemistry 2013

Made available in DSpace on 2018-06-27T18:56:29Z (GMT). No. of bitstreams: 2 Marcos Andre Pereira dos Santos.pdf: 1399807 bytes, checksum: 95e887716f9af791512ffaa6d1d92dc6 (MD5) Marcos Andre Pereira dos Santos.pdf.jpg: 3031 bytes, checksum: 1f26d8583aceb9e2d84423f908d4f9bd (MD5) Previous issue date: 2009-03-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work presents a study on the interaction of TiCl4 and TiCl3 molecules with a single- and bi-layer graphene by using the density functional theory. It is evaluated the changes in structural, electronic and magnetic properties of graphene after the adsorption of these molecules, that can behave donors or acceptors of electronic charge. We evaluated the arrangements for TiCl4 and TiCl3 molecules at three different sites using one molecule per unit cell, making use of periodic boundary conditions. In both cases the molecules have more stable chemical configuration when they are placed in the center of a hexagonal site. Significant changes in the electronic properties of the systems are observed. The molecule TiCl4 behaves like a Lewis acid (electronic charge acceptor), while in the case of the confined molecule between two graphene planes the molecule behaves as a Lewis basis (electronic charge donor). The TiCl4 on graphene remains non-magnetic in all configurations studied here. The adsorption of the molecule causes distortions in the graphene sheet, they originating a small change in the original electronic states, as a gap opening. TiCl3 molecule presents a Lewis basics behavior, which is seen directly on the lifting of the Fermi level. The TiCl3 presents a stronger binding energy when compared to the TiCl4 in graphene; this fact also shows that the electronic properties also change. It is observed that the magnetic moment of the adsorbed TiCl3 graphene associated with the spin polarization, dependent on the adsorption configuration. Therefore, this innovative work predicts the properties associated with the interaction of molecules with donors or accepted electronic charge behavior adsorbed on graphene this contributing for optimizing and developing potential application of this novel material. / Nesta Dissertação apresentamos um estudo da interação das moléculas TiCl4 e TiCl3 com grafeno de camada simples e bicamada fazendo uso da teoria do funcional da densidade. Avaliamos as alterações nas propriedades estruturais, eletrônicas e magnéticas do grafeno interagindo com as moléculas, as quais podem se comportar como sistemas doadores ou aceitadores de carga eletrônica. Foram avaliadas as configurações mais estáveis para as moléculas de TiCl3 e TiCl4 em três sítios utilizando uma molécula por célula unitária e fazendo uso de condições periódicas de contorno. Em ambos os casos, as moléculas possuem maior estabilidade química na configuração em que essas são colocadas no centro de um hexágono do grafeno. Após a simulação foram observadas alterações significativas nas propriedades eletrônicas do sistema. A molécula TiCl4 apresenta caráter de ácido de Lewis (receptor de carga), enquanto, no caso da molécula estar confinada entre dois planos de grafeno, comporta-se como uma base de Lewis (doador de carga). Também se observa que o TiCl4 permanece não magnético em todas as configurações estudadas. A adsorção da molécula causa distorções geométricas na folha de grafeno, originando uma pequena mudança nos estados eletrônicos do grafeno, visto na abertura de um gap de energia. Por outro lado, a molécula de TiCl3 apresenta caráter de base de Lewis, visto diretamente no aumento do valor do nível de Fermi. O TiCl3 apresenta energia de ligação bem superior quando comparado ao TiCl4 no grafeno, este fato mostra também que as propriedades eletrônicas também se modificam. Observa-se que o momento magnético do TiCl3 adsorvido no grafeno apresenta polarização de spin, dependente da configuração estrutural. Desta forma, este trabalho busca, de forma inédita, demonstrar as propriedades associadas com a interação de moléculas doadoras e aceitadoras de carga eletrônica adsorvidas no grafeno fomentando o potencial de aplicação tecnológica deste material inovador.

Introdução: O transplante renal é amplamente reconhecido como a melhor forma de tratamento para os pacientes que necessitam de terapia de substituição renal, e isso o torna vítima de seu próprio sucesso. Apesar de ser considerado um triunfo clínico-cirúrgico, é igualmente uma fonte de frustração, pela carência de doadores de órgãos e pelo crescimento das listas de espera. Há necessidade do desenvolvimento de estratégias que tornem maior o número de rins disponíveis para transplante. A idade do doador, o tempo de isquemia fria (TIF) e o HLA (antígeno leucocitário humano de histocompatibilidade) são fatores que estão relacionados à sobrevida do enxerto e, desta forma, envolvidos na diminuição da demanda e no aumento da oferta de órgãos. Objetivos: Avaliar o impacto das variáveis, idade do doador, compatibilidade HLA e tempo de isquemia fria, na sobrevida do enxerto de pacientes submetidos à alotransplantes renais com doador cadáver. Materiais e Métodos: Foram analisados retrospectivamente 454 pacientes submetidos a transplantes renais com doadores cadáveres realizados de abril de 1987 a dezembro de 2003, no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo (HCFMRP-USP). Resultados: Das variáveis analisadas, a idade do doador foi a de maior impacto estatístico na sobrevida dos transplantes renais estudados. A sobrevida dos enxertos oriundos de doadores com idade de 16 a 40 anos (143,30 meses em média), foi significativamente maior em comparação com a sobrevida nos grupos com doadores acima de 40 anos (66,46 meses) (p= 0,005) e nos doadores de 0 a 10 anos de idade (63,29 meses) (p= 0,01). A compatibilidade HLA e o TIF não interferiram de modo significativo na sobrevida dos transplantes estudados (p= 0,98 e p= 0,16 respectivamente). Conclusões: O tempo de isquemia fria (TIF) e a compatibilidade HLA não apresentaram impacto na sobrevida dos transplantes renais analisados. Os doadores cadáveres com idade de 16 a 40 anos podem ser considerados ideais, uma vez que demonstraram, de modo estatisticamente significativo, impacto favorável na sobrevida dos rins por eles doados. / Introduction: Renal transplantation is world wide recognized as the best therapy for patients that need kidney substitution treatment. It can be considered a clinical and surgical triumph and at the same time, its a form of frustration by the few organs donations available and by the growth of the waiting lists. New strategies are needed to be developed to increase the numbers of kidneys for transplantation. Donors aged, cold ischemia time (CIT) and human leukocyte antigen (HLA) compatibility are correlated with graft survival and consequently, with a diminished organs demand. Objectives: To study the impacts of donors age, cold ischemia time and HLA compatibility, on graft survival patients submitted to cadaver kidney allograft transplantation. Materials and Methods: Records from 454 patients, submitted to cadaver kidneys allograft transplantations between April/1987 and December/2003 by Division of Urology of the Clinical Hospital of Ribeirao Pretos School of Medicine at Sao Paulo University, were analyzed. Results: The age of kidneys donors was the most statistically significant data in the survival graft rate. Donors between 16 and 40 years old had a median survival rate about 143.30 months, that was significantly higher when compared with aged donors above 40 years (66.46 months) (p= 0.005) and below 10 years old (63,29 months) (p= 0.01). In our study, HLA compatibility and cold ischemia time didnt have statistically significant impact on renal allograft survival rate (p= 0.98 and p= 0.16, respectively). Conclusions: HLA compatibility and cold ischemia time didnt have significantly impact on renal allograft survival rate. Donors between 16 and 40 years old can be considered the ideal donors age, since they demonstrated statistically a favorable impact on grafts survival rate.

O óxido nítrico (NO) é o principal agente vasodilatador endógeno que regula o tônus e a homeostase vascular. Dentre os compostos doadores de NO, estão os complexos nitrosilos de rutênio. No presente estudo, o doador de NO estudado, RuBPY, não apresenta citotoxicidade para células do músculo liso vascular (MLV) ao contrário do NPS. O RuBPY apresenta eficácia semelhante ao NPS em relaxar o MLV de aorta de ratos, porém o NPS é mais potente. Ambos compostos liberam NO do tipo radicalar (NO) no meio intracelular, mas o NPS libera também íon nitroxil (NO-). O sequestrador da espécie NO (hidroxocobalamina) reduziu mais a resposta relaxante estimulada com RuBPY do que com o NPS. Nenhum dos dois compostos precisa ser reduzido quimicamente para liberar NO, uma vez que houve relaxamento quando utilizamos alta concentração de KCl como agente contrátil. Porém, este relaxamento foi inibido, o que mostra a importância dos canais para K+ no relaxamento induzido pelos doadores de NO. O bloqueador não seletivo de canais para K+ (TEA), inibiu somente o relaxamento ao RuBPY. A via NO-GCs-GK é ativada por ambos doadores de NO, para induzir relaxamento. A inibição da degradação do GMPc potencializou o relaxamento estimulado com RuBPY e NPS. O armazenamento de Ca+2 no retículo sarcoplasmático (RS) via ativação da SERCA é importante somente para o relaxamento induzido com RuBPY. O composto RuBPY inibiu a resposta contrátil estimulada com fenilefrina devido ao armazenamento de Ca+2 no RS e também por inibir o influxo capacitivo de Ca+2. A presença do endotélio vascular não alterou o relaxamento induzido pelo RuBPY, porém potencializou o relaxamento induzido pelo NPS. A análise da liberação de NO por amperometria demonstrou que o RuBPY libera NO somente em presença do tecido aórtico de ratos. Portanto, não houve liberação espontânea de NO, por fotólise pela luz visível ou por redução química. É necessária a presença de heme-proteínas como a guanilil-ciclase solúvel (GCs) inibida pelo ODQ, para haver a conversão do nitrito presente no RuBPY, a NO. Pela quantificação da fluorescência emitida pela sonda DAF-2DA, RuBPY liberou cerca de 3,5 vezes mais NO do que o NPS. Pela medida do potencial de membrana, demonstramos que o RuBPY induz hiperpolarização de membrana de células isoladas do MLV da aorta de rato. RuBPY tem efeito hipotensor dose-dependente, em ratos hipertensos renais, o que não ocorre em animais normotensos. A redução da pressão arterial em ratos hipertensos é maior do que nos normotensos. Em estudos iniciais de farmacocinética, verificamos que o composto RuBPY é absorvido por via oral e é distribuído entre alguns tecidos após ser administrado aos ratos, por gavagem. / Nitric oxide (NO) is the main endogenous vasodilator agent that regulates vascular tone. Among the compounds which are able of releasing NO, are the nitrosyl ruthenium complexes. The NO donor studied, RuBPY, does not present cytotoxicity in smooth muscle cells (SMC), in contrast to SNP. RuBPY has similar efficacy to SNP in inducing rat aorta relaxation, although SNP is more potent. Both compounds release intracellular radicalar NO (NO), and SNP also release ion nitroxyl (NO-). The NO scavenger (hydroxocobalamine) had greater effect on the relaxation induced by RuBPY than by SNP. Both compounds do not need to be chemically reduced to release NO, as demonstrated in aorta relaxation after pre-contraction with high concentrations of KCl. However, this relaxation was impaired, showing the importance of K+ channels to induce relaxation by NO released from these compounds. By using non-selective blocker for K+ channels (TEA), only the relaxation induced by RuBPY was inhibited. The NO-sGC-GK pathway is activated by NO donors to induce relaxation. Inhibition of cGMP degradation, potentiated the effect of RuBPY and SNP. Storage of Ca+2 in the sarcoplasmic reticulum (SR) via activation of SERCA is important only for the relaxation induced by RuBPY. The contractile response induced by phenylephrine was inhibited by RuBPY due to the storage of Ca+2 in RS and also by inhibiting the capacitive influx of Ca+2. The presence of endothelium had no effect on the relaxation induced by RuBPY, but it potentiated the relaxation induced by SNP. RuBPY released NO only in the presence of the rat aorta. The complex RuBPY did not spontaneously release NO, by photolysis by visible light, or by chemical reduction. RuBPY requires the presence of heme-protein such as guanylyl-cyclase, inhibited by ODQ, to convert nitrite to NO. The amount of NO released from RuBPY was about 3.5 times greater than that released from SNP. RuBPY induced membrane hyperpolarization of SMC. RuBPY has hypotensive effect in renal hypertensive rats in a dose-dependent way, which does not occur in normotensive rats. The decreased of blood pressure in hypertensive rats was greater than in normotensive rats. Initial studies of pharmacokinetics demonstrated that RuBPY is orally absorbed and it is also distributed in some tissues after being administered by gavage to rats.

Anticorpos antieritrocitários são produzidos contra um ou mais epítopos presentes na superfície dos glóbulos vermelhos. A pesquisa de anticorpos irregulares consiste em expor os anticorpos circulantes do indivíduo a antígenos conhecidos in vitro. Nos casos dos doadores de sangue, a legislação vigente determina a realização da pesquisa de anticorpo irregular (PAI), não exigindo, porém, a determinação da especificidade do anticorpo, e os componentes plasmáticos e os concentrados de plaquetas, cuja presença de anticorpos irregulares fora evidenciada, não devem ser utilizados para transfusão. O presente estudo caracterizou o perfil imunohematológico referente à presença de anticorpos irregulares em doadores de sangue do Instituto Paranaense de Hemoterapia e Hematologia Ltda., no período de abril de 2008 a dezembro de 2015, e avaliar: (i) a prevalência de doadores de sangue voluntários que apresentam PAI positiva, (ii) a especificidade dos aloanticorpos identificados, (iii) a prevalência dos aloanticorpos em doadores de acordo com o sexo e a faixa etária. O projeto de pesquisa foi aprovado pelo Comitê de Ética em Pesquisa. As informações sobre resultado da PAI, especificidade do anticorpo, sexo e idade disponíveis no sistema informatizado do Instituto Paranaense de Hemoterapia e Hematologia Ltda. foram acessadas. As análises estatísticas foram realizadas utilizando-se o teste exato de Fisher e valores de p<0,05 foram considerados estatisticamente significativos. Foram analisados 155.834 doadores. Em 474 (0,3%) doadores foram detectados anticorpos irregulares, dos quais, 358 doadores apresentaram resultados conclusivos (25% sexo masculino e 75% feminino). Os anticorpos de maior frequência foram anti-D (23%, n=83), anti-M (15%, n=55), anti-K (12,2%, n=44), anti-E (11,7%, n=42) e antiDia (11,7%, n=42). As associações de diferentes anticorpos ocorreram em 14,8% (48) da população estudada, sendo a mais frequente, anti-D + anti-C (n=19). A frequência de anticorpo irregular na população feminina foi significativamente maior que na masculina para as especificidades anti-D (p=0,0191) e anti-M (p=0,0005). O grupo de doadores com idade menor a 40 anos apresentou uma frequência estatisticamente maior de anticorpos irregulares com especificidades anti-M (p=0,0008) e anti-Dia (p=0,0176) quando comparado ao grupo de doadores com idade maior ou igual a 40 anos. Entre os 86 doadores do sexo masculino, a prevalência de anti-Dia foi significativamente maior na população masculina com idade inferior10 a 40 anos quando comparado aos doadores com idade maior ou igual a 40 anos (p=0,0273). O resultado obtido induz a suspeita de um provável IgM de origem natural considerando que esta população de doadores masculinos apresenta baixa chances de aloimunização por transfusão e não tem histórico de gestacional. A definição da prevalência e especificidade de aloanticorpos em doadores de sangue pode indicar a necessidade de um protocolo que defina a conduta frente ao doador com objetivo de proteção do doador de sangue e otimização do serviço de hemoterapia em relação às coletas de sangue realizadas / Anti-erythrocyte antibodies are produced against one or more epitopes present on the surface of red blood cells. The search for irregular antibodies consists in exposing the subject\'s circulating antibodies to known antigens in vitro. In the case of blood donors, the current Brazilian legislation determines that the irregular antibody test (PAI) must be checked in all blood donores, but does not require the determination of the specificity of the antibody, and the plasma components and the platelet concentrates from blood donors with irregular antibodies should not be used for transfusion. The present study characterized the immunohematology profile for the presence of irregular antibodies in blood donors from the Instituto Paranaense de Hemoterapia e Hematologia Ltda., from April 2008 to December 2015, in order to evaluate: (i) the prevalence of donors with irregular antibodies, (ii) the specificity of the alloantibodies identified, (iii) the prevalence of alloantibodies in donors according to gender and age group. The research project was approved by the Ethics Committee. PAI result, antibody specificity, sex and age data were collected from the system of the Instituto Paranaense de Hemoterapia e Hematologia Ltda. Statistical analyzes were performed using Fisher\'s exact test and values of p <0.05 were considered statistically significant. A total of 155,834 donors were analyzed. In 474 (0.3%) donors, irregular antibodies were detected, of which 358 donors presented conclusive results (25% male and 75% female). The most frequent antibodies were anti-D (23%, n = 83), anti-M (15%, n = 55), anti-K (12.2%, n = 44), anti-E (7%, n = 42) and anti-Dia (11.7%, n = 42). The associations of different antibodies occurred in 14.8% (48) of the study population, being the most frequent anti-D + anti-C (n = 19). The frequency of irregular antibody in the female population was significantly higher than in the male for the anti-D (p = 0.0191) and anti-M (p = 0.0005) specificities. The group of donors less than 40 years of age presented a statistically higher frequency of irregular antibodies with anti-M (p = 0.0008) and anti-Dia specificities (p = 0.0176) when compared to the group of donors aged 40 years or older. Among the 86 male donors, the prevalence of anti-Dia was significantly higher in the male population under 40 years of age when compared to those aged 40 years or older (p = 0.0273). This result induces the suspicion of a probable IgM of natural origin considering that this population of male donors has low chances of transfusion alloimmunization and has no gestational history. The definition of the prevalence and specificity of alloantibodies in blood donors may indicate the need for a12 protocol that defines the conduct towards the donor for the purpose of protecting the blood donor and optimizing the hemotherapy service in relation to the blood collections performed.

O principal objetivo deste trabalho foi a avaliação de estratégias que viabilizassem o uso do metano como doador de elétrons para desnitrificação em ambiente anóxico, visando à aplicação de tecnologia para pós-tratamento de efluente de reatores de manta de lodo e fluxo ascendente &#40;UASB&#41; aplicado no tratamento de esgoto sanitário. O trabalho foi dividido em cinco etapas, sendo que em todas elas o metano foi fornecido como única fonte de carbono orgânico. Na Etapa 1, foram comparados dois inóculos, em reatores em batelada sequenciais &#40;SBR&#41; com biomassa imobilizada. Como inóculo, foi utilizada mistura de sedimento marinho e sedimento de mangue &#40;SED-SBR&#41; ou lodo de reator UASB &#40;LAn-SBR&#41;. Na Etapa 2, foram avaliadas estratégias de enriquecimento da comunidade metanotrófica desnitrificante. Para tanto, foram operados dois SBRs: um com biomassa imobilizada &#40;Imob-SBR&#41; e outro com biomassa suspensa &#40;Susp-SBR&#41;. Na Etapa 3, foi operado reator contínuo de leito fixo e estruturado e fluxo ascendente &#40;Up-FSBR&#41;. Nas etapas 1 e 3, o metano suportou a ocorrência da desnitrificação e da manutenção celular. O consumo das formas nitrogenadas na Etapa 2 foi muito baixo e não foi possível concluir qual a melhor estratégia para enriquecimento da biomassa. Na Etapa 4, a biomassa retirada do Up-FSBR ao final da operação foi submetida a testes de endogenia. Foram operadas bateladas simples e, em cada uma, foi dosada apenas uma das formas nitrogenadas em condições com metano e sem metano. Os resultados mostraram que a redução das formas nitrogenadas é um processo paralelo à oxidação de metano. Na Etapa 5, foi avaliado o consumo de metano e de nitrato em SBR com biomassa suspensa em condições microaeradas e anóxicas. Foi observado que tanto a oxidação do metano quanto a redução do nitrato ocorrem em maior intensidade quando expostos a condições de microaerofilia. A análise dos resultados obtidos permite concluir que os organismos responsáveis pela redução de nitrato &#40;MD&#41; estabeleceram uma relação sintrófica com as bactérias oxidadoras de metano &#40;MM&#41;. Observou-se que, quando as bactérias oxidadoras de metano foram estimuladas, a desnitrificação ocorreu em maior intensidade. / This work aimed to develop the methane &#40;CH4&#41; application as electron donor to denitrification under anoxic conditions for post-treatment of Upflow Anaerobic Sludge Blanket &#40;UASB&#41; effluent, applied in sewage treatment systems. The study was conducted in five stages, in which methane was provided as the sole organic carbon source. In Stage 1, methanotrophic denitrification was assessed in Sequencing Batch Reactors &#40;SBR&#41; inoculated with a blend of marine and mangrove sediments &#40;SED-SBR&#41; or anaerobic sludge &#40;LAn-SBR&#41;. For both inocula. In Stage 2, strategies for the enrichment of methanotrophic-denitrifying community were evaluated. Two SBRs were operated: in one of them, biomass was immobilized &#40;Imob-SBR&#41; while the other was inoculated with suspended biomass &#40;Susp-SBR&#41;. In Stage 3, methanotrophic denitrification was addressed in an Upflow Fixed-Structured Bed Reactor &#40;Up-FSBR&#41;. In Stages 1 and 3, methane supported denitrification and cellular maintenance. In Stage 2, the consumption of oxidized nitrogen compounds was too low, and it was not possible to conclude which is the better strategy for enrichment of the methanotrophic-denitrifying community. In Stage 4, the biomass withdrawn from Up-FSBR was subjected to endogeny tests, in batch reactors. In each batch reactor, only nitrite or nitrate was provided, and both methane and methane-free conditions were tested. It was observed that nitrite and nitrate reduction occurred as a marginal process. In Stage 5, methane oxidation coupled to denitrification was assessed under both anoxic and microoxic conditions and both processes were stimulated under the latter. The obtained data suggest denitrifiers established a syntrophic relationship with methane-oxidizers, and when the latter was stimulated, the denitrification also occurred in greater extent.

Orientador: Paulo Eduardo Neves Ferreira Velho / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-24T03:36:54Z (GMT). No. of bitstreams: 1 Pitassi_LuizaHelenaUrso_D.pdf: 10118032 bytes, checksum: 2717c0557147ebb4a9a080297c902cd6 (MD5) Previous issue date: 2013 / Resumo: O gênero Bartonella constitui um grupo de patógenos zoonóticos re-emergentes e negligenciados que têm sido associados a um amplo espectro de doenças humanas graves. As Bartonella spp. são bacilos gram-negativos que infectam hemácias e células endoteliais, que podem causar infecção crônica ou mesmo fatal. As bartoneloses humanas mais conhecidas são a doença de Carrión, febre das trincheiras, doença da arranhadura do gato e angiomatose bacilar. Como existem espécies de Bartonella intraeritrocitárias e que podem causar infecção persistente e assintomática em humanos, objetivou-se avaliar o risco potencial de transmissão de Bartonella spp. por transfusão sanguínea. No primeiro estudo, os eritrócitos humanos foram infectados com B. henselae em 30 min e 1, 5, 10, e 72 h para a avaliação ultraestrutural por microscopia eletrônica de transmissão (ME). A B. henselae foi vista aderida a eritrócitos humanos 10 h após a inoculação e no interior de eritrócitos após 72 h. A localização intraeritrocitária de B. henselae havia sido mostrada em eritrócitos de gatos, mas nunca antes identificada em eritrócitos humanos. Foi realizado um segundo estudo para determinar as características ultraestruturais da Bartonella spp. nas células do sangue, pois estas informações eram muito limitadas. Este trabalho definiu os parâmetros para se caracterizar a bactéria no interior de células sanguíneas à ME. Um terceiro estudo, utilizando a ME e cultura de bactérias, demonstrou a capacidade da B. henselae sobreviver em uma unidade de concentrado de hemácias armazenada a 4° C durante 35 dias. A viabilidade de B. henselae em unidades de células vermelhas do sangue define o potencial de infecção associada à transfusão de doadores de sangue assintomáticos. Os receptores de transfusão de sangue muitas vezes são ou podem tornar-se imunodeficientes, apresentando o risco de desenvolver formas graves de bartonelose. Utilizando-se parâmetros ultraestruturais da infecção por Bartonella sp., relatou-se um caso de bartonelose como causa de morte 72h após uma transfusão. Um jovem com anemia aplástica, após receber várias transfusões sanguíneas, apresentou estado de choque e morreu após transfusão de um concentrado de hemácias. Os achados da ME permitiram o diagnóstico de infecção por Bartonella sp.. Foi investigada nesta tese a prevalência da Bartonella spp. em uma amostra de 500 doadores de sangue voluntários de Campinas, estado de São Paulo, Brasil, utilizando sorologia, cultura líquida e sólida, PCR e sequenciamento como métodos de identificação destes patógenos. Os anticorpos contra B. henselae e B. quintana foram detectados em 16,2% e 32%, respectivamente. O DNA das Bartonella spp. foi sequenciado e demonstrado em 3,2% do sangue doado (B. henselae em 15 amostras e B. clarridgeiae em uma) e em 1,2% foi documentada, por meio do isolamento do agente, bacteremia por B. henselae. Este estudo revelou a presença de patógenos no sangue de doadores assintomáticos que não estão sendo rastreados no sangue doado e que podem ser responsáveis por doenças não diagnosticadas em pacientes imunodeficientes. Os resultados apresentados estabelecem a necessidade de uma reavaliação por autoridades de saúde pública dos riscos e do impacto da transmissão da Bartonella spp. através de transfusões sanguíneas, especialmente para os pacientes imunodeficientes. Esforços devem ser feitos para a melhoria dos métodos de diagnóstico com sensibilidade suficiente para excluir a Bartonella spp. em doadores de sangue e para proteger os receptores de transfusão de sangue / Abstract: The genus Bartonella is a group of re-emerging zoonotic and neglected pathogens that have been associated with a wide range of serious human diseases. Bartonella spp. are gram-negative bacilli that infect red blood cells and endothelial cells, which can cause chronic infection or even fatal. In humans, Bartonella spp. are known causative agents of Carrion's disease, trench fever, cat scratch disease and bacillary angiomatosis. As they are intra-erythrocytic bacteria and can cause persistent infection and asymptomatic in humans, our objective for this proposal was evaluate the potential risk of transmission of Bartonella spp. by blood transfusion. In the first study, human erythrocytes were infected with B. henselae in 30 min and 1, 5, 10, and 72h for ultrastructural evaluation by transmission electron microscopy (TEM). B. henselae was seen adhering to human erythrocytes 10h after inoculation and inside the erythrocyte after 72 h. Intraerythrocytic localization of B. henselae has been demonstrated in cat erythrocytes, but never before identified in human erythrocytes. A second study was conducted to determine the ultrastructural features of the Bartonella spp. on blood cells because this information was very limited. This paper set the parameters to characterize the bacteria within the blood cell using TEM. A third study using TEM and bacterial culture demonstrated the ability of B. henselae to survive in a unit of red blood cells stored at 4°C for 35 days. The viability of B. henselae in units of red blood cells defines the potential for infection associated with transfusion in asymptomatic blood donors. Recipients of blood transfusions often are or may become immunodeficient, with the risk of developing severe forms of bartonellosis. Using the ultrastructural findings for the diagnosis of Bartonella sp. infection, we reported a case of bartonellosis as cause of death 72 h after transfusion. A young man with aplastic anemia who had received multiple blood transfusions went into shock and died after transfusion of a red blood cell. The findings of TEM allowed the diagnosis of infection with Bartonella sp.. Was investigated in this thesis the prevalence of Bartonella spp. in 500 volunteer blood donors from Campinas, São Paulo State, Brazil, using serology, solid and liquid culture, PCR and sequencing as methods for identifying these pathogens. Antibodies against B. henselae and B. quintana were detected in 16.2% and 32%, respectively. Bartonella sp. DNA was demonstrated in 3,2% of donated blood (15 infected with B. henselae and one with B. clarridgeiae) and 1.2% had B. henselae documented bacteremia by isolation of the agent. This study reveals the presence of pathogens in asymptomatic blood donors who are not being screened in donated blood and may be responsible for undiagnosed diseases in immunodeficient patients. The results presented establish the need for re-evaluation by public health officials of the risks and impact of Bartonella spp. transmission through blood transfusions, especially to immunocompromised patients. Efforts should be made towards improve diagnostic tests with sufficient sensitivity to rule out Bartonella ssp. infection in blood donors to protect transfusion recipients / Doutorado / Clinica Medica / Doutora em Clínica Médica