The customer journey is a re-submerged subject which has become increasingly relevant. The focus derives from the increased focus on customer experience as touch points are increasing as well as becoming more accessible. This is creating problems for companies to allocate their resources in marketing. Previous literature on customer journeys are also limited, mostly originating from research on customer experience. This motivates research within the field with the purpose of exploring the customer journey and connected phenomena.    The basis of this thesis was the customer journey which derives from customer experience. Further, the theoretical framework presents theories about customer experience, consumer buying decisions, customer journey and its touch points. The theoretical framework also presents two conceptual models concerning customer journey and customer experience. Focus was here on exploring the formulation of the customer journey, its steps and components. But also, on investigating how customer experiences affects customer journeys. This thesis had a qualitative research strategy and empirical material was collected through interviews. The empirical findings and the analysis resulted in several conclusions as this was an exploratory study. Reason for use of route, use of different touch points, customer experience impact and were found. Furthermore, the customer journey itself was identified as a reason for conducting a purchase. The result of this thesis might help companies allocate their resources more efficiently between touch points. As well as understanding how to create positive customer experience and the importance of it.

Os familiares são importantes cuidadores de pessoas com transtorno bipolar, podendo estar vulneráveis à sobrecarga, alterações na qualidade de vida e à utilização de psicofármacos. Os objetivos neste estudo foram verificar a prevalência do consumo de psicofármacos e fatores associados em familiares cuidadores de pessoas com transtorno bipolar e compreender o significado que eles atribuem para o próprio consumo do psicofármaco. Trata-se de estudo misto, realizado em duas etapas e em três serviços de um município paulista. A etapa quantitativa foi transversal analítica, para avaliar fatores associados ao consumo de psicofármacos entre familiares cuidadores. Participaram dessa etapa 100 familiares cuidadores, mediante aplicação da Escala de Avaliação da Sobrecarga Familiar; Escala de Avaliação da Qualidade de Vida e questionários para verificar o consumo de psicofármacos e dados demográficos e socioeconômicos. Para análise da associação entre variáveis demográficas e socioeconômicas e o consumo de psicofármacos foram utilizados os testes qui-quadrado e teste exato de Fisher. Para comparação dos escores médios das escalas de qualidade de vida e de sobrecarga familiar foram utilizados o teste U de Mann-Whitney, para as subescalas e domínios sem distribuição normal, e t de Student, para aqueles com distribuição normal. Foi realizado modelo de regressão logística para identificar os preditores de uso de psicofármacos. O consumo de psicofármacos foi de 32%, com predomínio de prescrições feitas por psiquiatras (41%), de antidepressivos (65,6%) e ansiolíticos (59,4%), e diferença estatisticamente significante em relação ao consumo quando comparado às variáveis sexo (p=0,031) e ocupação (p=0,006). A sobrecarga não apresentou associações estatisticamente significantes, enquanto a qualidade de vida foi melhor entre familiares cuidadores que não consumiam psicofármacos. As variáveis domínio social (OR=0,9530; IC 95% 0,9270-0,9760) da escala de qualidade de vida e unidade HC (OR=0,2674; IC 95% 0,0764-0,8141) contribuíram para o modelo de regressão. Na etapa qualitativa do estudo enfocou-se o significado que o familiar cuidador atribui ao próprio consumo de psicofármaco, utilizando-se o Interacionismo Simbólico como referencial teórico e a Teoria Fundamentada nos Dados como referencial metodológico. Os dados qualitativos foram obtidos por meio de entrevistas abertas gravadas com 32 familiares cuidadores e foram analisados em três etapas: codificação aberta, axial e seletiva. Os familiares cuidadores aprendem a conviver com a pessoa com transtorno bipolar e buscam o psicofármaco como forma de autocuidado para assumir a tarefa de ser cuidador. As crenças, medos e percepções relacionadas ao consumo, somadas à falta de informações e à presença de efeitos colaterais levam o familiar cuidador a repensar a importância do consumo. Contudo, a utilização do psicofármaco não é interrompida para a maioria em razão dos benefícios percebidos e da possibilidade de seguir a vida com mais equilíbrio e qualidade de vida. Os resultados desta pesquisa oferecem maior compreensão do fenômeno estudado e subsídios para a melhoria da assistência para essa clientela / Family members are important caregivers of people with Bipolar Disorder (BD) and may be vulnerable to overload, changes in quality of life and use of psychotropic drugs. This study aimed to verify the prevalence of consumption of psychotropic drugs and associated factors in family caregivers of people with BD and to understand the meaning they attribute to the consumption of psychotropic drugs. This mixed study was carried out in two stages and in three services of a city of São Paulo state. The quantitative stage used the analytical cross sectional method to evaluate factors associated to the consumption of psychotropic drugs by family caregivers. The total of 100 family caregivers participated in this stage and the Family Overload Evaluation Scale (FBIS-BR); the quality of life evaluation scale - WHOQOL-Bref; and questionnaires to verify the consumption of psychotropic drugs and demographic and socioeconomic data were used to collect data. The Chi-square test and Fisher\'s exact test were used to analyze the association between demographic and socioeconomic variables and consumption of psychotropic drugs. The Mann-Whitney U test for the subscales and domains with non-normal distribution and Student\'s t test for those with normal distribution were used to compare the mean scores of the quality of life and family overload scales. A logistic regression model was developed to identify the predictors of use of psychotropic drugs. Consumption of psychotropic drugs was of 32%, with predominance of prescriptions provided by psychiatrists (41%), antidepressants (65.6%) and anxiolytics (59.4%), and a statistically significant difference when consumption was compared with the sex variables (p=0.031) and occupation (p=0.006). The overload did not present statistically significant associations, whereas the quality of life was better among family caregivers who did not consume psychotropic drugs. The social domain variables (OR=0.9530; IC 95% 0.9270-0.9760) of the quality of life scale and HC unit (OR=0.2674, IC 95% 0.0764-0.8141) contributed for the regression model. The qualitative stage of the study focused on the meaning that the familiar caregiver attributes to the own consumption of psychotropic drug and used Symbolic Interactionism as theoretical reference and the Grounded Theory as a methodological reference. Qualitative data were obtained through recorded open interviews with 32 family caregivers and were analyzed in three steps: open, axial and selective coding. Family caregivers learn to live with the person with BD and take psychotropic drugs as a form of self-care related to the task of being a caregiver. Beliefs, fears and perceptions regarding the consumption coupled with lack of information and the presence of side effects, lead the family caregiver to rethink the importance of consumption. However, the use of psychotropic drugs is not interrupted for the majority due to perceived benefits and the possibility to continue life with balance and quality of life. The results showed a better understanding of the studied phenomenon and subsidies for the improvement of care for this clientele

A fasciite plantar, terceira lesão mais comum em corredores, apresenta como principais fatores etiológicos o alinhamento do retropé, o arco longitudinal plantar e a carga mecânica dos pés. Os únicos dois estudos que investigaram estes fatores, durante a corrida, permanecem controversos e ainda não claros, principalmente, em relação ao efeito da dor. Outra questão importante é o suporte teórico da associação entre as medidas clinicas dos pés com a carga mecânica no calcâneo, porém sem evidência científica comprovada. A compreensão dos padrões dinâmicos da carga plantar e a sua associação com as medidas clínicas do pé poderão perpetuar uma maior efetividade de recursos terapêuticos como calçados e palmilhas direcionadas a essa população. Portanto, o objetivo geral desse estudo foi avaliar o padrão de carga plantar e impacto dos pés em contato com o solo durante a corrida de corredores com fasciite plantar aguda e crônica e sua associação com o arco longitudinal medial e ângulo do retropé. Foram estudados 75 corredores adultos de ambos os sexos entre 20 a 55 anos. Destes 45 apresentavam fasciite plantar (30 com dor - FPA e 15 sem dor - FPC) e 30 eram corredores controles - GC. Para responder a questões específicas foram realizados dois experimentos. O primeiro teve como objetivo analisar e comparar as taxas de impacto estimadas e as cargas em três regiões distintas dos pés de corredores com fasciite plantar na fase aguda e crônica e corredores controle. A pressão plantar foi avaliada por meio de palmilhas capacitivas (Pedar X System) durante uma corrida de 40m a uma velocidade de 12±5%km/h, utilizando um calçado esportivo padrão. A dor foi mensurada pela escala visual analógica. As taxas de impacto e a carga plantar em retropé, mediopé e antepé foram analisadas em série temporal. Os dados foram processados no Matlab e comparados por ANOVAs (p<0,05). Os principais resultados indicam que a força máxima e integral da força no retropé e as taxas de impacto (20-80%; 10-100%) apresentam-se maiores em corredores com fasciite plantar em relação aos corredores controle (p<0,01). Porém, corredores com fasciite plantar na fase aguda apresentaram menores taxas de impacto e cargas plantares no retropé em relação à fase crônica (p<0,01). O segundo experimento teve como objetivo investigar a relação entre o arco longitudinal medial, o alinhamento do retropé e a dinâmica da pressão plantar em corredores com fasciite plantar: aguda e crônica. O índice do arco plantar e o alinhamento do retropé foram calculados no AutoCAD por meio de imagem fotográfica digital. Para análise da pressão plantar foi utilizado os dados previamente coletados no experimento 1 pelo sistema de palmilhas capacitivas (Pedar X System) durante a corrida. Uma análise de regressão múltipla foi realizada (p<0,05). Os resultados indicam que o arco plantar elevado pode predizer a integral da força (R=0,35, R2=0,15) e a força máxima (R=0,59, R2=0,35) no antepé na fase aguda e crônica, respectivamente. O alinhamento valgo do retropé prediz a força máxima no retropé na fase aguda (R=0,42, R2=0,18) e crônica (R=0,67, R2=0,45), além de predizer o aumento das taxas de impacto do pé na fase crônica da fasciite plantar, 20-80% (R=0,44, R2=0,19) e 10-100% (R=0,63, R2=0,40). Conclui-se que corredores com fasciite plantar aguda e crônica apresentam maiores cargas plantares no retropé e aumento das taxas de impacto do pé no solo. No entanto, a condição fasciite plantar aguda mostrou-se com menores taxas e cargas plantares no retropé em relação à fase crônica, possivelmente, devido ao mecanismo de proteção a dor na região do calcâneo. Além disso, o arco plantar prediz as cargas plantares do antepé de corredores com fasciite plantar e o alinhamento em valgo do retropé demonstrou ser uma medida clínica de fundamental importância para avaliação de corredores com fasciite plantar, pois permitiu predizer tanto o aumento das cargas e taxas de impacto na região do calcâneo e com isso, prevenir os sintomas e a progressão da fasciite plantar / The plantar fasciitis, the third most common injury in runners, presents as the main etiological factors rearfoot alignment, the longitudinal arch and mechanical load on the feet. The only two studies have investigated these factors during running and the results remain controversial and still not clear, specifically regarding the pain symptoms. Another important question is the theoretical support of the association between clinical measurements of the feet with the mechanical load on the heel, but without proven scientific evidence. Understanding the dynamic patterns of plant load and its association with clinical measures of foot may perpetuate more effective therapeutic resources, such as footwear and insoles that target this population. Therefore, the general objective of this study was to evaluate the load pattern and impact of plantar foot in contact with the ground during running in runners with acute and chronic plantar fasciitis and its association with the medial longitudinal arch and rearfoot angle. We studied 75 adult runners of both sexes between 20 and 55 years. Of these 45 had plantar fasciitis (pain-APF 30 with and 15 without pain - CPF) and 30 controls were runners - CG. To answer specific questions two experiments were conducted. The first aimed to analyze and compare the estimated impact rates and the plantar loads in runners with both acute and chronic PF, compared to controls. Seventy-five runners with heel contact running patterns were evaluated and divided into three groups: Acute PF (n=30); chronic PF (n=15); and controls (n=30). Pain was assessed by the Visual Analogue Scale. The plantar pressures was measured by X Pedar system during 40-meter running sessions at speeds of 12±5% Km/h with standard sport footwear. The impact rates and the loads over the rearfoot, midfoot, and forefoot were analyzed based upon temporal series. The data were processed in Matlab and compared by ANOVAs (p <0.05). The main results indicate that the maximum force and integral force in the rearfoot and impact rates (20-80%, 10-100%) were higher in runners with plantar fasciitis when compared with control runners (p <0.01). However, runners with plantar fasciitis in the acute phase showed lower impact rates and loads plantar on rearfoot in relation to chronic phase (p <0.01). The second experiment aimed to investigate the relationship between the medial longitudinal arch, rearfoot alignment and dynamic plantar pressure in runners with plantar fasciitis: acute and chronic. The plantar arch index and rearfoot alignment were calculated in AutoCAD using digital photographic image. For analysis of plantar pressure was used previously collected data in experiment 1 by the system of capacitive insoles (Pedar X System) during the running. A multiple regression analysis was performed (p <0.05). The results indicate that high plantar arch can predict the increase: integral force (R = 0.35, R2 = 0.15) and maximum force (R = 0.59, R2 = 0.35) in the forefoot in acute and chronic phase, respectively. The alignment of the rearfoot valgus predict the maximum force on rearfoot in the acute phase (R = 0.42, R2 = 0.18) and chronic (R = 0.67, R2 = 0.45), and predict increased rates impact of the foot during the chronic plantar fasciitis, 20-80% (R = 0.44, R2 = 0.19) and 10-100% (R = 0.63, R2 = 0.40). We conclude that runners with plantar fasciitis acute and chronic have higher plantar loads on rearfoot and increased rates of impact of the foot on the ground. However, the condition plantar fasciitis acute proved loads with lower impact rates and plantar load on rearfoot in relation to chronic phase, possibly due to the protective mechanism pain in calcaneal region. Moreover, the plantar arch predicts loads plantar in forefoot of the runners with plantar fasciitis and alignment in valgus rearfoot proved to be a measure of fundamental importance for clinical evaluation of runners with plantar fasciitis because it can predict the increase of the plantar loads and impact rates of calcaneal region and thus prevents the symptoms and progression of plantar fasciitis.

Este estudo teve como objetivo avaliar a partida de reator anaeróbio de fluxo ascendente e manta de lodo (UASB), em escala real, no tratamento de esgoto sanitário, sob condições hidráulicas desfavoráveis. O reator UASB faz parte do sistema de tratamento da ETE Água Vermelha, e é seguido de um biofiltro aerado submerso e de uma unidade de desinfecção por ultravioleta. Foi utilizado metade do volume do reator (117 \'M POT.3\'), possibilitando menor tempo de detenção hidráulica (TDH). A pesquisa foi dividida em duas fases, uma sem a adição de inóculo (fase I) e outra com inoculação (fase II). Na fase I, foram aplicados vazão de 480 \'M POT.3\'/dia e carga orgânica volumétrica (COV) de 1,74 kgDQO/\'M POT.3\'.dia com TDH de 6 horas. Para a fase II, estes valores foram de 600 \'M POT.3\'/dia, 2,88 kgDQO/\'M POT.3\'.dia e 5 horas. Os resultados mostraram que as excessivas velocidades ascensionais não permitiram que o reator tivesse desempenho satisfatório. Ocorreu remoção de matéria orgânica e sólidos suspensos apenas na fase II, com exceção dos sólidos suspensos voláteis, que apresentaram remoção nas duas fases. Os lançamentos indevidos de lodo de fossas à que a ETE está submetida, provocaram aumento de sólidos dissolvidos e alcalinidade no efluente. Os leitos de lodo formados nas fases I e II possuíam concentrações semelhantes de sólidos, porém o segundo apresentou maior eficiência de tratamento. Na fase I a remoção de matéria orgânica foi pequena e não ocorreu a formação da manta de lodo. Na fase II ocorreu o provável início da estabilização do processo, aproximadamente 60 dias após a inoculação, a partir de quando a remoção de matéria orgânica dissolvida foi considerada constante e foi observada a formação da manta de lodo. O descarte de 8,5 \'M POT.3\' de lodo pode ser feito a cada 30 dias. / The objective of this study was to evaluate the start-up of a full-scale upflow anaerobic sludge blanket (UASB) reactor, treating domestic sewage, under unfavorable hydraulic conditions. The UASB reactor is part of the wastewater treatment plant of Água Vermelha (São Carlos, SP, Brazil), and it is followed by a submerged aerated biofilter and an ultraviolet radiation disinfection unit. Half of the reactor volume was used (117 \'M POT.3\'), making it possible to work with a low hydraulic retention time (HRT). The research was divided in two phases, one without reactor seeding (phase I), and another when the reactor was seeded (phase II). At phase I, a flow rate of 480 \'M POT.3\'/d and a volumetric loading rate of 1,74 kgDQO/\'M POT.3\'.d were applied, with 6 hours of HRT. At phase II, this values were 600 \'M POT.3\'/d; 2,88 kgDQO/\'M POT.3\'d and 5 hours. The results showed that excessive upflow velocities prevented satisfactory performance of the reactor. Removal efficiencies of COD, BOD and TSS were obtained only in phase II, except for VSS, which showed removal in both stages. Unauthorized inputs of septic tanks sludge caused increase in dissolved solids and alkalinity in the effluent. The sludge beds obtained in phases I and II had similar solids concentration; however, the second presented better treatment efficiency. At phase I, organic matter removal efficiency was low and the blanket sludge was not observed. At phase II, the process likely stabilized 60 days after inoculation (operations\' day 240). From then on, dissolved organic matter digestion was constant and the sludge blanket was observed. The removal of the excess sludge can be done every other 30 days.

Este trabalho tem por meta focalizar a adaptação necessária na metodologia de planejamento atual, norteada segundo uma ótica determinística e conservadora, afim de permitir a elaboração de Programas de Obras de Transmissão/Subtransmissão, condicionados por limitações de teto no montante de investimento a curto e médio prazos. Para tanto, como tema central, busca-se desenvolver uma base conceitual e metodológica, para permitir uma exploração maximizada das possibilidades de carregamento de equipamentos e instalações do sistema de transmissão, contemplando a especificação das ferramentas computacionais necessárias para dar consequência prática à proposta. Considerando o potencial de análise proporcionado por métodos e técnicas de natureza probabilística, aplicados ao planejamento de Sistemas Elétricos, a espinha dorsal da evolução metodológica delineada no texto repousa sobre a avaliação de confiabilidade global e quantificação de riscos operativos. O texto foi estruturado em 8 capítulos onde, a partir de um capítulo introdutório, enfoca-se sequencialmente os temas: * Conceitos e Metodologias para Maximização do Carregamento de Transformadores de Potência. * Conceitos e Metodologias para Maximização do Carregamento de Linhas aéreas de Transmissão. * Impacto das Restrições Financeiras no planejamento e possibilidade de atuação a nível técnico para administrar o conflito entre qualidade de serviço e recursos limitados. * Conceitos e Metodologias para relaxação dos Critérios limitantes para o carregamento máximo de componentes do sistema, em ambiente de severas restrições financeiras, no horizonte de curto prazo. * Resultados obtidos em aplicações concretas a situações do Setor Elétrico, visando caracterizar o alcance do ferramental preconizado nos capítulos anteriores. * Critérios e Procedimentos para Padronização de Subestações e Linhas de Transmissão , coerentes com os conceitos de maximização de carregamento introduzidos nos dois primeiros capítulos. Ao final do texto, incluiu-se um capítulo de \"Conclusões\", com o objetivo de sumarizar os aspectos mais relevantes abordados no texto, assim como indicar linhas de ação e novos desenvolvimentos, necessários para permitir a efetivação prática da metodologia proposta. / An important problem posed to power system planners in developing countries is now becoming commonplace, even in developed countries, and concerns on how to fit an expansion plan into a tight budget, with the least reduction in reliability levels. This problem is known as power system planning under financial constraints and a series of methods is now available to deal with it. So, this work addresses the necessary adjustments in order to permit the Transmission / Sub-transmission Expansion Program establishment when there are investments constraints in a medium/short term range. The main emphasis is on the development of a methodological and conceptual framework, aiming at a maximizes exploration of the loading capability of transformers and transmission lines of the power network. In this context, a new methodological framework for transmission system planning in a financial constrained environment is presented. The proposed approach combines transmission projects priority evaluation and ranking as well as criteria flexibilization, being particularly suitable for short-term planning. The problem to be faced is to operate the system after a project postponement, while the service quality is kept so adequate as it is possible. One of the main issues to be focused is the transmission system uprating since a maximized equipment utilization can contribute to avoid a more significant restriction to the costumers. Therefore, a detailed description of the models for transmission lines and transformers loading limits assessment is presented, with emphasis on the main concepts and the application of probabilistic tools to circumvent the inherent conservatism of deterministic methods. A summary of the contents is presented in what follows: * Concepts and Methodology for Transformers loading maximization. * Concepts and Methodology for Transmission Lines loading maximization. * Financial Restrictions impacts on Expansion Planning Process and the possibilities to manager the conflict between service quality and investment constraints. * Concepts and Methodologies to the relaxation of Criteria that limits the loading capability of transmission lines, aiming at short term Planning under severe financial restrictions, * Obtained results in concrete applications of the proposed methodology and computational tools to Brazilian Electric Power System. * Methods and Criteria to standardization of Substations and Transmission Lines, according the main issues formerly introduced. * Conclusion and Future Research.

Submitted by Mariana Dornelles Vargas (marianadv) on 2015-06-01T13:54:05Z No. of bitstreams: 1 planejamento_portfolio.pdf: 2983834 bytes, checksum: 3f3b3523fda84404f3af648b65221e56 (MD5) / Made available in DSpace on 2015-06-01T13:54:05Z (GMT). No. of bitstreams: 1 planejamento_portfolio.pdf: 2983834 bytes, checksum: 3f3b3523fda84404f3af648b65221e56 (MD5) Previous issue date: 2013-04-26 / Nenhuma / A sobrecarga de escolha é um fenômeno que trata das consequências da diversidade de opções na experiência de escolha do usuário. Pesquisas mostram que o aumento do número de opções causa consequências adversas à experiência de escolha, como diminuição do prazer e da satisfação com a escolha. Com base no mercado de venda direta de higiene e beleza, buscou-se compreender a experiência de escolha do usuário frente à diversidade de alternativas de produtos do segmento, como shampoo, condicionador e sabonete líquido. Dois experimentos foram realizados para entender a experiência do usuário: no primeiro (n=103) a quantidade de produtos foi manipulada através de diferentes cores; e, no segundo (n=96), a quantidade foi manipulada utilizando diferentes formas de embalagens. Ainda, um survey (n=101) foi aplicado para compreender alguns hábitos e preferências do consumidor de higiene e beleza. Como resultado destas pesquisas, foi encontrado o contrário do que apontam as pesquisas na área de sobrecarga de escolha: os usuários suportam a quantidade de opções (50% acima da média do mercado) e têm preferência por produtos específicos para seu uso. Os resultados obtidos, sobre a experiência de escolha do consumidor de higiene e beleza, dão subsídios para o planejamento de portfólio de produtos no segmento, um elemento estratégico do sistema-produto de uma empresa, na perspectiva do design estratégico. O planejamento de portfólio de produtos, com base na experiência de escolha do usuário, auxilia a tornar a experiência de compra positiva para o consumidor, ao mesmo tempo em que beneficia o produtor. / Choice overload is an effect related to great variety of options in user's choice experience. Other research efforts have shown that, when the number of options is increased, there occur adverse consequences to choice experience, such as decreased pleasure and decreased satisfaction. Based on the direct sale market for hygiene and beauty products, our aim was to understand user choice experience in relation to diversity of product alternatives for the shampoo, conditioner and liquid soap segments. Two experiments were conducted to understand user experience: in the first one (n = 103), the amount of products was manipulated through different colors; in the second one (n = 96), the amount was manipulated using different types of packaging. A survey (n = 101) was also applied to understand some habits and consumer preferences for hygiene and beauty products. As a result, it was found the opposite of other research efforts on choice overload: users support the amount of options (50% above the market average) and have a preference for specific products for their use. The results obtained on the consumer choice experience for the hygiene and beauty segment provide a basis for planning of product portfolios on this segment, a strategic product-system element of a company from the strategic design perspective. Planning the product portfolio based on user's choice experience helps creating a positive shopping experience for consumer and benefits the producer.

A hemocromatose hereditária é caracterizada pelo aumento da absorção intestinal de ferro, acarretando progressivo acúmulo no organismo. Os objetivos foram: 1- determinar as frequências das mutações p.C282Y, p.H63D e p.S65C no gene HFE e avaliar os efeitos nas concentrações dos parâmetros do ferro em doadores de sangue; 2- pesquisar mutações nos genes: 2.1- HFE, 2.2- HJV e HAMP, 2.3- TFR2 e SLC40A1, em pacientes portadores de sobrecarga de ferro primária. Participaram 542 doadores de sangue provenientes do Hemocentro da Santa Casa de São Paulo. Foram incluídos 51 pacientes que apresentavam saturação de transferrina &#8805; 50% (para mulheres) e &#8805; 60% (para homens) e ausência de causas secundárias. Os genótipos para as mutações nos genes HFE foram avaliados pela PCR-RFLP. Foi realizado sequenciamento direto bidirecional para cada éxon dos genes, utilizando o sequenciador Genetic Analizer 3500XL®. Nos doadores de sangue, as frequências dos alelos HFE 282Y, HFE 63D e HFE 65C foram 2,1, 13,6 e 0,6%, respectivamente. Os homens que doaram pela primeira vez, portadores do genótipo HFE 282CY, apresentaram maiores valores de saturação de transferrina; e também os portadores dos genótipos HFE 63DD e 63HD apresentaram maiores concentrações de ferritina sérica, em relação aos de genótipo selvagem. Para os pacientes, 72,5% (37/51) apresentaram ao menos 1 alteração no gene HFE e 11 foram identificados como homozigotos para a mutação p.C282Y. Uma mutação não descrita na literatura (p.V256I) foi identificada no gene HFE e a modelagem molecular (análises de ligação e estrutural) detectou que a mutação não reduziu a afinidade entre as proteínas HFE e &#946;2-microglobulina. No sequenciamento dos éxons dos genes HJV e HAMP foram identificadas as alterações já descritas: HJV p.E302K, HJV p.A310G, HJV p.G320V e HAMP p.R59G. Para o gene TFR2, foram identificados 3 polimorfismos já descritos (p.A75V, p.A617A e p.R752H). No gene SLC40A1 foram observados 6 polimorfismos (rs13008848, rs11568351, rs11568345, rs11568344, rs2304704 e rs11568346) e 1 alteração não descrita previamente na literatura (p.G204S). As conclusões foram: 1- em relação aos doadores de sangue, a presença dos alelos HFE 282Y e HFE 63D foi associada ao maior aporte de ferro nos homens que não doaram sangue anteriormente. 2.1- Para os pacientes com sobrecarga de ferro, a mutação p.C282Y em homozigose, ou em heterozigose composta com a p.H63D, foi a mais frequente alteração encontrada (33,3%). 2.2- O diagnóstico molecular de hemocromatose juvenil (HJ) no Brasil (HJV p.G320V em homozigose) foi relatado. As mutações funcionais HJV p.E302K e HAMP p.R59G foram identificadas, sendo possível que estas alterações possam estar contribuindo para consequências fenotípicas juntas as outras mutações em regiões intrônicas ou regulatórias dos genes. 2.3- Mutações funcionais nos genes TFR2 e SLC40A1 não foram identificadas. / Hereditary hemochromatosis (HH) is characterized by increased intestinal iron absorption, which leads to a progressive accumulation of iron in the body. The aims were: 1- to assess the frequency of HFE gene mutations (p.C282Y, p.H63D and p.S65C) and to identify their relationship to iron status in blood donors; 2- to search in primary iron overload patients: 2.1- HFE, 2.2- HJV and HAMP, 2.3- TFR2 and SLC40A1 gene mutations. Blood donors (n=542) were recruited from Hemocentro of Santa Casa Hospital, Sao Paulo, Brazil. The study included 51 patients with transferrin saturation &#8805; 50% (women) and &#8805; 60% (men) and absence of secondary causes. The genotypes for HFE mutations were evaluated by PCR-RFLP. Subsequent bidirectional sequencing for each gene was performed using the Genetic Analizer sequencer 3500XL®. The frequencies of HFE 282Y, HFE 63D and HFE 65C alleles were 2.1, 13.6 and 0.6% in blood donors, respectively. The first time male donors carrying heterozygous genotype for the p.C282Y mutation had higher transferrin saturation values; and men carrying HFE 63DD and 63HD genotypes had higher serum ferritin concentrations when compared to the wild genotype. Thirtyseven (72.5%) out of the 51 patients presented at least one HFE mutation and 11 were identified as homozygous for the mutation p.C282Y. One novel mutation (p.V256I) in the HFE gene was indentified and molecular modeling (free energy and structural analysis in silico) showed that p.V256I mutation did not reduce the affinity binding between HFE and &#946;2-microglobulin. Sequencing in the HJV and HAMP genes revealed HJV p.E302K, HJV p.A310G, HJV p.G320V and HAMP p.R59G alterations. Sequencing in the TFR2 gene observed 3 polymorphisms (p.A75V, p.A617A e p.R752H); and sequencing in the SLC40A1 gene identified 6 polymorphisms (rs13008848, rs11568351, rs11568345, rs11568344, rs2304704 e rs11568346) and 1 p.G204S non-described mutation. The conclusions were: 1- for blood donors, the presence of HFE 282Y and HFE 63D alleles were associated with alterations on iron status only in first time male blood donors. 2.1- For patients with iron overload, the p.C282Y mutation in homozygous or in compound heterozygous with p.H63D, was the most frequent molecular change found (33.3%). 2.2- The molecular diagnosis of Juvenil Hemochromatosis (JH) in Brazil (homozygous genotype for the HJV p.G320V) was reported. The HJV p.E302K and HAMP p.R59G functional mutations were found and, it is conceivable that they may be contributing to phenotypic consequences together to other mutations in intronic or regulatory regions. 2.3- Functional mutation in the TFR2 and SLC40A1 genes were not identified.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive form of cancer with poor prognosis and limited treatment options. Since many patients present with metastatic disease and are thus ineligible for surgical resection, PDAC is almost ubiquitously fatal; new treatment options are therefore needed to combat this disease. A key hallmark of many cancers, including PDAC, is metabolic reprogramming and a shift towards a high glycolytic rate, known as the Warburg effect. This allows cancer cells to generate ATP in the face of hypoxia and to meet the increased metabolic requirements associated with rapid proliferation. We hypothesised that this shift towards glycolytic metabolism has important implications for the regulation of cytosolic Ca2+ ([Ca2+]i) in PDAC, since the plasma membrane Ca2+ ATPase (PMCA), which is critical for maintaining low [Ca2+]i and thus cell survival, is dependent on ATP to extrude cytosolic Ca2+. The relative contributions of mitochondrial vs glycolytic ATP in fuelling the PMCA in human PDAC cell lines (PANC-1 and MIA PaCa-2) were therefore assessed. Moreover, the effects of numerous mechanistically distinct metabolic inhibitors on key readouts of cell death, [Ca2+]i and ATP were investigated. Treatment with glycolytic inhibitors induced significant ATP depletion, PMCA inhibition, [Ca2+]i overload and cell death in both PANC-1 and MIA PaCa-2 cells, while mitochondrial inhibitors had no effect. Subsequently, these experiments were repeated on PDAC cells cultured in media formulated to "switch" their highly glycolytic phenotype back to one more reliant on mitochondrial metabolism. Culture in nominal glucose-free media supplemented with either galactose (10 mM) or alpha-ketoisocaproate (KIC, 2 mM) resulted in a switch in metabolism in MIA PaCa-2 cells, where proliferation rate and glycolysis were significantly decreased, and in the case of cells cultured in KIC, oxidative phosphorylation rate was preserved (assessed using Seahorse XF technology). Following culture of MIA PaCa-2 cells in either galactose or KIC, glycolytic inhibition failed to recapitulate the profound ATP depletion, PMCA inhibition and [Ca2+]i overload observed in glucose-cultured MIA PaCa-2 cells. These data demonstrate that in PDAC cells exhibiting a high rate of glycolysis, glycolytically-derived ATP is important for fuelling [Ca2+]i homeostasis and thus is critical for survival. Finally, using a cell surface biotinylation assay, the keyglycolytic enzymes LDHA, PFKP, GAPDH, PFKFB3 and PKM2 were all found to associate with the plasma membrane in MIA PaCa-2 cells, possibly in a tyrosine phosphorylation-dependent manner. To investigate whether the dynamic membrane-association of glycolytic enzymes provides a privileged supply of ATP to the PMCA in PDAC, the effects of tyrosine kinase inhibitors was assessed on PMCA activity. However, while these inhibited PMCA activity, this occurred without accompanying global ATP depletion. These data indicate that glycolytic ATP is critical for the regulation of [Ca2+]i by the PMCA in PDAC, and that the glycolytic regulation of the PMCA may be an important therapeutic locus. However, further research is required to determine whether membrane-bound glycolytic enzymes regulate its activity.

Este estudo teve como objetivo avaliar a partida de reator anaeróbio de fluxo ascendente e manta de lodo (UASB), em escala real, no tratamento de esgoto sanitário, sob condições hidráulicas desfavoráveis. O reator UASB faz parte do sistema de tratamento da ETE Água Vermelha, e é seguido de um biofiltro aerado submerso e de uma unidade de desinfecção por ultravioleta. Foi utilizado metade do volume do reator (117 \'M POT.3\'), possibilitando menor tempo de detenção hidráulica (TDH). A pesquisa foi dividida em duas fases, uma sem a adição de inóculo (fase I) e outra com inoculação (fase II). Na fase I, foram aplicados vazão de 480 \'M POT.3\'/dia e carga orgânica volumétrica (COV) de 1,74 kgDQO/\'M POT.3\'.dia com TDH de 6 horas. Para a fase II, estes valores foram de 600 \'M POT.3\'/dia, 2,88 kgDQO/\'M POT.3\'.dia e 5 horas. Os resultados mostraram que as excessivas velocidades ascensionais não permitiram que o reator tivesse desempenho satisfatório. Ocorreu remoção de matéria orgânica e sólidos suspensos apenas na fase II, com exceção dos sólidos suspensos voláteis, que apresentaram remoção nas duas fases. Os lançamentos indevidos de lodo de fossas à que a ETE está submetida, provocaram aumento de sólidos dissolvidos e alcalinidade no efluente. Os leitos de lodo formados nas fases I e II possuíam concentrações semelhantes de sólidos, porém o segundo apresentou maior eficiência de tratamento. Na fase I a remoção de matéria orgânica foi pequena e não ocorreu a formação da manta de lodo. Na fase II ocorreu o provável início da estabilização do processo, aproximadamente 60 dias após a inoculação, a partir de quando a remoção de matéria orgânica dissolvida foi considerada constante e foi observada a formação da manta de lodo. O descarte de 8,5 \'M POT.3\' de lodo pode ser feito a cada 30 dias. / The objective of this study was to evaluate the start-up of a full-scale upflow anaerobic sludge blanket (UASB) reactor, treating domestic sewage, under unfavorable hydraulic conditions. The UASB reactor is part of the wastewater treatment plant of Água Vermelha (São Carlos, SP, Brazil), and it is followed by a submerged aerated biofilter and an ultraviolet radiation disinfection unit. Half of the reactor volume was used (117 \'M POT.3\'), making it possible to work with a low hydraulic retention time (HRT). The research was divided in two phases, one without reactor seeding (phase I), and another when the reactor was seeded (phase II). At phase I, a flow rate of 480 \'M POT.3\'/d and a volumetric loading rate of 1,74 kgDQO/\'M POT.3\'.d were applied, with 6 hours of HRT. At phase II, this values were 600 \'M POT.3\'/d; 2,88 kgDQO/\'M POT.3\'d and 5 hours. The results showed that excessive upflow velocities prevented satisfactory performance of the reactor. Removal efficiencies of COD, BOD and TSS were obtained only in phase II, except for VSS, which showed removal in both stages. Unauthorized inputs of septic tanks sludge caused increase in dissolved solids and alkalinity in the effluent. The sludge beds obtained in phases I and II had similar solids concentration; however, the second presented better treatment efficiency. At phase I, organic matter removal efficiency was low and the blanket sludge was not observed. At phase II, the process likely stabilized 60 days after inoculation (operations\' day 240). From then on, dissolved organic matter digestion was constant and the sludge blanket was observed. The removal of the excess sludge can be done every other 30 days.

The current investigation was an examination of the repetition-to-repetition magnitudes and changes in kinetic and kinematic characteristics of the back squat using accentuated eccentric loading (AEL) and cluster sets. Trained male subjects (age = 26.1 ± 4.1 years, height = 183.5 ± 4.3 cm, body mass = 92.5 ± 10.5 kg, back squat to body mass ratio = 1.8 ± 0.3) completed four load condition sessions, each consisting of three sets of five repetitions of either traditionally loaded straight sets (TL), traditionally loaded cluster sets (TLC), AEL cluster sets (AEC), and AEL straight sets where only the initial repetition had eccentric overload (AEL1). Eccentric overload was applied using weight releasers, creating a total eccentric load equivalent to 105% of concentric one repetition maximum (1RM). Concentric load was 80% 1RM for all load conditions. Using straight sets (TL and AEL1) tended to decrease peak power (PP) (d = −1.90 to −0.76), concentric rate of force development (RFDCON) (d = −1.59 to −0.27), and average velocity (MV) (d = −3.91 to −1.29), with moderate decreases in MV using cluster sets (d= −0.81 to −0.62). Greater magnitude eccentric rate of force development (RFDECC) was observed using AEC at repetition three (R3) and five (R5) compared to all load conditions (d = 0.21–0.65). Large within-condition changes in RFDECC from repetition one to repetition three (∆REP1–3) were present using AEL1 (d = 1.51), demonstrating that RFDECC remained elevated for at least three repetitions despite overload only present on the initial repetition. Overall, cluster sets appear to permit higher magnitude and improved maintenance of concentric outputs throughout a set. Eccentric overload with the loading protocol used in the current study does not appear to potentiate concentric output regardless of set configuration but may cause greater RFDECCcompared to traditional loading