Much experimental evidence shows that the cytoskeleton is a downstream target and effector during cell death in numerous neurodegenerative diseases, including Parkinson's, Huntington's, and Alzheimer's diseases. However, recent evidence indicates that cytoskeletal dysfunction can also trigger neuronal death, by mechanisms as yet poorly understood. We studied a mathematical model of cytoskeleton-induced neuron death in which assembly control of the neuronal cytoskeleton interacts with both cellular stress levels and cytosolic free radical concentrations to trigger neurodegeneration. This trigger mechanism is further modulated by the presence of cell interactions in the form of a diffusible toxic factor released by dying neurons. We found that, consistent with empirical observations, the model produces one-hit exponential and sigmoid patterns of cell dropout. In all cases, cell dropout is exponential-tailed and described accurately by a gamma distribution. The transition between exponential and sigmoidal is gradual, and determined by a synergetic interaction between the magnitude of fluctuations in cytoskeleton assembly control and by the degree of cell coupling. We concluded that a single mechanism involving neuron interactions and fluctuations in cytoskeleton assembly control is compatible with the experimentally observed range of neuronal attrition kinetics. We also studied the transit of neurons through states intermediate between initial viability and cell death. We found that the stochastic flow of neuron fate, from viability to cell death, self-organizes into two distinct temporal phases. There is a rapid relaxation of the initial neuron population to a more disordered phase that is long-lived, or metastable, with respect to the time scales of change in single cells. Strikingly, cellular egress from this metastable phase follows the one-hit kinetic pattern of exponential decline now established as a principal hallmark of cell death in neurodegenerative disorders. Intermediate state metastability may therefore be an important element in the systems biology of one-hit neurodegeneration. Further, we studied the full spatiotemporal dynamics of death factor pulses released from dying neurons to emphasize the effects of the cell-to-cell coupling strength on neuron death rates. The rate of neuron cell loss monotonically increased with increased diffusion-dependent intercellular communication. Death factor diffusion effects may therefore be important moderators of one-hit neurodegeneration.

Much experimental evidence shows that the cytoskeleton is a downstream target and effector during cell death in numerous neurodegenerative diseases, including Parkinson's, Huntington's, and Alzheimer's diseases. However, recent evidence indicates that cytoskeletal dysfunction can also trigger neuronal death, by mechanisms as yet poorly understood. We studied a mathematical model of cytoskeleton-induced neuron death in which assembly control of the neuronal cytoskeleton interacts with both cellular stress levels and cytosolic free radical concentrations to trigger neurodegeneration. This trigger mechanism is further modulated by the presence of cell interactions in the form of a diffusible toxic factor released by dying neurons. We found that, consistent with empirical observations, the model produces one-hit exponential and sigmoid patterns of cell dropout. In all cases, cell dropout is exponential-tailed and described accurately by a gamma distribution. The transition between exponential and sigmoidal is gradual, and determined by a synergetic interaction between the magnitude of fluctuations in cytoskeleton assembly control and by the degree of cell coupling. We concluded that a single mechanism involving neuron interactions and fluctuations in cytoskeleton assembly control is compatible with the experimentally observed range of neuronal attrition kinetics. We also studied the transit of neurons through states intermediate between initial viability and cell death. We found that the stochastic flow of neuron fate, from viability to cell death, self-organizes into two distinct temporal phases. There is a rapid relaxation of the initial neuron population to a more disordered phase that is long-lived, or metastable, with respect to the time scales of change in single cells. Strikingly, cellular egress from this metastable phase follows the one-hit kinetic pattern of exponential decline now established as a principal hallmark of cell death in neurodegenerative disorders. Intermediate state metastability may therefore be an important element in the systems biology of one-hit neurodegeneration. Further, we studied the full spatiotemporal dynamics of death factor pulses released from dying neurons to emphasize the effects of the cell-to-cell coupling strength on neuron death rates. The rate of neuron cell loss monotonically increased with increased diffusion-dependent intercellular communication. Death factor diffusion effects may therefore be important moderators of one-hit neurodegeneration.

Advances in molecular methods for analyzing DNA, RNA and proteins in humans as well as in other animals, plants, fungi, bacteria or viruses have greatly increased the resolution with which we can study life’s complexity and dynamics on earth. While genomic, transcriptomic and proteomic laboratory tools for molecular diagnosis of disease are rapidly becoming more comprehensive, the access to such advanced yet often expensive and centralized procedures is limited. There is a great need for rapid and comprehensive diagnostic methods in low-resource settings or contexts where a person can not or will not go to a hospital or medical laboratory, yet where a clinical analysis is urgent. In this thesis, results from development and characterization of novel technologies for DNA and protein microarray analysis are presented. Emphasis is on methods that could provide rapid, cost-effective and portable analysis with convenient readout and retained diagnostic accuracy. The first study presents a magnetic bead-based approach for DNA microarray analysis for a rapid visual detection of single nucleotide polymorphisms. In the second work, magnetic beads were used as detection reagents for rapid differential detection of presence of pestiviral family members using a DNA oligonucleotide microarray with read-out by means of a tabletop scanner or a digital camera. In paper three, autoimmune responses from human sera were detected on a protein autoantigen microarray, again by means of magnetic bead analysis. Here, special emphasis was made in comprehensively comparing the performance of the magnetic bead detection to common fluorescence-based detection. In the fourth study, an immunochromatographic lateral flow protein microarray assay is presented for application in the classification of contagious pleuropneumonia from bovine serum samples. The analysis could be performed within 10 minutes using a table top scanner, and the performance of the assay was shown to be comparable to that of a cocktail ELISA. In the fifth paper, the lateral flow microarray framework is investigated in further detail by means of experiments and numerical simulation. It was found that downstream effects play an important role, and the results further suggest that the downstream binding profiles may find use in simple affinity evaluation. / QC 20100713

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The immunostimulatory properties of inactivated parapoxvirus ovis (iPPVO) have been investigated in different animal species and experimental settings. This study investigated the effects of administration of iPPVO on selected events of the innate response in mice. Neutrophil activation, phagocytic activity of macrophages, serum bactericidal activity, induction and antiviral activity of interferon type I (IFN - I) and expression of several classes of cytokines were assayed following intraperitoneal inoculation of Mus musculus with iPPVO (107 TCID50). Serum from iPPVO-treated animals showed IFN-I activity against murine encephalomyocarditis virus (EMCV) between 6 and 12 hours post infection (hpi), as shown by plaque reduction. A significant activation of neutrophils at 6hpi was observed by NBT reduction test in animals treated with the iPPVO. Peritoneal macrophages from mice treated with iPPVO demonstrated a significant increase (p<0.01) in phagocytic activity against Candida albicans both in vivo (between 12 and 96 hpi) and in vitro (24 and 72 hpi). iPPVO treated mice showed increased serum bactericidal activity against Escherichia coli (p<0.05) at two periods (24 and 72 hpi). A second study evaluated the expression of cytokines in response to inoculation of iPPVO. For this, spleens and serum samples were collected from mice treated with iPPVO at different intervals after inoculation and subjected to quantification of messenger RNA (mRNA) by real time PCR (qRT-PCR) and detection/quantification of serum cytokines by ELISA. Quantification of mRNA identified a significant and transient increase in the expression of various cytokines, with variable magnitude and kinetics. mRNA expression of proinflammatory cytokines (IL-1β, TNF-α and IL-8) peaked at 24 hpi (5.4 times increase) and 48 hpi (3 and 10 times, respectively). A 15-fold increase in expression of INF-γ and 6-fold for IL-12 was observed at 48 and 24 hpi, respectively. An increase in the expression of self-regulatory cytokines (Th2) cells, especially IL-10 and IL-4 was detected at later periods (72 and 96 hpi) with peaks of 4.7 and 4.9 fold, respectively. The determination of the concentration of serum cytokines by ELISA showed an increase in IL-1β, TNF-α, IL- 12, IFN-γ and IL-10 with kinetics similar to that observed by qPCR, especially for IL-1 and INF-γ. In summary, these results demonstrate that inoculation with iPPVO stimulates transiently a number events associated with cellular and humoral innate immune responses. If taken together, these effects would likely contribute for the enhanced resistance to certain pathogens observed in animals treated with iPPVO. / As propriedades imunoestimulatórias do Parapoxvirus ovis inativado (iPPVO) têm sido verificadas em diferentes espécies animais e condições experimentais. No presente trabalho foram investigados os efeitos da administração do iPPVO sobre eventos da resposta inata de camundongos. Ativação de neutrófilos, atividade fagocítica de macrófagos, atividade bactericida do soro, indução e atividade antiviral do interferon tipo I (INF-I) e expressão de várias classes de citocinas foram investigados em diferentes intervalos após inoculação de Mus musculus pela via intraperitonial com iPPVO (dose 107 TCID50). O soro de animais tratados com iPPVO apresentou atividade de INF-I frente ao vírus da encefalomiocardite murina (EMCV) entre 6 e 12 horas pós inoculação (hpi), como demonstrado pela redução significativa de formação de placas virais. Uma significativa ativação dos neutrófilos circulantes foi observada pela técnica de redução do NBT em animais tratados com o iPPVO às 6 hpi. Macrófagos peritoneais de camundongos tratados com iPPVO demonstraram um aumento significativo (p<0,01) na atividade fagocítica frente a Candida albicans tanto in vivo (entre 12 e 96 hpi) quanto in vitro (24 e 72hpi). Camundongos tratados com iPPVO apresentaram aumento na atividade bactericida do soro frente à Escherichia coli (p<0,05) em dois períodos avaliados (24 e 72 hpi). Um segundo estudo avaliou a expressão de citocinas em resposta à inoculação do iPPVO. Para isso, amostras de baço e soro foram coletados de camundongos tratados com iPPVO em diferentes intervalos após a inoculação e submetidas a quantificação de RNA mensageiro (RNAm) por PCR em tempo real (qRT-PCR) e detecção/quantificação de citocinas no soro por ELISA. A quantificação de RNAm permitiu detectar um aumento significativo e transitório da expressão de várias citocinas, com magnitude e cinética variáveis. A expressão de RNAm das citocinas pró-inflamatórias (IL-1β, TNF-α e IL-8) atingiu o pico às 24 hpi (aumento de 5,4 vezes), 48 hpi (3 e 10 vezes, respectivamente). Um aumento de 15 vezes na expressão gênica do INF-γ, e de 6 vezes para a IL-12 foi observado às 48 e 24 hpi, respectivamente. Um incremento na expressão das citocinas auto-regulatórias (Th2), principalmente IL-10 e IL-4, foi detectado em períodos mais tardios (72 e 96 hpi) com picos de 4,7 e 4,9 vezes, respectivamente. A determinação da concentração das citocinas séricas por ELISA revelou um aumento nos níveis de IL-1β, TNF- α, IL-12, INF-γ e IL-10, com uma cinética similar à observada pela técnica de qPCR, especialmente para IL-1β e INF-γ. Em resumo, esses resultados demonstram que o tratamento com iPPVO estimula de forma significativa e transitória uma série de eventos celulares e humorais ligados à resposta imune inata. Esses efeitos, se considerados em conjunto, provavelmente contribuem para o aumento da magnitude da resposta imunológica a certos patógenos observada em animais tratados com o iPPVO.

Humant immunbristvirus (HIV), hepatit B och C är blodsmittor som finns världen över. Dessa tre sjukdomar anses som allmänfarliga och som anmälningspliktiga sjukdomar. Livet med en smittsam blodsjukdom kan vara påfrestande för en patient, både fysiskt och psykiskt. Syftet med denna litteraturstudie var att beskriva patienters upplevelse av att leva med en smittsam blodsjukdom. Studien är en allmän litteraturstudie, där kvalitativ forskning användes för att få fram resultatet. Resultatet består av tre teman och sju subteman. De teman som skapades var: rädsla, känslan av att vara osedd och känslan av att vara sedd. Livet med en smittsam blodsjukdom kan vara påfrestande för en patient. Rädslan för sjukdomens konsekvenser är stor. Socialt stöd från anhöriga och sin omgivning har stor påverkan på patientens liv. Brist på socialt stöd kan bidra till att patienten drar sig undan, tar till alternativa hjälpmedel såsom alkohol och narkotika samt till att patienten utvecklar depression. Som vårdpersonal är det viktigt att se patienten bakom den smittsamma blodsjukdomen, då patienter ibland upplever mötet med sjukvården som något negativt. Vårdpersonalen som är rädda för sjukdomen, är även rädda för att bli smittade. Det bidrar till att patienten känner sig dömd och väljer att avstå från behandlingar. / Human immunodeficiency virus (HIV), hepatitis b and c are contagious blood diseases that exists worldwide. These three diseases are considered to be generally dangerous and as a notifiable disease. Living with a contagious blood disease can be stressful to patients both physically and mentally. The purpose of this literature study was to describe patients' experience of living with contagious blood diseases. The study is a general literature study, where qualitative research was used to produce the result. The result consists of three themes and seven different subthemes. That resulted in the creation of three themes: fear, the feeling of being unseen and the feeling of being seen. Living with a contagious blood disease can be stressful to a patient. There is a great fear for the consequence of the disease. Getting social support from relatives and their social environment has a major impact on the patient's life. The lack of social support can make the patient avoid social contact. Patients’ take alternative means such as alcohol, drugs and the patient can develop depression. As a formal caregiver it is important to see the person behind the infection, as the patient experience the meeting with formal caregivers as negative. The formal caregivers are afraid of the disease and to be infected. The patient feels judged and chooses to avoid treatment.

Made available in DSpace on 2014-06-11T19:35:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-09Bitstream added on 2014-06-13T21:07:33Z : No. of bitstreams: 1 siqueira_ak_me_botfmvz.pdf: 464792 bytes, checksum: aae10b408540ec568f7000a9c01f5a54 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Escherichia coli e considerado o principal agente causal de infeccao de trato urinario (ITU) e piometra em caes. A patogenicidade das linhagens esta relacionada a presenca de adesinas e diferentes fatores de virulencia. Foram avaliadas alteracoes hematologicas e diferentes fatores de virulencia em 51 linhagens de E. coli isoladas de ITU, 52 de piometra e 55 de fezes de caes sem sinais entericos. A producao de ¿-hemolisina foi verificada em 26 (51,0%) das estirpes de ITU e em 20 (38,5%) de piometra. Exames hematologicos revelaram principalmente anemia, trombocitopenia e leucocitose por neutrofilia e monocitose nos caes com ITU e piometra. Os maiores indices de sensibilidade nas 158 estirpes foram observados para norfloxacina, ciprofloxacina e enrofloxacina em mais de 60% dos isolados. Os maiores indices de resistencia foram encontrados em 60% ou mais das estirpes com o uso de sulfametoxazole/trimetoprim. Linhagens resistentes a tres ou mais antimicrobianos foram constatadas em 24 (47,1%) de ITU, 7 (13,5%) de piometra e 4 (7,3%) das fezes, das quais respectivamente, 17 (33,3%), 1 (1,9%) e 3 (5,5%), com resistencia multipla a cinco ou mais drogas. fimH foi observado em mais de 90% dos isolados. papC foi detectado em 12 (23,5%) linhagens de ITU, 19 (36,5%) de piometra e 10 (18,2%) das fezes. papGI nao foi detectado, enquanto papGII foi observado em 3 (5,8%) isolados de piometra. papGIII foi expressado em 10 (19,6%) linhagens de ITU, 15 (28,8%) de piometra e 9 (16,4%) das fezes. sfaS foi encontrado em 22 (43,1%) de ITU, 24 (46,1%) de piometra e 19 (34,5%) das fezes. afa foi detectado em 1 (1,9%) linhagem de ITU e de piometra... / Escherichia coli is considered the more important microrganism in urinary tract infection (UTI) and pyometra in dogs. The pathogenicity of strains is associated with different adhesins and virulence factors. Haematological exams and different virulence factors was evaluated in 51 E. coli strains isolated from UTI, 52 from pyometra and 55 from feces of dogs without enteric signs. Alpha-haemolysin was verified in 26 (51.0%) strains from UTI and 20 (38.5%) from pyometra. Haematological exams revealed mainly anaemia, thrombocytopenia and leucocytosis by neutrophilia and monocitosis in dogs with UTI and pyometra. Norfloxacin, ciprofloxacin and enrofloxacin were the most-effective drugs (>60%) for 158 E. coli strains. High rates of E. coli resistance to antimicrobials were observed in 60% or more of strains using sulfametoxazole/trimetoprim. Multiple drug resistance for three or more antimicrobials was observed in 2 (47.1%) strains isolated from UTI, 7 (13.5%) from pyometra and 4 (7.3%) from feces. From these, 17 (33.3%), 1 (1.9%) and 3 (5.5%), respectively, showed multiple resistance to five or more drugs. fimH was observed in 90% or more of 158 isolates. papC was detected in 12 (23.5%) strains isolated from UTI, 19 (36.5%) from pyometra and 10 (18.2%) from feces. None strain expressed papGI, while papGII was observed in 3 (5.8%) strains of pyometra. papGIII was detected in 10(19.6%) strains of UTI, 15 (28.8%) from pyometra and 9 (16.4%) from feces. sfaS was observed in 22 (43.1%) strains of UTI, 24 (46.1%) of pyometra and 19 (34.5%) of feces. afa was identified in 1 (1.9%) strains isolated from UTI and pyometra...(Complete abstract, click electronic address below)

Orientador: Márcio Garcia Ribeiro / Banca: Antonio Carlos Paes / Banca: Domingos da Silva Leite / Resumo: Escherichia coli e considerado o principal agente causal de infeccao de trato urinario (ITU) e piometra em caes. A patogenicidade das linhagens esta relacionada a presenca de adesinas e diferentes fatores de virulencia. Foram avaliadas alteracoes hematologicas e diferentes fatores de virulencia em 51 linhagens de E. coli isoladas de ITU, 52 de piometra e 55 de fezes de caes sem sinais entericos. A producao de ƒ¿-hemolisina foi verificada em 26 (51,0%) das estirpes de ITU e em 20 (38,5%) de piometra. Exames hematologicos revelaram principalmente anemia, trombocitopenia e leucocitose por neutrofilia e monocitose nos caes com ITU e piometra. Os maiores indices de sensibilidade nas 158 estirpes foram observados para norfloxacina, ciprofloxacina e enrofloxacina em mais de 60% dos isolados. Os maiores indices de resistencia foram encontrados em 60% ou mais das estirpes com o uso de sulfametoxazole/trimetoprim. Linhagens resistentes a tres ou mais antimicrobianos foram constatadas em 24 (47,1%) de ITU, 7 (13,5%) de piometra e 4 (7,3%) das fezes, das quais respectivamente, 17 (33,3%), 1 (1,9%) e 3 (5,5%), com resistencia multipla a cinco ou mais drogas. fimH foi observado em mais de 90% dos isolados. papC foi detectado em 12 (23,5%) linhagens de ITU, 19 (36,5%) de piometra e 10 (18,2%) das fezes. papGI nao foi detectado, enquanto papGII foi observado em 3 (5,8%) isolados de piometra. papGIII foi expressado em 10 (19,6%) linhagens de ITU, 15 (28,8%) de piometra e 9 (16,4%) das fezes. sfaS foi encontrado em 22 (43,1%) de ITU, 24 (46,1%) de piometra e 19 (34,5%) das fezes. afa foi detectado em 1 (1,9%) linhagem de ITU e de piometra...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Escherichia coli is considered the more important microrganism in urinary tract infection (UTI) and pyometra in dogs. The pathogenicity of strains is associated with different adhesins and virulence factors. Haematological exams and different virulence factors was evaluated in 51 E. coli strains isolated from UTI, 52 from pyometra and 55 from feces of dogs without enteric signs. Alpha-haemolysin was verified in 26 (51.0%) strains from UTI and 20 (38.5%) from pyometra. Haematological exams revealed mainly anaemia, thrombocytopenia and leucocytosis by neutrophilia and monocitosis in dogs with UTI and pyometra. Norfloxacin, ciprofloxacin and enrofloxacin were the most-effective drugs (>60%) for 158 E. coli strains. High rates of E. coli resistance to antimicrobials were observed in 60% or more of strains using sulfametoxazole/trimetoprim. Multiple drug resistance for three or more antimicrobials was observed in 2 (47.1%) strains isolated from UTI, 7 (13.5%) from pyometra and 4 (7.3%) from feces. From these, 17 (33.3%), 1 (1.9%) and 3 (5.5%), respectively, showed multiple resistance to five or more drugs. fimH was observed in 90% or more of 158 isolates. papC was detected in 12 (23.5%) strains isolated from UTI, 19 (36.5%) from pyometra and 10 (18.2%) from feces. None strain expressed papGI, while papGII was observed in 3 (5.8%) strains of pyometra. papGIII was detected in 10(19.6%) strains of UTI, 15 (28.8%) from pyometra and 9 (16.4%) from feces. sfaS was observed in 22 (43.1%) strains of UTI, 24 (46.1%) of pyometra and 19 (34.5%) of feces. afa was identified in 1 (1.9%) strains isolated from UTI and pyometra...(Complete abstract, click electronic address below) / Mestre

Made available in DSpace on 2015-05-08T14:47:24Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2465894 bytes, checksum: 24b77b39c7f8b72bb257b954159f5b13 (MD5) Previous issue date: 2012-03-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Introduction: in Brazil, the infectious and contagious diseases still represent a public health problem, mainly by the meaningful morbidity indexes, when compared to the ones of mortality. Thus, it is observed the importance of qualification of the nurse in service to the patient with infectious and contagious disease, observing his autonomy to evaluate the assistential necessities of the client in an efficient way.It is important that the care of nursing be based in a theoretical model and in the scientific methodology of the process of nursing, as well as registered, being a source of information.In this study, we chose using the concept model Human Basic Needs of Horta, which besides being one to the most spread in allthe country, it is adjustable to the service in the clients chosen, and it is the theoretical model adopted by the hospital where the study was developed. Objective: build up instruments for documentation of the processof nursing in a clinic with infectious and contagious diseases. Methodology: it is a methodological research, performed in the Clinic of Infectious and contagious diseases of the University Hospital LauroWanderley, in the city of João Pessoa PB. It was developed into three phases: identification of the empiric indicators of Basic Human Needs; validation of content of the empiric indicators for construction of the instrumentof data collection; and relationship of empiric indicators of the instrument of data collection with the nomenclature of diagnostic, results and intervention for the construction of the instrument of planning of the assistance. Results: the empiric indicators of the Basic Human Needs were identified from the bank of terms, of the nomenclature of diagnostic, results and interventions of nursing of the clinic of infectious and Contagious Diseases and of the literature updated in the area. The validation of the content in the empiric indicators happened with assistance nurses and professors who work in the Clinic of Infectious and Contagious Diseases.With the validated indicators, a instrument of data collection was build, organized according to the Basic Human Needs of Horta. The empiric indicators validated were also correlated with the nomenclature of the clinic, aiming to identify diagnostics, results and interventions of nursing for the construction of the instrument of planning of the assistance. Conclusion: this research brought big contribution for the Clinic of Infectious and contagious diseases, once there were not instruments that registered the process of nursing. Itis an obligation to point that these instruments will provide a support for the communication and the register of the assistance, making it more effective and promoting a bigger visibility by this part of the team of nursing and of the professionals of other areas. / Introdução: no Brasil, as doenças infectocontagiosas ainda representam um problema de saúde pública, principalmente pelos índices de morbidade significativos, quando comparados aos de mortalidade. Assim, observa-se a importância da qualificação do enfermeiro no atendimento ao portador de doenças infectocontagiosas, tendo em vista a sua autonomia para avaliar as necessidades assistenciais do cliente de maneira eficaz. É importante que o cuidado de enfermagem esteja embasado em um modelo teórico e na metodologia científica do processo de enfermagem, bem como documentado, sendo fonte de informação. Neste estudo, optou-se pela utilização do modelo conceitual das Necessidades Humanas Básicas de Horta, o qual, além de ser um dos mais difundidos em todo o país, ajusta-se ao atendimento da clientela escolhida e é o modelo teórico adotado pelo hospital onde foi desenvolvido o estudo. Objetivo: construir instrumentos para documentação do processo de enfermagem em uma clínica de doenças infectocontagiosas. Metodologia: trata-se de uma pesquisa metodológica, que foi realizada na Clínica de Doenças Infectocontagiosas do Hospital Universitário Lauro Wanderley, na cidade de João Pessoa - PB. Desenvolvida em três etapas: identificação dos indicadores empíricos das Necessidades Humanas Básicas; validação de conteúdo dos indicadores empíricos para a construção do instrumento de coleta de dados; e relacionamento dos indicadores empíricos do instrumento de coleta de dados com a nomenclatura de diagnósticos, resultados e intervenções para a construção do instrumento de planejamento da assistência. Resultados: os indicadores empíricos das Necessidades Humanas Básicas foram identificados a partir do banco de termos, da nomenclatura de diagnósticos, resultados e intervenções de enfermagem da Clínica de Doenças Infectocontagiosas e da literatura atualizada da área. A validação de conteúdo dos indicadores empíricos aconteceu com enfermeiros assistenciais e docentes que atuam na Clínica Doenças Infectocontagiosas. De posse dos indicadores validados, foi construído um instrumento de coleta de dados, organizado de acordo com as Necessidades Humanas Básicas de Horta. Os indicadores empíricos validados também foram correlacionados com a nomenclatura da clínica, a fim de identificar diagnósticos, resultados e intervenções de enfermagem para a construção do instrumento de planejamento da assistência. Conclusão: esta pesquisa trouxe grandes contribuições para a Clínica de Doenças Infectocontagiosas, uma vez que na mesmainexistiam instrumentos que documentavam o processo de enfermagem. Cumpre assinalar que estes instrumentos irão fornecer um suporte para a comunicação e o registro da assistência, tornado-a mais efetiva e promovendo uma maior visibilidade desta por parte da equipe de enfermagem e dos profissionais de outras áreas.

Os objetivos do presente estudo foram os de verificar a validade do método de reação em cadeia da polimerase em tempo real (qPCR) para detectar e quantificar o Staphylococcus aureus em amostras de leite conservadas com bronopol oriundas de quartos mamários bovinos subclinicamente infectados, e de avaliar os efeitos da presença e da quantidade de células da bactéria sobre a contagem de células somáticas (CCS), a composição do leite (lactose, gordura, proteína bruta, proteína verdadeira e caseína), e a produção de leite de quartos mamários bovinos subclinicamente infectados pelo patógeno. Para a quantificação do S. aureus e das células somáticas bovinas por meio do qPCR, foi utilizado leite cru bovino para o preparo dos padrões como meio de diluição da inoculação seriada de células somáticas e do S. aureus ATCC 29213, e construídas as equações log10UFC = 37,86 23,54 log10CtSAU e log10CCS = 49,3 - 34,0 log10CtBMCB, com base nos resultados obtidos pelas metodologias de referência para cada procedimento. Para testar a equivalência dessas equações aos respectivos métodos de referência, determinar a sensibilidade e especificidade analíticas e a repetibilidade do método proposto, foram coletadas amostras de leite dos quartos mamários de 60 animais de 2 rebanhos leiteiros da região de Pirassununga dos quais se determinou previamente a ocorrência de casos subclínicos de mastite por S. aureus. Dos quartos mamários também foram mensuradas as produções e coletadas amostras de leite para análise de composição, diagnóstico da mastite, e determinação da sensibilidade e especificidade diagnósticas do procedimento de qPCR estabelecido no estudo. Cada amostra foi submetida à análise de composição, CCS, cultura microbiológica, contagem em placas do S. aureus, processadas para a extração do DNA genômico bovino e do S. aureus, e submetida à reação de qPCR. Para análise da concordância entre os resultados obtidos pelos métodos de referência para o diagnóstico da mastite por S. aureus e o de qPCR foi utilizado o teste Kappa. Para avaliação da equivalência das contagens obtidas pelos métodos de referência do S. aureus e de células somáticas bovinas, foi utilizado o teste das diferenças de Bland-Altman. Para a identificação do efeito da infecção subclínica pelo S. aureus sobre a composição e produção de leite do quarto mamário afetado foi utilizada a análise da variância num delineamento em parcelas subdivididas em faixas. Para estimar o grau de relação entre as contagens de S. aureus, a CCS, produção e composição do leite produzido pelo quarto mamário afetado foi utilizado o coeficiente de correlação de Pearson. A correlação entre os resultados de contagem de células somáticas bovinas determinados pelos métodos de rotina e de qPCR para a quantificação de células somáticas apresentou coeficiente r = - 0,978 (P < 0,001). A correlação entre os resultados da contagem do S. aureus ATCC 29213 determinados pelos métodos de rotina e de qPCR para a quantificação do patógeno apresentou coeficiente r = - 0,989 (P < 0,001). A especificidade analítica do qPCR para a detecção do S. aureus em amostras de leite frente a Escherichia coli, Enterococcus sp., Pseudomonas aeruginosa, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, os estafilococos coagulase-negativa, e as espécies coagulase-positiva Staphylococcus hyicus e Staphylococcus intermedius foi de 100%. O método de qPCR aplicado à detecção de Staphylococcus aureus ATCC 29213 em amostras de leite é replicável e apresentou sensibilidade analítica com limite de detecção para a faixa de 10 UFC/mL à 4,2 x 106 UFC/mL. Em amostras de leite conservadas com bronopol provenientes de quartos mamários subclinicamente infectados, o S. aureus pôde ser detectado, mas não pôde ser quantificado pelo método de qPCR. Nessas amostras, a CCS pôde ser determinada de forma equivalente ao método de rotina. A CCS independe da contagem de S. aureus viáveis, mas foi observada correlação linear e negativa entre o número total de células do patógeno e a CCS. A mastite subclínica pelo S. aureus aumentou a CCS nos quartos mamários, mas não alterou a composição do leite. A doença diminuiu a produção de leite e de gordura dos quartos mamários anteriores acometidos pela infeção, mas não se observou efeito da interação entre o posicionamento da glândula e a infecção sobre a produção de leite. Houve correlação entre as concentrações de lactose (r = 0,42; P = 0,0051), de gordura (r = 0,46; P = 0,0016), de produção de gordura (r = 0,49; P = 0,001), e de leite com produção ajustada para o teor de 3,5% de gordura (r = 0,41; P = 0,006), e o número de S. aureus presentes na amostra de leite. / The objectives of this study were to verify the validity of the real-time polymerase chain reaction (qPCR) to detect and quantify Staphylococcus aureus in bronopol-preserved milk samples from subclinically infected mammary quarters, and to assess the effects of the presence and amount of the pathogen on the somatic cell count (SCC), the composition of milk and milk yield of bovine mammary quarters subclinically infected by the pathogen. In order to quantify S. aureus and bovine somatic cells through qPCR, raw bovine milk was used as a means of serial inoculation media of somatic cells and S. aureus ATCC 29213. From that, equations based on the reference methods for each procedure were built, log10UFC = 37,86 23,54 log10CtSAU and log10CCS = 49,3 - 34,0 log10CtBMCB, respectively. To test their equivalence with the reference methods, determine the analytical sensitivity and specificity, and repeatability of the proposed method, milk was sampled from quarters of 60 animals from two dairy herds in Pirassununga, where subclinical S. aureus mastitis cases had been previously diagnosed. Also, quarter milk yield had been measured and samples collected for milk composition analysis, diagnosis of mastitis, sensitivity and specificity of the procedure established in the study had been determined. Each sample was subjected to composition analysis, SCC, microbiological culture, plate counting of S. aureus, DNA extraction, and subjected to qPCR reaction. Agreement between results from reference methods and qPCR for the diagnosis of mastitis by S. aureus was assessed by Kappa test. Equivalence between S. aureus, SCC scores obtained by reference and qPCR was assessed with Bland-Altman procedures. The effect of S. aureus subclinical infection on milk composition and milk yield of affected quarters was measured using a strip plot design. To estimate the degree of relationship between the counts of S. aureus, SCC, yield and composition of the milk from affected quarters was assessed by the Pearson Correlation. Correlation between SCC determined by routine methods and qPCR was r = - 0.978 (P <0.001). Correlation between S. aureus ATCC 29213 determined by routine methods and qPCR was r = - 0.989 (P <0.001). Analytical specificity of qPCR to detect S. aureus in milk samples against Escherichia coli, Enterococcus sp., Pseudomonas aeruginosa, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, coagulase-negative staphylococci and coagulase-positive species, Staphylococcus hyicus and Staphylococcus intermedius was 100%. The use of the qPCR to detect S. aureus ATCC 29213 in milk samples is replicable. Analytical sensitivity detection limit of the method ranged from 10 CFU/mL to 4.2 x 106 CFU/mL. S. aureus could be detected, but not quantified by qPCR in bronopol-conserved milk samples from subclinically infected quarters. In these samples, SCC could be determined by qPCR as it had been done by routine method. SCC was not dependent on S. aureus viable cells, but a negative linear correlation between the total number of cells of the pathogen and SCC was observed. S. aureus subclinical mastitis increased quarters SCC, but did not change milk composition. The disease decreased quarter milk and fat yield, but no interaction effect was observed between the gland positioning and S. aureus subclinical infection on milk production. Correlations between lactose (r = 0.42, P = 0.0051), fat (r = 0.46, P = 0.0016), fat yield (r = 0.49, P = 0.001), and 3.5% fat adjusted milk yield (r = 0.41, P = 0.006), and the number of S. aureus present in the milk sample were observed.

Submitted by Maria Santos (mariajoaosantos26@gmail.com) on 2018-07-20T11:46:18Z No. of bitstreams: 1 2018_DISSERATION_MARIAJOAO_FINAL (1).pdf: 985094 bytes, checksum: ae65eaf60f5eb75bcd74e76b10b71d79 (MD5) / Approved for entry into archive by Janete de Oliveira Feitosa (janete.feitosa@fgv.br) on 2018-07-27T18:45:46Z (GMT) No. of bitstreams: 1 2018_DISSERATION_MARIAJOAO_FINAL (1).pdf: 985094 bytes, checksum: ae65eaf60f5eb75bcd74e76b10b71d79 (MD5) / Made available in DSpace on 2018-08-08T13:19:44Z (GMT). No. of bitstreams: 1 2018_DISSERATION_MARIAJOAO_FINAL (1).pdf: 985094 bytes, checksum: ae65eaf60f5eb75bcd74e76b10b71d79 (MD5) Previous issue date: 2018-06-20 / Objetivo – O objetivo desta tese é examinar a influencia que gênero e diferentes estímulos emocionais exercem sobre as intenções de partilha online das pessoas: (1) se os indivíduos partilham mais conteúdos positivos ou negativos; (2) quem, de entre homens ou mulheres, reage mais fortemente a estímulos emocionais; e (3) se a valência emocional e o género têm algum tipo de correlação. Metodologia - Esta dissertação utiliza um estudo experimental: 2 (valência emocional: positiva vs. negativa) x 2 (género: masculino vs. feminino) entre sujeitos. Resultados – Os resultados mostram que a valência emocional afeta, de facto, as intenções de partilha de conteúdo online e, mais importante, que as emoções negativas levam a uma maior predisposição para partilhar. Além disso, os resultados demonstram que as mulheres partilham mais conteúdos online quando comparadas com os homens, contudo, não foram observados efeitos heterogêneos de valência emocional. Limitações- A principal limitação desta pesquisa é o fato de ser baseada em respostas próprias a cenários hipotéticos, uma vez que pretendemos medir uma intenção de partilha. Isso significa que, embora a intenção de um indivíduo de se comprometer com um determinado comportamento seja o indicador mais adequado para o comportamento do próprio, isso ultrapassa o âmbito deste estudo. Aplicabilidade do trabalho - Esta pesquisa fornece informações relevantes sobre como criar conteúdo com grande probabilidade de ser partilhado online, ao contrário do conteúdo regular que a maioria das agências e profissionais de marketing utilizam para promover os seus produtos / Purpose – The purpose of this work is to examine the influence that gender and different emotional stimuli exert on people’s online sharing intentions. Precisely, we take a closer look at whether (1) people share more positive or negative content; (2) men or women react more strongly to these emotional stimuli; and (3) emotional valence has a heterogeneous effect across genders. Design/Methodology - This dissertation employs an experimental study: 2 (emotional valence: positive vs. negative) x 2 (gender: male vs. female) between-subjects design. Findings – Results show that emotional valence does indeed influence sharing intentions, and more importantly, that negative emotions lead to a higher willingness to share. Further, results demonstrated that women share more content online when compared to men but no heterogeneous effects of emotional valence were observed. Research limitations - The main limitation of this research is that it is based on self-reported answers to hypothetical scenarios, since we intend to measure potential behavior of sharing. Meaning that, although an individual’s intention to commit to a certain behavior is the most adequate predictor of one’s behavior, it is beyond the scope of this study to measure actual sharing behavior. Practical implications - This research produced valuable insights by providing relevant information on how to create content that will probably be highly shared, on the opposite to the regular content that most agencies and marketer use to promote their products.